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“BLENDING OF SOYMILK WITH COWMILK AND BUFFALO MILK TO PREPARE PANEER”21526505715ATHESIS SUBMITTED TOJIWAJI UNIVERSITY, GWALIORSUBMITTED FOR THE DEGREE OFDOCTOR OF PHILOSOPHYINFOOD TECHNOLOGY2014BYNEHA CHAUDHARYDedicated to my familyDeclaration of the candidateI hereby declare that the thesis entitled “Blending of soymilk with cow and buffalo milk to prepare paneer.” is my own work conducted under the supervision of Dr. M.P. Gupta, Retd. Professor, Dairy Technology, Deen Dayal Upadhyay Veterinary University, Mathura (U.P.) and approved by Research Degree Committee. I have put more than 200 days of attendance with the supervision at the centre.I further declare that, to the best of my knowledge the thesis does not contain any part of any work which has been submitted for the award of any degree either in this University or in any other University / Deemed university without proper citation Signature of the Supervisor Signature of the CandidateDr. M.P. Gupta, Neha ChaudharyDairy Technology,Deen Dayal UpadhyayaVeterinary University,Mathura.Signature of the Co- SupervisorDr. Avinash TiwariSchool of Studies in Botany,Jiwaji University, Gwalior. Signature of the co- ordinatorCertificate of the SupervisorCERTIFICATEThis is to certify that the work thesis entitled “Blending of soymilk with cow and buffalo milk to prepare paneer” is a piece of work done by Neha Chaudhary under my guidance and supervision for the degree of Doctor of philosophy of Jiwaji University, Gwalior. (M.P.), India. That the candidate has put in an attendance of more than 200 days with me. To the best of my knowledge and belief the thesis:embodies the work of the candidate herself,has duly been completed,fulfils the requirements of the ordinance relating to the Ph.D. degree of the University andis up to the standards both in respect of contents and language for being referred to the examiners.Signature of the Supervisor Signature of the Co - SupervisorDr. M.P. Gupta, Dr. Avinash Tiwari, Dairy Technology, School of Studies in Botany,Deen Dayal UpadhyayaJiwaji University,Veterinary University, Gwalior. Mathura. ForwardedSignature of the Co - ordinatorCONTENTSACKNOWLEDGEMENTSABBREVATIONSCHAPTERSPAGE NO.CHAPTER -1IntroductionCHAPTER -2Review of LiteratureCHAPTER -3Material and method CHAPTER -4Result and DiscussionCHAPTER -5SummaryCHAPTERS -6BibliographyACKNOWLEDGEMENTThe successful completion of this thesis can undoubtedly be considered as one of the happiest moment of my life.All the prayers and good wishes of my family members, friends and well wishers have been instrumental in the final culmination of this compilation.At this moment when I took back in the past it seems that whatever I have been able to achieve, would not have been possible without generous contribution from individuals who went beyond their limits to help me make this dream of mine convert into a reality today.First of all ,I convey my thanks to Almighty God for all his blessings at every step providing me the capacity to complete this piece of work.It is my pleasure to express my deepest sense of gratitude and heartfelt respect to my respected supervisors Dr. M.P.Gupta, Retd. Professor, Deen dayal upadhyay veterinary university,Mathura (U.P.) and Dr.Avinash Tiwari,Professor, Jiwaji university, Gwalior,men of impeccable integrity, unassailable height, and the guide for this research work; their dexterity with the subject, motivation, benevolence, magnanimity and their unparallel belief in me made the compilation of this research work possible. I do not have words to express my gratitude and praise for these gentleman for fatherly affection, which inspired me to bring this research to a successful completion. I shall always be obliged for their constant encouragement and persuasive direction towards achieving my goal.I thank them from the bottom of my heart, for all the faith he had in me and for all his affection, untiring ever willing help, valuable suggestions, constant encouragement and enduring guidance.It is my honour to be involved with such a stalwart.My profound respect and sincere gratitude is extended to Prof Sangeeta Shukla, Vice chancellor, Jiwaji university, gwalior for her whole-hearted cooperation for the work.I also extend sincere thanks from the bottom of my heart to Dr. R.M.Agarwal Coordinator,Food Technology, Jiwaji University, Gwalior for his necessary help during my research work. I express my deep sense of regard to all the staff members of Institute of Food Technology , Jiwaji university, Gwalior for their relentless help throughout the research work.I am deeply indebted to my friends, Mrs. Rolly kanchan, and Mrs. Mamta Shankar, Richa Kashyap who not only gave their time but also shared their thoughts with me. I salute all those whose perceptions, observations and inputs have helped me directly or indirectly.No words in this world can ever express the love and encouragement given by my parents ,Prof. Dheer Singh and Mrs Suman Singh.I am thankful to God for giving me such nice parents. This work is not complete without the blessings and deep affection of my Father in law and mother in law (Shri Ravi Shankar and Smt. Shubhra).I am deeply indebted to my Husband (Dr. Sachin), my loving son Akshat andmy brother Amit and sister Silky. for all the love ,support , encouragement given by them during my tough days of thesis. Words are insufficient to express my overwheleming sense of gratitude and internal indebtness to my loving son (Akshat) who suffered a lot during research days. (NEHA CHAUDHARY)Chapter -1 INTRODUCTION INTRODUCTIONMilk is an almost ideal food. It has high nutritive value. It supplies body building proteins, bone forming minerals and health giving vitamin and furnishes energy giving lactose and milk fat. Besides supplying certain essential fatty acid, it contain the above nutrients in an easily digestible and assimiable form. The milk is so much nutritionally rich that it can be said as “nutritional bank” (Ganguli, 1974). Milk contains mostly Ca, P, and constitutes the most important source of bio available Ca in our diet. Milk and Dairy products are part of a healthy diet (Soliman, 2005). Five hundred ml milk supplies 70 percent calcium and riboflavin and half of vitamin A and thiamine to be required daily by 5 years old child. Milk is a poor source of nicotinic acid, but milk protein is a good source of amino acid tryptophan from which nicotinic acid is formed in the body. The nicotinic acid of milk is fully available whereas that from cereal sources is in bound form and hence not available to humans. Milk for the young ones contains almost all the quality amino acids in the milk casein, fat and fatty acids, milk sugar, minerals like Ca, P, K, Mn, Mg, Zn, and Co etc, and the vitamins with the exception of vitamin C and Iron. India stands at the top most position at global level in terms of total milk production with 81 million tones per annum during 2000- 2001 and it is estimated that about five percent of the total milk produced in India is converted into paneer (Makhal and Sen, 2001) which accounts for about 3, 00,000 tones of paneer annually valued at rupees 1050 crores. It is estimated that Annual production of paneer was 4,496 MT in year 2003-04 (Dairy India, 2007). Worldwide, buffalo milk is the second most produced milk after cow’s milk, with 82 billion litres produced each year and constituting 12.5% of all milk consumed by the human population (Ménard et al., 2010). According to Bureau of Indian Standards, paneer is an indigenous milk product prepared by the combined action of acid coagulation and heat treatment of cow or buffalo milk or a combination of them both, having white spongy body, close knit structure and primarily used as a base material in preparation of culinary dishes (Rao et al., 1992). Paneer is highly nutritious since it remains about 90 per cent fat and protein, 50 per cent minerals and 10 per cent lactose of the original milk (David, 2012). Paneer has a fairly high level of fat and protein content, low level of sugar and it also contains some minerals and vitamins. It’s nutritive value is fairly high, Singh and Kanawji,1988. Paneer is recommended for patients of diabetes, coronary heart disease, dental caries and pregnant women (Chopra and Mamtani, 1995). Soybean is one of the nature’s wonderful nutritional gifts. Soybean provides high quality protein with minimum saturated fat. Soybean contains all the three nutrients viz., carbohydrate, protein and fat required for good nutrition, as well as fiber, vitamins and minerals. It has high PUFA content. Soybean has more than twice the amount of minerals, especially calcium, iron, zinc and phosphorus than any other legume (Itapu, 2003 and Venter, 2004). Soy protein is highly digestible (92 to 100 per cent) and contains all essential amino acids. Soy protein products contain high concentration (up to 1 g/kg) of isoflavones that exerts protective properties against breast, prostate, colon and lung cancers (Kirupa et. al.,2011). Soybean protein provides all the essential amino acids in the amounts needed for human health. Soy protein is generally regarded as particles called protein bodies, which are estimated to contain at least 60–70 per cent of the total soybean protein Soybeans also contain biologically active or metabolic proteins, such as enzymes, trypsin inhibitors, hemagglutinins, and cysteine proteases very similar to?papain.?The amino acid pattern of Soy protein is virtually equivalent to that of milk and egg protein in terms of quality. The 1990’s FAO/WHO protein evaluation committee put Soy protein at par with egg and milk protein and ahead of beef protein. Soy milk, like cow's milk, varies in fat content, but the most commonly sold varieties have less fat than whole milk, similar fat content to 2 per cent milk, and more fat than skim/nonfat milk. It has low saturated fat content with high amount of polyunsaturated fatty acid and is readily available source of essential fatty acids. Soybean is very good source of minerals, especially calcium, iron, Phosphorus, and Zinc and vitamin C and vitamin E. Soy protein has received increased attention in recent years among consumers, researchers, and the media. A report released in 1995 estimated that over 12,000 food products were available that contained soy protein (Anderson et al., 1995).Soy milk?(also called?soya milk,?soymilk,?soybean milk, or?soy juice?and sometimes referred to as?soy drink/beverage) is a beverage made from?soybeans In addition to being a rich source of nutrients, soybean has a number of phytochemicals, which offer health benefits such as cancer prevention, cholesterol reduction, combating osteoporosis and menopause regulation, Although Many soy products have limited human use in the Western hemisphere due to undesirable off flavors (Favaro Trindade, 2001). soybeans are high in protein. . Since soy doesn't contain?galactose, a product of lactose breakdown, soy-based infant formulas can safely replace breast milk in children with?galactosemia.?Like lactose-free cow's milk, soymilk contains no lactose, which makes it a good alternative for lactose-intolerant. Today, researchers are interested in both the nutritional value and the potential health benefits of soy.The investigators have shown that consumption of soy protein selectively decreases total and LDL Low density level (bad) cholesterol and maintains HDL High density level (good) cholesterol in individuals with elevated normal blood cholesterol level. Soybean is considered as a natural alternative for hormone replacement therapy for treating women who are in menopause. Increasingly, more studies have revealed the protective role of soy foods in a wide range of health conditions including cardiovascular diseases, cancer and osteoporosis (Messina and Setchell, 1994). Protein, with high biological value, lecithin and unsaturated fats, non metabolizable carbohydrates (stachyose and raffinose) and biologically active compounds like isoflavons, phytoestrogens (genistein and daidzein), give to soy milk the ability to prevent especially the diseases of the digestive tube (Tham et al., 1998). Researchers are studying one supplement, ipriflavone, to see if it may help strengthen bones. Soy protein has received increased attention in recent years among consumers, researchers, and the media. Soy protein products offer benefits to women in various life stages. Benefits include improved diet and cardiovascular status, prevention of certain types of cancer, improved health following menopause, obesity prevention/control, and more options for food variety. The area of soy protein research has increased in popularity in recent years among multiple health disciplines. Future research efforts are likely to include more scientific advances in the use of soy in the diet of Americans. As more is learned about the health benefits of soy, additional foodstuffs will likely be available to meet the community's needs for soy products.A report released in 1995 estimated that over 12,000 food products were available that contained soy protein (Anderson et al., 1995), and sales of soy beverages rose more than 82% in 1999 (Nestle, 2002). : "25 grams of soy protein a day, as part of a diet low in saturated fat and cholesterol, may reduce the risk of heart disease." One serving, (1 cup or 240 ml) of soy milk, for instance, contains 6 or 7?grams of soy protein.Soy milk is one of the novel products of Soybean, which is rich in protein, vitamins, and minerals and free from lactose and cholesterol. Plain soy milk is unsweetened, although some soy milk products are sweetened. Salted soy milk is also consumed in China. In the West, soymilk has become a popular alternative to cow's milk, with a roughly similar protein and fat content.?Soy milk is commonly available in?vanilla?and?chocolate?flavors as well as its original unflavored form. In some Western countries where?‘veganism’ has made inroads, it is available upon request at?cafés?and?coffee?franchises as a cow's?milk substitute Soy milk has about the same amount of?protein?as cow's milk, though the?amino acid profile?differs. Natural soy milk contains little digestible?calcium?as it is bound to the bean's pulp, which is indigestible by humans. To counter this, many manufacturers enrich their products with?calcium carbonate?available to human digestion. Unlike cow's milk, it has little?saturated fat?and no?cholesterol.It is low in saturated fats and contains all Soy Oligosaccharides. A recent study from Europe found that individuals with a habitually health-conscious lifestyle (e.g., individuals who did not eat meat, but did eat fish, or were vegetarians or vegans) were more likely to consume soy foods than the average person (Keinan-Boker et al., 2002). The recent increase in soymilk and tofu (coagulated soymilk) consumption especially in western countries is due to the recognition of the health benefits of soy foods.Vegetarians are individuals who, for various reasons, do not eat meat. Vegans, in comparison, are individuals who do not eat any products from animals, including eggs, milk, and cheese. Vitamin B12?is only found in animal products and, therefore, may be lacking in the diet of vegans. Use of soymilk is one way to obtain this essential vitamin (Dudek, 2001). Cereals and meat substitutes are other options. Infants who are not able to tolerate lactose formulas (those based on cow's milk, casein/whey-based formulas; e.g., Similac, Enfamil, Carnation) may be prescribed soy-based formulas if they are not breastfed (Wardlaw, 2000).Soy milk can be made from whole soybeans or full-fat soy flour. The dry beans are soaked in water overnight or for a minimum of 3 hours or more depending on the temperature of the water. The rehydrated beans then undergo wet grinding with enough added water to give the desired solids content to the final product. The ratio of water to beans on a weight basis should be about 10:1. The resulting slurry or purée is brought to a boil in order to improve its nutritional value by heat inactivating soybean trypsin inhibitor, improve its flavor and to sterilize the product. Heating at or near the boiling point is continued for a period of time, 15–20 minutes, followed by the removal of an insoluble residue (soy pulp fiber or?okara) by filtration.In India Soybean cultivation has reached to 65.0 lakh hectares recording production of 76.1 lakh tones with an average of 1171kg/ha, because the soybean plant is a?legume, it also replenishes the nitrogen content of the soil in which it is grown. In the West, soymilk has become a popular alternative to cow's milk, with a roughly similar protein and fat content. (McGee, Harold 2004). The coagulated protein from soy milk can be made into?tofu, just as dairy?milk?can be made into paneer. In general, whole soybean-based foods, such as tofu and soy milk, have the highest protein and isoflavone content. The soy germ in whole soybeans is particularly high in isoflavones.Paneer provides an easy means of conserving and preserving valuable milk solids during the flush season or in high milk producing areas and transporting it to the regions where either milk production is scanty or the terrain is hard for easy transportation of milk (Sachdeva et al., 1985). The blending of soymilk with cow and buffalo milk will reduce the cost and enhance the nutritional quality of the product as Soymilk is a rich source of vitamin C and Iron which will supplement the cow and buffalo milk as it is poor in vitamin C and Iron content. Therefore, formulation changes that enhance the overall flavor and textural characteristics of soy beverages may be necessary to further increase soy consumption (Potter et al., 2007)Using?soybeans?to make milk instead of raising cows may be?ecologically?advantageous, because the amount of soy that could be grown using the same amount of land would feed more people than if used to raise cows ( Shurtleff and Aoyagi, 2000 ). Fermented soy milk products may provide economic and nutritional benefits, because they can be prepared at higher protein levels at comparable or lower cost than regular fermented milk products (Karleskind et al., 1991) Sensory characteristics of a given food cover the perception of appearance, texture, odor, flavor and mouth feel which in turn influence the acceptability and intake of the product (Cardello et al., 2000).The information on optimization of blending of soy milk with cow milk and buffalo milk to prepare paneer, it’s nutritional value, therapeutic value and its effect on quality of paneer are meager. Considering these facts, the present investigation “Blending of soymilk with cowmilk and buffalo milk to prepare paneer” was undertaken with the following objectives:ObjectivesTo study the physico-chemical characteristics of raw material.To optimize the processing variables for the manufacture of soy milk.To optimize the level of soymilk in cow and buffalo milk to prepare paneer.To study the effect of blending of soymilk on the physicochemical, and sensory characteristics of paneer.To study the changes in physicochemical, textural and sensory characteristics of paneer during storage at refrigeration temperature and room temperature.Chapter -2Review of Literature REVIEW OF LITERATUREMilk and its products are an excellent source of vital nutrients and functional constituents. It is described as nature’s nearly perfect food and some leading experts recognize milk and its product as important constituents of a well balanced, nutritionally adequate diet. Indigenous dairy products have played an important role in the socioeconomic life of our people since time immemorial. India is emerging as the largest milk producing country in the world with the milk production of about 94.60 million tonnes in 2008 (Bhasin, 2008). Paneer is one of the popular indigenous dairy products in which the technique of acid coagulation of milk at high temperature is used to conserve wholesome and unique nutrients of milk. Paneer is a heat and acid coagulated traditional milk product, mainly consisting of milk solids obtained from the coagulation of hot milk with permitted organic acid and subsequent drainage of whey followed by pressing (Reeta and Kumar, 2013).2.1 HistoricalIt is believed that the nomads of south west Asia were the first to develop several distinctive heat and acid coagulated varieties of cheese (Mathur et al.,?1986). The people of?Kusana?and?Saka Satavahanaperiods (AD 75-300) used to consume the solid mass prepared from mixture of warm milk and curd, which resembles paneer (Mathur,?1991).?Kradi?cheese—a semisoft dried cheese found in Jammu and Kashmir is quite similar to paneer (Punoo et al.,?2007). It is estimated that about five percent of the total milk produced in India is converted into paneer (Makhal and Sen, 2001) which accounts for about 3, 00,000 tones of paneer annually valued at rupees 1050 crores. It is estimated that Annual production of paneer was 4,496 MT in year 2003-04 (Dairy India, 2007). Paneer has a fairly high level of fat and protein, low level of sugar and contains some important minerals (e.g. calcium) and vitamins. In general paneer contains 55-70 percent moisture, 22-27 percent fat , 17-18 percent protein , 2.0-2.5 percent lactose , and 1.5- 2.0 percent minerals (Kanawjia et al., 1990). According to Prevention of Food Adulteration Act (PFA ,1976), paneer shouldcontain not more than 70 percent moisture and its fat content should not be less than 50 percent of dry matter where as skim milk paneer should contain not more than 70 percent moisture and the fat content should not be more than 13 percent of dry matter (Kanawjia et al., 1990). Paneer should contain not more than 50,000 total bacterial counts, not more than 90 coliform counts and 250 yeast and mould count per gram of paneer (ISO: 1084, 1983). Paneer is recommended for diabetic patients, dental carries, growing children and pregnant women due to high calcium and protein (Chopra and Mamtani, 1995). However, short shelf life is one of the most serious problems faced in marketing and distribution of paneer to therestaurants and fast food centers, which consume a huge quantity of this product as a base material for patties, sandwiches, culinary dishes, etc. Paneer has a good market value, but it is not able to find its rightful place in Indian market due to its short life of about a week under refrigeration and one day at room temperature. Various chemical preservatives were tried by different scientists singly or in combination for the preservation of paneer and they succeeded in extending the shelf life to some extent.There is a long history of people safely consuming soya beans, dating back to the 11th century BC (3,000 years ago) in the eastern part of Northern China. The period from the first century AD to the 15th-16th century saw the introduction of soya beans in many parts of Asia, including Japan and India, and in 1765 the soya bean was introduced to the USA (JHCI, 2002). Since then, it has become an important part of the diets of many populations and in more recent years has found favour with vegetarians and vegans because of its many nutrients and health benefits. However, as the popularity of soya has grown, so has the number of critics questioning the benefits of this humble bean. Soybean was first domesticated in the eastern half of north China in the 11th century B.C. It was grown mainly for the seeds that were used for fresh, fermented, and dried food products (Whigham, 2004). The first introduction of soybean into the USA occurred when Henry Yonge, the Surveyor General of the Colony of Georgia, planted soybean on his farm at the request of Samuel Bowen in 1765 (Hymowitz, 2004). Bowen, a former seaman employed by the East India Company, brought soybean to Savannah from China via London. In 1766, Bowen planted soybean on his plantation “Greenwich” located at Thunderbolt, a few miles east of Savannah, Georgia (Hymowitz, 2004)2.2 Soyabean And Its Products2.2.1 Chemical Composition of SoyabeanTogether,?soybean oil?and protein content account for about 60 per cent of dry soybeans by weight (protein at 40 per cent and oil at 20 per cent). The remainder consists of 35 per cent carbohydrate and about 5 per cent?ash.Table 2.1 Soybean, mature seeds, raw Nutritional value per 100?gEnergy1,866?kJ (446?kcal)Carbohydrates30.16 gSugars7.33 gDietary fiber9.3 gFat19.94 g?saturated2.884 gmonounsaturated4.404 g?polyunsaturated11.255 gProtein36.49 g?Tryptophan0.591 g?Threonine1.766 g?Isoleucine1.971 g Leucine3.309 g?Lysine2.706 gMethionine0.547 gCystine0.655 gPhenylalanine2.122 g?Tyrosine1.539 g?Valine2.029 gArginine3.153 g?Histidine1.097 g?Alanine1.915 g?Aspartic acid5.112 g?Glutamic acid7.874 g?Glycine1.880 g?Proline2.379 g?Serine2.357 gWater8.54 gVitamin A?equiv.1 μg (0%)Thiamine (vit. B1)0.874 mg (76%)Riboflavin (vit. B2)0.87 mg (73%)Niacin (vit. B3)1.623 mg (11%)Pantothenic acid?