. 1 1 1 1 ' 1 1 1 1111;1'-,11j 1 .‘ 1 1 . ~- ' : 1 , 1 1 I._ 1 . 1 1 1 , 1 ‘ ,1 ' ' , V _ ,1 1 ' 1"1- 1 _ “"1: ‘ , 1 , ,, ,1,,‘1.1,. f . 1‘ 1 . 1 1 1'1 1 V .1 1'-'~.-’1""‘:,r" ‘l ' ‘ , L‘ . 1 ' ' 1 '11 ' 1 - 1 "“ ' . 1 ' 1. 1 " 1 1 1- H j . ,‘ 1 ' ,1 1 11 .1 1 . 1 1 1. 11 ' 1~ .1 - 1. 1 11 1 1 ’ 1--111 , 1 '5 .1 I " ' ‘1'1 . - 1‘1, ,1 ‘1 . ‘ ,1.,,,... v 1 '. 1 . ' ' n .1 _ £ 1 , 1 A~ ' ‘ ‘ .‘ ‘4 1- 1 1 1. 1 . , ,1 1 . ,. A _ 1 . 1 , , . 1 1 . 4 1 . 1‘1 .1 .1 , 1 ‘ . ‘ ‘ 1, ; ‘ N 11 ‘I - '11:“‘1931‘1' 11 A.“ THESIS 300999! 8596 JVJJV Umfi’we 1111111111111 1111 11111111111? This is to certify that the thesis entitled The Effect of Mechanical Treatment of Meat Pieces on Sensory Parameters of Sectioned and Formed Processed Meats presented by Jorge Fuentes Zapata has been accepted towards fulfillment of the requirements for Food Science and mg:— James F. Price Doctor of Philosophy degree in Human Nutrition Dr. Major professor Date July 2, 1981. 0-7639 3'“ ‘ w» _ 1-13 , \ ~11 4? ' 5 fl , ' 1"! g J V b)" , 1 ' ~ 1‘? Mr A. »_ t," 4 fl MSU RETURNING MATERIALS: P.3te in book drop to LIBRARIES remove this checkout from your record. fiNES will “— be charged if beck is returned after the date Stamped below. {FEB Q 6199?} 037 ', THE EFFECT OF MECHANICAL TREATMENT OF MEAT PIECES ON SENSORY PARAMETERS OF SECTIONED AND FORMED PROCESSED MEATS By Jorge Fuentes Zapata A DISSERTATION Submitted to Michigan State University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Food Science and Human Nutrition 1981 ER: @ ABSTRACT THE EFFECT OF MECHANICAL TREATMENT OF MEAT PIECES ON SENSORY PARAMETERS OF SECTIONED AND FORMED PROCESSED MEATS By Jorge Fuentes Zapata A study was designed and conducted to determine the effects of tumbling time (60, 120 and 180 minutes); pressure during tumbling (vacuum and non vacuum); condition of the meat (fresh and frozen and thawed meat); and brine injection level (16% and 32%) on the nature of the exudate after tum- bling and the quality parameters of sectioned and formed hams. Results indicated that protein extraction from frozen meat was faster than that from fresh meat with tumbling time. Pat was extracted rapidly after a short period of tumbling, and the use of vacuum during tumbling did not affect protein and fat extraction. Longer tumbling periods and absence of vacuum during tumbling increased lipid oxidation, with the effect being more evident with frozen meat. After brine pumping salt and nitrite were retained better by fresh meat than frozen meat. Frozen meat tended to absorb much of the cure during tumbling. The use of vacuum did not contribute to myosin extrac- tion when the meat was tumbled for 60 minutes. However, the use of vacuum resulted in hams with good color distribution and better tenderness and texture characteristics than those tumbled without vacuum. Jorge Fuentes Zapata No differences in yields, calculated according to Federal regualtions, were evident in hams from fresh and frozen meat, indicating that frozen meat is quite suitable for this type of processing. Nitric oxide pigment content in hams was adversely affected by vacuum during tumbling. However, color intensity of the hams was not different. Microsc0pic study showed a common pattern of increased fiber disrupture in the tissue with tumbling time and in a single muscle chunk going from the interior part to the peripheral part. Fibers from frozen meat showed more damage after tumbling than those from fresh meat. The use of vacuum during tumbling eliminated presence of air bubbles in the exudate. There were some discrepancies in the direction of the effects of the treatments on binding strength evaluated by the Instron and by taste panel. However, both methods indi- cated that hams pumped 16% bound significantly better than those pumped 32%. The use of vacuum during tumbling improved tenderness of hams but not color distribution. Durante los dias gastados escribiendo este trabajo han venido recuerdos y vivencias a mi mente con desusual claridad. He recordado mi in- fancia y los nifios pobres de la tierra donde na- ci. Aquellos nifios de piel oscura de sol, sala- da de sudor, quemada de frio y salpicada de pol- vo de los terrosos callejones donde viven y jue- gan. Aquellos nifios de mirar desconfiado, de ac- titudes candorosas, de cuerpos humildes pero re- sistentes, de lenguaje limitado pero de profun- dos y nobles sentimientos en sus interiores. He sofiado que algfin dia todos ellos tendrah la for- tuna y oportunidad que yo he tenido en la Vida para alcanzar educacidn superior. Para algunos vendra'pronto; para la mayoria restante, aque- llos nifios que llevaran su pureza y humildad junto a sus vidas pobres, yo les dedico esta tesis. ACKNOWLEDGMENTS The author wishes to express his thankful appreciation to Dr. James F. Price for his continued guidance, interest and valuble suggestions throughout this study. The ready assistance and suggestions of the guidance committee: Drs. Loran L. Bieber, Lawrence E. Dawson, Albert M. Pearson, Ian J. Gray and Robert A. Merkel is acknowledged. Appreciation is extended to Rose M.Gartner for contin- uous advice and assistance in the processing room and labora— tories around the Department facilities used in this project. Gratitude is extended to Dr. Bruce R. Harte from the Department of Packaging for his help in the texture analyses. For the friendly words of wisdom from Don, John, Tom, Jan, Paco and Nacho, my fellow graduate students in the Meat Laboratory, an appreciative thank—you and a note of encouragment are extended. To the more permament staff of the Meat Laboratory, Al Booren, Jean McFadden, Tom Forton, Dora Spooner and Bea Eichelberger, thank-you for your patience with my poor English. Lastly, to my parents and parents—in-law who have encouraged me to ardently pursue my education throughout my life a grateful thank-you for your patience. And to my wife Fatima and son André, whose patience, encouragment and ii sacrifice through our graduate study made this accomplish- ment possible, my endless love and gratitude. iii TABLE OF CONTENTS LIST OF TABLES .................................. LIST OF FIGURES .................................. INTRODUCTION .................................... LITERATURE REVIEW ............................... The Protein System in Pork Muscle .......... Extracellular components ............ Intracellular proteins .............. The Conversion of Pork Muscle to Meat ..... Myofibrillar Proteins and Functional PrOperties of the Meat .................... The Process of Binding of Meat Pieces ..... The Technique of Tumbling and Massaging Meat Pieces ..................... Non-meat Proteins in the Binding of Meat Pieces ............................... New Trends in the Acceptance by Consumers of Sectioned and Formed Meats MATERIAL AND METHODS ............................ Description of the Experiment ............. Statistical Design ........................ Source of Meat ............................ Preconditioning of Fresh Hams ............. Processing and Sampling Operations ........ Methods of Analysis ....................... RESULTS AND DISCUSSION .......................... Chemical Composition of the Raw Meat ...... Changes in Chemical Composition Through Processing ........................ The Parameters of the Exudate ............. Composition of the Soluble Phase .......... Parameters Related to the Final Product iv 46 46 47 66 8O Page SUMMARY AND CONCLUSIONS .......................... 108 APPENDIX A Taste Panel Score Sheets ................... 113 APPENDIX B Chemical Analyses .......................... 116 APPENDIX C Analysis of Variance Tables ................ 122 LIST OF REFERENCES ........ ....................... 138 Table 2a 2b 10 11 LIST OF TABLES Description of processing treatments used in the manufacturing of boneless hams ............ p ......................... Processing treatments as arranged for statistical analysis by 3-way ANOVA ...... Processing treatments as arranged for statistical analysis by 2-way ANOVA ....... Brine composition as used in the manufacturing of boneless hams ............ Smokehouse cooking schedule for boneless hams ............................ Proximal composition and TBA values in raw pork used in the manufacture of boneless hams .......................... Means of soluble phase volume of the exudate formed during tumbling of hams Means of protein content (mg/ml) in the soluble phase as determined by Biuret method ............................. Relative mobility of the major myo- fibrillar protein bands identified on SDS-PAGE ............................... Estimated yields of hams as calculated by Federal inspection procedures, using 3.79 as k factor .......................... Tensile strength values (g/cmz) mea- sured to separate pieces of ham by the seam or binding area .................. Evaluation of the overall appearance of ham slices by a visual inspection panel Vi Page 28 29 29 33 35 46 66 7O 76 83 99 102 Table 12 13 Evaluation of binding strength between pieces of meat in a ham slice by semi- trained panelists OOOOOOOOOOOOOOOOOOOOOOOOO Evaluation of meat tenderness in ham slices by taste panel ooooooooooooooooooooo vii Page 105 106 LIST OF FIGURES Figure 1 Processing flow chart and sampling points in the manufacture of bone- less hams ................................. 