Composition of hydrolysates from meat

JOURNAL OF FOOD COMPOSITION AND ANALYSIS 12, 219}225 (1999) Article No. jfca.1999.0815 Available online at http://www.idealibrary.com on Composition of Hydrolysates from Meat Maria Elisabeth M. Pinto E Silva, Rosa N. Mazzilli, and Fabiana Cusin Department of Nutrition of the School of Public Health USP, Brazil Received 11 February 1998, in revised form 17 November 1998, and accepted 27 July 1999 Vegetal enzymes, such as bromelin found in pineapple (Ananas comosus L.), are e$ciently employed in meat hydrolysis. Fractional beef, turkey and chicken were hydrolysed with bromelin under ordinary household conditions. The hydrolysates were analysed for protein, water, fat, and fatty acids and acidity. The protein contents of the hydrolysates (per 100 g of meat) were 11 g for beef, and 12 g for both turkey and chicken. The beef hydrolysate had 1.4 g of fat per 100 g of meat. After hydrolysis, 90% of the fatty acids remained in the beef hydrolysate, whereas 50% remained in the turkey hydrolysate and 70% in the chicken hydrolysate. The acidity of the fresh meats was slightly lower (pH"5.7) than the acidity of the hydrolysates (pH"4.9) due to the presence of pineapple, which is a positive aspect for preserving the hydrolysates. It was concluded that these hydrolysates are important low cost sources of nutrients, and that they can be very well employed in all kinds of diets. The cost of hydrolysates produced under ordinary household conditions is 50% lower than manufactured products.  1999 Academic Press Key =ords: meats; hydrolysates; composition. INTRODUCTION Because of metabolic or clinical conditions, some people require partially or totally digested nutrients in order to facilitate assimilation (Alpers, 1978; Burini and Cam- pana, 1985; Schimidl et al., 1994). Proteins are an indispensable component of all diets, and therefore warrant careful examination of the best forms for consumption and for e!ective assimilation. Depending on the origin of the protein, 10}20% of absorption will occur in the duodenum, 60% in the jejunum and the remainder in the ileum (Alpers, 1978; TomeH , 1994). Minor unhydrolysed peptides represent 50% of the "nal products of protein digestion in the lumen (Alpers, 1978; Silk et al., 1985; TomeH , 1994). Enzymes, acids and alkalis are used to hydrolyse proteins. Proteolytic enzymes produced the best results in retaining the nutritional value in several preparations (Donzelli et al., 1990; Fox et al., 1982). Research has shown that puri"ed proteolytic enzymes, such as papain (from papaya) and bromelin (from pineapple) produce good results in digestion of meat products (Asakura, 1982; Stabile, 1989). The concentration of soluble solids obtained was signi"cant. Once protein properties (stability, solubility, texture and water retention) are modi"ed by heating, the e!ect of hydrolysis is greater during cooking (50}603C) when vegetable enzymes (papain, bromelin, "cin) are added to the meat. These enzymes are active even at 803C as their inactivation occurs only at higher temperatures (Fox et al.,1982; Park and Draetta, 1971). Park and Draetta (1971), Asakura (1982) and Stabile (1989) studied the e$ciency of papain, bromelin, "cin, trypsin and microbial protease in the hydrolysis of meats, and 0889}1575/99/030219#07 $30.00/0  1999 Academic Press analysed the temperature, pH and concentration of soluble solids. Takeda and Okeda (1970) and Trigo and Sakaki (1975) veri"ed therapeutic applications of hydrolysates in premature children with satisfactory results. Some authors used puri"ed isolated enzymes in their reports (Asakura, 1982; Lahl and Braun, 