(B5)0.793 mg (16%)Vitamin B60.377 mg (29%)Folate?(vit. B9)375 μg (94%)Vitamin B120 μg (0%)Choline115.9 mg (24%)Vitamin C6.0 mg (7%)Vitamin E0.85 mg (6%)Vitamin K47 μg (45%)Calcium277 mg (28%)Iron15.7 mg (121%)Magnesium280 mg (79%)Manganese2.517 mg (120%)Phosphorus704 mg (101%)Potassium1797 mg (38%)Sodium2 mg (0%)Zinc4.89 mg (51%)Source:?USDA Nutrient Databaseg – gram, mg - milligram, % - per cent 2.2.2NutritionalValueSoya (Glycine max) is a particularly good source of protein as it contains the eight essential amino acids which the human body needs. Soya milk and other soya products provide a rich source of polyunsaturated fatty acids (including the ‘good’ fats – omega-3) and are free of cholesterol. Compared to cow’s milk, soya milk contains lower levels of saturated fat and higher levels of unsaturated essential fatty acids, which can lower cholesterol levels.Soya products provide an excellent source of disease-busting antioxidants, B vitamins (including folate) and iron. Calcium-fortified soya products such as soya milk and tofu provide a valuable source of this important mineral without the saturated animal fat, animal protein (casein) and cholesterol found in dairy products. One serving of 200ml soya with added calcium and vitamins contains 30 per cent of the recommended daily amount (RDA) of calcium – equivalent to cow’s milk. It is also fortified with vitamin B12 and 200ml provides 100 per cent of the RDA of this important nutrient.Many soya foods also contain valuable fibre which is important for good bowel health and can also lower cholesterol. Soya foods, particularly those made from whole soya beans, offer a wide range of nutritional and health benefits.?Soy protein is a major component of the diet of food-producing animals and is increasingly important in the human diet. However, soy protein is not an ideal protein because it is deficient in the essential amino acid methionine. Methionine supplementation benefits soy infant formulas, but apparently not food intended for adults with an adequate nitrogen intake. Soy protein content of another essential amino acid, lysine, although higher than that of wheat proteins, is still lower than that of the milk protein casein. Adverse nutritional and other effects following consumption of raw soybean meal have been attributed to the presence of endogenous inhibitors of digestive enzymes and lectins and to poor digestibility. To improve the nutritional quality of soy foods, inhibitors and lectins are generally inactivated by heat treatment or eliminated by fractionation during food processing. Although lectins are heat-labile, the inhibitors are more heat-stable than the lectins. Most commercially heated meals retain up to 20 per cent of the Bowman-Birk (BBI) inhibitor of chymotrypsin and trypsin and the Kunitz inhibitor of trypsin (KTI). To enhance the value of soybeans in human nutrition and health, a better understanding is needed of the factors that impact the nutrition and health-promoting aspects of soy proteins. This paper discusses the composition in relation to properties of soy proteins. Also described are possible beneficial and adverse effects of soy-containing diets. The former include soy-induced lowering of cholesterol, anticarcinogenic effects of BBI, and protective effects against obesity, diabetes, irritants of the digestive tract, bone, and kidney diseases, whereas the latter include poor digestibility and allergy to soy proteins (Friedman and ?Brandon,(2001). Health Benefits2.2.3.1 Heart healthScientists agree that soya protein can promote heart health – a fact supported by dozens of controlled clinical trials. In 1995, a review published in the New England Journal of Medicine investigated the effect of soya protein on cholesterol levels (Anderson et al., 1995). In 34 of the 38 studies reviewed, replacing animal protein with soya protein decreased cholesterol levels (in the remaining four trials the subjects had low cholesterol levels to start with). The role of soya protein in heart health has since been widely accepted and approved by many different health bodies.The UK government’s Joint Health Claims Initiative (JHCI) offers market advice and a code of practice for both the UK food industry and consumers to ensure that health claims on foods are both scientifically truthful and legally acceptable. In 2002 the JHCI approved the health claim: “the inclusion of at least 25 grams soya protein per day as part of a diet low in saturated fat can help reduce blood cholesterol” (JHCI, 2002a).In 2005, a review of 23 trials, published in the American Journal of Clinical Nutrition, examined the cholesterol-lowering effects of soya protein containing isoflavones (a phytoestrogen or plant hormone). It confirmed that soya protein significantly lowers cholesterol levels but suggested that isoflavones alone did not (Zhan and Ho, 2005).Another study looked at the cholesterol-lowering effects of soya proteins in healthy young men, a sub-group that has been somewhat overlooked. It showed that soya protein, regardless of its isoflavone content, reduces cholesterol (McVeigh et al., 2006).These studies suggest that this effect involves a combination of factors in soya, including: isoflavones, soya protein peptides (small chains of amino acids – the building blocks of protein) and its amino acid content (the sequence of amino acids that make up soya protein and which may differ significantly to that of animal protein). These factors appear to work together to lower cholesterol and so reduce the risk of heart attack and stroke.Menopausal symptomsIn Japan, where soya consumption is higher than most other places in the world, the incidence of menopausal hot flushes is much lower than in the West. However, within Japan there are variations. A six-year study of over 1,000 Japanese women showed that those who consumed the most soya foods had less than half the number of hot flushes compared to women consuming the least amount of soya (Nagata, 2001).There are also many studies showing that supplementing the diet with soya foods or soya protein isolates can substantially reduce the frequency or severity of hot flushes and other menopausal symptoms in some women.It was shown in 1995 that the incidence of hot flushes was reduced by 40 per cent in women consuming 45 grams of soya flour for 12 weeks compared to a 25 per cent reduction among those consuming wheat flour (Murkies et al., 1995). In 1997, another study found that hot flushes were reduced by 54 per cent in women consuming tofu, soya milk and miso, compared to a 35 per cent reduction in the control group (Brzezinski et al., 1997). A third study, in 1998, showed that 60 grams of an isolated soya protein supplement containing 76mg total isoflavones significantly reduced hot flushes by 45 per cent compared to a 35 per cent reduction in the control group (Albertazzi et al., 1998).More recently, Faure et al.,2002 showed that a soya isoflavone extract reduced the incidence of hot flushes by 61 per cent after 12 weeks compared to a 21 per cent reduction in the control group (Faure et al., 2002). Not surprisingly, some of these researchers suggest that soya protein may provide an alternative to hormone replacement therapy (HRT) for reducing menopausal symptoms.The safety of specific amounts of isoflavones has not yet been established and so it is better to obtain isoflavones from whole soya foods rather than as isolates extracted from soya.BonehealthThe first published human trial investigating the effects of soya foods on bone health and osteoporosis indicated that soya protein may be effective in reducing the risk of this debilitating disease (Potter et al., 1998). It was found that supplementing the diet of postmenopausal women with 40 grams of soya protein a day (containing 90mg of isoflavones) for six months significantly increased both the bone mineral content and density of the lumbar spine.These findings were supported by a later study that looked at the effect of 80mg of soya isoflavones a day on bone density (Alekel et al., 2000). This also showed that isoflavones reduced bone loss from the lumbar spine of women who may otherwise be expected to lose two to three per cent of bone per year. In 2003, a review of the published research concluded that diets rich in phytoestrogens (and therefore isoflavones) were likely to benefit bone health (Setchell and Lydeking-Olsen, 2003).Studies from Japan and China show that postmenopausal women with the highest intake of isoflavone-rich soya foods have the highest bone mineral density in the lumbar spine compared with women with low intakes of soya (Somekawa et al., 2001 and Mei et al., 2001). A more recent study of Chinese women, published in the European Journal of Nutrition, confirmed a strong link between soya isoflavones and a reduction of bone loss in postmenopausal women who were not obese (Ye et al., 2006).In this study, a total of 90 women aged 45-60 years were randomly assigned to one of three treatment groups – taking daily doses of 0 (placebo), 84 and 126mg of isoflavones for six months. Bone mineral density of the spine and hip was measured at the start of the trial and when it ended after six months. It showed that as the intake of soya isoflavones increased, the degree of bone loss decreased, possibly caused by soya isoflavones inhibiting the process where minerals are leached from the bone (bone resorption).?ThecalciumparadoxAs food is digested, acids are released into the blood in varying proportions, depending upon the food. The body tries to neutralise this acid by drawing calcium from the bones and the calcium is then excreted in the urine (the calciuric response). Animal protein from cow’s milk and dairy products, as well as meat, fish and eggs, has a particularly strong acidic effect compared to plant protein because of the sulphur-containing amino acids it contains. As increasing amounts of animal products are eaten, the sulphur content of the diet increases and so does the level of calcium in the urine. This increases calcium loss and may be a risk factor for the development of osteoporosis.This so-called calciuric response is well-documented in the scientific literature – the more cow’s milk, meat, fish and eggs you eat, the more calcium is lost from the body. The irony is that cow’s milk is promoted as the best source of calcium but it is also a powerful source of calcium loss. The World Health Organisation (WHO) calls this the ‘calcium paradox’, where the damaging effects of animal (but not vegetable) protein may outweigh the positive effects of the calcium it contains (WHO, 2004). In other words, you’re better off getting calcium from plant foods such as cereals, nuts, seeds and pulses, which don’t cause such huge losses of calcium from the bones.The evidence indicates that soya isoflavones are good for bone health. Importantly, some researchers suggest that soya foods could provide an alternative treatment to HRT for women at risk of osteoporosis and this may offer a cheap, drug-free prevention for this debilitating condition (Lydeking Olsen et al., 2004).Cancer riskThe low rates of breast and prostate cancers seen in Asian countries have encouraged scientists to investigate the role of soya foods on these and other hormone-related cancers.Breast cancerThere is some evidence that soya intake during adolescence may reduce the risk of breast cancer later in life. The Shanghai Breast Cancer Study investigated 1,400 breast cancer cases in China (Shu et al., 2001) and found that women who consumed the most soya as teenagers had half the risk of breast cancer as adults.A year later, scientists investigated the link between adolescent soya intake and breast cancer in Asian-American women (Wu et al., 2002). They found that women who consumed soya at least once a week during adolescence had a significantly reduced risk of breast cancer. Over 1,000 women, including 501 breast cancer patients, were asked how often they ate soya foods such as tofu, soya milk and miso.Results showed that those who were high soya consumers as both adolescents and adults had a 47 per cent reduction in risk of breast cancer. Those who ate little soya as adults but had eaten it regularly during adolescence showed a 23 per cent reduction in risk. Women who were low consumers during adolescence and high consumers during adulthood showed little reduction in risk. These studies suggest that high soya intake during adolescence reduces breast cancer risk and the risk continues to fall if people continue to eat soya as an adult. Drawing the evidence together, Trock et al, 2006, performed a review of 18 studies on soya exposure and breast cancer risk published between 1978 and 2004. Results showed a modest association between a high soya intake and a reduced breast cancer risk. The low rate of breast cancer in Japan and the high rate of survival amongst those who are affected is often used to promote soya foods as being beneficial – or at least, not being harmful – for breast cancer patients. Some researchers, however, are cautious and think that even the small oestrogen-like effect of soya foods may be detrimental for women who have gone through the menopause, whose natural oestrogen levels have dropped and who have been diagnosed with oestrogen-receptor positive (hormone-sensitive) breast cancer (PCRM, 2002).Their concern is that the weak oestrogen activity of soya isoflavones may stimulate the growth of tumours which are sensitive to oestrogen. This is not a concern for premenopausal women, who have much higher levels of oestrogens which are many times more potent than phytoestrogens. These concerns are based largely on the results of in vitro (test tube) and animal studies but as these have produced mixed results their relevance to human breast cancer patients remains unclear. On the whole, the evidence suggests that consuming moderate amounts of soya foods is much more likely to benefit health rather than harm it, both in terms of breast cancer risk and other chronic diseases.Prostate cancer?Prostate cancer rates also vary widely around the world, tending to peak in developed, wealthy countries. Japan is the exception, where prostate cancer rates are surprisingly low, despite its high standard of living, and some evidence suggests that soya may be responsible.There are a limited number of studies investigating the role of soya in prostate cancer but in 1998 a large-scale study in 59 countries found that, overall, soya products were significantly protective (Hebert et al., 1998). It showed that death from prostate cancer increases with an affluent diet, specifically dairy milk, meat and poultry, while a diet based on cereals, soya beans, nuts and oilseeds reduces the death risk. Cognitive effectsSeveral studies indicate that soya intake may improve both short-term and long-term memory, mental flexibility and planning. Researchers at the Centre for Neuroscience at King’s College, London, investigated the effects of a high soya diet (100mg isoflavones per day) compared to a low soya diet (0.5mg isoflavones per day) in student volunteers (File et al., 2001).After just 10 weeks, those receiving the high soya diet exhibited significant improvements in short-term and long-term memory and in mental flexibility.A second study investigated the effects of soya isoflavones (60mg per day) in the cognitive ability of a group of postmenopausal women aged between 50 and 65 years of age (Duffy et al., 2003). After 12 weeks, significant improvements were seen in the soya group, including their recall of pictures and in a sustained attention task. Although the groups did not differ in their ability to learn rules, the soya group showed significantly greater improvements in tests learning rule reversals and planning. Its conclusion was that significant cognitive improvements in postmenopausal women can be gained from 12 weeks of consumption of soya isoflavones.Another study investigated the effects of soya supplements (60mg isoflavones per day) on postmenopausal women and found that after just six weeks, the soya group showed a greater improvement of nonverbal (identifying objects, for example) short-term memory than the placebo group (File et al., 2005). Those on soya produced significantly better performances in mental flexibility and planning ability. There were, however, no improvements in long-term memory, category generation or sustained attention.These studies show that soya isoflavones can have a significant positive effect on cognitive ability but the benefits may be restricted to people under the age of 65 (Kritz-Silverstein et al., 2003; Kreijkamp-Kaspers et al., 2004). Certainly, further investigation is warranted.2.2.4 Uses of SoyabeanApproximately 85 per cent of the world's soybean crop is processed into soybean meal and vegetable oil (canola oil).?Soybeans can be broadly classified as "vegetable" (garden) or field (oil) types. Vegetable types cook more easily, have a mild, nutty flavor, better texture, are larger in size, higher in protein, and lower in oil than field types.?Tofu?and?soy milk?producers prefer the higher protein cultivars bred from vegetable soybeans originally brought to the United States in the late 1930s. The "garden" cultivars are generally not suitable for mechanical combine harvesting because there is a tendency for the pods to shatter upon reaching maturity.Among the legumes, the soybean, also classed as an oilseed, is preeminent for its high (38–45 per cent)?protein?content as well as its high (approximately 20 per cent) oil content. Soybeans are the second-most valuable agricultural export in the United States behind corn. The bulk of the soybean crop is grown for oil production, with the high-protein defatted and "toasted" soy meal used as livestock feed. A smaller percentage of soybeans are used directly for human consumption.2.2.4.1 OilSoybean seed contains about 19 per cent oil. To extract soybean oil from seed, the soybeans are cracked, adjusted for moisture content, rolled into flakes and solvent-extracted with commercial hexane. The oil is then refined, blended for different applications, and sometimes hydrogenated. Soybean oils, both liquid and partially hydrogenated, are exported abroad, sold as "vegetable oil", or end up in a wide variety of processed foods. The remaining soybean meal is used mainly as animal feed Lusas and Riaz, (1995). 2.2.4.2 MealSoybean meal is the material remaining after solvent extraction of oil from soybean flakes, with a 50 per cent?soy protein?content. The meal is 'toasted' (a?misnomer?because the heat treatment is with moist steam) and ground in a?hammer mill. Soybean meal is an essential element of the American production method of growing farm animals, such as poultry and?swine, on an industrial scale that began in the 1930s; and more recently the?aquaculture?of?catfish. Ninety-eight percent of the U.S. soybean crop is used for livestock feed ?( HYPERLINK "" "Soybean".?Encyclopedia Britannica Online?) Soybean meal is also used in lower-end?dog foods (Lusas and Riaz, (1995)).2.2.4.3 FlourSoy flour refers to soybeans ground finely enough to pass through a 100-mesh or smaller screen where special care was taken during desolventizing (not toasted) to minimize?denaturation?of the protein to retain a high?protein dispersibility index, for uses such as?food extrusion?of?textured vegetable protein. It is the starting material for production of soy concentrate and soy protein isolate.Soy flour is made by roasting the soybean, removing the coat, and grinding into a flour. Soy flour is manufactured with different fat levels ( Lim, 2012).?Alternatively,?raw?soy flour omits the roasting step.Defatted soy flour is obtained from?solvent?extracted?flakes, and contains less than 1 per cent oil. ( Lim, 2012).?"Natural or full-fat soy flour is made from unextracted, dehulled beans, and contains about 18 per cent to 20 per cent oil. ( Lim, 2012).??Its high oil content requires the use of a specialized Alpine Fine Impact Mill to grind rather than the usual?hammer mill. Full-fat soy flour has a lower protein concentration than defatted flour.Low-fat soy flour is made by adding some oil back into defatted soy flour. Fat levels range from 4.5 per cent to 9 per cent.( Lim, 2012).?High-fat soy flour can also be produced by adding back soybean oil to defatted flour at the level of 15 per cent.Soy lecithin?can be added (up to 15 per cent) to soy flour to make lecithinated soy flour. It increases dispersibility and gives it emulsifying properties. ( Lim, 2012).?Soy flour has 50 per cent protein and 5 per cent fiber. It has higher levels of protein, thiamine, riboflavin, phosphorus, calcium, and iron than?wheat flour. It does not containgluten. ( Lim, 2012).??As a result,?yeast-raised?breads?made with soy flour are dense in texture. Among many uses, soy flour thickens sauces, prevents?staling?in baked food, and reduces oil absorption during frying. Baking food with soy flour gives it tenderness, moistness, a rich color, and a fine texture. ( Lim, 2012).?Soy grits are similar to soy flour except the soybeans have been toasted and cracked into coarse pieces.( HYPERLINK "" \o "Kinako" Kinako?is a soy flour used in?Japanese cuisine).2.2.4.4 Infant formulaSoy-based?infant formula?(SBIF) is sometimes given to infants who are not being strictly breastfed; it can be useful for infants who are either allergic to pasteurized cow milk proteins or who are being fed a?vegan?diet. It is sold in powdered, ready-to-feed, and concentrated liquid forms.Some reviews have expressed the opinion that more research is needed to determine what effect the phytoestrogens in soybeans may have on infants.( Miniello et al., 2003)?Diverse studies have concluded there are no adverse effects in human growth, development, or reproduction as a result of the consumption of soy-based infant formula. Giampietro et,al.,(2004) Strom et,al., (2001), and Merritt and Belinda (2004). One of these studies, published in the?Journal of Nutrition, Merritt and Belinda (2004) concludes that there are:... no clinical concerns with respect to nutritional adequacy, sexual development, neurobehavioral development, immune development, or thyroid disease. SBIFs provide complete nutrition that adequately supports normal infant growth and development. FDA has accepted SBIFs as safe for use as the sole source of nutrition.2.2.4.5 Meat and dairy alternatives and extendersSoybeans can be processed to produce a texture and appearance similar to many other foods. For example, soybeans are the primary ingredient in many?dairy product?substitutes (e.g.,?soy milk,?margarine,?soy ice cream,?soy yogurt,?soy cheese, and soy cream cheese) and meat alternatives (e.g.?veggie burgers). These substitutes are readily available in most supermarkets. Soy milk does not naturally contain significant amounts of digestible?calcium. Many manufacturers of soy milk sell calcium-enriched products, as well. Soy is also used in?tempeh: the beans (sometimes mixed with grain) are fermented into a solid cake.Soy products also are used as a low-cost substitute in meat and poultry products. Hoogenkamp, (2005) and Endres (2001)?Food service, retail and institutional (primarily school lunch and correctional) facilities regularly use such "extended" products. Extension may result in diminished flavor, but fat and cholesterol are reduced. Vitamin and mineral fortification can be used to make soy products nutritionally equivalent to animal protein; the protein quality is already roughly equivalent. The soy-based meat substitute?textured vegetable protein?has been used for more than 50?years as a way of inexpensively extending?ground beef?without reducing its nutritional value. Riaz,( 2006) and Liu, (1997)2.2.4.6 Cattle feedCattle are often fed soy.?Spring grasses?are rich in omega-3 fatty acids, whereas soy is predominantly omega-6.2.2.5Soyabean Products2.2.5.1 Soy milkSoy milk?