2 The effect of tumbling time on the protein content of the exudate of fresh meat (a) and frozen meat (b) and the interactions due to the use of vacuum during tUmbling ................. 3 Protein content (Z) in the exudate (a) and in the finished hams (b) as affected by tumbling time and level of pumping ................................ 4 The effect of tumbling time on fat content in the exudate from fresh meat (a) and frozen meat (b) .............. 5 Fat content in the exudate (a) and finished hams (b) as affected by tumbling time and brine pumping level ..................................... 6 Moisture content in the exudate of fresh (a) and frozen (b) meat as affected by tumbling time and pres- sure during tumbling interactions ......... 7 Moisture content in the exudate (a) and finished hams (b) as affected by tumbling time and brine pumping level ..................................... 8 TBA number (mg malonaldehyde/lOOO g sample) in the exudate of pork mus- cle as affected by condition of the meat and tumbling time .................... 9 TBA number (mg malonaldehyde/lOOO g sample) for exudate (a) and finished hams (b) as affected by tumbling time and pumping levels ................... viii Page 31 48 49 51 52 53 55 57 58 Figure 10 ll 12 13 14 15 16 17 18 19 20 Salt content in the exudate of fresh (a) and frozen (b) meat as affected by tumbling time and pressure during tumbling ................................. Salt content, percent for exudate (a) and finished hams (b) as affected by tumbling time and pumping levels ......... Nitrite content in the exudate from fresh meat (a) and frozen meat (b) as a function of tumbling time ........... Soluble phase volume (ml) averaged over the pressure factor, as a function of tumbling time ................ Soluble phase volume (ml) in the exudate as a function of tumbling time and pumping level ................... Protein content in the soluble phase for fresh (a) and frozen (b) meat as affected by 3-way interactions of the major effects ....................... Protein content (mg/ml) in the exudate soluble phase as affected by tumbling time and pumping level ................... Standard protein mix in SDS—PAGE as a function of molecular weight and rela- tive mobility ............................ Scanning of a SDS-PAGE gel with the major bands assigned to 8 of the most common myofibrillar proteins ............. Relative concentration of myosin (a); M-line protein and Z—protein (b); @- actinin (c); and Tropomyosin complex (d) in the exudate soluble phase as a function of tumbling time ................ Relative concentration of G-actin (a); Troponin-T and high molecular weight Tropomyosin monomer (b); low molecular weight Tropomyosin monomer (c); and Myosin light chains (d) in the exudate soluble phase as a function of tum- bling time ............................... ix Page 60 62 65 67 68 71 72 74 75 77 78 Figure Page 21 Processing factors affecting final yields in the process of manufacture of boneless hams .......................... 81 22 Percent conversion of pork meat into boneless ham.as a function of tum- bling time. Yields calculated from actual processing losses .................. 82 23 Cooking losses (%) of the meat as a function of tumbling time and pumping level ..................................... 85 24 Nitric oxide pigment content (a) and percent pigment conversion (b) in hams as a function of tumbling time ............ 87 25 Nitric oxide pigments (a) and percent pigment conversion (b) as a function of tumbling time and pumping level ........... 88 26 Color parameters L (I), a (II) and b (III) in ham slices as a function of tumbling time ............................. 89 27 Microphotograph of a cross section of bicep femoris fibers in ham fresh meat, vacuum, 60 min tumbling (X 80) ............ 91 28 Microphotograph of a cross section of bicep femoris fibers in ham from fresh meat, vacuum, 120 min tumbling (X 80) ..... 91 29 Microphotograph of a cross section of bicep femoris fibers in ham from fresh meat, vacuum, 180 min tumbling (X 80) ..... 92 30 Microphotograph of a longitudinal cut of bicep femoris fibers in ham from fresh meat, non vacuum, 60 min tum- bling (X 80) .............................. 92 31 Microphotograph of a longitudinal cut of bicep femoris fibers in a ham from fresh meat, non vacuum, 120 min tum- bling (X 64) .............................. 94 32 Microphotograph of a longitudinal cut of bicep femoris fibers in ham from fresh meat, non vacuum, 180 min tum- bling (X 80) .............................. 94 Figure 33 Microphotograph of a seam or binding junction area in ham from fresh meat, vacuum and 120 min tumbling (X 64) ... 34 Microphotograph of a seam or binding junction area in ham from fresh meat, vacuum, 120 min tumbling (X 64) ...... 35 Microphotograph of a seam or binding junction area in ham from fresh meat, non vacuum, 120 min tumbling (X 64) .. 36 Microphotograph of a cross section of bicep femoris fibers in ham from frozen meat, vacuum, 120 min tumbling (X 80) 37 MicrOphotograph of a cross section of bicep femoris fibers in ham from fresh meat, vacuum, 120 min tumbling (X 80) Xi ..... Page 95 96 96 98 98 INTRODUCTION Ancient processing of meat products evolved as an art and only in recent history have scientific principles and advanced technologies been applied in meat processing. Today approximately one out of seven pounds of meat produced in most of the developed countries around the world is consumed as sausage or other processed meat items. Since meat and meat products play a key role in the diets of most cultures by providing high quality proteins, minerals and vitamins and a high satiety value to consumers, the demand for these foods will no doubt remain high. Although the origin of meat processing has been lost in history, it most likely began when primitive man first dis- covered that salt is an effective preservative and that cooking prolongs the keeping quality of fresh meat. Today processed meats are highly regarded for the convenience and variety they provide to the meat portion of the diet. More— over, increasing consumer demand for leaner meat, milder fla- vor, tender texture and low levels of additives in cured meats has encouraged the industry to experiment with new processing developments. Recently developed techniques in the production of sectioned and formed meat products allow the retention of the structural integrity of the original muscle source and result in a greater uniformity than in the original pro- duct. Such techniques have become widely used in the meat industry in several European countries and in the United States. Although these processes are considered to be innova- tive ones, they actually are adapted applications of ancient principles. They attempt to form a stable heat set protein gel which will effectively bind legal limits of fat and water in an attractive meat product packed so as to maintain wholesomeness, appeal and palatability for a maximum length of time. Two of the most popular techniques used in the pro- duction of sectioned and formed meats are massaging and tum- bling. In both cases brine-injected muscle chunks are placed in massagers or tumblers and subjected to various mechanical treatments. Mechanical work is then imparted to the chunks of meat through a process of mixing, churning and pounding in such a manner that the pieces of muscle become soft and pliable and develop a creamy, tacky exudate on their surfaces in the form of a protein coat. The protein coat is then heat-coagulated by cooking to form a binding matrix between muscle chunks which allows the product to possess the look of ”intact" muscle foods, such as roast or hams. The purpose of massaging and tumbling meat is to ensure a quality finished product and to obtain the following objec- tives: to maximize yields, impose color and binding, reduce cooking time and loss, control added substances and reduce inventory. The resultant uniformity of the brine distribu— tion, the shortened curing time and the saved pickle are equally important factors to consider in using these two processsing techniques. This study was designed to assess the mechanical effect of tumbling meat pieces on the nature of the exudate and on texture, cure distribution and acceptability parameters of sectioned and formed boneless hams. Three specific objec- tives were emphasized (l) the determination of the optimum tumbling sequence of meat pieces for optimum bind, texture and cure distribution characteristics, (the effect of vacuum during the tumbling operation is also assessed at this point); (2) the determination of the effect of the nature of the meat source, (e.g. fresh hams versus frozen and thawed hams) on bind, texture and cure distribution characteristics; and (3) the determination of the effect of the pickle cure level on the acceptance characteristics of the final product. LITERATURE REVIEW The Protein System in Pork Muscle Muscle proteins, as they are organized and distributed within the muscle, have traditionally been classified into two main groups: extracellular and intracellular proteins. The former occur outside the sarcolemmal membrane and the latter are contained inside that membrane (Asghar and Pear- son, 1980). A. Extracellular components: The connective tissue and the proteins of the interstitial space constitute the extra- cellular components. Morphologically, connective tissue comprises three distinct components. 