1994; Mahmoud, 1994; Schimidl et al., 1994). However, the high cost of this process restricts its practical application in household diets. Stabile et al. (1992) studied meat (beef, chicken and "sh) hydrolysates produced using fresh pineapple juice in households, and demonstrated that both household and manufactured formulas are similar in osmolarity, energy content and composition. Stabile (1991) optimized the process of meat hydrolysis with fresh pineapple juice, and showed that the juice-produced hydrolysate retained a higher percentage (50%) of soluble solids relative to that produced with puri"ed enzyme. This study presents new ways of preparing hydrolysates with high nutritional value that can bene"t people of any socio-economic class. The study describes the composi- tion of beef, turkey and chicken hydrolysates produced by digestion with pineapple juice. MATERIAL AND METHODS Material Slabs of meat readily available, such as &&coxa o mole'' (slab constituted muscles from the internal side of cattle's leg), turkey and chicken chest were used in this study. The bromelin source was Pearl and Hawaii pineapple (Ananas comosus L.) fresh juice, as recommended by Stabile (1991). Methods Meat hydrolysis. The pineapple fruit was carefully washed, peeled and fragmented. The juice was extracted by a domestic centrifuge (Phillips do Brasil } Walita 170W, 14 000 rpm). Approximately 600 ml of juice can be obtained from 1000 g of pineapple. Tendons, nerves, skin and visible fat were removed from the meat, which was then fragmented and ground in a food processor (Walita Master Plus Phillips do Brasil 400 W, 17 000 rpm). Meat and juice were mixed in equal proportions (1 kg of meat/1 kg of juice) in a bowl, manually homogenized and kept in hot water (603C) for 30 min. Then the mixture was heated to the boiling point, which was reached after 8 min. This temperature was maintained for 5 min. The mixture was then poured into a sieve to remove insoluble residue, following Stabile's methodology (1991). Analytic methods. Moisture was determined with gravimetric analysis by oven drying at 1053C to stable weight (Inst. Adolfo Lutz, 1985). Fat content was determined by continuous extraction in Soxhlet equipment (Horwitz, 1980). To determine fatty acids, the fat was extracted using petroleum ether and diethyl ether (1: 1) plus sodium sulphate solution (Bligh and Dyer, 1959). Methylation and saponi"cation were carried out with methanol, sodium hydroxide, ammonium chloride and sulphuric acid. Extraction was done using petroleum ether. The methylated samples were analysed in a gas chromatograph HP 5890 (CA, U.S.A.), containing #ame ionization detector and integrator HP 3396, with a capillary column (50 m;0.32 mm;0.17 mm). HP-20M carbowax. The initial temperature was 1303C and the "nal was 2003C, 220 PINTO E SILVA, MAZZILLI AND CUSIN TABLE 1 Percentage composition of raw beef (R) and its hydrolysate (H)*, g/l Constituent RH Water 74.3 (0.8) 80.7 (0.6) Ash 1.3 (0.2) 0.8 (0.0) Carbohydrate * 5.1 (0.8) Protein 22.5 (1.6) 12.0 (1.2) Total fat 1.6 (0.6) 1.4 (0.3) Saturated** 0.7 (0.0) 0.7 (0.0) Monounsaturated** 0.7 (0.0) 0.6 (0.0) Polyunsaturated** 0.1 (0.0) 0.1 (0.0) * Values are means (S.D.); n"6. ** Values are means (S.D.); n"5. For the raw beef, the pattern is g/100g and for the hydrolysed g/100 ml. increasing 53C/min. The fatty acids were identi"ed by tests with methyl ester pattern (Sigma RM3 and RM6). Protein content was determined by micro-Kjeldahl method, employing 6.25 as calculating factor (Inst. Adolfo Lutz, 1985). Carbohydrates were not determined analytically but for purposes of crude comparison, total carbohydrates were