(soymilk?or?soya milk) is a?plant milk?produced by soaking dry?soybeans?and grinding them with water.2.2.5.1.1 Health and nutritionSoy milk is a?complete protein?and has about the same amount of?protein?as cow's milk; it can replace animal protein and other sources of?dietary fiber, vitamins and minerals. (Sacks et al., 2006)It contains little digestible?calcium?because it is bound to the bean'spulp, which is indigestible by humans. To counter this, manufacturers enrich their products with?calcium carbonate.?Unlike cow's milk, soy milk has little?saturated fat?and no?cholesterol.Soy products contain?sucrose?as the basic?disaccharide, which breaks down into?glucose?and?fructose. Since soy doesn't contain?galactose, a product of lactose breakdown, soy-based infant formulas can safely replace breast milk in children withgalactosemia.?Like lactose-free cow's milk, soymilk contains no lactose, which makes it a good alternative for?lactose-intolerant?people.It has been suggested that soy consumption is associated with a reduction in?low-density lipoprotein?("bad cholesterol") and triglycerides Anderson et al., (1995)?Research has refuted claims that soy affects bone mineral density. (Darling et al., 2009)?Research has found no link between soy and increased estrogen levels in men, although studies thus far have been limited in duration ( Messina, (2010). For people who suffer from?gout,?purine?in soy can make the condition worse. The?US National Institutes of Health?(NIH) recommends gout sufferers limit consumption of soy products (although also suggests that soy may have health benefits by reducing the risk for heart disease).Table 2.2 - Soy milk, unsweetened, with added calcium and vitamins Nutritional value per 100?gEnergy138?kJ (33?kcal)Carbohydrates1.74 g?Sugars0.41 gDietary fiber0.5 gFat1.61 gsaturated0.206 gmonounsaturated0.392 g ?polyunsaturated1.009 gProtein2.86 gWater93.14 gThiamine (vit. B1)0.154 mg (13%)Riboflavin (vit. B2)0.207 mg (17%)Niacin (vit. B3)0.165 mg (1%)Pantothenic acid?(B5)0.082 mg (2%)Vitamin B60.049 mg (4%)Folate?(vit. B9)1.5 μg (0%)Vitamin B121.11 μg (46%)Vitamin C0 mg (0%)Vitamin D1.2 μg (8%)Calcium124 mg (12%)Iron0.46 mg (4%)Magnesium16 mg (5%)Phosphorus32 mg (5%)Potassium120 mg (3%)Sodium37 mg (2%)Source:?USDA Nutrient Databaseg – gram, mg - milligram, % - per cent , vit.-vitamin2.2.5.2 TofuTofu, also known as?bean curd, is a food made by?coagulating?soy milk?and then pressing the resulting curds into soft white blocks. Tofu is made by coagulating?soy milk?and pressing the resulting curds. Although pre-made soy milk may be used, some tofu producers begin by making their own soy milk, which is produced by soaking, grinding, boiling and straining dried (or, less commonly, fresh)?soybeans.2.2.5.2.1 Nutrition and health informationProteinTofu is relatively high in?protein, about 10.7 per cent for firm tofu and 5.3 per cent for soft "silken" tofu with about 5 per cent and 2 per cent fat?respectively as a percentage of weight (Ang et al., 1999).In 1995, a report from the?University of Kentucky, financed by?The Solae Company?St. Louis, Missouri?(the PTI division of DuPont), concluded that soy protein is correlated with significant decreases in serum cholesterol, Low Density Lipoprotein?LDL?(bad cholesterol) and triglyceride concentrations (Anderson et al., 1995).?However, High Density Lipoprotein?HDL?(good cholesterol) did not increase. Soy?phytoestrogens?(isoflavones: genistein and daidzein) absorbed onto the soy protein were suggested as the agent reducing serum cholesterol levels. On the basis of this research, PTI, in 1998, filed a petition with?Food and Drug Administration?for a health claim that soy protein may reduce cholesterol and the risk of heart disease.The FDA granted this health claim for soy: "25 grams of soy protein a day, as part of a diet low in saturated fat and cholesterol, may reduce the risk of heart disease."?For reference, 100?grams of firm tofu coagulated with calcium sulfate contains 8.19?grams of soy protein.?In January 2006, an?American Heart Association?review (in the journal?Circulation) of a decade-long study of soy protein benefits showed only a minimal decrease in cholesterol levels, but it compared favorably against animal protein sources Sacks et al.,2006.TempehTempeh?, is a traditional soy product originally from Indonesia. It is made by a natural culturing and controlled?fermentation?process that binds?soybeans?into a cake form, similar to a very firm vegetarian?burger patty. 2.2.5.3.1NutritionThe soy carbohydrates in tempeh become more digestible as a result of the fermentation process. In particular, the?oligosaccharides?associated with gas andindigestion?are greatly reduced by the?Rhizopus?culture. In traditional tempeh-making shops, the starter culture often contains beneficial bacteria that produce vitamins such as?B12 ( Liem et al.,1977 and Delores?1987), though it is uncertain whether this B12?is always present and?bioavailable (Allison, 2000).?In western countries, it is more common to use a pure culture containing only?Rhizopus oligosporus, which makes very little B12?and could be missing?Klebsiella pneumoniae, which has been shown to produce significant levels of B12?analogs in tempeh when present. Whether these analogs are true, bioavailable B12, has not been thoroughly studied yet.?The fermentation process also reduces the?phytic acid?in soy,which in turn allows the body to absorb the minerals that soy provides.2.3 Chemical Composition of PaneerPaneer is a rich source of animal protein available at a comparatively lower cost and forms an important source of animal protein for vegetarians. Over and above its high protein content and digestibility, the biological value of protein in paneer is in the range of 80 to 86 (Shrivastava and Goyal, 2007). In addition, paneer is a valuable source of fat, vitamins and minerals like calcium and phosphorus (khan and pal., 2011).The chemical composition of paneer given in literature has wide variation. Paneer contains 58.6 percent moisture,16.45 percent fat,19.94 percent protein, 2.51 percent lactose, 1.5-2.0 percent minerals (Reeta and Kumar, 2013).These differences in paneer composition might be attributed to the differences in initial composition of milk, method of manufacture and losses of milk solids in whey (Rao et al., 1992).Wide variations have been reported in moisture, fat and mineral content of market samples of paneer. The proximate composition of paneer samples as reported by various researchers is given in Table 2.3 Table 2.3 Proximate composition of paneerMilk fat (per cent) Constituents in Paneer (per cent )ReferencesMoistureFatProteinLactoseAsh Buffalo milk3.556.9918.1018.43----Chawla et al.,?(1987)?5.056.7722.30--------Bhattacharya et al.,?(1971)?5.056.4322.50--------Shukla et al., (1984)5.555.1923.8017.99----Chawla et al., (1987)5.850.7227.1317.991.87Rajorhia et al., (1984)5.854.1023.5018.201.80Sachdeva and Singh (1987)5.951.1226.8617.382.00Pal and garg (1989)5.955.1223.4719.922.43Pal et al., (1999)51.9326.1715.742.711.80Pal and Yadav (1991)53.5124.1216.442.601.88Pal and Kapoor (2000)47.0523.0019.77----2.75Masud et al., (2007)58.616.4519.942.51----Reeta and kumar (2013)The nutritional and textural properties of low fat paneer using soy protein isolate (SPI) as fat replacer was investigated. The physico-chemical and sensory characteristics of 4 types of paneer made of low-fat milk (3 per cent milk fat (MF) and 10 per cent solids-not-fat (SNF)) and SPI of 0 (T1), 0.1 (T2), 0.2 (T3) and 0.3 per cent SPI (T4) were compared with high fat paneer (TC) made of high fat milk (6 per cent MF and 9 per cent SNF). CaCl2 (0.2 per cent, w/v) was used as coagulant at 75?±?1°C. Increased level of SPI in paneer increased yield, protein, ash, moisture content and decreased fat, moisture protein ratio, lactose and calorie contents. Titratable acidity and pH varied in narrow range (kumar etal., 2011).2.4 Legal Standard of Paneer:According to food safety and standards (food products standards and food additives) regulations, (2011), paneer should not contain more than 70 percent moisture and its fat content should not be more than 15 percent of the dry matter. This Indian Standard was adopted by the Bureau of Indian Standards, after the draft finalized by the Food Analysis and Nutrition Sectional Committee had been approved by the Food and Agriculture Division Council.Paneer and Chhana are Indian varieties of fresh cheese, the former being used in cooked vegetable dishes and other culinary items, whereas the latter for preparation of certain sweets. Selection of these products for their respective use is greatly influenced by their sensory attributes. This standard is complementary to IS5162 :1980 ‘Specification for Chhana @-st revision)’ and IS 10484:1983 ‘Specification for Paneer’.Statistical Evaluation of ResultsTable 1 Evaluation Card for Paneer/Chhana(Clauses 7.3 and 7.4))Name --------------- Date-----------------BatchorcodeNo.------------- Time--------------Section AScore the sample using the guideline given in Section B for deducting for perceived defects. S1 Characteristics Maximum Score Sample Score i) Flavour 50 ii) Body and texture 35iii) Colour and appearance 10iv) Package 5Section B S1 Characteristics Defects Degree of DefectsNo. Slight Definite Pronounced i) Flavour Bitter, foreign, feedy/weedy, 8 13 21 flat, musty, putrid, cheesy, smoky, stale, yeasty ii) Body and texture Crumbly hard mealy, 3 5 9 coarse open pasty, rubbery, chewy, weakiii) Colour and Dull, dry, mouldy, foreign material, 1 2 4 appearance visible dirtiv) Package Damaged soiled, greasy 1 2 32.5 Raw materials for paneerThe main raw material which is used for preparation of paneer is milk. Now-a-days some additives are also used to improve the quality characteristics of paneer.2.5.1 Type of milkVarious types of milk have been used for the manufacture of paneer. The quality of paneer is determined by the quality of milk from which it is produced (Nayak and Bector?1998).2.5.1.1Buffalo milkFor making good quality paneer, buffalo milk is considered more suitable than cow milk . Ghodekar (1989) reported that higher amounts of casein and minerals (calcium, phosphorus) were responsible for imparting firm and rubbery body to buffalo milk paneer. Fat globules and casein micelles of bigger size and higher concentration of fat, casein, calcium, phosphorus and lower voluminosity and solvation properties of casein micelles in buffalo milk compared to cow milk makes it better suited to paneer making with spongy character .Masud (2002) advocated use of buffalo milk having 6 per cent fat for preparation of best quality paneer. Several workers recommended buffalo milk standardized to 5–6 per cent fat for paneer manufacture (Kumar et al.?2008a).2.5.1.2 Cow milkGood quality paneer can be obtained from cow milk using certain modifications in the manufacturing process or through use of additives (Jadhavar et al.?2009a). Vishweshwaraiah and Anantakrishnan (1986) reported that paneer obtained from cow milk standardized to 4.5 per cent fat conformed to the PFA standards. Pruthi and Koul (1989) also found that paneer made from crossbred cows (HF x Sahiwal) milk having 3.7 per cent fat and 8.25–8.42 per cent SNF conformed to the PFA standards. However, cow milk yields inferior quality paneer especially in sensory characteristics compared to buffalo milk. Such effect could be ascribed to different make-up of casein micelles and lower protein and calcium contents in cow milk compared to buffalo milk.2.5.1.3 Mixed milkMixed cow and buffalo milk (1: 1) with 5 per cent fat yielded a superior paneer than cow milk alone (Shukla et al.,?1984). On the other hand, Sachdeva et al.,?(1985) suggested substitution of one third of buffalo milk with cow milk without any adverse effect on the sensory quality of paneer.. Pal and Yadav (1991) suggested use of buffalo: cow milk (1:1) with fat level of 3.5 per cent for production of low-fat paneer. However, Chavan et al.,?(2007) recommended addition of 20 parts of buffalo skim milk to 80 parts of cow whole milk for production of better quality paneer.2.5.1.4 Goat milk/sheep milkUse of goat milk resulted in paneer that lacked compactness (Shukla et al.,?1988). Prasad et al.,?(1990) made paneer from goat milk with acceptable characteristics and without any goaty odour. Agnihotri and Pal (1996) prepared good quality creamy white paneer, free from ‘goaty’ smell or salty taste, from Barbari goat milk with 4.86 per cent fat and 8.96 per cent SNF employing coagulation temperature of 87–88?°C using 0.15 per cent citric acid. Sheep milk could be used to manufacture paneer which resembled buffalo milk paneer (Kale et al.,?2008). Pal et al.,?(2008) standardized the processing variables (heat treatment of 90?°C, coagulation temperature of 90?°C and coagulant strength of 2 per cent citric acid) for the manufacture of paneer from ewe’s milk with 6.94 per cent fat.2.5.1.5 Low fat milk (Low fat paneer)Acceptable quality low fat paneer with 42 per cent fat on dry matter (FDM) was made from buffalo milk standardized to 3.5 per cent fat. Cow or buffalo milk with fat levels lower than 3.5 per cent resulted in product with unacceptable flavour (not having typical flavour) and body and texture (lacked softness) (Chawla et al.,1985,?1987 Arya and Bhaik?1992). Sanyal and Yadav (2000a,?b) proposed certain modifications like coagulation temperature of only 60?°C and addition of cultured skim milk (2.5 per cent) and use of additives like NaCl (0.25 per cent) prior to coagulation of buffalo milk (2 per cent fat, 9 per cent SNF) to obtain reduced fat paneer (30 per cent FDM) with acceptable quality. Good quality paneer can be made from milk standardized to even 3.5 per cent fat without resorting to addition of any additives (Chandan?2007b).Low fat paneer of acceptable quality can be produced from cow milk standardized to a fat content of 3.5 per cent (Vishweshwaraiah and Anantakrishnan?1986). Aneja (2007) and Chandan (2007a) reported that acceptable quality paneer with 24% per cent FDM is available in western countries.2.5.1.6 Dried milkChawla et al.,?(1987) used buffalo milk fortified with not fat dry milk (NFDM) for paneer manufacture to increase its yield to 24.4 per cent from 18.3 per cent. Nakazawa et al.,?(1989) used unfermented reconstituted skim milk and fruit juices for manufacture of ‘fruit paneer’. Singh and Kanawjia (1991) standardized the process for production of paneer from recombined milk using cow skim milk powder and butter oil. They recommended addition of 0.15 per cent calcium chloride to recombined milk for paneer manufacture. Singh and Kanawjia (1992) manufactured paneer from buffalo whole milk powder reconstituted to different levels of total solids (15–25 per cent TS) and concluded that paneer obtained from reconstituted whole milk with 15 per cent TS was the best in terms of yield, TS recovery and sensory characteristics of paneer.2.5.2 AdditivesVarious types of food grade additives have been incorporated into the milk during paneer production in order to improve a few parameters such as yield, sensory characteristics and shelf life as well as to reduce the cost of production.2.5.2.1 Calcium compoundsCalcium helps in building the cross linkages during the formation of curd and thus helps in increasing the recovery of milk solids, yield and improves body and texture and overall acceptability scores of paneer. Cow milk paneer has softer body than buffalo milk paneer since cow milk is lower in calcium content. In order to produce good quality cow milk paneer, calcium chloride at the rate of 0.08–0.15 per cent was used to get better quality paneer (Sachdeva et al.?1991; Arya and Bhaik?1992). Singh and Kanawjia (1988) observed that use of 0.1 per cent CaCl2?to milk prior to coagulation increased total solids recovery, yield and all the sensory characteristics. Singh and Kanawjia (1991) recommended addition of 0.15 per cent CaCl2 for paneer to be made from recombined cow milk. A combination of di sodium phosphate and calcium chloride has been used in western countries for preparation of low-fat cheese. It increased the softness and elasticity of curd due to the formation of colloidal calcium phosphate (Teknotext?1995). Calcium phosphate addition to the milk can help in the coagulation of whey proteins thereby increasing the yield of curd (Dybing and Smith?1998). Calcium ions help in neutralization of milk protein charges and induce aggregation and precipitation of casein. The calcium phosphate microcrystals formed when using phosphate salt in milk, provides a substrate for protein adsorption, with subsequent cross-binding of the casein micelles to form sturdy aggregates of co-precipitated calcium phosphate and casein (Guo et al.,?2003). Hill et al.,?(1982) recommended use of high temperature and CaCl2?for getting better yield through co-precipitation of casein and whey proteins. Arora et al., (1996) observed that addition of CaCl2?increased fat, protein, TS, pH and TS recovery and thus yield of paneer made from diluted milk which is most commonly encountered in un-organized sector. Kanawjia and Rizvi (2003) recommended use of 0.15 per cent CaCl2?to microfiltered milk retentate prior to acidification in paneer manufacture.2.2.5.2 Herb impregnationKaur et al.,?(2003) and Bajwa et al., (2005) reported that incorporation of coriander and mint at level of 10 per cent by weight in paneer improved the overall acceptability score and yield of product.2.2.5.3 Vegetable oilLow-cost, low-calorie, health promoting paneer can be made using skim milk added with vegetable oil. Hydrogenated vegetable oil and groundnut oil proved to be better than soya oil; the later product was unacceptable. Higher fat level (i.e. 5.5 vs. 3.5 per cent) resulted in better acceptability of the resulting filled-paneer (Roy and Singh?1999). Kanawjia and Singh (2000) found that paneer obtained from skim milk and vanaspati (HVO) resulted in quite acceptable paneer.2.2.5.4 Coconut milkVenkateswarlu et al., (2003) obtained that addition of 10 per cent of coconut milk (25 per cent fat) to skim milk resulted in highly acceptable quality paneer.2.2.5.5 Protein enrichmentThe protein content of paneer can be increased using non-conventional low cost proteins to improve its nutritional value and to achieve economy. They were also used to improve the sensory characteristics of low fat paneer thus making it ideally suited for dietary management of consumers suffering from protein malnutrition and coronary complications. Incorporation of whey solids raised the yield of paneer by 20.9 per cent, though recovery of milk solids decreased (Singh et al.,?1991b). Kanawjia and Singh (2000) found that incorporation of low cost calcium salt of groundnut protein isolates to skim milk and vegetable fat mixture produced nutritionally superior paneer than the conventional paneer. Salve et al.,(2007) advocated use of 2.0 per cent whey protein concentrate (72 per cent protein) to buffalo milk with only 4 per cent fat for improving the quality attributes of low fat paneer (<50 per cent FDM). Sivakumar et al.,(2007) found that the inclusion of 0.2 per cent soy protein isolate (SPI) to buffalo milk with 4 per cent fat increased the yield of low-fat paneer containing around 40 per cent FDM, at the same time improving its texture, juiciness and overall acceptability when compared to paneer devoid of added SPI. SPI was mainly added as a fat replacer. Paneer was prepared by incorporating 3, 5 or 10 per cent of either soymilk, buttermilk or skimmed milk to buffalo milk. In all cases, oil uptake on frying with groundnut oil was not appreciably different from that of the control up to the 5 per cent level of incorporation but, at 10 per cent, the oil uptake was higher than the control. Paneers with buttermilk and soy milk had softer textures than the control (Sharma et al.,?1998).2.2.5.6 ButtermilkButtermilk, a by-product of butter industry which can be utilized in the manufacture of paneer. Buttermilk is of two types, sweet and sour buttermilk. Pal and Garg (1989) proposed two additional manufacturing steps for the manufacture of paneer from sour buttermilk, i.e., neutralization of sour butter milk to 0.15 per cent titratable acidity by sodium bicarbonate and washing of curd with hot water (72?°C) before pressing to mitigate the problems of self-coagulation of milk during heating, development of acidic smell, sour taste and grainy texture in paneer. Shoekand et al., (1990) found that fresh cream buttermilk can be used for standardizing the buffalo milk upto casein to fat ratio of 0.70 (4.47 per cent fat, 3.97 per cent protein) without affecting its quality and acceptability.2.2.5.7 Sodium compoundsChawla et al., (1987) found that addition of 0.1 per cent sodium citrate or 0.5 per cent sodium chloride to milk helped in increasing the moisture content of low fat paneer and thereby yield of the product. Yadav et al., (1994) found that use of common salt (0.5per cent) in milk led to an improvement in the body and texture characteristics and yield of low-fat buffalo milk paneer, besides enhancing its shelf life (i.e. 2?days at room temperature). Incorporation of small amounts of cultured skim milk (2.5 per cent) and salt (0.05per cent) helped to improve the moisture, yield and sensory quality of reduced-fat paneer (Sanyal and Yadav?2000a,?b; Mendiratta et al.,?2004). Kaur et al., (2003) found that dipping of paneer blocks in brine solution (1–5 per cent) decreased its moisture and water activity while the flavour and overall acceptability were enhanced when 3 per cent brine solution was used.2.2.5.8 Emulsifying saltsPal and Kapoor (2000) used various emulsifying salts namely monosodium phosphate, disodium phosphate, trisodium citrate, tetrasodium pyrophosphate, sodium tripolyphosphate and sodium hexametaphosphate (1–3 per cent) for preparation of processed paneer. None of the emulsifying salts had any significant influence on the chemical composition in the product. Although, such salts are not permitted by PFA in India but they are permitted by codex alimentarius in directly acidified cheeses and other similar type of dairy products. These were used to make processed paneer which could be similar to processed cheese, which is very popular in western countries. But the addition of such salts proved to be of no significance in paneer.2.2.5.9 HydrocolloidsSachdeva and Singh (1988a) observed increase in moisture retention and thus yield of paneer when sodium alginate, carrageenan or pre-gelatinized starch at levels of 0.10, 0.15 and 0.15 per cent respectively were used as hydrocolloids. Roy and Singh (1994) reported that addition of 0.1 per cent pre-gelatinized starch coupled with use of higher coagulation temperature (90°C) improved the body and texture as well as yield of filled paneer; sodium alginate at 0.1 per cent level did not exert any beneficial effect. Sharma et al., (1999) used carboxymethyl cellulose (CMC) as an additive for oil reduction in deep-fat fried paneer and found that it appreciably reduced the oil uptake by paneer on frying.2.2.5.10 Dietary fibreThe use of soy fibre and inulin (1 per cent) resulted in an improvement in the sensory, rheological and nutritional properties of low-fat paneer which otherwise was criticized for having hard, coarse, rubbery and chewy body and texture (Kanawjia and Khurana?2006). Kantha and Kanawjia (2007) utilized 0.56 per cent of soya fibre in the manufacture of low-fat paneer.2.2.5.11 Fruit juicesNakazawa et al.,?