1. Fibrous proteins: The major fibrous proteins in the extracellular spaces include collagen, elastin and reticulin (Forrest 3E gl., 1975). 2. Ground substance: The ground substance occupies the extracellular space of the connective tissue and is a viscous fluid derived from the plasma. It is com- posed of globular muc0protein (protein associated with mucopolysaccharides), tr0pocollagen and tr0po- elastin (Asghar and Pearson, 1980). 3. Cells: Two types of cells are recognized: fixed cells and wandering cells, the former include the fibroblasts, undifferentiated mesenchyme cells and adipose or fat storage cells (Forrest gt gt., 1975). Intracellular proteins: Pork muscle cells contain a large variety of proteins, many of which are involved in the glycolytic pathway of muscle metabolism and the con- traction relaxation process. These are the so-called intracellular proteins, and they are further classified into two main groups: the sarcoplasmic and the myo- fibrillar proteins. 1. Sarcoplasmic proteins: Sarcoplasmic proteins are the soluble proteins of the sarcoplasm located within the sarcolemma. These proteins are soluble at ionic strengths of 0.05 or less (Goll gt gl., 1974). They comprise about 30 to 35% of the total muscle proteins. They include a nuclear fraction, a mitochondrial fraction, a microsomal fraction and a cytoplasmic fraction, based on ultracentrifugation studies @sghar and Pearson, 1980). As many as 50 to 100 different proteins are known to constitute the sarco— plasm (Goll gt gt., 1970). Some of these proteins are the nucleoproteins and lipoproteins, the TCA cycle and the electron transport chain enzymes, myoglobin, as well as protein component of the microsomes, sarcoplasmic reticulum, the T-system and the lyzosomes. Myofibrillar proteins are those components of the unique myofibrillar system within muscle fibers. They are further divided into two subclasses: (l) the myofilamentous proteins, including myosin and actin, and (2) the regulatory proteins, including the tropomyosin-troponin complex, a- and B-actinins, M-protein and C-protein (Maruyama and Ebashi, 1979). According to Asghar and Pearson (1980) all these proteins are involved either in muscle contraction or in its regulation. A detailed discussion on each of the contractile protein has been made by Gergely (1966) and Briskey and Fukazawa (1971). The Conversion of Pork Muscle to Meat The conversion of muscle to the component tissue of a cut of meat can be summarized as being the effects of the degradation of ATP in the period from death to postrigor It is true that commercial handling practices after slaughter can influence the subsequent quality of meat, but they can only do this within limits set by the physiological and bio- chemical characteristics of an animal before and at the time of slaughter (Lister, 1970). According to Kastenschmidt (1970) the variable rate of postmortem metabolism has important implications in the ultimate usefulness of muscle as food. According to this author "fast glycolyzing" muscle are those having a pH of 5.5 or less at 30 min. postmortem. "Slow glycolyzing” muscle have a pH of 6.0 or higher at 60 min. postmortem. "Stress resistant" animals are those which can withstand antemortem stress and whose muscles after death are usually slow glyco- lyzing. Finally, "stress susceptible" animals are those which cannot tolerate antemortem stress. They usually have fast- glycolyzing muscle or expire before they can be exsanguinated. It is generally accepted that the deficient water-binding capacity of the pork meat is associated with a rapid pH fall after slaughter due to rapid glycolysis . This type of meat has been found less suitable for sausage mamfihcflue and detri- mental for the quality of canned hams (Wismer-Pedersen, 1969). Numerous research efforts have been made to relate live animal parameters to a judgment of the quality of postmortem meat. A color and structure score (Wisconsin system) ranks porcine meat from 1 being pale, soft and exudative (PSE) to 3 being normal to 5 being dark, firm and dry (DFD), (Cassens gt gl., 1975). It is now known that meat from stress-suscep- tible animals may be PSE, DFD or even normal in appearance, depending on the handling of the animal before, during and after slaughter. Cooper gt gt. (1969) made an attempt to explain the cause of PSE condition in porcine muscle. These authors found that stress-susceptible animals present skel- etal muscle with a large number of intermediate fibers which are dependent upon aerobic metabolism, but unlike typical red fibers they have especially high ATPase and phosphorylase activity, breaking down ATP and accelerating glycolysis to trigger a rapid glycolytic rate in the entire muscle. Additionally, even the regular white, and to a lesser extent the regular red fibers have rather intense ATPase and phosphorylase activity and further contribute to the accel- eration of these metabolic phenomena in the muscles of stress- susceptible animals. Merkel (1971a) found fewer capillaries per square millimeter in PSE muscle. Thefibers of PSE muscle were also significantly larger. He concluded that PSE muscle would be more predisposed to the development of anoxia. There seems to be little doubt that PSE meat is less desirable for certain processing procedures than is normal meat. PSE hams have been reported to produce gelatinous cookout losses with poor color and texture when compared to normal hams (Cassens gt gt., 1975; Merkel, 1971b). Myofibrillar Proteins and Functional Pr0perties of the Meat The myofibrillar proteins and the connective tissue proteins are fibrous and elongated and form viscous solutions with large shear resistance. These properties together with other lines of indirect evidence (Marsh, 1970; Marsh, 1972), have led to the axiom that variation in meat tenderness is directly and almost entirely the result of variations in the state of myofibrillar and connective tissue protein fractions (Goll gt gl., 1974). Although tenderness is an important factor in processed meat production, heat-gelling and emulsification properties are critical characteristics in some types of processed meats such as comminuted sausage, fine cut sausage and sectioned and formed meat products. Again, myofibrillar proteins, especially myosin, play a fundamental functional role (Briskey and Fukazawa, 1971). According to these authors, myosin appears to have a major influence, whereas actin has little influ- ence on gelation. They also reported that when actin and myosin are combined, however, gel strength is improved and the complexbinds more water than myosin alone. According to Hamm and Hofmmn (1965) the heat coagulation of myofibril- 1ar proteins is attributable to intermolecular associations of side groups (other than sulfhydryl groups) on the mole- cules. The experiments of Fukazawa gt gt. (1961a, 1961b, 1961c) show myosin to be a key constituent of the desirable binding quality in experimental sausage. Trautman (1966) reported that muscle protein character- istics and their food manufacturing properties are decidedly influenced by the rate, temperature and extent of postmortem pH decrease. Decreasing pH reduces salt-soluble protein solubility and heat gelling prOperties. It also reduces the solubility of water soluble proteins and releases free heme from,myoglobin. The effect of heating on muscle systems, particularly on myofibrillar proteins, has been studied by Hamm (1966). He reported that changes in myofibrillar proteins at 30-50°C include two steps: (1) an unfolding of peptide chain and (2) the formation of relatively unstable cross linkages result- ing in a tighter network of protein structure within the 10 isoelectric range of pH. At 50-55°C a rearrangement of the myofibrillar proteins occurs causing a delay in the changes of water-holding capacity. At these temperatures new cross- linkages begin to form. They are quite stable and cannot be split by addition of weak base or acid. At 55-80°C most of myofibrillar proteins are coagulated. Above 80°C disulfide bonds form.by oxidation of the sulfhydryl groups of acto- myosin. Above 90°C H28 splits off from the sulfhydryl groups of actomyosin. Some other influences of heating on muscle systems include changes in digestibility, a decrease in vitamins, the development of the flavor and color of cooked meat, and the change in tenderness, resulting from changes in collagen molecules rather than changes in muscle proteins (Hamm, 1966). Goll gt gt. (1964) studied solubility of myofibrillar proteins after death. The authors found that significantly greater amounts of protein could be extracted from bovine muscle which had been excised immediately postmortem than from muscles left attached to the skeleton, even after 312 hours postmortem. However, the excised muscles were the least tender, these findings are in contradiction to those of Hegarty gt gt. (1963), who found a positive relation between myofibrillar protein solubility and tenderness. Sayre and Briskey (1963), studying porcine muscle myofibrillar proteins reported results similar to those by Goll gt gt. (1964). They demonstrated that myofibrillar protein solubility ranged from no reduction during the first 24 hours after 11 death when pH