calculated by di!erence. Finally, potentiometric DMPH-2 (pH 0.01 Digimed, Sa o Paulo, Brasil) was used to determine acidity. RESULTS AND DISCUSSION The average of the compositions of six samples of beef hydrolysate (g/100 g) is presented in Table 1. Water makes up approximately 80% of the hydrolysate. The volume of the hydrolysate produced from a mixture of 1 kg of meat with 1 kg of fresh pineapple juice was 900 ml. The hydrolysate has a protein content that is 50% lower than that of the original raw meat. The decrease in protein content in the hydrolysates corresponds to the dilution by the addition of the pineapple juice. This dilution also explains the higher water percentage. Asakura (1982) and Stabile (1991) demonstrated that the concentration of soluble solids in hydrolysates increased with temperature (at constant pH"5) until it reached a maximum value at 603C for 30 min. There was no further increase in soluble solids at higher temperatures, and the enzymatic activity ceased when the temperature reached the boiling point. Park and Draetta (1971) veri"ed that bromelin is inactive at temperatures higher than 903C (Donzelli et al., 1990; Schimidl et al., 1994). There were no di!erences in the total fat and fatty acid contents of the raw beef samples and their hydrolysates. Although external fat was removed before hydrolysis, the meat samples still contained internal fat. The similarity in fat content of the hydrolysates and the raw meat samples suggests that most of the internal fat became dissolved in the hydrolysates. Di!erences in fat content between various hydrolysates could arise from di!erences in the internal fat contents, of the meat samples from which they were produced. The national tables of percentage composition (Fundaia o IBGE, 1985) report the total fat content of beef as 6.1g/100g. The fat content in the samples studied here is much lower, probably due to the removal of all external fat before measurement (Table 1). Analysis of the composition of the fatty acids of beef and its hydrolysate showed that oleic acid (C18:1) was in the highest proportion, followed by palmitic acid COMPOSITION OF HYDROLYSATES FROM MEAT 221 TABLE 2 Composition of raw turkey meat (R) and its hydrolysate (H)*, g/100 g edible portion Constituent RH Water 73.4 (1.4) 80.9 (0.8) Ash 1.5 (0.3) 0.8 (0.0) Carbohydrate * 5.0 (1.2) Protein 24.5 (1.4) 12.4 (1.1) Total fat 1.5 (0.7) 0.7 (0.4) Saturated** 0.5 (0.0) 0.3 (0.0) Monounsaturated** 0.5 (0.0) 0.2 (0.0) Polyunsaturated** 0.4 (0.1) 0.2 (0.0) * Values are means (S.D.); n"6. ** Values are means (S.D.); n"5. For the raw beef, the units are g/100 g and for the hydrolysate g/100 ml. (C16:0) and stearic acid (C18:0). Similar results were observed by Julla (1988). Less than 10% of the fatty acids were polyunsaturated. The percentage of saturated fatty acids was 45.8%, which is lower than the values of 52.7% reported by Pitre, and 48.0% reported by Girard et al., both cited in Julla's revision (1988). The average percentage composition of turkey meat and its hydrolysate is present- ed in Table 2. Signi"cant reductions (of approximately 50%) are observed in protein, ash and total fat. The water content has increased in the hydrolysate. These di!erences are due to the dilution by the pineapple juice in the process of hydrolysis. With respect to fat composition, it is important to consider the preparation procedures. Tendons, nerves and external fat were manually removed, always by the same person. After sifting the meat, fat can be retained with the small remaining residue (probably elastin and collagen). Since turkey fat is not found internally to the same extent as