(1989) added fruit juices to reconstituted skim milk for obtaining ‘fruit flavoured paneer’ having desired sensory characteristics.2.6 Processing parameters2.6.1 Heat treatment of milkHeat treatment of milk has a profound effect on physico-chemical, sensory and microbiological properties of paneer. It also affects TS recovery and thus yield of paneer. Heat treatment of milk is essential to destroy the pathogenic as well as spoilage micro-organisms. It also denaturates whey proteins, reduces solubility of colloidal calcium phosphate, thus co-precipitating them along with the casein upon acidification of milk. These constituents increase the yield of curd, which are otherwise lost in whey (Walstra and Jenness?1983). Heat treatment at 90?°C for 10–15?min was necessary to achieve desired yield. Different time-temperature combinations adopted by various workers are detailed in Table?2.4Table 2.4 Recommended time-temperature combinations for heating milk prior to acid coagulationTemperature-time combinationEffect on paneerReference80?°C/No holdSuitable for paneer making from cow milkVishweshwaraiah and Anantakrishnan (1985a)85?°C/No holdPaneer from crossbred cow milkArya and Bhaik (1992); Bajwa et al. (2005)85?°C/No holdSuitable for paneer from buffalo milkRao et al. (1984)85?°C/5?minSuitable for paneer from buffalo milkMasud et al. (2007)95?°C/10?minSuitable for buffalo milk paneerChawla et al. (1985)90?°C/No holdSuitable for buffalo milk paneerSachdeva and Singh (1988b)Heating momentarily to 90o?or 118?°CSuitable for low-fat paneer from mixed (cow : buffalo; 1:1) having 3.5% fatPal et al. (1991)96?°C/No holdSuitable for skim milk paneerMendiratta et al. (2007)2.6.2 Type and strength of coagulantPaneer manufacture involves the coagulation of milk proteins to form curd. During this process large clumps of proteins are formed in which fat and other colloidal and dissolved solids get entrapped. The coagulation of milk occurs when pH of milk reaches 4.6 which is the isoelectric point of its major protein, casein. The type and concentration of the acid and the mode of delivery into the hot milk influence the moisture level and product yield.Several coagulants have been tried namely lemon juice, citric acid, tartaric acid, lactic acid, malic acid, hydrochloric acid, phosphoric acid, acetic acid, fermented milk, sour/cultured whey, yoghurt and lactic cultures. Calcium lactate has also been used as coagulant (Sachdeva and Singh?1987; Kumar et al.,?1998; Deshmukh et al.?2009).The concentration of coagulant has a profound effect on the body and texture of paneer. Low acid strength results in soft body and smooth texture, while high acid strength results in hard body. The strength of coagulant adopted by different workers for paneer manufacture is delineated in Table 2.5Table 2.6 Strength of coagulants used for paneer makingStrength of coagulant solutionReference1 per cent citric acid for buffalo milk cow milkBhattacharya etal. (1971);Chawla et al. (1985); Singh and Kanawjia (1988);0.6 per cent (HCl/phosphoric acid/citric acid in sour whey) for buffalo milk. First two were economicalSachdeva and Singh (1988b)1.0 per cent (tartaric acid/citric acid/lactic acid) and acidophilus whey (1.22 per cent TA) for buffalo milk. Acidophilus whey resulted in highest TS recovery (66.4 per cent)Sachdeva and Singh (1987)2.0 per cent citric acid for cow milk and low fat milk (2.2 per cent fat)Vishweshwaraiah and Anantakrishnan (1985a); Arya and Bhaik (1992)2.0 per cent citric acid for cow and buffalo milk mix (1:1) 3.5 per cent fatPal et al.?(1991)2.0 per cent of citric or malic acid for buffalo milkPal et al.?(1999)2.5 per cent citric acid solution for cow milkSharma et al.?(2002)1 per cent citric acid solution and cultured whey in ratio of 3:1 for skim milk paneerMendiratta et al.?(2007)10.0 per cent lactic acid for buffalo milkMasud et al.?(2007)2 per cent citic acid for ewe’s milkPal et al.?(2008)1 per cent citric acid for soy and cow milk mix (1:1)Jadhavar et al(2009a)The amount of coagulant required for coagulation of milk depends upon the type of milk, buffering capacity of milk, type of coagulant and the coagulation temperature employed.2.6 Level of addition of acid per 1.0?kg of milk for paneer makingQuantity of coagulant per 1.0?kg milkReference3?g citric acid for buffalo milkRao et al. (1984)2.34?g citric or lactic acid for cow milkVishweshwaraiah and Anantakrishnan (1985a)2.04?g citric acid (homogenized) and 1.90?g citric acid (unhomogenized) cow milksVishweshwaraiah and Anantakrishnan (1985a)1.95?g citric acid for buffalo milkChawla et al. (1987)1.53?g HCl, 1.14?g phosphoric acid and 2.00?g citric acid for buffalo milkSachdeva and Singh (1987)205?ml of sour whey with citric acid (1.79 per cent TA) and 190?ml of acidophilus whey (1.22 per cent TA)Sachdeva and Singh (1987)2.1?g lactic acidSachdeva and Singh (1987)2.1?g citric acid for buffalo milkSachdeva and Singh (1988b)1.41?g citric acid (buffalo) and 1.52?g citric acid (cow milk)Pal and Yadav (1991)2.03?g of citric acid and 2.00?g of malic acid for buffalo milkPal et al. (1999)2.27?g of lactic acid for buffalo milkMasud (2002)1.68?g citric acid for buffalo skim milkSivakumar et al. (2005)0.2 per cent citric or lactic acid by weight of buffalo milkKumar et al.?(2007,?2008b)?A study was conducted to determine the effect of different types of acids viz., citric acid, tartaric acid and malic acid each at 2, 3 and 5 per cent concentrations on the quality of paneer made using reconstituted milk. The moisture, total solid recovery and yield and sensory scores for flavour, body and texture and overall acceptability of paneer decreased with the increasing strength of acid. However, these parameters for paneer made using coagulants at 2 and 3 per cent levels were statistically comparable (P?>?0.05). Fat and protein per cent increased with the increase in the concentration of the acid. No difference was observed in the levels of ash and fat on dry matter basis and pH and appearance scores at all the three concentrations of the coagulants. The type of coagulant also elicited variations in most of the constituents of paneer. The paneer samples made with citric acid and tartaric acid had significantly higher (P?≤?0.05) values for fat, protein, ash, total solids recovery, fat on dry matter basis, body and texture and overall acceptability scores than paneer made with malic acid at all concentrations. No significant difference was seen in appearance and flavour scores among all the samples. In order to produce paneer with the most desirable characteristics from reconstituted milk, it is suggested citric acid and tartaric acid at 2 per cent concentration can be utilized as coagulants Khan et al.,(2014).2.6.3 Temperature of coagulationThe temperature and pH of coagulation have a significant effect on the body and texture, TS recovery and yield of paneer. The optimum temperature of coagulation differs for different types of milk and their composition, including fat. Coagulation temperature influences moisture retention in paneer. An increase in temperature of coagulation from 60 to 90?°C decreased the moisture content of paneer from 59.0 to 49.0 per cent. Paneer obtained by coagulating milk at 70?°C had the best organoleptic quality and had desired frying quality namely integrity/shape retention and softness (Sachdeva and Singh?1988b; Chandan2007b).A coagulation temperature of 70?°C has been recommended for paneer making from buffalo milk (Bhattacharya et al.,?1971;?Sachdeva and Singh?1988b). Temperatures higher than this resulted in dry and hard paneer while lower temperature yielded product having very moist surface (Sachdeva and Singh?1988b). Masud (2002) and Bajwa et al., (2005) recommended use of higher (85?°C) and lower (72?°C) coagulation temperature for buffalo milk paneer. Chawla et al., (1985) recommended coagulation temperature of 85?°C for low-fat buffalo milk.To obtain good quality paneer, most workers recommended higher coagulation temperature for cow milk. The suggested coagulation temperature for obtaining good quality paneer from cow milk was 80–85?°C (Vishweshwaraiah and Anantakrishnan?1985a; Mistry et al.,?1992; Arya and Bhaik?1992; Sharma et al.?2002). Coagulation temperature of 90o?and 70?°C has been recommended when preparing paneer from ewe’s milk and mixed milk (cow: buffalo; 1:1) respectively (Pal et al.,?1991,?2008). Singh and Kanawjia (1991) suggested 90?°C of coagulation temperature for making paneer from recombined cow milk. Low coagulation temperature of 60?°C has been used by Sanyal and Yadav (2000a) for preparing reduced-fat paneer.2.6.4 pH of coagulationVariation in the pH of coagulation has a significant effect on the body and texture, flavour, quality and yield of paneer. According to De (1980) and Sachdeva and Singh (1988b), with the fall in pH (5.5-5.0), the moisture retention and yield of paneer decreased. Paneer made from cows’ milk coagulated at pH 5.0 was sensorily superior to the one coagulated at pH 5.5 (Vishweshwaraiah and Anantakrishnan1985a). However, at coagulation pH of 5.0 the moisture, TS recovery and yield were lower. The moisture content and yield of paneer increased from 50 to 58.6 per cent and from 20.8 to 24.8 per cent respectively, when coagulation pH increased from 5.1 to 5.4. Sensory quality was best at pH 5.3–5.35 which is recommended for paneer making from buffalo milk (Sachdeva and Singh?1988b). Sachdeva et al,.?(1991) recommended the pH range of 5.20–5.25 for cow milk paneer.2.6.5 Whey drainageAfter coagulation of milk, the curd is allowed to settle down for 5?min without stirring. During this period the temperature should not be allowed to drop below 63?°C. Thereafter, the curd along with the whey was transferred in a hoop lined with muslin cloth to remove the whey (Bhattacharya et al.,?1971).2.6.6 Hooping and pressingThe curd is transferred to hoops lined with muslin cloth and subjected to pressing to obtain a compact block of paneer. Different workers have used different pressure for varied time period for paneer manufacture. Bhattacharya et al., (1971) and Sachdeva et al., (1991) applied pressure of 40–45?kg for 10–15?min for paneer hoop sized 35x28x10 cm for buffalo milk paneer with moisture around 56%. De et al., (1971) and Vishweshwaraiah (1986) employed a pressure of 2?kg/cm2?for 25?min on wooden hoop (4x4x4 inches) to obtain paneer with 55.0% moisture, while Kulsheshtha et al., (1987) suggested applying a pressure of 1?kg/cm2?and found moisture level in paneer was inversely related to the pressure applied. Kumari and Singh (1992) used 0.08?kg/cm2?for paneer preparation from cow and buffalo milk which resulted in paneer with 47.9 and 42.7% moisture respectively. Aneja et al., (2002) recommended higher weights of 70–100?kg on hoops for 10–15?min.2.7 PreservationPaneer is a highly perishable product. It was reported that the freshness of paneer remains intact only for 3?days at refrigeration temperature (Bhattacharya et al.,HYPERLINK "" \l "CR12"1971). At room temperature paneer does not keep good for more than one day. In order to increase the shelf life of paneer, additives, modification in paneer manufacturing process, surface treatments and packaging materials have been recommended by various workers. Singh et al. (1988) dipped paneer blocks of 1.5?kg each in 5 per cent brine, chilled water and acidified water (pH 5.0) to enhance their shelf life (up to 12?days at refrigeration temperature). Singh et al., (1989) found that addition of 0.15 per cent sorbic acid to milk for paneer preparation or wrapping the paneer in sorbic acid-coated butter paper (2?g/m2) extended its shelf life to 30?days at ambient temperature.Vishweshwaraiah (1986) observed that dehydration of paneer cubes (2 cm3) to 15 per cent moisture by keeping them in hot air drier at 75?°C for 4?h and deep freezing at ?9 and ?15?°C led to shelf life of 2?months and 8?days respectively. A low shelf life of only 8?days on deep freezing (at ?15?° C) was probably due to surface drying, which limited its usage, as samples were kept without packaging material. Reduced fat paneer (30–42 per cent FDM) had higher shelf life than full fat (>50 per cent FDM) paneer as paneer with lower fat content underwent less deteriorative changes due to lipolysis (Ghodekar?1989). Sachdeva and Singh (1990) found that dipping of paneer in 5 per cent brine, acidified brine (5 per cent NaCl, pH 5.5) and hydrogen peroxide solution (0.2 per cent, v/v) with or without delvocid (0.5 per cent, w/v) extended the shelf life of paneer cubes of small size (1.0×0.25×0.5 inches) to 22, 20, 32 and 22,?days respectively compared to 10?days for control at 8–10?°C; smaller paneer size helped in better diffusion of the solution and thus longer shelf life.Rao et al., (1992) utilized hurdle technology involving mild heat treatment, minor reduction in water activity and acidification (pH 5.0) to extend their shelf life of paneer to 14?days at 30?°C. Pal et al., (1993) observed that paraffining of low-fat paneer cubes (1 inch cubes) increased the shelf life by over 10?days compared to unparaffined ones. Rao and Patil (1999a) used 1 per cent each of sodium chloride, sucrose and glycerol to decrease the water activity of paneer which led to shelf life extension. Rao and Patil (1999b) developed paneer curry using hurdle technology. The product was so formulated as to have a water activity of 0.95, pH of 5.0, potassium sorbate content of 0.1 per cent. The product kept well for about 1?month and had better quality than the heat-sterilized product stored under similar conditions (30?°C). Singh and Rai (2004) used hot (60?°C for 5?min) and cold (8–10?°C for 6?h) diffusion of paneer cubes with sodium chloride and potassium sorbate and subsequent microwave drying to extend their shelf life. Singh et al., (1991a) found that use of 0.10% sorbic acid in milk coupled with irradiation of the product at 2.5 KGy increased the shelf life of paneer to 30?days at ambient temperature. However, irradiation is not permitted so far by the PFA. In case of paneer, PFA only allows use of sorbic acid and its sodium, potassium or calcium salts at the rate of 2,000?ppm and nisin at the rate of 12.5?ppm for preservation purposes. Punjrath et al., (1997) used blast freezing (?20?°C) to enhance the shelf life of paneer blocks (1.5 cm3) to more than 1?year at a storage temperature of below ?19?°C.2.8 Yield of paneerThe yield of paneer is dictated by the composition of milk used (type of milk, standardization for fat or fat and SNF), heat treatment given to milk, type and strength of coagulant, losses incurred after coagulation (based on pH and temperature of coagulation) and moisture content of resultant paneer after pressing (Bhattacharya et al.,?1971; Chawla et al.,?1985,?1987; Sachdeva and Singh?1987,?1988a,?b; Sharma et al.,?2002). According to Chandan (2007a), the yield of paneer was dependent on the fat and SNF content of the milk used, fat and protein recovered in paneer and its moisture content. A yield of around 21–23 per cent for paneer containing 51–54 per cent moisture can be obtained from buffalo milk, while yield from cow milk is about 17–18 per cent.Sharma et al.?(2002) reported that the composition of milk, which changes with the seasons of the year, had profound effect on the yield of paneer. Milk obtained during winter season gave highest yield (15.5 per cent) while rainy season led to least yield (14.79 per cent). Co-precipitation of casein and whey protein through acidification in presence of calcium chloride (0.15 per cent) led to enhance yield of paneer (Hill et al.,?1982; Singh and Kanawjia?1988; Arora et al.,1996; Makhal and Kanawjia?2005). The yield of paneer from buffalo milk decreased with an increase in coagulation temperature (Sachdeva and Singh?1988b), while for cow milk it increased (Singh and Kanawjia?1988).Rao et al.,(1984) suggested that higher yield of paneer could be obtained by heating milk to 85?°C and coagulating milk using 0.3 per cent citric acid solution at 70?°C. Sachdeva and Singh (1987) found that the yield of paneer was 21.8, 21.7, 22.6, 22.8, 23.4, 23.2 and 22.7 per cent, respectively, when using 1.0 per cent citric acid, 1.0 per cent tartaric acid, 1.0 per cent lactic acid, 0.6 per cent phosphoric acid, 0.6 per cent HCl and 0.6 per cent citric acid in naturally soured whey and whey cultured with?Lactobaccillus acidophilus. Higher fat levels in initial milk gave higher yield of paneer (Chawla et al.,?1987; Pal and Yadav?1991). Yield of paneer was greater when using 2 per cent malic acid rather than 2 per cent citric acid solution as coagulant (Pal et al.,?1999). Jadhavar et al.?(2009b) obtained 14.2 per cent yield of paneer from cow milk, standardized to 4 per cent fat. Deshmukh et al.,?(2009) observed that use of acidified and cultured whey helped in increasing the yield of paneer.Homogenization increased the yield of paneer significantly (Vishweshwaraiah and Anantakrishnan,HYPERLINK "" \l "CR133"1985a). Chawla et al.,?(1987) observed that the incorporation of sodium chloride (0.5 per cent) and NFDM dramatically improved the yield of paneer. Sanyal and Yadav (2000b) found that addition of 0.25 per cent common salt along with 2.5 per cent cultured skim milk increased the yield of reduced-fat paneer. Sanyal et al.,( HYPERLINK "" \l "CR105" 2004) observed that the addition of common salt at the rate of 0.75 per cent to milk increased the yield of reduced-fat paneer by 19.59 per cent above that of control. Addition of 7.5 per cent of whey solids (obtained by heating cheese whey to 85–87?°C at pH 4.8 followed by filtration) to milk increased yield by about 21.0 per cent (Singh et al.?1991b). Incorporation of hydrocolloids like 0.10 per cent Na-alginate, 0.15 per cent carrageenan or 0.15 per cent pre-gelatinized starch helped in increasing the yield of paneer by retaining more moisture (Sachdeva and Singh?1988a). Gupta and Pal (1995) observed that paneer made from reverse osmosis retentate (25 and 33 TS per cent) resulted in higher moisture retention, culminating in higher yield by 2–3 per cent on original milk quantity basis compared to control. Kaur and Bajwa (2003) found that reduction of the size of leaves by chopping or grinding and stage of incorporation significantly affected the yield and moisture content of herb impregnated paneer. Kantha and Kanawjia (2007) observed that both fat level in milk and incorporation of soy fibre had a significant effect on the yield of paneer; fat level of 2.5 per cent and soy fibre level of 0.56 per cent was found to be optimal.2.9 Quality characteristics of paneer2.9.1 Sensory quality of paneerThe ultimate aim of any food item is not only to provide nutrients but also to give sense of delight to consumers by virtue of desired color, flavour and texture. The quality of paneer depends upon the quality of milk from which it was made. Milk fat exerts significant effect on the organoleptic quality of paneer. The sensory score increased with increasing fat (4 to 6 per cent) levels (Arora and Gupta,?1980). Low fat paneer with acceptable organoleptic quality was made from cow milk with 3.5 per cent fat (Vishweshwaraiah and Anantakrishnan,?1986). Chawla et al.?,(HYPERLINK "" \l "CR18"1985) reported that acceptable quality paneer could be obtained from milk possessing 3.5–6.0 per cent fat. Singh and Kanawjia (1988) observed that the sensory score of cow milk paneer improved with increase in coagulation temperatures (i.e. 75–90?°C). Such high temperature of coagulation also held true for paneer obtained from recombined milk added with 0.15% CaCl2?(Singh and Kanawjia?1991). A coagulation temperature of 85?°C has been recommended for paneer making from reconstituted milk (15.0 per cent TS) (Singh and Kanawjia?1992).Sachdeva et al.,?(1991) observed that incorporation of 0.08 per cent CaCl2?in manufacture of cow milk paneer helped in improving the sensory score for appearance, flavour, body and texture compared to milk without added calcium salt. They reported maximal sensory score when pH of coagulation was 5.20-5.25. Pal et al.?,(HYPERLINK "" \l "CR74"1991) observed that the sensory score of low-fat paneer was greater when milk was heat treated at 118?°C rather than at 90?°C. Pal and Yadav (1991) observed that paneer made from mixed milk of buffalo and cow in the ratio of 3:1 or 1:1 had superior sensory characteristics than that made using 1:3 ratio or cow milk. Singh et al.?,(1991b) observed that the overall acceptability scores of paneer made with 0, 5, 7.5 and 10 per cent whey solids (which were obtained by heating cheese whey to 85–87?°C at pH 4.8 followed by filtration) were 8.4, 8.1, 7.2 and 6.4 for raw paneer, and 8.7, 8.4, 7.7 and 7.6 for fried paneer respectively. Kumar et al., (1998) found that the body and texture and overall acceptability scores of paneer made using calcium lactate coagulant were better than those obtained for the product made using citric acid or sour whey.Arya and Bhaik (1992), found that paneer made from cow milk (2.2 per cent fat) resulted in a product lacking in softness and typical flavour. Syed et al.,?(1992) found that the total sensory score of paneer made using different milks tended to decrease in the following order: Paneer from buffalo milk with 6.0 per cent fat (93.33 score)>Paneer from cow milk with 4.5 per cent fat (88.97 score)>Paneer from skim milk with 0.1 per cent fat (84.87). Arora et al.,?(1996) observed that use of 0.05 per cent CaCl2?in milk diluted with water to 4.6 per cent fat and 8.0 per cent SNF resulted in paneer comparable to that made from normal milk (5.5 per cent fat and 9.0 per cent SNF).Citric acid yielded sensorily superior paneer compared to malic acid; the body and texture of paneer obtained using malic acid was quite poor (Pal et al.,?1999). The sensory attributes of paneer made by coagulating milk at 80?°C using either cultured whey of?Lactobacillus acidophilus?and?Streptococcus lactis or acidified whey at 1.0 per cent did not differ from one another; however, use of cultured whey withStreptococcus lactis resulted in paneer with lower scores for body and texture. The use of whey cultured with?Streptococcus lactis?resulted in paneer with a soft and loose body and open texture, which was unsuitable for sale (Deshmukh et al.,?2009).Kaur et al.?(2003) found that paneer dipped in 3 per cent brine had a good sensory score. They also reported that the use of mint (Mentha spicata) and coriander (Coriandrum sativum) improved the flavour of paneer. Sanyal et al. (2004) observed that desired sensory quality of reduced fat paneer could be obtained through use of 0.25 per cent sodium chloride in milk for paneer making. Bajwa et al.?(2005) advised heating temperature of 85?°C, coagulation temperature of 72?°C and immediate straining to yield highly acceptable paneer at all the levels of herb impregnation. The overall acceptability score was the highest at 10.0 per cent incorporation of coriander and mint leaves. Paneer made from buffalo milk heated at 85?°C yielded sensorily superior product then when heated at 80 or 90?°C (Masud et al.,?2007). Kantha and Kanawjia (2007) reported that acceptable paneer could be obtained from milk having 3.0 per cent fat and 0.25 per cent soy fibre. Chavan et al.,?(2007) found that paneer made from blend of buffalo skim milk and cow milk (20:80) was comparable to that of standard buffalo milk containing 5.5 per cent fat. Kumar et al.,?(2007, HYPERLINK "" \l "CR54" 2008b) found that the sensory score of paneer decreased with an increase in the level of incorporation of the coagulant i.e. from 0.2 to 0.6 per cent. Pal et al., (2008) found that acceptable paneer could be obtained from ewes’ milk by coagulating the milk at 90?