remained high at rigor onset to 75% reduction in muscle with low pH and high temperature at the onset of rigor mortis. They also suggested that muscle protein solu- bility appeared to be one of the major factors affecting the juice-retaining properties of muscle. The Process of Binding of Meat Pieces Although an invention related to binding of chunks of meat was patented in the early 1960's (Maas, 1963), little work is found in the literature on the binding of pieces of meat and the mechanism underlining such binding before 1970. At this time,this type of binding became extremely important for the poultry industry, expecially with the advent of new products such as turkey loaves and rolls. In 1970 it was estimated that 2 Z of all turkey meat was used in the pro- duction of these convenience items, (Vadehra and Baker, 1970). These authors found the binding of meat pieces,when apprOpri— ately heated,to be complex and involve the following factors: (1) water-holding capacity, (2) cell disrupture and breakage, (3) release of intracellular material, (4) the myofibrillar and connective tissue proteins, and (5) extraneous sources of protein. Maesso gt gt. (1970a) reported no difference in bind- ing in turkey and broiler meat pieces (1 inch cubes). How- ever, breast muscle was found to give better binding than leg muscle. The difference in pH in these muscles was reported to have some practical implications. 12 Acton (1972a) reported a significant decrease in cook- ing loss along with an increase in binding strength as meat particle size become smaller in poultry loaves. Acton, (1972b) also reported an increase in cooking loss as the internal temperature of poultry loaves increases above 55°C. Acton and McCaskill (1972) found that salt—soluble rather than the water-soluble proteins in poultry meat are respon- sible for increased meat binding and cooking yield. Maesso gt gt. (1970b) reported that mechanical beat- ing of meat releases the intracellular content of broken muscle cells and causes a significant increase in binding. They also reported an increase in binding by NaCl, Kena (Na-tripolyphosphate, tetra-Na-pyrophosphate and Na-acid- pyrophosphate) and hexametaphosphate. When NaCl was combined with Kena they observed a significant additive effect. MacFarlane gt gt. (1977) studied the ability of isolated muscle proteins, actomyosin and myosin, to bind pieces of meat together. They found that myosin is able to bind meat pieces not previously subjected to mechanical agitation or having salt added. Actomyosin was found to match myosin in this respect only at high salt concentrations (1.2 and 1.4M). Schnell gt gt. (1970) have clearly demonstrated the importance of salt-soluble proteins in binding and reduction of cook loss in chunk-type products. Moreover, these authors concluded that salt-soluble proteins are not the only source of binding materials. Bard (1965) reported that extraction yields of salt 13 soluble proteins are influenced by NaCl concentration, ex- traction time, extraction temperature and the extent of rigor development in the muscle tissue. The author stated that there may be other factors of equal or even greater impor- tance than those reported. Pepper'auischmidt (1975) showed that both salt and phosphates increase the binding strength and cook yield of beef rolls, and that binding strength is higher in the salt-phosphate than in the salt treatments. Similar results were reported by Moore gt gt. (1976) with beef rolls. Furthermore, these authors reported that the cook yield is closely associated with binding strength. The effect of phosphates on salt-soluble protein ex- tractability and binding strength of the sausages has been studied by Fukazawa gt gt. (1961c). They concluded that the ionic strength of the cured meat maintains a condition such that the muscle structural protein is drawn to the outside through the sarcolemma of the muscle cell and that such action may be promoted by the use of phosphates. Further— more, they stated that the binding quality of sausage has a close relationship to the myosin A (myosin protein) content and to the dissociable components of myosin B (actomyosin complex) with phosphates having the effect of contributing the dissociation of the complex. Fukazawa gt gt. (1961b) pointed out the importance of suitable amounts of remaining native myosin in fibrils for good binding properties. The fact that the mechanism of binding between chunks of meat is a heat initiated reaction, as described by Schnell 14 gt gt. (1970) and Vedehra and Baker (1970) has led several authors to investigate the gelation properties of myosin. Ishioroshi gt gt. (1979) reported that the heat-induced gela- tion of myosin is optimally developed at temperatures between 60 and 70°C and at pH.6.0 in 0.6 M KCl. Yasui gt gt. (1979) showed similar results to those obtained by Ishioroshi gt gt. (1979). Furthermore, these authors pointed out that the heat-induced gelation of myosin may be the result of the deve10pment of a three-dimensional network structure which holds water in a less mobilized state. Samejima gt gt. (1969) reported that heavy and light meromyosin fragments have little influence on binding properties. They further concluded that an intact molecule of myosin is required for development of binding prOperties upon heating. Schmidt gt gt. (1981) point out that the prOperties characteristic of myosin gels suggest that the mechanism behind the gelation of myosin involves the formation of fairly stable bonds by irreversible changes in its quaternary structure that are caused by heating. Siegel and Schmidt (1979) found that the binding abil- ity of crude myosin preparations are significantly greater than the binding ability of either a muscle homogenate free of fat and sarcoplasmic proteins (a total muscle homogenate) or a non-protein control consisting of salt, phosphate and water. They suggested that ionic interactions are implicated in the binding phenomena. Turner gt gt. (1979) reported that crude myosin 15 extracted from postrigor bovine muscle has a potential use as a meat binding agent, since no myosin was extracted from muscles in either prerigor and postrigor state. They also reported 1 M salt and 0.25% tripolyphosphate in the extracting solution as the best concentrations to obtain maximum yields. Ford gt gt. (1978) found significant correlations between overall acceptability of restructured beef steak- ettes containing added myosin and the flavor, juiciness, tenderness and objective measurements in binding strength. Significant correlations were also found between the objec- tive and subjective assessments of binding strength. Reynolds gt gt. (1978) studied the effects of ultra- sonic treatment on binding strength in cured ham rolls. They found that ultrasound causes changes in muscle micro- structure, increases breaking strength, decreases cooking loss and increases the extractability of salt-soluble pro- tein. The Technique of Tumbling and Massaging Meat Pieces The success of meat processing into sectioned and formed meats has been reported by Schmidt (1978). He pointed out that more than 284 million pounds of sectioned and formed hams were produced under federal inspection in 1977. In addition, the same author lists nineteen patents on sectioned and formed meat processes granted since 1963. 16 Almost all these procedures included tumbling or massaging procedures. Anonymous (1981), reported that according to the U.S. Department of Agriculture, about 2 billion pounds of boneless ham products were manufactured in 1979. About 50% of that tonnage was produced as smoked or cooked bone- less or sectioned-and-formed hams (including water/added), and 14% as canned products. Tumbling, typically used in the domestic cured meat industry, includes both tumbling and massaging action. Tum- bling, per se, involves the result of "impact energy" influ- ences on muscle such as would occur in allowing meat to fall frmm the upper part of a rotating drum or striking it with paddles or baffles. Such action leads to the transfer of kinetic energy to the muscle mass and a resultant tempera- ture rise of the processing material. Massaging is a less physically rigorous process and involves "frictional energy" resulting from the rubbing of one meat surface on another or on a smooth surface of a container (Weiss, 1974). In both cases brine-injected muscle chunks are placed in massagers or tumblers and subjected to various mechanical treatments. Mechanical work is imparted to the chunks of meat through a process of mixing, churning and pounding in such a manner that the chunks of muscle become soft and pli- able and develOp a creamy, tacky exudate on their surfaces in the form of a protein coat. The protein coat is then heat coagulated by cooking to form a binding matrix between muscle chunks which allows the product to possess the look 17 of "intact" muscle foods such as roasts or hams (Theno gt gt. 1977). Thus, the binding between meat chunks is concluded to be a heat-mediated phenomenon which causes a structural rear- rangement of the solubilized meat proteins, and renders them more susceptible to essential protein binding. The forma- tion of the protein matrix is therefore essential to Optimal binding in sectioned and formed products (Theno gt gt., 1976). According to Schmidt (1979) the