beef fat, it is found mainly as insoluble elements in the "nal hy- drolysate. The national table of percentage composition (Fundaia o IBGE, 1985) report the fat content of turkey meat was 6.6 g/100 g, which is higher than the values of the samples here (Table 2). This di!erence could be explained by the removal of external fat from the samples since the national tables do not report their technique of preparation. The composition of fatty acids in the raw turkey meat was the same as its hydrolysate. The oleic acid (C18:1) was in the highest concentration, followed by linoleic acid (C18: 2) and palmitic acid (C16:0). Approximately 30% of the fatty acids were polyunsaturated. The results obtained for the chicken meat hydrolysate are shown in Table 3. Reductions of approximately 50% for ash, protein and total fat in the hydrolysate were observed. The water content increased as a result of the dilution with pineapple juice, which also accounts for the presence of carbohydrates. The reduction in fat content was a consequence of dilution. The fat content di!ers from the value of 3.3 g/100 g reported in the national tables of percentage (Fundaia o IBGE, 1985). This is probably due to the removal of skin, tendons and external fat before preparation of the hydrolysates. The national tables do not report whether or not these elements had been removed. There was no di!erence in the pattern of fatty acids (Table 3); the percentage of oleic acid (C18: 1) was higher than the palmitic (C16:0) and linoleic (C18:2) acids. A small percentage of the fatty acids was polyunsaturated. Julla's (1988) value of 28% for the percentage of saturated fatty acids in relation to total fatty acids (from Girard et al.) is 222 PINTO E SILVA, MAZZILLI AND CUSIN TABLE 3 Composition of raw chicken (R) and its hydrolysate (H)*, g/100 g edible portion Constituent RH Water 74.4 (1.4) 81.1 (0.6) Ash 1.7 (0.6) 0.8 (0.1) Carbohydrate * 4.6 (1.0) Protein 23.9 (0.8) 12.6 (0.4) Total fat 1.0 (0.3) 0.8 (0.1) Saturated** 0.3 (0.0) 0.3 (0.0) Monounsaturated** 0.4 (0.0) 0.4 (0.0) Polyunsaturated** 0.2 (0.0) 0.2 (0.0) * Values are means (S.D.); n"6. ** Values are means (S.D.); n"5. For the raw turkey, the pattern is g/100g and for the hydrolysed g/100 ml. lower than the value of 32.9% reported here. The beef hydrolysate showed a higher percent of saturated fat than the turkey and chicken hydrolysates, which agrees with the report presented by Julla (1988). The pH data presented in Table 4 shows values of approximately 5.7 0.1 for the raw meats and 4.8 0.1 for the hydrolysates. The acidity of the hydrolysates is signi"cant since it makes them more resistant to Bacillus stearothermophillus FS 1518,as reported by Stabile et al. (1990). These pH values promote myoglobin fading, and preserve the meta-myoglobin. This accounts for the stronger brownish colour of the beef hydrolysates since beef is the most pigmented of the three meats (Livingston and Brown, 1981). Nevertheless, this did not impair the products'#avour and solubility. The stable pH preserved its microbiological quality when kept under refrigeration for one week (Stabile, 1990). It has been noted in the literature (Pederson, 1994; Phillips and Beuchat, 1981) that protein hydrolysis results in a bitter #avour due to the release of the amino histidine, tryptophan, isoleucine and phenylalanine. This bitterness can be partially masked with spices or other #avoured ingredients. The presence of starch is reported to reduce bitterness (Fox et al., 1982; Pederson, 1994; Phillips and Beuchat, 1981). All three kinds of meat hydrolysate presented mild residual #avour with the beef hydrolysate as the most