°C using 2.0 per cent strength of citric acid solution.Verma, 2013 compared specially four types of foods, i.e. cutlet, bread rolls, sandwiches and burgers that we make as for school lunch boxes or fast food corners prepared by incorporating buffalo milk paneer and soy substituted buffalo milk paneer in the ratio of 30:70, (30 per cent buffalo milk and 70 per cent soy milk ) were compared for their protein contents and sensory properties. On dry weight basis soy substituted buffalo milk paneer incorporated foods contents high amounts of protein than those incorporated with milk paneer, comparative sensitive evaluation showed that incorporation of soy substituted buffalo milk paneer did not affect sensory characteristics of all the products, except for flavor in bread rolls ,after taste in cutlets and sandwiches. The mean score for all the sensory attribute ranked between 6.6 and 8.6 on a nine point hedonic scale. Addition of mixed spices improved the sensory attributes of soy substituted buffalo milk incorporated foods. 2.9.2 Microbiological quality of paneerThe microbiological quality of paneer depends on the microbiological quality of milk, and the hygiene exercised during manufacture of paneer and its subsequent handling, packaging and storage. Microorganisms such as coliforms, yeasts and moulds that might be present in raw milk get destroyed completely, when milk is heated at 82?°C for 5?min. But these microbes may contaminate the product through a number of sources like air, water, equipment, knife, muslin cloth and persons handling the products (Aggarwal and Srinivasan?1980). These microbes can cause proteolytic and lipolytic changes, discolouration and other defects in the product (Thakral et al.,?1986). Vishweshwaraiah and Anantakrishnan (1985b) suggested that SPC lower than 5,000/g and greater than 2 lakhs/g could be rated as ‘Excellent’ and ‘Poor’ respectively.Bureau of Indian Standards (BIS?1983) set limits for microbial count viz., total plate count <5×105/g, yeast and mould count <250/g, and coliform count of <90/g. Rajorhia et al. (1984) observed that paneer made at pilot scale level at National Dairy Research Institute, Karnal had lower yeast and mould, and coliform count than paneer obtained from market (Delhi, Karnal). Such a finding for laboratory and market paneer was also observed by several workers (Vishweshwaraiah and Anantakrishnan?1985b; Das and Ghatak?1999; Goyal et al.,2007; Singh and Singh?2000). Such a difference in SPC was also noted in products sold by local and standard shops in India (Kumari and Kalimuddin?2002). Higher counts in the market samples points at lack of adequate sanitary practices during manufacture and storage. Kalhan and Grover (1984) observed that fresh paneer prepared under hygienic conditions did not show any pathogenic microorganisms. Singh et al?,(1989) reported microbial profile of fresh paneer as SPC—2.3×103, proteolytic bacteria—7.4×102, lipolytic bacteria—4.9×102and fungi—10/g. Pal et al.?(1993) found that the SPC, yeast and mould and coliform count of paneer were 3.03, 1.90 and 0.86 log cfu/g, respectively. Nath et al.,?(2007) observed SPC of 5.4×103?and yeast and mould count of 260/g in freshly prepared paneer. Ghodekar (1989) observed that the market paneer was contaminated with yeast and mould which led to deterioration in sensory quality of paneer during storage due to proteolytic and lipolytic changes. Such products become unacceptable and potentially injurious to the health of consumers. Kumar and Sinha (1989) observed that more than 60.0 per cent of paneer samples from organized dairies and markets in India were contaminated with coliforms.Vaishnavi et al.,?(2001) obtained SPC in the range of 3×102?to 9.7×1010?cfu/g in paneer samples sold in Chandigarh. A few workers reported that majority of market paneer (Ranchi, Gwalior) samples were contaminated with?Bacillus cereus?and?Pseudomonas aeruginosa?respectively (Kumari and Kalimuddin 2003; Bharadwaj et al,.HYPERLINK "" \l "CR10"2007);?aflatoxin contamination in market panner was also observed (Choudhary et al.,HYPERLINK "" \l "CR21"2007). Chhabra (2008) observed yeast count in the range of 24–180?cfu/g in paneer. Bacteriocins action as biopreservative helps in increasing the shelf life of the food products by providing safety from food pathogenic and spoilage microorganisms (O’Sullivan et al., 2002; Settanni and Corsetti, 2008; Settanni and Moschetti, 2010). The effectiveness of bacteriocin isolated from E. faecium BS 13 strain to act as biopreservative and its role in increasing shelf life of paneer and khoya was checked in presence of bacteriocin as compared to control and addition of chemical preservative (sorbic acid). In case of control sample, a slight increase in microbial count was observed up to 5 days followed by sharp increase in the count after 10 days of storage period, which could be due to the initiation of spoilage of paneer at 4oC . In case of bacteriocin supplemented paneer samples, microbial cell load was found to be 14 × 106 cfu ml/1 after treatment with bacteriocin which was less by factor of 104 as compared to the control samples (untreated). This could be due to bacteriocidal effect of bacteriocin on the microorganisms initially present in the sample (Samelis et al., 2003). After 10 days of storage, the microbial load was observed to be 56 × 1010 cfu ml-1. The results depicted that the microbial load of bacteriocin treated samples after 10 days were comparable to initial control sample (zero hrs). The large increase in number of L. monocytogenes was observed in nisin free cheese sample as reported in earlier studies too (Govaris et al., 2001). Even after 15 days of storage, only 40 × 1011 microbes were observed as compared to 190 × 1015 in control. It has been previously reported that bacteriocin at concentration of 320 IU ml-1 and 160 IU ml-1in pasteurized milk showed lowest total bacterial count and increased the shelf life upto 12 days of refrigerated storage (Ibrahim and Elbarbary, 2012). Lower concentration (40Uml-1) is ineffective in preventing the spoilage of milk after 6 days of storage. Similar studies on decreased viable cell count in milk and soft-cheese had been reported by other authors (Bizani et al., 2008; Sable et al., 2000). 2.9.3 Textural propertiesDesai et al., (1991) studied the textural properties of paneer procured from six different sources in Karnal. Instron Universal testing machine fitted with a 100?N load cell was used for the study. They observed that the hardness, gumminess and chewiness of raw paneer varied significantly, whereas cohesiveness and springiness did not differ among the market paneer samples. They concluded that the hardness was inversely related to the moisture content and directly related to the calcium content of paneer.Syed et al., (1992) observed that the hardness of paneer was highest for skim milk paneer when compared to cow and buffalo milk paneer. Kumari and Singh (1992) found that cow milk paneer had higher values for cohesiveness, gumminess and chewiness than buffalo milk paneer, whereas the hardness and springiness were greater in buffalo milk paneer. However, the paneer or channa from buffalo milk have been found to produce harder and chewy texture due to higher concentration of casein in the micelle state with bigger size, harder milk fat due to larger proportion of high melting triglycerides in it and higher content of total and colloidal calcium (Sindhu?1996). Kanawjia and Singh (1996) observed values of 13.2?N, 0.68, 7.8?mm, 9.8?N and 71?N-mm respectively for hardness, cohesiveness, springiness, gumminess and chewiness when measured using Instron Texture machine using 1000?N load cell.The textural properties of low fat paneer using soy protein isolate (SPI) as fat replacer was investigated. The physico-chemical and sensory characteristics of 4 types of paneer made of low-fat milk (3 per cent milk fat (MF) and 10% solids-not-fat (SNF)) and SPI of 0 (T1), 0.1 (T2), 0.2 (T3) and 0.3 per cent SPI (T4) were compared with high fat paneer (TC) made of high fat milk (6 per cent MF and 9 per cent SNF). CaCl2 (0.2%, w/v) was used as coagulant at 75?±?1°C. The gumminess, chewiness and firmness showed the same trend. Resilience and cohesiveness values showed no significant difference among the samples. Hunter colour L values showed a decreasing, and a and b values increasing trend with increasing levels of SPI. Sensory appearance and colour scores were lower (p?≤?0.05) for T1-T4 than TC. More than 0.2 per cent SPI imparted beany flavour to paneer.(kumar et al., 2011)Paneer is highly perishable at ambient conditions and its shelf-life is very low. At high temperature, it develops a sour smell, and bitter taste. Drying can be one of the methods to increase shelf-life. Drying experiments were conducted at 62, 72 and 82°C and 10, 14 and 18 kPa absolute pressures with superheated steam. The product quality was judged by instrumental texture profile. Hardness, adhesiveness, gumminess and chewiness increased with increase in temperature and decreased with increase in pressure. Springiness and cohesiveness, however, decreased with increase in temperature and pressure. Textural properties of fresh and rehydrated paneer indicated large variation on hardness, adhesiveness and resilience whereas springiness, cohesiveness and chewiness had marginal variation as compared to fresh paneer. Shrivastav and Kumbhar (2010).Karadbhajne and Bhoyarkar (2010) prepared paneer using citric acid as coagulant. The paneer prepared from 2 per cent and 4 per cent coagulant was stored for 8 day and its texture was analyzed on TA-XT ANALYZER for alternate days which shows different texture for different days. The paneer prepared from 2 per cent coagulant having firmness (2578) and its springiness was (38.672). On third day the firmness was (5712) similarly on fifth and seventh day it was (3069) and (9280). The graph shows on first, third and fifth day the force required were same. Till 5 days the firmness, hardness, and springiness remains same whereas on eight day the firmness was decreased. The paneer prepared from 4 per cent coagulant having firmness (2412) and springiness (36.11). On fifth day and seventh day the firmness gets decreased to (1903) and (38.95) respectively. It shows graphically that on first, third and fifth day the firmness, hardness, and springiness of the paneer remains same whereas on eight day it was decreased. This study shows that the paneer can be consumed or acceptable for five days and its texture remains good.2.9.4 Shelf lifeA shelf stable, convenience product egg yolk paneer (EYP) was developed by incorporation of optimized quantities of binders, salt, natural antioxidants and egg yolk. Dehydrated EYP was packed in metalised polyester pouches, stored at ambient temperature (27?±?2?°C) for 6?months and sampled periodically for quality evaluation. The protein and fat content of dehydrated EYP was 26.2?±?1.75 per cent and 36.1?±?2.46, respectively. The shelf stability of the product was achieved by keeping a moisture content (5.6?±?0.50 per cent) and water activity (0.43?±?0.05) low. An excellent rehydration capacity (64.8?±?5.39 per cent) was observed in the EYP, whereas, the rehydration ratio of the product was 1:2.7. Changes in Free Fatty Acids, Thiobarbituric acid, textural profile analysis and Hunter colour units (L, a and b) during storage did not affect the quality characteristics of the product. About 38 per cent loss in carotenoid content was recorded during storage of the product. Staphylococcus aureus, E coli, Salmonella and Shigella, however, were not detected in any sample throughout the storage period. Sensory evaluation revealed that rehydrated yolk paneer had excellent texture and was very close to fresh ones (before drying) during storage for 6?months(Pawar 2012).Paneer tikka, a popular dish for vegetarians, is a tongue tingling favourite of Indian governments. It is a perishable commodity which requires more than five hours for its preparation. The shelf life of paneer tikka is hardly one day at room temperature which impedes its proper marketing. In order to enhance the shelf life of paneer tikka, the product was vacuum packed in two high barrier packages, viz., LLDPE(*)/BA(?)/Nylon-6/BA/LDPE(?) (110?μ, (*)linear low density polyethylene, (?)binding agent, (?)low density polyethylene) and metallized polyester/LDPE (20/75?μ) along with LDPE (100?μ) as control, and stored at 3?±?1?°C. The stored samples were evaluated at 0, 10, 20, 30 and 40?day(s) for changes in chemical characteristics, namely moisture, pH, titratable acidity, free fatty acids content, tyrosine content and water activity. The analysis of variance of the data revealed that type of packages had significant influence on all the above parameters excluding water activity during storage of paneer tikka. (Ahuja KK,?Goyal GK, 2013). Chapter -3Material and method MATERIALS AND METHODSIn this chapter the materials used, the processing techniques, the analytical methods and sensory evaluation methodology at various stages of the research to meet the objectives have been discussed.3.1 Procurement of Materials3.1.1 Soyabean Soy beans were procured from the local market of Jhansi.3.1.2 Cow and Buffalo milkCow milk and Buffalo milk were collected from the local area of Jhansi.3.1.3 Coagulating agentLaboratory grade citric acid was used as a coagulating agent in paneer making process.3.2 Preparation of soyamilk Soyabean grains were sorted and cleaned with potable water. Cleaned soybean was soaked in water overnight and hulls were removed by rubbing with clean hands and washing with clean water. The soaked soybean was ground with hot water in the ratio of 1:8 and filtered through muslin cloth. The filterate was boiled for 20 minutes and cooled to obtain plain milk. Whole soybean Sorted and cleaned with potable water Soaked in water overnight Removed hulls using clean hands Washed to remove hulls Ground soyabean with hot water in the ratio 1:8 Filtered through muslin cloth Boiled the filtrate for 30 minutes Plain soymilk Figure 1: Processing of soymilk3.3 Preparation of Blended PaneerSoy paneer was prepared in laboratory from the aforementioned soy milk. For coagulation of soy milk one per cent concentration of citric acid solution was used as coagulant. Soymilk sample was then heated to a coagulation temperature about 70°C in big size container and citric acid was added slowly with gentle and continuous stirring. After completion of coagulation, stirring was stopped and the contents were left undisturbed at room temperature and strained through a clean muslin cloth. The coagulum thus obtained was pressed and soaked in chilled water for 30 minutes. The blended paneer was removed from chilled water and packed in polythene bags after removing surface water. Soymilk was blended with cow and buffalo milk in different ratios (0 to 100 ) Blended milk was then heated at standardized Coagulationtemperatureof70°C Citric acid solution (one per cent) was added slowly with gentle and continuous stirring After completion of coagulation, stirring was stopped The contents were left undisturbed at room temperature and strained through a clean muslin cloth. Coagulum was obtained The coagulum thus obtained was pressed and soaked in chilled water for 30 minutes. The blended paneer was removed from chilled water and packed in polythene bags after removing surface water 3.4 Analytical Procedures3.4.1 Physico-chemical composition of cow milk, buffalo milk, and soy milkChemical components such as fat, protein, carbohydrate, total ash, total solids and minerals were determined according to methods given in AOAC (2000).3.4.1.1 Moisture The moisture content of samples was determined by oven drying method (Ranganna, 1991). Aluminum dish was heated and weighed (W1). 5 g sample was added in the dish and weighed (W2). Dish was heated in hot air oven at 70±1?C until the constant weight was obtained; Dish was weighed (W3). Moisture content was calculated by using the formula 3.1. (W2-W3) Moisture (per cent) = x 100……...... (3.1) (W2-W1)3.4.1.2 ProteinThe protein content of sample was estimated by Kjeldahl method (using Pelican Equipment). Sample of 0.20 g was transferred into the protein digestion tube. Ten milliliters of concentrated sulphuric acid and 3 to 4 g of digestion mixture (Potassium sulphate and copper sulphate in the ratio of 5:1) was added to the tube. Digestion was carried out in the Kjeldahl unit at 400?C till the contents become clear. The digested samples were cooled and diluted with some quantity of distilled water and tube was fitted into distillation unit. Digested sample was distilled for about 6 min with 40 per cent NaOH. Liberated ammonia was absorbed in 4 per cent boric acid solution containing a few drops of mixed indicator (one part of methyl red and one part of bromocresol green). The distillate was titrated against 0.1 N HCl until the brown blue colour changes to pink. A blank determination was carried out using all the reagents except sample (Ranganna, 1991). Nitrogen per cent was calculated by the following formula. Calculated Nitrogen percent was multiplied by the conversion factor to know the protein percent. (Titre value – Blank) x 14 x Normality of acid x 100 Per cent Nitrogen = Sample Weight x 1000 Per cent protein = % N x 6.25 …… (3.2) 3.4.1.3 Fat Fat content of the sample was estimated using soxhlet extraction method (Socsplus equipment of Pelican Equipment). 2g of sample was weighed and transferred into the thimble that was inserted into the thimble holder. About 50 to 75 ml of petroleum ether (BP 60? to 80?C) was taken in beaker and the thimble is placed in the beaker. Beaker was placed in the system and the water tap was opened for condensation. The solvent was preheated at 40 to 60? C, solvent was boiled and allowed to stand for half an hour. When boiling was complete the temperature was increased by 20?C, depending upon the solvent temperature (80±20?C). Solvent was condensed in the beaker and allowed the condensed solvent to flow through the thimble to perform reclamation. When the reclamation process was complete, beakers were transferred to oven, maintained the temperature at 80?C to evaporate all the solvent from the beaker. The beakers were finally kept in desiccator, allowed to cool and weighed with fat . Fat percentage was calculated by using the formula 3.3. W3-W2Fat (per cent) = x 100 ………. (3.3) W1 Where, W1 = Weight of the sample W2 = Weight of the dried beaker, and W3 = Weight of beaker + fat Milk were analyzed for fat by Gerber’s method (Ranganna, 1991)3.4.1.4 Ash Ash content of the sample was estimated by incinerating the sample at 550?C in muffle furnace (Ranganna, 1991). Empty silica dish was weighed and approximately 5 g sample was taken in the same silica dish. Dish along with sample was placed over an electric heater and the product was allowed to charr and continued heating till fumes came out of it. Cooled silica dish was placed in a muffle furnace, and the temperature was raised to 550-600 ?C and maintained for at least four hours, switched off and cooled for overnight. The dish was cooled in a desiccator if its temperature was higher than room temperature and then weighed (Ranganna, 1991). Ash percentage was calculated by the following formula 3.4. (W3 – W1) Ash (per cent) = x 100 ………. (3.4) (W2 - W1) Where,W1 = Weight of empty silica dish W2 = Weight of the sample + silica dish W3 = After ashing, weight of silica dish + ash3.4.1.5 CarbohydrateCarbohydrate content was obtained by difference method (Ranganna, 1991). The carbohydrate content was calculated by using formula 3.5.Carbohydrates (per cent) = 100 - (moisture + ash + protein + fat) …… (3.5)3.4.1.6 Crude fiberThe crude fiber content in various samples was estimated using AOAC (1976) method. Two g of defatted sample was digested with 200 ml of 1.25 per cent sulphuric acid for 30 min. After filtration through a linen cloth, the residue was washed with boiled distilled water until it was free from acid. The acid free residue was digested with 200 ml of 1.25 per cent sodium hydroxide for 30 min. The contents were filtered hot through linen cloth. The residue was transferred to a gooch crucible, which had been prepared previously with a thin but close layer of ignited asbestos and washed with boiled distilled water until the residue was alkali free. Finally, the residue was washed with 15 ml of 95 per cent ethyl alcohol. The contents of crucible were dried to a constant weight at 100° C. The dried residue was ignited in a muffle furnace at 550°C ±15°C for 30 min. The per cent loss in weight was expressed as crude fiber. 3.4.2 Rheological Analysis3.4.2.1 Texture profile analysisThe texture analyzer (model TAXT2i) was calibrated at the beginning of each testing session using a 25 kg load cell. A cutting probe (HDP/LKB light knife blade perspex) and compression plate were used in conjunction with a texture analyzer (Boune,1982) for determination of cutting and pressing strength. Cutting and pressing strength were measured with pre test speed at 2.0 mm for a total travel of 10.0 mm for paneer samples by keeping the samples horizontally and cutting with vertical blade. The force required was noted. 