goal of these proce- dures is the formation of a stable heat set protein gel that will effectively bind legal limits of fat and water in an attractive and palatable meat product packed in such a way to remain wholesome, attractive and palatable for a maximum length of time. According to Starr (1979), in practice, the purpose of massaging and tumbling meat are to ensure a quality finished product and to obtain the following objectives: (1) maxi- mizing yields, (2) impose color and binding, (3) reduce cooking time, (4) control added substances, (5) reduce inven- tory and (6) save curing brine. As stated by Woolen (1971) perhaps the most important effect of mechanical working imparted to the meat, other than high yield and homogeneous appearance, is the evening out of the brinecfistrflxfiiqn and shortening of curing time. This author also suggested that application of tumbling to curing is best achieved by injecting the brine before the first tumble. This process allows the absorption and distribution of the brine. It is followed by a maturing 18 period, often ending with a second tumble, which is used for the extraction of the salt-soluble proteins to provide for the bonding of meat surfaces when meat is thermally processed. This procedure has led to the development of automated tumr blers in which a programmable system allows the meat chunks to be tumbled under vacuum for predetermined intervals and then to equilibrate before tumbled again (Anonymous, 1971). In some equipment tumbling and massaging are combined. Addis and Schanus (1979) reported on a vacuum massage tum- bler designed in Europe. According to these authOrs, massag- ing treatment is applied for 10 to 20 hours. Any brine not absorbed by the meat during stitch pumping can be added to the massaging vats and eventually incorporated during massag- ing. Weiss (1974) summarized the advantages and disadvan- tages of tumbling and massaging. He lists the following advantages: (1) improved brine penetration and uniformity of dispersion; (2) uniform color development; (3) improved release of salt-soluble protein enhancing product bind and coherency; (4) development of a more uniform fine texture; (5) improved yield during processing; (6) reduced product weight loss during consumer preparation; (7) production of a finished product with very desirable slicing characteristics. The many disadvantages he lists include: (1) the initial skinning, boning and defatting procedures require expertise and precision; (2) the considerable massaging time required to develop the qualitative aspects associated with the l9 technique; (3) excessive massaging results in tissue integ- rity destruction and adverse temperature rise; (4) excessive moisture absorption adversely influencing finished product coherency, bind and appeal; (5) massaging and tumbling equip— ment primarily EurOpean in origin; (6) the technique employs batch production units to produce desirable results; (7) batch production units limited to 1500 pounds or less to facilitate manufacture of finished products with superior quality and consumer appeal. Research in the United States on the effects of tum- bling and massaging started in the 1970's. Siegel gt gt. (1976 and 1978b) showed that as the massaging time increases so does the level of fat and protein in the exudate of hams, although these increases are more pronounced in the presence of salt and phosphate. The influence of tumbling and sodium tripolyphosphate (Na-TPP) on salt and nitrite distribution in porcine muscle was investigated by Krause gt gt. (1978a). The results indi- cated that both Na-TPP and tumbling significantly increase the migration of salt and nitrite and result in an increase in cure color deve10pment. These observations agree with those made by Okerman and Organisciak (1978). The results by Krause gt gt. (1978a) also indicated that Na-TPP and tum- bling increase the level of residual nitrite content, al- though the tumbled hams have higher levels of cured meat pig- ments formed. 20 Krause gt gt. (1978b) studied the influence of tumbling, tumbling time, trim and Na-TPP on quality and yield of cured hams. They reported that tumbling has a significant influ- ence on external appearance, internal ham color, slicability, taste, yield and aroma. The most dramatic effect, however, is on sliceability and yield. The authors also reported a significant improvement in external color, sliceability, taste and aroma and yield of cured hams independent of the tumbling effect. Rejt gt gt. (1978) used massage under vacuum in the elaboration of canned hams. They observed that massaged muscles show a definite change of structure, particularly of surface layers, and an increased water-holding capacity. After heat treatment hams show higher tenderness and lesser cooking loss than the non-massaged meat. Siegel gt gt. (1978b) reported that the massaging process involves great degrees of tissue destruction at the cellular level which aids in the extraction, solubilization, concentration and distribution of the major myofibrillar proteins on surfaces and interiors of muscle chunks. All these results of massag- ing are beneficial to the improvement of binding. Theno gt gt. (1978a, 1979b and 1978c) reported the observation of light and scanning electron microscope microphotographs of tumbled ham material. These authors indentified the pres- ence of fiber fragments in the exudate of hams tumbled for 24 hours regardless of whether salt and phosphate were added to the meat. The treatments with salt and phosphates showed 21 clouds of solubilized protein. The length of massaging en- hanced the effects in all treatments. Further massaging re- sulted in longitudinal disruption of the fibers shown under the scanning electron microscope. They also reported that at low salt concentrations in the brines, the junctions exhibited poor binding characteristics with high levels of fat and cellular fragments as seen under the light microscope. Junctions from rolls with adequate salt (22%) and phosphate (0.5%) exhibited good binding characteristics. Cassidy gt gt. (1978) made similar observations. In addition, however, they reported that intermittent tumbling resulted in more alterations in cell structure than continuous tumbling. Ockerman gtht. (1978) found increased cohesiveness values in canned hams tumbled for 30 min. when meat was cured with salt and tripolyphosphate. They also stated that tum- bling for 30 min. is not sufficiently long to increase yield, texture or sensory characteristics of hams. Knipe gt gt. (1981) studied the effect of intermittent tumbling and tumbling temperature on total aerobic plate counts (ATPC) and quality of boneless, cured hams. They showed that a significant rise in internal temperature of the meat can be observed after 3 hours tumbling (10 min. tum- bling, every hour, for 18 hours). They also reported that the exudate ATPC is significantly reduced after 18 hours tumbling. Solomon t l. (1980) studied the effect of vacuum and rigor condition on cure absorption in tumbled porcine 22 muscles. Their results indicate that vacuum.and prerigor state independently increase the absorption of NaCl. They also pointed out that vacuum is implicated in increased binding functionality, since breaking strengths of ham slices were found to be greater when vacuum tumbling was used. Non-meat Proteins in the Binding of Meat Pieces Hawley (1977) reported the use of non-meat proteins along with the pumping brine as a technique for augmenting intact muscle protein in hams. They recommended pumping to 145% of green weight in order to obtain finished hams with approximately a 130% yield when cooked (89% smokehouse yield). The procedure also includes massaging or tumbling to assure distribution and equilibration of the brine and vacuumrmixing after tumbling to remove entrapped air from the muscle. Siegel _t _t. (1979b) studied the effects of various levels of isolated soy protein (ISP) in combination hams. They reported that massaging and ISP improves both binding and cook yield. Increased levels of injection decrease bind- ing strength and cooking yield. Massaging improves uniform- ity, textural appeal and overall acceptability, but it de- creases tenderness and does not effect juiciness and flavor. In a similar study with ISP Siegel gt gt. (1979c) reported that ISP occupies primarily perimysial spaces and that 23 massaging acts to incorporate these proteins into the endo- mysial spaces and mix them with extracted myofibrillar pro- teins. According to the authors the ISP appears to enhance myofibrillar protein extraction by binding water, thus in- creasing the effective concentration of salt and phosphate. Kardouche gt gt. (1978) used ISP at different levels up to 3% with pre- and postrigor turkey in the preparation of rolls. They concluded that as the level of ISP increases the flavor, tenderness, texture and acceptability scores in- crease, and the shear values decrease. They also reported that the level of ISP has greater influence on the shear value than the rigor state of the meat. Siegel gt gt. (1979a) ranked the binding abilities of several non-meat proteins in the presence of 8% salt and 2% sodium tripolyphosphate from highest to lowest as wheat glu- ten, egg white, corn gluten, calcium reduced dried skim milk, bovine blood plasma, ISP and