intense. Pinto e Silva et al. (1998) used hydrolysates as ingredients in several dishes, such as soup, dumpling, mousse and mixed vegetable and fruit juice. In sensory evaluations, the tasters registered good acceptance. A comparative study of costs between manufactured diets (with casein hydrolysate, whey and soybean watery liquid) and household beef hydrolysate showed that the hydrolysate is approximately 50}70% more economical. Manufactured diets cost between US$ 5.00 and US$ 8.00 (hydrolysate formulation based on Oliveira et al., 1992). The puri"ed costs about US$ 80.00/kg, and is often not a!ordable. Pineapple is available throughout the year and costs only US$ 1.00/kg. Domeni and Galeazzi (1997) tested the use of hydrolysates in enterstomies in hospital nutrition departments. They showed that there was no signi"cant di!erence in e$ciency between manufactured and household formulas, despite the large di!er- ence in their costs. Beef hydrolysate has the major advantage that it can be employed in the various phases of enterostomies until patients have totally recovered and can return to their usual diets. COMPOSITION OF HYDROLYSATES FROM MEAT 223 TABLE 4 pH of raw beef, turkey and chicken (R) and their respective hydrolysates (H)* Beef Turkey Chicken Meat Samples RHRHRH 1 5.6 4.9 5.7 4.9 5.8 4.8 2 5.4 5.0 5.7 5.2 5.7 5.2 3 5.7 4.2 5.9 4.3 5.8 4.9 4 5.5 4.4 5.8 4.7 5.5 4.6 5 5.7 4.6 5.5 4.6 5.7 4.5 6 5.6 5.0 5.9 5.0 5.7 5.0 Average 5.6 4.7 5.8 4.8 5.7 4.8 S.D. 0.1 0.3 0.1 0.0 0.1 0.3 * Values are means: n"6. It is appropriate to continue research into low-cost dishes which are easy to prepare in household conditions, and which can be used in various di!erent types of diets and nutritional supplementation. ACKNOWLEDGEMENTS To the Central Laboratory of FrigobraH s, for the analysis; To the Technical Session of Mercado In- stitucional Sadia Comercial, for donating the meats. REFERENCES Alpers, D. H. (1978). Digestion and absorption of carbohydrates and protein. In Physiology of the Gastrointestinal ¹ract (L. R. Johnson, Ed.), 2nd edn., pp. 1469}1487. Raven Press, New York. Asakura, Y. (1982). HidroH lise de protemHnas da carne bovina pela aia o da papaina, bromelina e "cina. Master thesis, Faculdade de Cie( ncias Formace( uticas, Universidade de Sa o Paulo, Sa o Paulo, Brasil [In Portuguese]. Bligh, E. G., and Dyer, W. J. A. (1959). A rapid method of total lipid extraction. Can. J. Biochem. Physiol. 37, 911}917. Burini, R. C., and Campana, A D. (1985). Digesta o, absoria o, circulaia o utilizaia o de nutrientes. In Suporte nutricional, parenteral e enteral (M. C. Riella, Ed.), pp. 162}190. Guanabara, Rio de Janeiro, Brasil [In Portuguese]. Domeni, S. M. A., and Galeazzi, M. A. M. (1997). Prescriia o e uso de formulados para nutriia o enteral pelos serviios de nutriia o hospitalares do municmHpio de Campinas } Sa o Paulo, Brasil Rev. Nutr. P;CCAMP 10(2), 114}119 [In Portuguese]. Donzelli, F., Dalt, L., Scchiavona, A., Baraldi, E., and Muraro, M. (1990). Clinical experience with a hydrolysed soy formula in infants with protracted enteritis and atopic eczema. Pediatr. Med. Chir 12, 71}79. Fox, P. F., Morrissey, P. A., and Mulvihll, D. M. (1982). Chemical and enzymatic modi"cation of food proteins In: Developments in Food Proteins } 1, (B. F. J. Hudson, Ed.), pp. 1}60. Applied Science Publ., London. Fundaia o Ibge (1985). ¹abelas de composimaJ o de alimentos: Estudo Nacional da Despesa Familiar. Vol. 3, Rio de Janeiro, Brasil [In Portuguese]. Horwitz, W. (1980). O.cial Methods of Analysis of the Association O.cial Analytical Chemists, 13th edn., AOAC, Washington. Instituto Adolfo Lutz (1985). Normas analnHticas do Instituto Adolfo ¸utz. 3a edn., of Vol. 1, Sa o Paulo, Brasil [In Portuguese]. Julla, B. (1988). QualiteH s nutritionelles et organoleptiques des viandes bovines. Cah. Nutr. Die` t. 23(1), 27}33 [In French]. 