3.4.3 Microbiological AnalysisTotal plate count, yeast and mould count and coliform count of paneer samples were determined as per the standard methods given in APHA (1992). Samples were inoculated in duplicate plates of suitable media and incubated at the recommended temperature (Table 3.1). At the end of incubation period, the plates were counted for number of colonies.Table 3.1: Media and incubation condition for microbial examinationDeterminationMediumIncubationTypepHTemperature (?C)Period (hr)Total plate countPlate Count Agar 7 3724-48Yeast and mold countPotato Dextrose Agar 3 2272-110Coliform countViolet Red Bile Agar 7 3724-483.4.4 Sensory AnalysisA panel consisting of 10 members was selected for sensory evaluation. Blended milk and paneer samples were presented to panelists drawn from the faculty members and students of the department for evaluation. The panelists were asked to judge the samples for color, taste, flavor and overall acceptability using a 9-point hedonic scale rating (Amerine et al., 1965) as per the performa (Appendix).3.4.5 Statistical AnalysisThe data obtained were analysed statistically on a completely randomized design using analysis of variance technique (Snedecor and Cochran,1968) to find if the differences were significant or not. 3.4.6 Storage StudiesSamples were analysed for changes in moisture, protein, fat, ash, Carbohydrate and titratable acidity. Microbiological and sensory attributes were also evaluated. Chapter -4Result and DiscussionResult & DiscussionThe experimental findings of utilizing soymilk for developing highly nutritious paneer by blending it with cow milk and buffalo milk are presented and discussed in this chapter. Blends of soymilk with cow milk and soymilk with buffalo milk were heated and coagulated to prepare paneer. The results showing the effect of blending on chemical and sensory characteristics of cow milk and buffalo milk and their paneer are presented. The presentation in this chapter is divided into following sections:Chemical composition of soybean, soymilk, cow milk and buffalo milk.Effect of blending of soymilk with cow milk on physico-chemical characteristics, sensory scores of milkEffect of blending of soymilk with buffalo milk on physico-chemical characteristics, sensory scores of milkOptimization of processing variables for the manufacture of soy milkEffect of blending of soymilk with cow milk on physico-chemical characteristics, sensory scores and textural properties of soy cow milk paneer.Effect of blending of soymilk with buffalo milk on physico-chemical characteristics, sensory scores and textural properties of soy buffalo milk paneer.4.1 Chemical composition of raw material The chemical composition of Soyabean, Soymilk, Buffalo Milk and Cow Milk are presented in table 4.1.From the results it can be revealed that moisture, fat, protein, ash and carbohydrate content of soyabean were 7.60, 19.68, 34.76, 4.47 and 33.49 per cent, respectively. The results showed that the Soyabean contained 277 mg/100g and 704 mg/100g of calcium and phosphorus respectively. The values are similar to those reported by Rehman etal.,(2007). The result presented in table 4.1 revealed that the moisture, fat, protein, ash and carbohydrate content in soyamilk were 92.12, 2.28, 4.03, 0.58 and 0.99 percent, respectively. The results showed that soyamilk contained 25 mg/100g and 52 mg/100g of calcium and phosphorus respectively. It was observed that values obtained in the present investigation are similar to those reported by Rehman etal.,(2007), Rivas etal., (2002), Hajirostamloo,(2009) and Chen (1983). The results noted in table 4.1 showed that, 164.19 mg calcium and 113.0 mg phosphorus per 100 gram of buffalo milk were detected. The moisture, fat, protein, ash and carbohydrate content of buffalo milk were 83.0, 6.0, 4.0, 0.8 and 6.2 per cent, respectively. The values are similar to those reported by Han et al. (2012), Menard et al. (2010), Ahmad et al. (2008), and Miller et al. (1999) and Posati and Orr,(1976). The moisture, fat, protein, ash and carbohydrate content in cowmilk were 87.0, 3.8, 3.30, 0.7 and 4.3 per cent, respectively. Cowmilk contained 119.83 mg calcium and 94.03 mg phosphorus per 100 gram The values are similar to those reported by Hajirostamloo,(2009),Rehman et al.,(2007), and Rivas et al., (2002), and Posati and Orr,(1976). The results reported in Table 4.1 showed that the richest source of fat, protein and ash are buffalo milk, soyamilk and cow milk respectively. The calcium and phosphorus content are maximum in buffalo milk followed by cow milk and soy milk. Table 4.1 - Chemical composition of Soyabean, Soyamilk, Buffalomilk and Cowmilk.Attribute (per cent)SoyabeanSoymilkBuffalo MilkCow MilkMoisture7.6089.6083.087.1Fat19.68 (db)2.286.03.8Protein34.76(db)4.034.03.30Ash4.47 (db)0.580.800.70Carbohydrate(by difference)33.493.516.25.10Calcium (mg)27725164.19119.83Phosphorus (mg)70452113.094.03 db=dry basis *Values are average of three determinations4.2 Blending of soymilk with cow milk and buffalo milk4.2.1Chemical composition of blends of soymilk with cow milkThe results of effect of blending of soyamilk with cow milk on chemical composition like total solids, protein, fat, ash, , pH and acidity of blends are presented in table 4.2. Soy milk was incorporated in cow milk at 5, 10, 15, 20, 25,50, 75, and 100 percent to produce different blends of milk.From the results presented in Table 4.1 it is revealed that cow milk and soymilk contained 12.9 per cent and 10.4 per cent total solids, respectively. The total solid content of raw blends varied proportionately. The variation was non significant up to 15 per cent proportion of soyamilk. This indicated that the blending of soymilk with cow milk reduced the amount of total solids.The variation in the amount of total solids was significant (P ≤ 0.05) at 20 per cent and above proportion of soyamilk blend with respect to cow milk, however it was non significant at lesser than 20 per cent, level of soymilk with cow milk blends.The values for protein content in cow milk and soy milk obtained in the present investigation were 3.3 per cent and 4.03 per cent respectively. The protein content in blends varied proportionately.This indicated that the blends of soy milk with cow milk increased the amount of protein significantly (P ≤ 0.05) in the blends containing more than 15 per cent soymilk, however the variation was non significant upto 15 per cent soymilk, however the variation was non significant upto 15 per cent level of soyamilkThe values for fat content in cow milk and soy milk obtained in the present investigation were 3.80 per cent and 2.28 per cent respectively. The fat content in blends was found to be proportionately decreased with an increase in the amount of soymilk. The decrease in the fat content was significant ((P ≤ 0.05) in blends containing 15 per cent and more amount of soymilk.Table 4.2 –Effect of blending on chemical composition of soymilk with cow milk Proportionof soymilk, (%)Total solids (%)Protein (%)Fat (%)Ash (%)pHAcidity (%)012.903.303.800.706.900.21512.783.343.720.696.890.211012.653.383.650.696.880.211512.533.413.570.686.880.212012.403.453.500.686.860.222512.283.483.420.676.850.225011.653.673.040.656.820.237511.033.862.660.626.780.2310010.404.032.280.586.740.24CD at 5%0.500.120.190.030.040.01% = per cent *Values are average of three determinationsFig.1. Effect of blending on chemical composition of soymilk with cow milk In the present study, the value of 0.7 per cent for ash content was obtained in cow milk and 0.58 per cent in soy milk. The variation in ash content was non significant upto 20 per cent leval of soymilk, however the variation was significant (P ≤ 0.05) at 25 per cent and more soymilk proportion with cow milk. The enhancement in protein amount and decrease in total solids, fat and ash content might be due t the variation in the proximate composition of soymilk and cow milk.The values of pH for cow milk and soymilk were 6.9 and 6.74, respectively, in the present investigation. The pH decreased slightly and proportionately in the blends with an increase in the amount of soymilk. The variation in Ph was non significant below 20 percent level of soymilk in blends thereafter a significant (P ≤ 0.05) change in Ph was observed. The variation in pH might be due to the pH of water used in preparation of soymilk and it may have slightly lower pH in comparison of cow milk.The value of acidity per cent for cow milk and soymilk were 0.21 and 0.24, respectively in the present investigation. The acidity increased slightly and proportionately of the blends with an increase in the amount of soymilk. The variation was non significant in the blends below 20 per cent level of soymilk and thereafter a significant (P≤0.05) increase was observed. The changes in pH and acidity might be due to the use of slightly acidic water to prepare soymilk because of which the pH of soymilk was lower and acidity was higher than cow milk.4.2.2 Sensory Evaluation of blended cow milk with soy milk A laboratory panel consisting of 10 untrained members evaluated soy cow milk blends for sensory attributes such as colour, flavour, taste and overall acceptability. Sensory scores along with statistical analysis of soy cow milk blends are presented in Table 4.3 to 4.6 Table 4.3 : Sensory scores for colour of blends of soy milk with cow milk PanelistsProportion of soymilk, per cent0 55 510152025507510018777776552888877655388887765548887766555888877655688777765578888776558888877655988887765410777777655Mean7.97.87.77.676.9654.9CD at 5% level: 0.31Table 4.4: Sensory scores for flavour of soy milk and cow milk blendsPanelistsProportion of soymilk, per cent055 510152025507510018778765552888776655387887655448888766555878876555688877665578878765548787776555988877665410787776555Mean7.87.77.67.5765.454.7CD at 5% level: 0.38Table 4.5: Sensory scores for taste of soy milk and cow milk blendsPanelistsProportion of soymilk, per cent055 510152025507510018888776552888877655387777775548888766555888877655688777775578788776558887777655988878765410788887755Mean7.97.87.77.67.26.96.354.9CD at 5% level: 0.37Table 4.6: Sensory scores for overall acceptability of soy milk and cow milk blendsPanelistsProportion of soymilk, percent055 510152025507510018877776552888876655388787775548888766555878877655688777775578888776558888877755988878765410778787755Mean7.97.87.77.67.26.86.454.9CD at 5% level: 0.37Results presented in Table 4.3 showed that the mean sensory score for colour of cow milk was maximum. However, the inclusion of soymilk up to 25 percent did not decrease the colour significantly (p≤0.05) and thereafter the increased levels of soymilk lowered the colour of blend significantly (p≤0.05). The mean score for colour of milk obtained from the blends containing 75 and 100 per cent soymilk were in acceptable range on a 9 point hedonic scale, and did not differ significantly (p≤0.05) from each other. However, the blend containing 50 per cent soymilk was in an acceptable range but was rated significantly (p≤0.05) lower than all other blends containing less than 50 percent soyamilk and significantly (p≤0.05) higher than those having 75 and 100 percent soymilk. The effect of soymilk inclusion with cow milk on the flavour presented in Table 4.4 showed that the cow milk scored highest for flavour on 9 point hedonic scale. It was found that the incorporation of soymilk up to 15 per cent did not lower the mean scores for flavour significantly (p≤0.05). The increase in soymilk proportion from 15 to 20 had no significant (p≤0.05) variation in flavour. Further increase in the proportion of soymilk lowered the mean sensory score for flavour significantly (p≤0.05). The variation in flavour between the blends with 20 and 25 per cent soymilk and 25 and 50 per cent were non- significant. The blend consisting 50, 75 and 100 per cent proportion of soymilk were in acceptable range. The result of the effect of soymilk inclusion on the taste of blends showed that cow milk scored highest for taste on 9 point hedonic scale. It was observed that the incorporation of soymilk up to 15 percent did not lower the mean sensory scores for taste significantly (p≤0.05) and thereafter it had significant (p≤0.05) effect. However raising of soymilk from 15 to 20 per cent did not alter the taste significantly (p≤0.05).The blend prepared with 20 per cent soyamilk differ significantly (p≤0.05) from cow milk. But further increase in the proportion of soymilk lowered the score significantly (p≤0.05).The differences in taste between the products made from blends having 20 and 25 per cent soymilk were non significant and those between 25 and 50 per cent soyamilk were non significant. The blend having 50 per cent soymilk was in acceptable range and thereafter it was in unacceptable range. The cow milk scored highest for mean overall attributes but the inclusion of sensory scores of soymilk up to 15 percent did not lower the mean overall sensory scores significantly (p≤0.05).The blend containing 50 percent soymilk were in acceptable range on a 9 point hedonic scale. The further increase in the proportion of soymilk in the blends lowered mean overall sensory scores below acceptable range on a 9 point hedonic scale.Statistical analysis of all the four attributes of sensory evaluation concluded that the incorporation of soymilk in cow milk up to 15 per cent proportion was found most suitable, however, soymilk may be included up to 25 per cent proportion in cow milk without significantly (P≤0.05) affecting any sensory attributes.4.3 Blends of soymilk and buffalo milk4.3.1Chemical composition of milk blendsThe result of chemical composition like Total solids, protein, fat, ash, pH and acidity of blends are presented in Table 4.7. Soy milk was blended with buffalo milk at 5, 10, 15, 20, 25, 50, 75, and 100 per cent to produce different milk blends.From the results presented in Table 4.7 it is revealed that buffalo milk and soymilk contained 17.0 per cent and 10.4 per cent total solids, respectively. The total solid content of blends decreased proportionately but the variation was non significant. This indicated that the blending of soymilk with buffalo milk reduce the amount of total solids.The values for protein content in buffalo milk and soy milk obtained in the present investigation were 4.0 percent and 4.03 per cent respectively. The protein content in blends was almost the same as both soymilk and buffalo milk had similar protein content. The values for fat content in buffalo milk and soy milk obtained in the present investigation were 6.0 per cent and 2.28 per cent, respectively. The fat content of raw blends was found to be proportionately decreased with an increase in the amount of soymilk. The fat content in blends decreased significantly (P≤0.05) with an increase in the amount of soymilk. The variation might be due to the lower amount of fat in soymilk. In the present study, a value of 0.8 per cent for ash content was obtained in buffalo milk and 0.58 per cent in soy milk. The ash content in blends decreased proportionately with the amount of soymilk. The ash content of blends varied in proportion with the amount of soymilk with buffalo milk in the blends. The values of pH for buffalo milk and soymilk were 6.7 and 6.6 respectively in the present investigation. The pH decreased slightly and proportionately in the blends with an increase in the amount of soymilk. The variation in pH might be due to the pH of water used in preparation of soymilk and it may have slightly lower pH in comparison of buffalo milk.The value of acidity percent for buffalo milk and soymilk were 0.21 and 0.24, respectively in the present investigation. The acidity increased slightly and proportionately of the blends with an increase in the amount of soymilk. The variation was non significant in the blends up to 50 per cent level of soymilk and thereafter a significant (P≤0.05) increase was observed. Table 4.7: Effect of blending on chemical composition of soymilk with buffalo milkProportionof soymilk, (%)Total solids (%)Protein (%)Fat (%)Ash (%)pHAcidity (%)017.004.006.0000.8006.7900.210516.674.015.8140.7896.7870.2111016.344.035.6280.7786.7840.2131516.034.045.4420.7676.7810.2142015.684.065.2560.7566.7780.2162515.354.075.0700.7456.7750.2175013.704.0154.1400.6906.7600.2257512.054.0223.2100..6356.7400.23210010.404.0302.2800.5806.7400.240CD at 5%0.480.100.250.020.030.01%= per cent*Values are average of three determinationsFig 2. Effect of blending on chemical composition of soymilk with buffalo milk 4.3.2 Sensory Evaluation of blended buffalo milk with soy milk A laboratory panel consisting of 10 untrained members evaluated soy buffalo milk blends for sensory attributes such as colour, flavour, taste and overall acceptability.Sensory scores along with statistical analysis of soy buffalo milk blends are presented in table 4.8 to 4.11. Results presented in Table 4.8 showed that the mean sensory score for colour of buffalo milk was maximum. However, the inclusion of soymilk up to 25 per cent did not decrease the colour significantly (p≤0.05) and thereafter the increased levels of soymilk lowered the colour of blend significantly (p≤0.05). The mean score for colour of milk obtained from the blends containing 75 and 100 percent soymilk were in acceptable range on a 9 point hedonic scale, and did not differ significantly (p≤0.05) from each other. However, the blend containing 50 per cent soymilk was in an acceptable range but was rated significantly (p≤0.05) lower than all other blends containing less than 50 per cent soymilk and significantly (p≤0.05) higher than those having 75 and 100 per cent soymilk. The effect of soymilk inclusion with buffalo milk on the flavour presented in Table 4.9 showed that the buffalo milk scored highest for flavour on 9 point hedonic scale. It was found that the incorporation of soymilk up to 15 percent did not lower the mean scores for flavour significantly (p≤0.05). The increase in soymilk proportion from 15 to 20 had no significant (p≤0.05) variation in flavour. Further increase in the proportion of soymilk lowered the mean sensory score for flavour significantly (p≤0.05). The variation in flavour between the blends with 20 and 25 per cent soymilk and 25 and 50 per cent were non- significant. All the blend consisting up to 100 per cent proportion of soymilk were in acceptable The result of the effect of soymilk inclusion on the taste of blends showed that buffalo milk scored highest for taste on 9 point hedonic scale. It was observed that the incorporation of soymilk up to 15 percent did not lower the mean sensory scores for taste significantly (p≤0.05) and thereafter it had significant (p≤0.05) effect. However raising of soymilk from 15 to 20 per cent did not alter the taste significantly (p≤0.05).The blend prepared with 20 per cent soyamilk differ Table 4.8 -: Sensory scores for colour of soy milk and buffalo milk blendsPanelistsProportion of soymilk, percent055 510152025507510018887777762888777776388877777648888787775888887777688888877778788888768887888876988888787610887888767Mean87.97.87.77.67.57.36.96.4CD at 5% level 0.40Table 4.9: Sensory scores for flavour of soy milk and buffalo milk blendsPanelistsProportion of soymilk, percent055 5 10152025507510018777876762888777766387887777648887877665887877666688888776678888778768787787676988877786610888877666Mean7.97.87.77.57.476.86.46CD at 5% level: 0.44Table 4.10: Sensory scores for taste of soy milk and buffalo milk blendsPanelistsProportion of soymilk, percent055 510152025507510018877776662888877766387787777748878776775878777776688778777678788777768788787866988878777610788887777Mean7.87.77.67.57.476.96.76.3CD at 5% level 0.44Table 4.11: Sensory scores for overall acceptability of soy milk and buffalo milk blendsPanelistsProportion of soymilk, percent055 510152025507510018877777662888888766387787777748878787775878877776688878777678888877778888877776988878777710788787776Mean7.97.87.77.67.57.276.86.4CD at 5% level: 0.41significantly (p≤0.05) from buffalo milk. But further increase in the proportion of soymilk lowered the score significantly (p≤0.05).The differences in taste between the products made from blends having 20 and 25 per cent soymilk were non significant (p≤0.05) and those between 25 and 50 per cent soyamilk were non significant. The blend having 50 per cent soymilk was in acceptable range and thereafter it was in unacceptable range. The buffalo milk scored highest for mean overall attributes but the inclusion of soyamilk up to 15 per cent did not lower the mean overall sensory scores significantly (p≤0.05).The blend containing soymilk upto 100 per cent proportion were in acceptable range on a 9 point hedonic scale. The further increase in the proportion of soymilk in the blends lowered mean overall sensory scores below acceptable range on a 9 point hedonic scale.Statistical analysis of all the four attributes of sensory evaluation concluded that the incorporation of soymilk in buffalo milk up to 15 per cent proportion was found most suitable, however, soymilk may be included up to 25 per cent proportion in buffalo milk without significantly (p≤0.05) affecting any sensory attributes.4.4 Optimization of processing variables for the manufacture of soymilk 4.4.1 Optimization of Coagulant concentrationOn the basis of information available in literature some preliminary trials were conducted, two per cent citric acid and five per cent calcium lactate and five per cent calcium acetate were selected for coagulation of milk for preparation of paneer. At these coagulant concentrations maximum yield of paneer were obtained.Similar coagulants were also used by Johari (1989) and Ahmed etal (1981).Sen and De (1984) used calcium lactate as coagulant in the form of four percent solution.Sen and Rajorhia (1985) reported that ten grams of calcium lactate was required to coagulate one kg of fresh sweet milk at 80°C for maximum recovery of milk solids. . Shelke et al., (2002) used citric acid solution of 1.5 per cent during the production of chhana for rasogolla preparation. The amount of coagulant required for coagulation of milk decreased by increasing coagulation temperature irrespective of the type of milks (Table 4.12).The highest amount of coagulant was required for coagulating blended soya cow milk followed by soya buffalo milk. Rao, 1991 observed that 3.5 gram of citric acid per kg of cow milk was required for producing best quality chhana for rasogolla making. The optimum amount of calcium acetate required to coagulate one liter of cow milk was found four gram (Johari et al, 1994). Karadbhajne and Bhoyarkar (2010) observed that in case of ascorbic acid, the paneer prepared from 2 per cent and 4 per cent ascorbic acid was found very good in terms of per cent yield, colour, flavour, taste, and even in terms of shelf life as compared to citric acid, lactic acid and tartaric acid..Table 4.12: Amount of coagulant required per liter of milk at different temperaturesType of milkCoagulant concentration Temperature of coagulation(°C)Coagulant required per litre milk(ml)Cow2 per cent citric acid606570752802301801825 per cent calcium lactate606570752001701501505 per cent calcium acetate606570751301088085Buffalo1 per cent citric acid606570752602101601625 per cent calcium lactate606570751901721501505 per cent calcium acetate606570751241108085Cow+Soya1 per cent citric acid606570752602402202255 per cent calcium lactate606570751901601401455 per cent calcium acetate60657075120997075Buffalo + Soya1 per cent citric acid606570752402001501555 per cent calcium lactate606570751801601401455 per cent calcium acetate606570751169880854.4.2 Standardization of Coagulation Temperature4.4.2.1 Effect on moisture content, paneer yield and recovery of milk solids Milk samples were coagulated at 60°C, 65°C, 70°C and 75°C by using one per cent citric acid, five per cent calcium lactate and five per cent calcium acetate solutions. The results obtained with regard to yield, moisture content and recovery of milk solids in paneer are presented in Table 4.13.The maximum yield of paneer was obtained by coagulating milk at 70°C irrespective of the type of milk and coagulant used. As the coagulation temperature was decreased from, 70 to 65 °C and from 65 to 60°C the yield of paneer decreased. Also, by increasing the coagulation temperature from 70 to 75 °C the yield of paneer remained same .However, the moisture content of paneer increased by lowering the coagulation temperature. Sachdeva and Singh (1988) also made similar observations. The recovery of milk solids in paneer also decreased with decrease in coagulation temperature. Murdia and Wadhwani (2010) observed that the texture and yield of tofu can be increased by increasing the coagulation temperature from 80 to 90°C.The yield of paneer from all the types of milk obtained in the present investigation were lower as compared to the earlier findings of Sen and De (1984), and Sachdeva and Singh (1988a).They prepared chhana and paneer by using citric acid and calcium lactate. These differences in the yields may be accounted for the loss of milk solids in whey, as milk were coagulated at low temperatures. Mathare etal.,(2009) observed that there is an increase in yield with the increase in coagulation temperature. Masud etal.,(2007) observed that paneer prepared at 95oC required minimum amount of coagulant, contributing low cost of production and paneer prepared at 85oC got maximum sensory score.The moisture content in paneer was found maximum at 60 oC which decreased with increase in temperature up to 70oC and thereafter constant moisture content was obtained in paneer irrespective of type of milk and type of coagulant. The recovery of milk solids was found maximum on 70 oC coagulation temperature which decreased with lowering the temperature and was almost the same on increasing the temperature above 70 oC irrespective of type of coagulant and type of milk.From the result presented in Table 4.13 it can be concluded that the yield and recovery of milk solids was maximum with minimum moisture content at 70oC temperature. Hence 70oC temperature was standardized for coagulation of paneer.Verma etal., (2013) used blended calcium sulphate and citric acid of 2 per cent by weight (1 per cent +1 per cent) concentration for coagulation of the soymilk (soymilk + cream). Blended milk was heated to coagulation temperatures of 820C/5 min, and the coagulants were added slowly with gentle and continuous stirring at 800C. The maximum yield and recovery of total solids in paneer were obtained when milk were coagulated at 70°C by five per cent calcium acetate solution in case of all the types of milk. Similar observations were made by Johari etal. (1994).Table 4.13:- Effect of coagulation temperature and coagulant on the yield, moisture content and recovery of milk solids in paneerType of milkCoagulantTemperature of coagulation(°C)Yield(per cent)moisture(per cent)Recovery of milk solids(per cent)Cow1 per cent citric acid6065707516.2217.5018.8518.8358.9259.1556.0656.0655.0256.1857.4957.485 per cent calcium lactate6065707519.4020.0021.2621.2556.9856.1657.0557.0359.1359.9860.8160.805 per cent calcium acetate6065707519.9120.1621.5621.5556.9256.0255.1255.1161.9162.0362.7862.77Buffalo1 per cent citric acid6065707516.0816.7218.7418.7458.0257.1756.0856.0659.0560.5058.2058.205 per cent calcium lactate6065707518.7421.1621.8621.8555.8954.1253.4553.4458.2059.9861.3461.325 per cent calcium acetate6065707518.9521.3222.0022.0056.8755.1554.7854.7762.2662.9455.1955.15Cow+Soya1 per cent citric acid6065707515.9816.4017.7217.7157.6858.1557.0657.0654.8955.7656.8856.875 per cent calcium lactate6065707519.9820.2321.6121.6056.7857.2957.6657.6658.6359.2259.9859.975 per cent calcium acetate6065707520.1220.9921.6721.6457.8857.4556.3356.3161.9561.4362.2062.19Buffalo + Soya1 per cent citric acid6065707515.5516.7717.5617.5657.6656.456.0856.0759.3260.4360.8860.845 per cent calcium lactate6065707517.4420.3221.8621.8554.5653.0952.2152.2058.1259.5460.2160.195 per cent calcium acetate6065707518.6518.2119.4819.4654.5553.3354.2654.2562.3262.2161.6661.644.4.2.2 Effect of coagulation temperature on sensory quality of paneer The paneer samples prepared by coagulation of various types of milk at 600C, 650C, 700C and 750C, using different coagulants were evaluated for flavor, colour and appearance, smoothness, body and texture and overall acceptability, and the results obtained are presented in the table 4.14.The panelists were asked to evaluate paneer samples keeping in mind its suitability for paneer preparation. The highest sensory scores were awarded for paneer obtained by coagulating all types of milk at 700C irrespective of type of coagulant used. As the temperature of coagulation was increased from 60 to 650C or from 65 to 700C and from 70 to 750C, the paneer samples obtained were awarded significantly (P≤0.05) higher sensory scores when compared with one prepared at 700C.Similar observations were made by Sachdeva and Singh?1988b; Chandan?2007b. ?Sachdeva and Singh?1988b; Chandan?2007b proved that Paneer obtained by coagulating milk at 70?