sodium caseinate. New Trends in the Acceptance by Consumers of Sectionediand Formed Meats ‘ Considerable concern has been expressed over the cur- rent dietary intake of fats and additives contained in processed meats. Kolari (1980) has discussed the salt dietary concern. He concluded that, although current evidence does not pro- vide the basis for drastically reducing salt dietary intake for the general population, moderation needs to be considered 24 for those at risk of develOping essential hypertension. Marsden (1980) reported that the contribution of the processed meats to the sodium level in the American diet is significant and the meat industry should be aware of its involvement in this controversy. He concluded that sodium- containing additives perform important technological func- tions in addition to their contribution to flavor. Conse- quently, if it becomes necessary to reduce the level of sodium in processed meats, the amount of the reduction should not be arbitrarily determined. Nitrites present in processed meats are thought to pose a health hazard by virtue of their ability to form N-nitro- samines. Many of these compounds are carcinogenic and, in addition, some exhibit mutagenic, embryopathic or teratogenic properties. Although there is no direct evidence the N-ni- troso compounds are carcinogenic to man, indirect proof from animal studies on 12 species would suggest this potential danger to man (Gray and Randall, 1979). The argument has been made that'discontinuing the use of nitrite as a food additive would greatly reduce or eliminate this risk (Tannen- baum, 1979). However, according to the same author, the risk that might exist from the use of nitrites according to present regulations would be minuscle compared to those resulting from the body's natural processes. The other point of controversy concerns the fat con- tent in meat and meat products as a major contributor to the development of such chronic diseases as cardiovascular 25 disease and cancer (Leveille, 1980). This author states that, although there is no scientific evidence to support the recommendation to reduce meat consumption, a challenge should be made to the meat industry to reduce the fat con- tent of both fresh and processed meats. These points are part of the reasons why today con- sumers exhibit new preferences related to processed meats. They look for leaner and milder products containing lower levels of fat and additives (salt, sodium, nitrite) than previous products have contained. The manufacture <1f sectioned and formed processed meats may prove to be a process in which fat and additives levels can be carefully controlled in order to produce a finished product widely accepted by every segment of the population. MATERIAL AND METHODS Description of the Experiment The study was conducted in two parts in order to ratio- nalize sample collection and duplicate processing yield data. In the first experiment, conducted in the fall of 1980, 60 hams were assigned to fifteen processing treatment groups. Four hams were used per treatment. The experiment was duplicated in the winter of 1981 with 30 hams assigned also to the processing treatment groups. However, only 2 hams (per treatment) were used in this second experiment. The following sources of variation were considered in the experiments: A. Tumbling or massaging sequences. Three tumbling se- quences were tested. 1. Sixty minutes of mechanical working of the meat was accomplished by keeping the meat for 4 hours inside the tumbler with 15 minutes tumbling and 45 minutes pausing in each hour. 2. One hundred and twenty minutes mechanical working of the meat was accomplished by keeping the meat for 8 hours inside the tumbler with 15 minutes tumbling and 45 minutes pausing in each hour. 3. One hundred and eighty minutes mechanical working of 26 27 the meat was accomplished by keeping the meat for 18 hours inside the tumbler with 10 minutes tumbling and 50 minutes pausing in each hour. B. Tumbling pressure effect: Two pressure conditions during tumbling of the meat were studied. 1. Vacuum: Meats were tumbled for a period of time given by the tumbling sequence treatment under 25 inches of Hg vacuum” 2. Non-vacuum: Meats were tumbled as long as required by the respective tumbling sequence at normal atmo- spheric pressure. C. Conditions of the meat: Two sources of meat were studied. 1. Frozen and thawed pork 2. Fresh pork D. Level of brine injection: Two levels of brine injection, based on raw meat weight, were studied. 1. Sixteen percent pumping 2. Thirty-two percent pumping Processing treatments identified by code numbers are shown in Table 1. Statistical Design The effects of tumbling sequence, tumbling pressure and condition of the meat were analyzed by a 3-way analysis of variance (ANOVA). This part of the design included treat- ments 1-12, as shown in Table 2a. 28 ><><><>< X XX ><>< x x x x x x x x x x x x Nam NS flea g a name «An mo Hm>mH coauommcfi mnfium wcaxuo3 Hmoficmnowe mo mafia ><><><>< x cmuoum Smoum x . UG>ICOZ EDSUGD mg «a mH Nfi Ha OH |\ LG umma ecu mo cowuficaoo umme mo.wdwanasu wcwusp mwsmmmum noses: ucmEumwuH .mEm: mmmamcon mo wswusuomwscme use :a new: mucmEumwuu waammooonm mo coaudfiuomwn I a manna 29 Table 2a - Processing treatments as arranged for statistical analysis by 3—way ANOVA. Tumbling Treatment Number Sequence Vacuum Non Vacuum min Fresh Frozen Fresh Frozen 60 10 9 12 11 120 6 5 8 7 180 2 1 4 3 The effects of tumbling sequence and level of brine injection were statistically analyzed by a 2-way ANOVA. This part of the design included treatments number 1, 5, 9, 13, 14 and 15, as shown in Table 2b. ANOVA was conducted at the MSU Computer Center using the Statistical Package for the Social Science (SPSS), version 8 (Nie gt gt., 1975). Table 2b - Processing treatments as arranged for statistical analysis by 2-way ANOVA. Tumbling Treatment Number Sequence 16% brine pumping 32% brine pumping (min) 60 10 15 120 6 14 180 2 l3 When significant differences were observed between more than two means, the Bonferroni t statistics for 30 nonorthogonal designed contrasts (Gill, 1978 and Neter and Wasserman, 1974) was performed to determine which means were significantly different. A part of the taste panel results was analyzed by the Chi square method according to Steel and Torrie (1960), and American Society for Testing and Materials (1968). Source of Meat Fresh pork was obtained from Peet Packing Co., Chesaning, Mi. Although the requested weight for hams was 16 to 18 lbs. per unit, the hams arriving to our laboratory weighed between 14 and 22 lbs. Fresh hams intended to be used as a fresh meat source were delivered in groups of 4 or 8 units, 1 0r 2 days before the date of processing. Fresh hams intended to be used as a frozen meat source were delivered as a single batch at the beginning of the experiment. Preconditioning of Fresh Hams Preconditioning of fresh hams is indicated in Figure l as the first stage of the processing flow chart. Fresh hams used as a fresh meat source were vacuum packaged upon deliv- ery into Cryovac (polyvinylidene chloride) bags and kept in a cooler at 2°C until processed. Fresh hams used as a frozen source of meat were individually weighed upon delivery, wrapped in butcher paper and vacuum packaged in Cryovac bags. The packaged meats were then frozen and stored at 31 'DELIVERING T F Iv] WEIGHING VAC. PACKAGING 1 VAC. PACKAGING FREEZING AT -29°C 1 THAWING AT 2°C 1 ‘FROZEN & THAWED MEAT I T STORAGEAT 2°C FRESH MEAT J— WEIGHING f MUSCLE SEPARATIOE] w MEA J SAMPLE L WEIGHING I PUMPING WITH BRINE I Q 'TUMBLING _J ll EXUDATE STUFFING IN CAS INGS SAMPLE l COOKING I COOLING HAM 1 SAMPLE 1 VACUUM PACKAGING Q ‘1 FREEZING STORAGE REFRIG. STORAGE FIRST STAGE PRECONDITIONING SECOND STAGE PROCESSING THIRD STAGE STORAGE FIGURE 1 - Processing flow chart and sampling points in the manufacturing of boneless hams. 32 -29°C. The frozen hams were taken out of the freezer as needed and allowed to thaw in the cooler at 2°C for five or six days before processing. Processingfiand Sampling Operations As the flow chart in Figure 1 indicates, the processing of the meat is the second stage in the operation. At this point fresh hams were individually weighed, skinned, boned and separated into muscle groups and the fat was trimmed to less than 1 mm thick. The weight of skin and fat, bones, fines and trimmed muscles were recorded at this stage. Five different muscle groups were identified and sepa- rated from.each ham: biceps femoris, semimembranous (with gracilis attached), semitendinous, the quadriceps group (commonly known as the knuckle of the ham) and the gastroc- nemius group (commonly known as the mouse of the ham). The trimmed ham muscles were then sampled (labeled as raw meat sample) and analyzed for moisture, fat, protein and lipid oxidation by the TBA method. The meat was then injected with brine at either 16% or 32% of the raw meat weight by using a stainless steel pickle pump equipped with a spray multiple needle injection system (Hubert Distributing Co., Cincinnati, OH. Catalog numbers 38233EC, 4NH and SNCHA). All brines contained high grade improved fine flake salt (Diamond Crystal Salt Co., St. Clair, MI. ) sugar (Monitor Sugar Company, Bay City, MI.), sodium tripolyphosphate 33 (FMC Corp., Phosphorous Chemical Div., Newark, CA.), sodium