224 PINTO E SILVA, MAZZILLI AND CUSIN Lahl, W. J., and Braun, S. D. (1994). Enzymatic production of protein hydrolysates for food use. Food ¹echnol. 48(10), 68}71. Livingston, D. J., and Brown, W. D. (1981). The chemistry of myoglobin and its reactions. Food ¹echnol. 244}252. Mahmoud, M. I. (1994). Physicochemical and functional properties of protein hydrolysates in nutritional products. Food ¹echnol. 48(10), 89}95. Oliveira, M. N., Baru!aldi, R., Pinto E Silva, M. E. M., Nieto, R., Alvares, E., and Nogueira, A. C. S. (1992). Development of diets based on digest meat II Congreso Internacional de QumH mica de la Anque Food Science and Technology: Industry and Distribution Vol. III Burgos } October, 1992 } Actas, pp. 49}54. Park, Y. K., and Draetta, I. S. (1971). Aplicaia o de enzimas proteoliticas no amaciamento de carne de boi. Rev. Bras de ¹ecnologia 2, 125}129 [In Portuguese]. Pedersen, B. (1994). Removing bitterness from protein hydrolysates. Food ¹echnol. 48(10), 96}98. Phillips, R. D., and Beuchat, L. R. (1981). Enzyme modi"cation of proteins. In Protein Functionality in Foods, J. P. Cherry (Ed.), pp. 274}298. American Chemistry Society Div. of Agric. and Food Chemistry (ACS Symposium Series Vol. 147), Washington. Pinto E Silva, M. E. M., Mazzilli, R. N., and Barbieri, D. (1998). Hidrolisado proteico como recurso dieteH tico. J. Pediatria 4(3), 217}221 [In Portuguese]. Schimidl, M. K., Taylor S. L., and Nordlee, J. A. (1994). Use of hydrolysate-based products in special medical diets. Food ¹echnol. 48(10), 77}85. Silk, D. B. A., Grimble, G. K., and Rees, R. G. (1985). Protein digestion and amino acid peptide absorption Proc. Nutr. Soc. 44, 63}72. Stabile, M. N. O. (1989). Hidrolisados de carne bovina: biotecnologia de obtenia o. Master thesis, Facul- dade de Cie( ncias Farmace( uticas Universidade de Sa o Paulo, Sa o Paulo, Brasil [In Portuguese]. Stabile, M. N. O. (1991). Otimizaia o do processo biotechnoloH gico de hidroH lise de carne bovina. Doctoral thesis. Faculdade de Cie( ncias Farmace( uticas Universidade de Sa o Paulo, Sa o Paulo, Brasil [In Por- tuguese]. Stabile, M. N. O., Baru!aldi, R., Stedefeld, E., Basilio, M. C., Pinto E. Silva, M. E. M., Nieto, R. A., and Alvares, E. D. (1990). Carne preH -digerida a partir de suco de abacaxi Aliment. ¹echnol. 32, 99}104 [In Portuguese]. Takeda, M., and Okeda, T. (1970). Utilizaia o da carne lio"lizada em dietoterapia Rev. Med. IAMSPE 1, 23}24 [In Portuguese]. TomeH , D. (1994). ProteH ines alimentaires et meH tbolisme des proteH ines 1 } Bases physiologiques Cah. Nutr. DieH t 29, 54}60 [In French]. Trigo, M., and Sakaki, M. (1975). Estudo da complementaia o alimentar proteica em prematuros (reH cem- nascidos de baixo peso): avaliaia o de 56 casos com controle domiciliar Rev. IAMSPE 6(2), 9}14 [In Portuguese]. COMPOSITION OF HYDROLYSATES FROM MEAT 225 . contents of the hydrolysates (per 100 g of meat) were 11 g for beef, and 12 g for both turkey and chicken. The beef hydrolysate had 1.4 g of fat per 100 g of meat. After hydrolysis, 90% of the. was in the highest proportion, followed by palmitic acid COMPOSITION OF HYDROLYSATES FROM MEAT 221 TABLE 2 Composition of raw turkey meat (R) and its hydrolysate (H)*, g/100 g edible portion Constituent. average of the compositions of six samples of beef hydrolysate (g/100 g) is presented in Table 1. Water makes up approximately 80% of the hydrolysate. The volume of the hydrolysate produced from