°C had the best organoleptic quality and had desired frying quality namely integrity/shape retention and softness .Among the coagulants , paneer samples prepared by using calcium acetate and citric acid were awarded significantly (P≤0.05) higher sensory scores followed by calcium lactate and calcium chloride. Similar observations were reported by Johari etal., (1994).The higher body and textural scores by the addition of calcium compound (CaCl2) have also been reported in the literature;?Kanawjia and Rizvi, 2003 and khan and Pal, 2010).Table 4.14 Effect of coagulation temperature on sensory attributes of cowmilk paneerCoagulantTemperature (°C)Scores for sensory attributesFlavourColour and appearanceSmoothnessBody and textureOverall acceptabilityCitric acid (1 per cent)606570757.07.58.06.97.07.58.07.06.06.57.55.96.57.07.56.36.57.58.56.3Calcium lactate solution (5 per cent)606570758.07.57.06.87.57.06.05.97.06.56.26.18.07.57.06.97.57.06.56.4Calcium acetate solution (5 per cent)606570758.58.07.57.28.07.06.56.37.06.56.06.07.57.06.56.38.07.06.56.3Table 4.15: Effect of coagulation temperature on sensory attributes of buffalo milk paneerCoagulantTemperature (°C)Scores for sensory attributesFlavourColour and appearanceSmoothnessBody and textureOverall acceptabilityCitric acid (1 per cent)606570757.07.58.56.96.07.57.57.07.06.58.05.97.07.07.06.47.07.56.56.3Calcium lactate solution (5 per cent)606570758.07.57.06.87.57.06.05.87.06.56.26.28.07.57.06.87.57.06.56.3Calcium acetate solution (5 per cent)606570758.58.07.57.18.07.06.56.17.06.56.05.77.57.06.56.48.07.06.56.0On the basis of sensory evaluation, it was revealed that among all the coagulation temperature, best paneer could be obtained by coagulating milk with citric acid at 70°C.Therefore, for all further studies 70 °C was taken as an ideal coagulation temperature for the preparation of paneer.4.5 Paneer made from blends of soymilk with cow milk (soy cow milk paneer)4.5.1Chemical composition of paneerThe result of chemical composition like total solids, protein, fat, ash, pH and acidity of paneer made from blends of soymilk with cow milk are presented in table 4.16. Cow milk was incorporated in soy milk at 5, 10, 15, 20, 25, 50, 75, and 100 per cent to blended paneer.From the results presented in Table 4.16 it is revealed that cow milk paneer and soymilk paneer contained 48.74 per cent and 45.51 per cent total solids, respectively. The total solid content in paneer made from different blends of soyamilk with cow milk varied proportionately but the variation was non significant. This indicated that the blending of soymilk with cow milk lowered the total solids in paneer by increasing the proportion level of soymilk as it had higher amount of moisture.The values for protein content in paneer made from cow milk and soy milk obtained in the present investigation were 21.23 percent and 27.67 percent respectively. Similar values are reported by Masud etal., 1992.The protein content in paneer made from blends varied proportionately and the variation was significant (P≤0.05).This indicated that the blending of soy milk with cow milk increased the amount of protein in paneer made from blends as the proportion level of soymilk increased. The values for fat content in paneer made from cow milk and soy milk in the present investigation were 22.46 per cent and 8.81 per cent respectively. The fat content in paneer prepared from the blends was decreased proportionately with an increase in the amount of soymilk in blends. Hence the fat content decreased significantly (P≤0.05) in paneer with an increase the level of soymilk during blending. Table 4.16: Effect of blending on chemical composition of paneer made from blends of soy milk with cow milkProportionof soymilk\soyapaneer(%)Total solids (%)Protein (%)Fat (%)Ash (%)pHAcidity (%)048.7421.2322.462.535.400 0.500548.5821.5421.782.445.3920.5011048.4221.8621.092.415.3840.5031548.2622.1920.412.355.3760.5042048.1022.5119.732.295.3600.5062547.9422.8319.042.235.3500.5075047.1224.4415.632.035.3200.5137546.3226.0512.211.685.2800.52210045.5127.678.812.925.2400.535CD at 5%0.120.340.380.030.020.01 % = per cent *Values are average of three determinationsFig 3 .Effect of blending on chemical composition of paneer made from blends of soy milk with cow milkIn the present study, a value of 2.53 per cent for ash content was obtained in cow milk paneer and 2.92 per cent in soy milk paneer. The ash content of both were similar to the values reported by Krupa et al (2011). The ash content in paneer made from blends varied proportionately and the variation was non significant upto 25 per cent proportion of soymilk in cow milk thereafter the increase in ash content was significant (P≤0.05).The values of pH for cow milk paneer and soymilk paneer were 5.400 and 5.240 respectively in the present investigation. The pH decreased slightly and proportionately in paneer made from the blends with an increase in the amount of soymilk. The variation in pH might be due to the pH of water used in preparation of soymilk as it may have slightly lower pH in comparison of cow milk paneer.The variation in pH was non significant. The value of acidity per cent for cow milk paneer and soymilk paneer were 0.500 and 0.535, respectively determined in the present investigation. The acidity increased slightly and proportionately in paneer made from blends with an increase in the amount of soymilk. The variation was non significant in the paneer up to 50 percent level of soymilk and thereafter a significant (P≤0.05) increase was observed. The variation might be because of the variation in milk used for blending. 4.5.2 Sensory Evaluation of blended paneer made from blends of soymilk with cow milk A laboratory panel consisting of 10 untrained members evaluated paneer made from blends of soymilk with cow milk for sensory attributes such as colour, flavour, taste and overall acceptability. Sensory scores along with statistical analysis of blended paneer are presented in table 4.17 to 4.20 Results presented in Table 4.17 showed that the mean sensory score for colour of cow milk paneer was maximum. However, the inclusion of soymilk up to 20 per cent did not decrease the colour significantly (p≤0.05) and thereafter the increased levels of soymilk paneer lowered the colour of blend significantly (p≤0.05). The mean score for colour of milk obtained from the blends containing 75 and 100 per cent soymilk Table 4.17-: Sensory scores for colour of paneer made from blends of soymilk with cow milk PanelistsProportion of soymilk/ soyapaneer, percent055 510152025507510018888777672888877766387887877748888877665888888776688788877778878877768888777776988878677710888787776Mean87.97.87.77.67.276.76.4CD at 5% level 0.41Table 4.18: Sensory scores for flavor of paneer made from blends of soymilk with cow milk PanelistsProportion of soymilk/soyapaneer, per cent055 510152025507510018878777762888877777387887787648888866675888887666688877777778778887768887778776988878877610788787766Mean7.97.87.77.67.57.26.96.76.3CD at 5% level 0.47Table 4.19: Sensory scores for taste of paneer made from blends of soymilk with cow milk PanelistsProportion of soymilk/ soyapaneer, percent055 510152025507510018887777662888877777388788776648878877675888788777688877877678878787778878877776988878777610778887776Mean7.97.87.77.67.57.376.76.4CD at 5% level 0.42Table 4.20: Sensory scores for overall acceptability of paneer made from blends of soymilk with cow milk PanelistsProportion of soymilk/ soyapaneer, percent0 55 510152025507510018877777772888878777387787777748888787675888787767688788777678788787768788887776988878777610888787776Mean7.97.87.77.67.57.376.86.5CD at 5% level 0.42were in acceptable range on a 9 point hedonic scale, and did not differ significantly from each other. However, the blend containing 50 percent soymilk was in an acceptable range but was rated significantly (p≤0.05) lower than all other blends containing less than 50 per cent soymilk and significantly (p≤0.05) higher than those having 75 and 100 per cent soymilk paneer.The effect of soymilk inclusion with cow milk on the flavour of paneer made from blends presented in Table 4.18 It showed that the cow milk paneer scored highest for flavour on 9 point hedonic scale. It was found that the incorporation of soymilk up to 20 per cent did not lower the mean scores for flavour significantly (p≤0.05). The increase in soymilk with cow milk proportion from 15 to 25 had no significant variation in flavour of paneer made from these blends. Further increase in the proportion of soy milk in cow milk lowered the mean sensory score for flavour significantly (p≤0.05) in paneer. The variation in flavour between the blends with 20 and 25 per cent soymilk paneer and 25 and 50 per cent were non- significant. The blend consisting 50, 75 and 100 per cent proportion of soymilk with cow milk were in acceptable range. The result of the effect of soymilk (Table 4.19) inclusion with cow milk on the taste of paneer showed that cow milk scored highest for taste on 9 point hedonic scale. It was observed that the incorporation of soymilk with cow milk to prepare paneer up to 20 per cent did not lower the mean sensory scores for taste of paneer significantly (p≤0.05) and thereafter it had significant (p≤0.05) effect. However raising of soymilk proportion from 20 to 25 and 25 to 50 per cent to prepare paneer did not alter the taste significantly (p≤0.05).The paneer made from blends containing 25 per cent soyamilk differ significantly (p≤0.05) from cow milk paneer. But further increase in the proportion of soymilk lowered the score significantly (p≤0.05).The differences in taste between the paneer made from blends having 20 and 25 per cent soymilk were non significant (p≤0.05) and those between 25 and 50 per cent soyamilk were non significant. All the blend having soymilk were reported in acceptable range by the panelist. The results presented in Table 4.20 showed that the paneer made from cow milk scored highest for mean overall acceptability .The inclusion of soymilk with cow milk to prepare paneer up to 20 per cent did not lower the mean overall sensory scores significantly (p≤0.05).The paneer made from blend containing 25 and 50 and 75 and 100 per cent soymilk did not differ significantly (p≤0.05) with each other for overall acceptability score on a 9 point hedonic scale. From the results of all the sensory attributes, it can be concluded that the paneer prepared from the blends containing 20 per cent proportion of soymilk did not defer significantly from the paneer made from cow milk. Hence soymilk may be included with cow milk upto 20 per cent proportion to prepare paneer satisfactorily. Statistical analysis of all the four attributes of sensory evaluation concluded that the incorporation of soymilk/soypaneer in cow milk paneer up to 15 per cent proportion was found most suitable, however, soymilk/soypaneer may be included up to 25 per cent proportion in cow milk without significantly (p≤0.05) affecting any sensory attributes4.6 Paneer made from blends of soymilk with buffalo milk4.6.1Chemical composition of blended paneerThe result of chemical constituents like total solids, protein, fat, ash, total solids, pH and acidity of paneer made from blends of soymilk with buffalo milk are presented in Table 4.22. Soy milk was incorporated in buffalo milk at 5, 10, 15, 20, 25, 50, 75, and 100 per cent level to prepare paneer. From the results presented in Table 4.21 it is revealed that buffalo milk paneer and soymilk paneer contained 49.93 per cent and 45.51 per cent total solids, respectively. The total solid content in paneer varied proportionately but the variation was non significant. This indicated that the blending of soymilk with buffalo milk decreased the amount of total solids because the coagulum of soy protein retain more moisture than buffalo milk protein coagulum.The values for protein in paneer made from buffalo milk and soy milk obtained in the present investigation were 16.09 per cent and 27.67 per cent respectively. Similar values are reported by Masud etal, 1992 and Gandhi (2009).The protein content in paneer varied proportionately to the amount of soymilk and buffalo milk in blends from which the paneer was made.The increase in protein was significant (P≤0.05) with the increase of the amount of soyamilk in blends. Table 4.21-: Effect of blending on chemical composition of paneer made from blends of soy milk with buffalo milk Proportionof soymilk\soyapaneer (%)Total solids (%)Protein (%)Fat (%)Ash (%)pHAcidity (%)049.9316.0928.862.675.20 0.50549.7116.6427.852.685.200.501049.4917.1526.852.695.190.501549.2817.7625.852.715.190.502049.0318.3224.842.735.160.512548.8318.9023.842.755.160.515047.7321.8618.832.805.150.517546.6322.7513.812.855.150.5210045.5127.678.812.925.140.54CD at 5%0.180.450.380.050.030.01 %= per cent *Values are average of three determinationsFig 4.Effect of blending on chemical composition of paneer made from blends of soy milk with buffalo milkThe values for fat content in paneer obtained in the present investigation from the blends of soy milk with buffalo milk decreased significantly (P≤0.05) with an increase of soymilk in blends from which paneer was made.It might be due to the lesser amount of fat present in soymilkIn the present study, a value of 2.67 per cent for ash content was obtained in paneer made from buffalo milk and 2.92 per cent in soy milk paneer. The ash content of both were similar to the values reported by Gandhi, 2009. The ash content of paneer showed a slight increase but the variation was non significantThe values of pH for buffalo milk paneer and soymilk paneer were 5.20 and 5.14, respectively in the present investigation. The pH decreased slightly and proportionately in the blends with an increase in the amount of soymilk. The variation in pH might be due to the pH of water used in preparation of soymilk and it may have slightly lower pH in comparison of buffalo milk paneer.The value of acidity percent for buffalo milk paneer and soymilk paneer were 0.50 and 0.54, respectively in the present investigation. The acidity increased slightly and proportionately in the blended paneer with an increase in the amount of soymilk in blends. The variation was non significant in the paneer up to 50 per cent level of soymilk and thereafter a significant (P≤0.05) increase was observed.The blending of soy milk with buffalo milk to prepare paneer showed a significant increase in protein and significant decrease in fat of paneer with the increase of soymilk in blends. 4.6.2 Sensory Evaluation of paneer made from blends of soy milk with buffalo milk Sensory scores along with statistical analysis of blended paneer are presented in table 4.22 to 4.25. Results presented in Table 4.22 showed that the mean sensory score for colour of paneer made from buffalo milk and 5 per cent inclusion of soymilk. Paneer made from blends consisting soymilk upto 20 was maximum. However, the inclusion of soymilk/soypaneer up to 25 per cent did not decrease the colour significantly (p≤0.05) and thereafter the increased levels of soymilk paneer lowered the colour of blend significantly(p≤0.05). The mean score for colour of paneer obtained from the blends containing 25, 50, 75 and 100 per cent soymilk Table 4.22: Sensory scores for colour of paneer made from blendsPanelistsProportion of soymilk/ soypaneer, percent055 510152025507510018887777762888777776387887777648887887775888888777688788777678878877678888878777988888777610788887766Mean7.97.97.87.77.67.376.86.4CD at 5% level 0.39Table 4.23: Sensory scores for flavour of paneer made from blendsPanelistsProportion of soymilk/ soyapaneer, percent055 510152025507510018877778772888877877387787777748878876765888788777688877777778888877778878878776988888777710788788877Mean7.97.87.77.67.57.37.276.8CD at 5% level 0.43Table 4.24: Sensory scores for taste of paneer made from blendsPanelistsProportion of soymilk/soyapaneer, percent055 510152025507510018887777772888877776388787776748878787765878778766678778777778878877778878787777988878877710778877777Mean7.87.77.67.57.47.376.86.7CD at 5% level 0.43Table 4.25: Sensory scores for overall acceptability of paneer made from blendsPanelistsProportion of soymilk/ soyapaneer, percent055 510152025507510018877777762888878676387787777648888887775888777766688888787678888877778888888767988888777710888787777Mean87.97.87.77.67.376.86.5CD at 5% level 0.41were in acceptable range on a 9 point hedonic scale, and did not differ significantly (p≤0.05) from each other. The effect of inclusion of soymilk with buffalo milk to prepare paneer on the flavour presented in table 4.23 showed that the buffalo milk paneer scored highest for flavour on 9 point hedonic scale. It was found that the incorporation of soymilk up to 30 per cent did not lower the mean scores for flavour significantly (p≤0.05). The increase in soymilk/soypaneer proportion from 20 to 50 had no significant (p≤0.05) variation in flavour. Further increase in the proportion of soymilk lowered the mean sensory score for flavour significantly (p≤0.05). The variation in flavour between the paneer made from blends 50, 75 and 100 per cent soymilk were non- significant. The paneer made from all the blends was in acceptable range. The result in Table 4.24.present the effect of soymilk inclusion on the taste of paneer made from different blends. The paneer made from buffalo milk scored highest for taste on 9 point hedonic scale. It was observed that the incorporation of soymilk with buffalo milk up to 20 per cent did not lower the mean sensory scores for taste significantly (p≤0.05) and thereafter it had significant (p≤0.05) effect. However raising of soymilk percent soyamilk with buffalo milk to prepare paneer from 20,25 and 50 per cent did not alter the taste significantly (p≤0.05).The blended paneer prepared from 25 per cent soymilk included blend differ significantly (p≤0.05) from buffalo milk paneer. But further increase in the proportion of soymilk lowered the score significantly (p≤0.05).The differences in taste between the paneer made from blends having 50 and 75 percent soymilk were non significant (p≤0.05) and those between 75 and 100 percent soyamilk were non significant. The results for mean sensory score for overall acceptability are presented in Table 4.25 showed that the buffalo milk scored highest for mean overall attributes but the inclusion of soymilk with buffalo milk to prepare paneer up to 20 per cent did not lower the mean overall sensory scores significantly (p≤0.05).The blend containing 20 to 25, 25 to 50, 50 to 75 and 75 to 100 per cent soymilk were non significant with each other and were in acceptable range on a 9 point hedonic scale. Statistical analysis of all the four attributes of sensory evaluation concluded that the incorporation of soymilk in buffalo milk to prepare paneer up to 20 per cent proportion was found most suitable with put significantly affecting the mean sensory scores of any attribute 4.7 Storage studySoymilk blended with cow milk (20:80) paneer was kept at refrigerated conditions at 4°C for 12 days. The samples were analysed for physico - chemical microbiological and sensory quality after an interval of every 3 days.4.7.1 Physico-chemical changesThe paneer samples were analysed for moisture, fat, protein, and free fatty acids, titratable acidity and pH at room temperature. The results obtained are presented in Table 4.26Table 4.26: Physico chemical changes in Paneer during storageType of paneerStorage period in daysMoisture (%)Fat (%)Protein (%)TitratableAcidity(%)FFA(%)pHSoy cow milk paneer (20:80)051.9022.5119.730.500.0825.37351.6522.5119.730.530.0975.26651.3022.4919.730.560.1085.15950.9722.4519.700.590.1205.081250.622.3519.700.620.1604.95CD at 5%0.0080.0080.0070.0080.0050.003Soya buffalo milk paneer (85:15)050.9724.8418.320.510.1365.16350.6224.8018.320.530.1455.05650.2724.7818.320.560.1514.86950.0024.7618.300.590.1594.801249.7824.7218.300.630.1664.65CD at 5%0.0080.0080.0070.0080.0010.003% = per cent *Values are average of three determinationsThe moisture content of paneer decreased significantly (p≤0.05) during storage in all samples studied. The fat content also decreased significantly (p≤0.05) during storage. This decrease in fat content may be due to lipolysis.The degradation of fat is primarily caused by the growth of yeast and molds (Kumar and Srinivasn,1983b).No significant difference could be established in protein content of paneer samples with respect of type of milk even during storage. Hence, no definite trend in protein content value could be observed during storage. The free fatty acid significantly (p≤0.05) increased during storage. This increase in FFA content in paneer has been attributed to hydrolysis of fat. Hasan (2001) also observed a significant increase in FFA content during storage of paneer spread. The titratable acidity significantly (p≤0.05) increased during storage at 4°C due to the activity of microorganisms. Similar observations were made by Khodke 2012 in soyapaneer during storageThe pH decreased significantly (p≤0.05)during storage of all the samples.Similar observations were made by Goyal and Srinivasn (1989) in channa samples during storage. According to Anbarasu and Vijayalakshmi (2007), pH of soypaneer increased or decreased on type of contaminating micro-organisms.The paneer samples were analysed for moisture, fat, protein, and free fatty acids, titratable acidity and pH at refrigerated storage (4°C). The results obtained are presented in Table 4.27Table 4.27Physico chemical changes in Paneer during storageType of paneerStorage period in daysMoisture (%)Fat (%)Protein (%)TitratableAcidity(%)FFA(%)pHSoyacow milk paneer (85:15)051.9021.2321.320.530.0825.37351.7521.2321.320.530.0925.37651.4521.2121.320.550.1025.32951.2021.1821.290.560.1155.301249.9021.1221.290.570.1505.26CD at 5%0.0080.0080.0070.0070.0080.003Soya buffalo milk paneer (85:15)050.9729.6218.160.530.1365.36350.6029.6218.160.530.1385.36650.3029.6018.160.550.1455.31950.0029.5818.130.560.1535.291249.7829.5518.130.570.1625.25CD at 5%0.0080.0080.0070.0070.0080.003% = per centThe moisture content of paneer decreased significantly (p≤0.05) during storage in all samples studied.The fat content also decreased significantly (p≤0.05) during storage.This decrease in fat content may be due to lipolysis.The degradation of fat is primarily caused by the growth of yeast and molds (kumar and Srinivasn,1983b).No significant difference could be established in protein and free fatty acid content of paneer samples with respect of type of milk even during storage.Hence, no definite trend in protein content and free fatty acids value could be observed during storage.The titratable acidity significantly (p≤0.05) increased during storage at 4°C due to the activity of microorganisms. Similar observations were made by Khodke 2012 in soyapaneer during storageThe pH decreased significantly (p≤0.05)during storage of all the samples.Similar observations were made by Goyal and Srinivasn (1989) in channa samples during storage. According to Anbarasu and Vijayalakshmi (2007), pH of soypaneer increased or decreased on type of contaminating micro-organisms.4.7.2 Microbiological quality of paneerMicrobiological changes in paneer were enumerated with respect to total plate count (TPC), yeast and mold count,and coliform count during storage at 30±5°C and 5±1°C.4.7.2.1 Total plate count (TPC)The Effect of storage temperature,and storage period of paneer were determined by assessing its TPC which is presented in Table 4.28.The data revealed that there was a rapid increase in TPC at room temperature than at refrigeration temperature. The results showed that the initial total plate count of paneer at room temperature on zero days was 3.5 × 102 cfu/g and at the end of 2nd day it was 4.6 × 103 cfu/g and at the end of 3rd day it was 5.2 × 103 cfu/g.At refrigeration temperature, paneer samples showed TPC 4.6 × 103 cfu/g on 6th day.Thus it can be stated that quality maximum shelflife of paneer was found 6 days at refrigeration temperature and 2 days at room temperature.Hasan (2001) also noted similar changes in total plate count of paneer spread during storage for Table 4.28 Microbiological changes in paneer (Total plate count) during storage at refrigeration temperature and room temperature.Type of paneerStorage period in daysTotal plate count at room temperature (cfu/g)Total plate count at refrigeration temperature (cfu/g)Soya cow milk paneer (85:15)03.5 × 102 3.5 × 10235.2 × 1034.0× 1036N.A.4.6 × 1039N.A.5.5 × 10312N.A.N.A.Soya buffalo milk paneer (85:15)03.5 × 102 3.5 × 10235.2 × 1034.0× 1036N.A.4.6 × 1039N.A.5.5 × 10312N.A.N.A. *Values are average of three determinationsN.A.