ascorbate (Permacurate Roche, Hoffman La Roche Inc., Nutley, NJ.) and sodium nitrite (analytical reagent, Mallinckrodt Inc., Paris, KY.). Brine compositions are shown in Table 3. The brines were analyzed for salt and nitrite just before the pumping of the meat. Table 3 - Brine composition as used in the manufacturing of boneless hams. Ingredient Concentration in the brine, pgrcent Brine 1 (16% pumping) Brine 2'(32% pumping) Salt 13.00 7.03 Sugar 4.87 2.63 Phosphate 1.62 .88 Ascorbate .36 .19 Nitrite .10 .05 Water 80.05 89.22 Pumped meat was then placed inside the tumbling machine and tumbled as required by the respective processing treat— ment in a cooler at 2°C. The tumbler used in this study was a Roschermatic mixing, curing and massaging machine, model MM 80 (Roscherwerke GmbH, Osnabrfick, W. Germany), equipped with a mixing arm rotating at 20 r.p.m. The drum was oper- ated at an angle of 40° so that the mixing arm could always grab the meat. Whenever vacuum was required, a Welch Duo- Seaal vacuum pump model 1405 (Sargent Welch Scientific Co., 34 Skokie, IL.) was used to pull 25 inches of Hg vacuum inside the tumbler. After mechanical working the meat was taken out of the tumbler, weighed and sampled from the cores of the mus- cles (labeled as tissue) and from the creamy exudate sur— rounding the meat pieces (labeled as exudate). Tissue and exudate samples were further analyzed for composition (pro— tein, fat and moisture) by proximate analysis, lipid oxida- tion, nitrite and salt. Exudate material was also analyzed for soluble phase volume, protein content in the soluble phase and character of the proteins in the soluble phase by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Tumbled meat was then stuffed into prestuck clear regular fibrous casings, 61 cm long and 14.2 cm diameter (Union Carbide Corp., Chicago, IL.), by using a hand oper- ated jiff net horn (Meat Packers and Butchers Supply Co., catalog number 81135, Los Angeles, CA). Full casings were then tightly sealed using a hand operated stretch clip machine, model J-l (Global Industrial Machinery Corp. Chicago, IL). After stuffing, the product was labeled, weighed and secured in tightly stretched stockinette clipped at either end before cooking in a smokehouse. Cooking of boneless hams was done in an Elek-Trol laboratory smokehouse (Drying Systems Inc., Chicago, IL) aczcording to schedule shown in Table 4. All hams were 35 cooked to an internal temperature of 68°C. Table 4 - Smokehouse cooking schedule for boneless hams. Temperature, °C Relative Time in Dry Bulb Wet Bulb Humidity, % (hours) 60.0 43.9 40 2 71.1 53.3 40 6 80.0 61.1 40 41 1 This value represents an average time needed to reach 68°C internal temperature in the finished product. After cooking, the temperature of the hams was brought down overnight in a cooler at 2°C. Then, fully cooked hams were sliced and sampled (labeled as ham). Hams were ana- lyzed for moisture, fat and protein levels, lipid oxidation (TBA), residual nitrite, salt, pigments (cured, total and conversion) and color parameters. Hams were also sampled for texture studies, taste panel and microscopy study on the biceps femoris part of the finished product. Methods of Analysis 1. Proximate analysis a) Protein content was determined by the microneldahl method for nitrogen according to AOAC (1965) proce— dure. Results were expressed as protein percent using 6.25 as a conversion factor for nitrogen values. b) Moisture content was determined by the air drying 36 method of the AOAC (1965) in convection oven at 102°C for 18 hours. Moisture was reported as weight loss percent. Dried samples were saved for fat determination. c) Fat content was determined by extracting dried sam— ples with anhydrous ether for 5 hours in Goldfisch apparatus, as described in AOAC (1965). Salt analyses were performed according to the official Volhard method by the AOAC (1965). Nitrite determination was made according to methods described on a technical report by the United States Department of Agriculture (USDA, 1979), as modified from the AOAC (1965) method. Lipid oxidation, as a measure of the rancidity of the meat, was determined acanxfing to the method by Tarladgis gt gt. (1960) , modified by Zipser and Watts (1962) , for cured meats. Cured pigments, total pigments and pigment conversion were determined by the method of Hornsey (1956), as described by Konieco (1979). Color determination was performed by using a Hunter Lab Color/Difference Meter, model D 52-2 (Hunter Associates Laboratory, Inc., Fairfax, VA.). The instrument was standardized against a pink tile with values L = 67.6; a = 21.4 and b = 11.9. Strength of the binding between pieces of meat was assessed in the final product by using a Universal Testing Instrument, model TTC, equipped with a tension 37 load cell B, which was implemented with the appropriate grip coupling (Instron Corp. Canton, MASS.). Ham pieces approximately 1 cm thick, 2 cm width and 8 cm long and containing a binding zone across the center of the ham piece, were mechanically pulled apart at a constant speed of 2.54 cm/min The force needed to separate the pieces of meat at the binding line was recorded in a chart running at 2.54 cm/minand calibrated for 2.12 kg force full scale deflection of the pen. Soluble phase volume determination was made by weighing 20 g of exudate and 10 g of 3.9% w/v NaCl solution in a 100 ml homogenizing flask. The mixture was then blended at a low speed in a Virtis "45" homogenizer (Virtis Research Equipment, Gardiner, N.Y.) for 30 seconds. Next, 10 g of the slurry were weighed in duplicate into 15 ml Corex centrifuge tubes and then centrifuged at 2°C for two hours at 40,000X g in a Sorvall refrigerated centrifuge model RC2-B, equipped with a SS-34 rotor (Ivan Sorvall, Inc., Norwalk, CONN.). After centrifugation 3 layers were clearly visible in the tubes: the upper layer or fat cap, the intermediate layer or soluble phase and the bottom layer composed mostly of connective tis- sue and muscle tissue fragments. The fat cap was then separated by a small spatula and the soluble phase was allowed todrain.into 15 m1 graduated conical tubes pro- vided with funnels with two layers of cheesecloth for 15 minutes inside a cooler room at 2°C. The collected 10. 38 fluid was eXpressed as soluble phase volume. One ml of soluble phase was next diluted with 1 ml glycerol, stirred in a Vortex tube mixer and stored in a freezer at -20°C for further electrophoretic study. Biuret analysis: Protein content in the soluble phase was made by the microBiuret method described by Goa (1953). Sodium dodecyl sulfate-polyacrylamide gel electropho- resis (SDS-PAGE) was done by the method of Weber and Osborne (1969), modified by Porzio and Pearson (1977), and adapted for pork muscle proteins as follows: a) Electrophoresis solutions (1) Tris-Glycine stock solution (0.5M Tris; 1.5 M Glycine) was prepared in a one-gallon plastic bottle and stored at 2°C. (2) 25% Acrylamide; 0.25% N,N-Methylenebisacrylamide (BIS) stock solution was prepared and stored at 2°C in plastic bottle. This solution was for 10% gels cross-linked with BIS. (3) 2.5% sodium dodecyl sulfate (SDS) solution was stored at room temperature. (4) 1% ammonium persulfate solution was prepared immediately before using. (5) Chamber buffer solution (0.1% SDS, 0.20 M Tris- Glycine, pH 8.8) was prepared by appropriate dilution of solutions 1 and 3 and adjusted to pH 8.8 with HCl or NaOH solutions. 39 (6) Tracking dye solution was made of 1.0% SDS, 0.05 M Tris-HCl; 0.5% mercaptoethanol, 20% glycerol and 0.01% Pyronin Y in distilled water. pH was adjusted to 7.2 with 6N HCL and the solu- tion stored in a plastic bottle in freezer at -29°C. (7) Staining solution was made of 50% methanol, 7% glacial acetic acid and .033% Coomassie bril- 1iant blue in distilled water. This solution was prepared immediately before use. (8) Destaining solution was made of 7.5% glacial acetic acid and 5% methanol in distilled water. b) Gel Preparation (1) 10 ml solution (2), 5 ml solution (1), 1.25 ml glycerol, 1.0 ml solution (3), 0.01 ml of N,N, N',N'-Tetramethylethylenediamine (TEMED), 6.75 ml of water and 1.0 ml solution (4) were com- bined in a beaker with permanent but soft stir- ring. The solution was then transferred to running tubes and filled to 8 cm of the tube length. Gels were then overlayed with water and allowed to polymerize for 2 hours. c) Sample preparation (1) Soluble phase samples stored in freezer in a 1:1 dilution with glycerol were apprOpriately diluted with solution (6) to contain 0.4 mg pro- tein/m1. Diluted samples were then heated in a 4O boiling water bath for 5 min. (2) A standard purified protein mix containing myo- sin (MW : 200,000), bovine serum albumin (BSA) (MW : 60,000), ovalbumin (MW : 45,000) and ly- sozyme (MW': 15,000) was prepared in the same way as the soluble phase proteins. These pro- teins were mixed in equal parts to make a total protein concentration of 0.4 mg protein/ml. d) ElectrOphoresis (l) The tubes containing the gels were placed in the electrOphoresis chamber. Next, the lower and upper buffer chambers were filled with solu- tion (5) and the gels loaded with 50 ul sample. The entry of the sample into the gels was con- ducted at a current of 0.2 mA per gel. After the dye had completely entered, the current was raised to 0.5 mA per gel and the migration con- tinued until the dye front reached the tube end (10 to 12 hours total run). ElectrOphoresis was run in a cell Model 150 A connected to a power supply Model 400 and the gels were further destained in a diffusion chamber Model 172 A. All these apparatuses were manufactured by Bio- Rad (Bio-Rad Laboratories, Richmond, CA.). e) Gel densitometry (1) Gels were scanned using a Beckman DU Spectro- photometer, Model 2400 (Beckman Instruments, 41 Inc., Fullerton, CA.) equipped with a gel scan- ner 2520 and a photometer 252 by Gilford (Gil- ford Instrument Laboratories, Inc., Oberlin, OH). This system was surfaced to an HP integrator Model 3380 S (Hewlett Packard, Avondale, PA). The gels were scanned at a rate of 1.0 cm/min. and a chart speed of 2.0 cm/min. Start delay and slope sensitivity settings were 0 and 3.0 mV/min., respectively. SDS-PAGE gels were scanned at a wavelength of 550 nm. The rela- tive areas of the individual protein peaks were recorded. The relative mobility of the bands was assessed from.the total length of the gel (or tracking dye migration distance) and from the distances migrated by individual proteins. 