= Not applicable (not fit for consumption)12 days at 7°C. Lamdande (2012) also noted similar changes in total plate count of paneer during storage for 14 days at 5°C and 30°C.4.7.2.2 Yeast and mold countTable 4.29 shows the yeast and mold count of paneer during storage at room temperature and refrigeration temperature.At room temperature , paneer samples showed yeast and mold count 4.8 × 103 cfu/g after 2 days of storage. At refrigeration temperature, paneer samples showed yeast and mold count 3.6 × 103 cfu/g on 6th day. Hasan (2001) also noted similar changes in yeast and mold of paneer spread during storage for 12 days at 7°C. Lamdande (2012) also observed same trend in yeast and mold of paneer during storage for 14 days at 5°C and 30°C.Table 4.29 :Microbiological changes in paneer (yeast and mold count) during storage at refrigeration temperature and room temperature.Type of paneerStorage period in daysYeast and mold count at room temperature (cfu/g)Yeast and mold count at refrigeration temperature (cfu/g)Soya cow milk paneer (85:15)02.5 × 102 2.5 × 10234.8× 1033.0× 1036N.A.3.6 × 1039N.A.5.5 × 10312 N.A.N.A.Soya buffalo milk paneer (85:15)02.5 × 102 2.5 × 10234.8× 1033.0× 1036N.A.3.6 × 1039N.A.5.5 × 10312N.A.N.A. *Values are average of three determinationsN.A.= Not applicable (not fit for consumption)4.7.2.3 Coliform countThe changes in coliform count of paneer are presented in Table 4.30.At room temperature, coliforms were detected after 3 days of storage in all the samples.Paneer had coliform count of 8.1× 101/g after 3 days of storage.At refrigeration temperature, coliform count was 8.1× 101/g after 9 days of storage. Hasan (2001) also noted similar changes in yeast and mold of paneer spread during storage for 12 days at 7°C.Table 4.30 :Microbiological changes in paneer (coliform count) during storage at refrigeration temperature and room temperature.Type of paneerStorage period in daysColiform count at room temperature (cfu/g)Coliform count at refrigeration temperature (cfu/g)Soya cow milk paneer (85:15)0N.D. N.D.38.1× 1011.3× 1016N.A.2.5 × 1019N.A.8.1 × 10112N.A.N.A.Soya buffalo milk paneer (85:15)0N.D. N.D.38.1× 1011.3× 1016N.A.2.5 × 1019N.A.8.1 × 10112N.A.N.A. *Values are average of three determinationsN.A.= Not applicable (not fit for consumption) 4.7.3 Changes in sensory scores of soy cow milk paneer and soy buffalo milk paneerThe results showing the effect of storage time and temperature on sensory scores for colour, flavour, taste and overall acceptability during storage in Table 4.31to 4.464.7.3.1 Colour/AppearanceThe sensory scores of blended paneer for colour and appearance obtained during storage are presented in Table 4.31 to 4.34 .The sensory scores for colour and appearance of paneer during storage decreased for all the samples.The sensory scores remained within the acceptable limit upto 6 days at refrigeration temperature.At room temperature, this limit was found 3 days.Similar findings were observed by Hasan (2001).Table 4.31: Effect on colour of blended paneer (soy cow milk paneer,85:15) during storage at room temperaturePanelistsNo. of days0 3 3691218.04.0NANANA28.05.0NANANA37.04.0NANANA48.04.0NANANA57.05.0NANANA68.05.0NANANA78.04.0NANAjNA87.04.0NANANA98.05.0NANANA108.04.0NANANAMean7.604.40NANANACD at 5% level: 1.05NA=Not AcceptableTable 4.32: Effect on colour of blended paneer (soy cow milk paneer, 85:15) during storage at refrigeration temperaturePanelists No. of days0 33691218.06.06.05.0NA28.06.06.04.0NA38.06.06.05.0NA47.07.05.04.0NA58.07.06.04.0NA68.07.05.04.0NA78.07.06.05.0NA88.07.06.05.0NA98.07.05.04.0NA108.08.06.05.0NAMean7.906.806.204.50NACD at 5% level: 0.71NA=Not AcceptableTable 4.33: Effect on colour of blended paneer (soy buffalo milk paneer,85:15) during storage at room temperaturePanelistsNo. of days0 33691218.04.0NANANA28.04.0NANANA37.04.0NANANA48.04.0NANANA57.05.0NANANA68.04.0NANANA78.05.0NANANA88.05.0NANANA98.04.0NANANA108.05.0NANANAMean7.804.40NANANACD at 5% level: 0.99NA=Not AcceptableTable 4.34: Effect on colour of blended paneer (soy buffalo milk paneer,85:15) during storage at refrigeration temperaturePanelists No. of days0 33691218.06.06.05.0NA28.06.05.04.0NA38.06.06.04.0NA47.07.05.04.0NA58.07.06.04.0NA68.07.05.04.0NA77.07.06.05.0NA88.07.05.05.0NA98.07.05.04.0NA108.07.06.05.0NAMean7.806.705.904.40NACD at 5% level: 0.70NA=Not Acceptable4.7.3.2 FlavourThe results showed a decreasing trend of sensory scores for flavour at refrigeration and room temperature during storage. The decline in flavour scores of paneer samples at refrigeration temperature was appreciably slower than in the paneer stored at room temperature. However, the sensory scores for flavour at refrigeration temperature were within acceptable range after 6 days in paneer.At room temperature paneer had sensory score for flavour with in acceptable limit only upto 3 days.Decrease in flavour was positively related with physico chemical changes.As titratable acidity, and free fatty acid values increased during storage period, the flavour score of paneer decreased correspondingly.Table 4.35: Effect on flavour of blended paneer (soy cow milk paneer,85:15) during storage at room temperaturePanelistsNo. of days0 33691218.04.0NANANA28.05.0NANANA37.05.0NANANA48.04.0NANANA58.04.0NANANA67.04.0NANANA78.04.0NANANA88.05.0NANANA98.04.0NANANA108.04.0NANANAMean7.804.30NANANACD at 5% level: 0.95NA=Not AcceptableTable 4.36: Effect on flavour of blended paneer (soy cow milk paneer,85:15) during storage at refrigeration temperaturePanelists No. of days0 33691218.07.06.05.0NA28.06.06.05.0NA38.06.06.05.0NA48.06.07.04.0NA57.07.07.04.0NA68.07.07.04.0NA77.07.06.05.0NA87.07.06.04.0NA98.07.07.04.0NA108.06.06.04.0NAMean8.247.126.404.80NACD at 5% level: 0.73NA=Not AcceptableTable 4.37: Effect on flavour of blended paneer (soy buffalo milk paneer,85:15) during storage at room temperaturePanelistsNo. of days0 33691218.04.0NANANA27.05.0NANANA37.05.0NANANA48.04.0NANANA57.04.0NANANA67.04.0NANANA78.04.0NANANA88.04.0NANANA98.04.0NANANA108.04.0NANANAMean7.704.20NANANACD at 5% level: 0.99NA=Not AcceptableTable 4.38: Effect on flavour of blended paneer (soy buffalo milk paneer,85:15) during storage at refrigeration temperaturePanelists No. of days0 33691218.07.06.05.0NA28.06.06.05.0NA38.06.05.05.0NA48.06.06.04.0NA58.07.07.04.0NA68.06.07.04.0NA77.07.06.05.0NA87.07.06.04.0NA98.07.07.04.0NA108.06.06.04.0NAMean7.806.506.204.40NACD at 5% level: 0.76NA=Not Acceptable4.7.3.3 TasteThe results showed a decreasing trend of sensory scores for taste at refrigeration and room temperature during storage.The decline in taste scores of paneer samples at refrigeration temperature was appreciably slower than in the paneer stored at room temperature.However, the sensory scores for taste at refrigeration temperature were within acceptable range after 6 days in paneer.At room temperature paneer had sensory score for taste with in acceptable limit only upto 3 days.Decrease in taste was positively related with physico chemical changes.As titratable acidity, and free fatty acid values increased during storage period, the taste score of paneer decreased correspondingly.Table 4.39: Effect on taste of blended paneer (soy cow milk paneer,85:15) during storage at room temperaturePanelistsNo. of days0 33691218.04.0NANANA28.05.0NANANA37.05.0NANANA48.04.0NANANA58.04.0NANANA68.04.0NANANA78.04.0NANANA88.05.0NANANA98.04.0NANANA108.04.0NANANAMean7.904.40NANANACD at 5% level: 0.86NA=Not AcceptableTable 4.40: Effect on taste of blended paneer (soy cow milk paneer,85:15) during storage at refrigeration temperaturePanelists No. of days0 33691218.07.06.05.0NA28.06.06.05.0NA38.06.05.04.0NA48.06.06.04.0NA58.06.07.04.0NA67.06.07.04.0NA78.07.06.05.0NA87.07.07.04.0NA98.07.07.04.0NA108.06.06.04.0NAMean7.806.406.304.30NACD at 5% level: 0.76NA=Not AcceptableTable 4.41: Effect on taste of blended paneer (soy buffalo milk paneer, 85:15) during storage at room temperaturePanelists No. of days0 33691218.04.0NANANA27.05.0NANANA37.04.0NANANA48.04.0NANANA57.04.0NANANA67.04.0NANANA77.04.0NANANA88.04.0NANANA98.04.0NANANA108.04.0NANANAMean7.604.10NANANACD at 5% level: 0.91NA=Not AcceptableTable 4.42: Effect on taste of blended paneer (soy buffalo milk paneer, 85:15) during storage at refrigeration temperaturePanelists No. of days0 33691218.07.06.05.0NA28.06.05.04.0NA38.06.06.05.0NA48.06.06.04.0NA57.07.07.04.0NA68.06.06.04.0NA77.07.06.04.0NA88.07.06.04.0NA98.07.07.04.0NA107.06.06.04.0NAMean7.706.506.104.20NACD at 5% level: 0.72NA=Not Acceptable4.7.3.4 Overall acceptabilityThe sensory scores of paneer for overall acceptability obtained during storage are presented in table 4.43 to 4.46.The overall acceptability scores of paneer during storage decreased for all the samples. The overall acceptability scores at refrigeration temperature were within acceptable range after 9 days but at room temperature , this acceptable period was reduced to 3 days.The reduction in overall acceptability scores might have been observed due to cumulative effect of change in flavor/taste,colour/appearance sensory scores of paneer during storage. Similar observations were made by Hasan (2001).Table 4.43: Effect on overall acceptability of blended paneer (soy cow milk paneer, 85:15) during storage at room temperaturePanelistsNo. of days0 33691218.04.0NANANA28.05.0NANANA38.04.0NANANA48.04.0NANANA58.04.0NANANA68.04.0NANANA77.05.0NANANA88.04.0NANANA98.04.0NANANA108.04.0NANANAMean7.904.20NANANACD at 5% level: 0.78NA=Not AcceptableTable 4.44: Effect on overall acceptability of blended paneer (soy cow milk paneer,85:15) during storage at refrigeration temperaturePanelists No. of days0 33691218.07.06.05.0NA28.06.05.04.0NA38.06.06.05.0NA48.06.06.04.0NA57.07.07.04.0NA68.06.06.04.0NA77.07.06.04.0NA88.07.07.04.0NA98.07.07.04.0NA108.06.06.04.0NAMean7.806.506.204.20NACD at 5% level: 0.73NA=Not AcceptableTable 4.45: Effect on overall acceptability of blended paneer (soy buffalo milk paneer, 85:15) during storage at room temperaturePanelists No. of days0 33691218.04.0NANANA27.04.0NANANA38.05.0NANANA48.05.0NANANA58.04.0NANANA68.05.0NANANA77.04.0NANANA88.05.0NANANA98.05.0NANANA108.05.0NANANAMean7.804.60NANANACD at 5% level: 0.99NA=Not AcceptableTable 4.46: Effect on overall acceptability of blended paneer (soy buffalo milk paneer, 85:15) during storage at refrigeration temperaturePanelists No. of days0 33691218.07.06.05.0NA27.06.06.04.0NA38.06.06.05.0NA48.06.06.04.0NA57.07.07.04.0NA68.07.06.05.0NA77.07.07.04.0NA88.07.07.04.0NA98.07.07.04.0NA108.06.06.04.0NAMean7.706.606.404.30NACD at 5% level: 0.71NA=Not Acceptable4.7.4 Textural analysisThe textural characteristics of the blended paneer samples were determined to evaluate the changes in the hardness characteristic of the blended paneer. Many instrumental measurements have been reported for the evaluation of paneer texture (Desai et al. (1991), Shrivastav and Kumbhar (2010) Karadbhajne and Bhoyarkar, (2010).. The hardness of the paneer was gradually increases during the storage period. The changes in the hardness strength of the paneer samples were assessed during the storage period of 12 days with an interval of 3 days (table 4.47 to 4.48). The initial maximum hardness of blended soy cow milk paneer samples and blended buffalo milk paneer samples were 157.7g and158.4 g respectively .This hardness strength was increased to the level of 158.9 g and 159.5 g at room temperature and 158.6 g and 159.6 g at refreigeration temperature respectively after 12 days of storage. Hardness of paneer increased with increase in temperature and pressure.Similar observations were made by Shrivastav and Kumbhar (2010) The change of hardness showed non significant difference (P≥0.05). Table 4.47 : Changes in texture of paneer during storage at room temperature Hardness testBlended sampleStorage period in days0 days3 days6 daysSoy cow milk paneer sample (85:15)157.7 g158.9 gNASoy buffalo milk paneer sample (85:15)158.4159.5 gNANot AcceptableTable 4.48 : Changes in texture of paneer during storage at refrigeration temperature Hardness testBlended sampleStorage period in days0 days3 days6 days9 days12 daysSoy cow milk paneer sample (85:15)157.7 g158.0 g158.2 g158.6 gNASoy buffalo milk paneer sample (85:15)158.7 g159.0 g159.3 g159.6 gNANA=Not Acceptable Chapter -5Summary SUMMARYThe present investigation was envisaged with the objective of utilizing soymilk for developing highly nutritious paneer by blending it with cow milk and buffalo milk. The effect of blending of soy milk with cow milk and buffalo milk on physicochemical characteristics and sensory scores of milk and their blends were studied. The proportion of soymilk blending with cow milk and buffalo milk to prepare paneer was optimized. The effect of blending of soymilk on physicochemical, textural and sensory characteristics of paneer was studied. The changes in physiochemical, textural and sensory characteristics of paneer during storage at refrigeration temperature and room temperature were studied. The extensive studies were made in respect of changes in color, flavor, taste and overall acceptability due to blending of soymilk with cow and buffalo milk and paneer made from these blends were studied. The results were summarized as below:The moisture, fat, protein, ash and carbohydrate content of soybean were 7.60, 19.68, 34.76, 4.47 and 33.49 percent. It contained 277 and 704 mg per 100 g of calcium and phosphorus, respectively. The soy milk prepared had 90.80, 2.28, 4.44, 0.58 and 3.1 percent moisture, fat, protein, ash and carbohydrate respectively. The amount of calcium and phosphorus was found 25 and 52 mg/100 g of soy milk respectively.The cow milk contained 87.10, 3.80, 4.00, 0.70 and 4.4 percent moisture, fat, protein, ash and carbohydrate. The moisture, fat, protein, ash and carbohydrate were 83.0, 6.5, 3.8, 0.8 and 5.9 percent respectively in buffalo milk. The cow milk had 119.843 and 94.03 mg per 100g of calcium and phosphorus, respectively. The buffalo milk had higher amount of calcium and phosphorus i.e. 164.19 and 113.0 percent respectively as compared to soymilk and cow milk.The soy milk was blended with cow milk and buffalo milk in proportion of 0, 5, 10, 15, 20, 25, 50, 75 and 100 percent. The blending of soymilk enhanced the level of protein and decreased the level of total solids, fat and ash in the blends with cow milk. The variation in proximate compositional constituents was proportional and significant (p≤0.05) the values of pH and acidity were almost same for cow milk and soymilk.The results of sensory evaluation showed that the incorporation of soymilk in blends of cow milk up to 15% proportion was found suitable without affecting the sensory characteristics significantly. The blends containing other proportions of soymilk in cow milk were in acceptable range. However, the blends containing soy milk up to 15% level were significantly superior to other blends, hence it was concluded that soymilk may be incorporated in cow milk up to 15 %.The blending of soymilk with buffalo milk increased the protein content proportionately and significantly (p≤0.05) and decreased the total solids, fat contents, and ash contents. The pH and acidity was more or less similar in all the blends of soy milk with buffalo milk. The results of the sensory evaluation showed that the mean sensory scores for color, flavor, taste and overall acceptability had no significant effect up to 15 % level of soymilk. Thereafter, a significant (p≤0.05) lowering in sensory attributes was observed in comparison to buffalo milk. All the blends were in acceptable range on a nine point headoinc scale, hence 15% soymilk may be included in buffalo milk suitably without lowering the sensory attributes significantly.The blends of soymilk with cowmilk were coagulated into paneer by using coagulant at 70oC temperature. The paneer made from cowmilk contained higher amount of total solids and fat, where as the paneer made from soymilk was rich in protein and ash content. The paneer made from the blends showed a diluting effect on fat and total solids contents and an enriching effect on protein and ash.The results of sensory evaluation showed that the blending of cow milk upto 20% showed no significant effect on colour, flavor, taste and overall acceptability of paneer. The paneer made from other blends was also in acceptable range; however, it was significantly (p≤0.05) lower with respect to the sensory attributes of cow milk paneer on nine point hedonic scale. The blending of soymilk in buffalo milk to prepare paneer was coagulated with suitable amount of coagulant. The paneer made from the above blends was studied for its chemical composition. The total solids reduced in paneer by increasing the amount of soymilk. The protein increased significantly (p≤0.05) by enhancing the incorporation of soymilk in the blends. The fat content in paneer decreased significantly (p≤0.05) with increase in the proportion of soymilk in blends. The ash content also increased by increasing the amount of soymilk in the blends. The paneer prepared from the blends was studied for its sensory characteristics such as color, flavor, taste and overall acceptability. The results of the present investigation showed that the paneer prepared from the blends containing soymilk upto 20% was significantly (p≤0.05) superior from the paneer prepared from other blends. However the paneer prepared from all the blends was in acceptable range though it scored significantly (p≤0.05) lower than the paneer prepared from buffalo milk. Hence it was concluded that soymilk may be incorporated in buffalo milk up to 20% to prepare paneer without affecting the sensory attributes significantly (p≤0.05) on a nine point hedonic scale.The paneer prepared from the blends of soymilk with cow milk and buffalo milk in the proportion of 20:80 was kept at refrigeration temperature (4oC) for storage studies. The samples were analyzed for physico-chemical, microbiological and sensory quality after an interval of every 3 days. The results showed that the moisture content of paneer decreased significantly (p≤0.05) during storage in all the samples. The fat content also decreased during storage. This decrease in fat content may be due to lipolysis. No significant difference was noted in protein content with respect to storage period. The free fatty acids significantly increased during storage. This increase in free fatty acid in paneer could be attributed to the hydrolysis of fat. The titrable acidity significantly increased during storage due to the activity of microorganisms.The effect of storage temperature and storage period on paneer was determined by assessing its total plate count. The results showed a rapid increase in TPC, yeast and mold and coliform count at room temperature than at refrigeration temperature. It was observed that the maximum shelf life of paneer was 6 days at refrigeration temperature and 2 days at room temperature. At room temperature coliforms were detected after 3 days of storage, however at refrigeration temperature coliform count was detected after 9 days of storage. The paneer stored at room and refrigeration temperature was studied for its sensory characteristics. The results showed that the color, flavor, taste and overall acceptability showed a decreasing trend which was rapid at room temperature than the refrigeration temperature. The paneer was within acceptable range for 2 days only at room temperature, however, at refrigeration temperature it was found in an acceptable range after 6 days also. Decrease in sensory attributes was related with physico-chemical changes as titrable acidity and free fatty acids values increreased during storage. All the sensory attributes decreased correspondingly.The textural characteristics of the paneer samples were determined to evaluate the changes in the hardness characteristics. The initial maximum hardness of blended soy-cow milk paneer and soy-buffalo milk paneer samples were 157.7 g and 158.4 g, respectively. The hardness strength was increased to the level of 158.9 g and 159.5 g at room temperature and 158.6 g and 159.6 g at refrigeration temperature, in soy-cow and soy- buffalo milk paneer after 2 and 9 days of storage, respectively. Hardness of paneer increased with increase in temperature and pressure. On the basis of the above results it is concluded that the highly nutritious paneer can be prepared by blending 20 per cent proportion of soymilk with cow milk and buffalo milk without significantly affecting the sensory characteristics of paneer.Chapter – 6Bibliography BIBLIOGRAPHYAACC, 2000. Approved Methods of American Association of Cereal Chemists. 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American Journal of Clinical Nutrition. 81 (2) 397-408APPENDIX-I SENSORY SCORE CARD FOR PANEERON 9-POINT HEDONIC SCALEName of the Panelist …………Faculty/StudentExperiment No. ………………Date …………….You are requested to evaluate the following sample of paneer.Paneer is a food product with a good source of high protein , high nutrition and energy food. It should be chewy with pleasant flavour and acceptable taste and texture. The colour of the product should be cremish white.Kindly evaluate the sample of paneer on the basis of following 9-point hedonic scale. Like extremely9Like very much8Like moderately7Like slightly6Neither like nor dislike5Dislike slightly4Dislike moderately3Dislike very much2Dislike extremely1Sample No.Sensory AttributesColourTexture Taste /FlavourOverall Acceptability?????????????????????????Remarks……………………………………………………………………………….…………………………………………………………………………… (Signature) ................
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