11. Microscopy study. a) b) Sample preparation and fixing: Finished hams were sampled from the biceps femoris muscle by cutting pieces of meat (approximately 20 mm long, 5 mm wide and 2 mm thick) and keeping them in a 10% neutral formalin solution. Dehydrating, clearing and infiltration: This proce- dure was carried out in an Autotechnicon Model 2 A instrument (the Technicon Company, Chauncey, NY). Fixed tissues first were placed in tissue buttons and then in a basket carrier for the following immer— sion schedule: 1 hour into each of two 70% ethanol C) d) e) 42 containers; 1 hour into an 80% ethanol container; 1 hour into each of two 95% ethanol containers; 1 hour into each of two 100% ethanol containers; 1 hour into a 50% ethanol - 50% xylene container; 1 hour into each of two 100% xylene containers; and 2 hours into each of two liquid paraffin containers. Paraffin used was "Paraplast", m.p. 56-57°C (Scien- tific Products, McGaw Park, IL) at about 60°C. Imbedding: The infiltrated tissue preparations were next imbedded into a plastic disposable boat (approx- imately 2.5 cubic cm. volume) with melted paraffin and allowed to cool down overnight at room tempera- ture. Then the plastic boats were removed and dis- carded. Sectioning: Paraffin blocks containing tissue mater- ial were mounted in a Minot-Mikrotome, Type 1212 (E. Leitz GMBH Wetzlar, Germany) and cut to a 6 micron thickness. Next, paraffin ribbons containing the sectioned tissue material were floated in a warm water bath containing 2% gelatin and pulled from the ends to remove the wrinkles by stretching the tissue material. The sections were then picked up on glass slides by using a camel hair brush. They were drained approximately 1 minute and finally dried on a light bulb until the paraffin melted down. Staining: Tissue samples were stained with Harris' Hematoxylin and Eosine-Phloxine solutions according 43 to Luna (1968), with the following schedule of slide immersion: 5 min. into each of two xylene cells; 2 min. into each of two 100% ethanol cells; 2 min. into a 95% ethanol cell; 2 min. into a 80% ethanol cell; 2 min. into a distilled water cell; 10 min. into a hematoxylin cell; 4 dips into a 1% HCl cell; 2 min. into a tap water cell, or until slide was blue; 2 min. into an eosin cell; 2 dips into a 95% ethanol cell; 2 dips into a 100% ethanol cell; 2 min. into a 100% ethanol cell; 2 min. into a 50% ethanol- 50% xylene cell; 2 min. into a xylene cell and, finally, 5 min. into a xylene cell. f) Mounting: Stained preparations were covered with l or 2 drops of Pro-Texx mounting medium (Scientific Products, McGraw Park, IL) and topped with a cover- slip glass. These slides were allowed to air dry overnight at room temperature. g) Microscopic observation: This procedure was done with either a Sterozoom microscope (Bausch and Lomb, Rochester, NY) with 10X and 1X to 7X magnification factors for ocular and objective lenses, respectively, or with a Zeiss photo-microscope III (Carl Zeiss, Oberkochen, West Germany) under 200X magnification factor. Pictures were taken through both microscopes. 12. Taste Panel a) A semi-trained taste panel was conducted in two sessions with 12 panelists. In the first session b) C) 44 panelists were instructed on the evaluation of slices of hams by visual inspection. Three types of defects were emphasized at this point: color uniformity, surface texture and presence of non-muscle material. The panelists were then asked to evaluate ham slices corresponding to the 15 processing treatments used in this study. The score sheet used in this trial is shown in Appendix A-l. It was then demonstrated to the panelists how to evaluate selected pieces of ham for strength of the binding at the junction line between two chunks of meat. In the second session panelists were asked to evalu- ate the binding strength by comparing pairs of sam- ples. Four variables were studied in this trial: times of tumbling (short tumbling time versus long tumbling time); use of vacuum during tumbling ( vac- uum versus non—vacuum); condition of the meat (fresh pork versus frozen and thawed pork); and level of brine pumping (16% pumping versus 32% pumping). Panelists were also asked to evaluate the tenderness or juiciness of the same samples by mouth feeling. The score sheet for this trial is shown in Appendix A—2. Taste panel sample preparation (1) Samples used in the first session for visual inspection were ham slices (15 cm average dia- meter and 1.5 cm average thickness). Ham slices 45 were shown at room temperature. Samples used in the second session of the taste panel for physical evaluation were cut as 8 cm ham line Samples long, 3 cm width and 0.6 cm thick average pieces containing a meat junction or seam across the length of the meat piece. were offered at room temperature. Samples used in the second session of the taste (3) panel for mouth feeling or tenderness were cut into 3 cm long by 3 cm width and 0.5 cm thick ham pieces from zones of plain muscle in the finished product. They were offered to the panelists at room temperature. RESULTS AND DISCUSS ION Chemical Composition of the Raw Meat Fresh pork and frozen and thawed pork were compared for moisture, protein and fat by proximate analysis and for ran- cidity by the TBA method. Mean and standard error values for these variables are shown in Table 5. No significant differences (P50.01) between fresh and frozen meat were detected at this point. Protein, fat and moisture content Of these meats are quite similar to those reported by Kramlich gt gt. (1973) for thin separable raw-lean of the pork ham. It is important to note that the low TBA values found in the meats reflect a very sound condition of the raw pork in terms of lipid oxidation. Table 5 - Proximate composition and TBA values in raw pork meat used in the manufacture of boneless hams. \ Condition Moisture Fat Protein TBA N9- of the meat % r’/.. % mg malonaldehyde \_ ger 1000g sample Fresh 71.60t1.3l 7.42:1.70 19.94:.62 .135:.050 FrozEn 70.02t1.65 8.91:2.07 20.43:.74 .101t.044 1N=18 46 47 Changes in Chemical Composition Through Processing Protein content of the meat in the tissue and exudate after tumbling and in the final product is shown in Appendix B-l. A significant increase (P50.01) in protein content with tumbling time was observed in the exudate from fresh meat tumbled with or without vacuum (Figure 2a), but not in the exudate of frozen meat (Figure 2b). Significant interactions between the three factors in study (tumbling time, condition of the meat and pressure during tumbling) are shown in the analysis of the variance (ANOVA) table (Appendix C-l). Figure 3 shows the effect of tumbling time on the protein content in both the exudate and the ham for the meat pumped with brine to 16% and 32%. Protein levels are significantly lower in the exudate from meat injected 32% with brine than those in the exudate from meat injected 16%. This is due to the dilution effect of the higher level of water in the meat system injected 32% with brine. The results also show a significant increase (P50.01) in protein in the exudate with tumbling time (Figure 3a). The significant effects of tum- bling time and pumping level on protein in the exudate as well as the absence of interactions between these two factors are shown in the corresponding ANOVA table (Appendix C-2). Protein content in the hams pumped 32% brine were lower than in those pumped 16% (Figure 3b). This was, probably, because the final moisture content in hams pumped 32% was higher than in those pumped 16%. 48 .wceanssu wcauav Eanom> we on: msu on one mcofiuomuwuce mtg new wumvnxm use mo acmucoo cfimuoue one :o mafia mafiHnESu mo uowmww 05% I N muswfim Sod m .3 2333.. hegemoeuwcmum mum mumuuou udwuomuoasm Esaom>ucoz 111.:u:-:. acouwuuec :ue3 w>u=o msmm mam ca nuseom "ouoz E==Um> IIIIIIIIIL :HE .mseu wcwfinESH ow~ Omd ow 0mg emu ow _ . 4 . . _ Z x l Nd . 2 N :«muoum x 1 «m 1 mg mewd mam mafia mafiaesau an mouommmw mm Any mam; cosmecflm use aw mam Amv mumvnxo msu cw ANV ucmucoo :fimuoum I m muzwfim .Coé w .3 3.88:3 haucmowuwcwfim mum wuouuwa unauomuweam acmummwwm sufiB m>uso mEmm may :0 nucwom ”ouoz E==Um> .ummE :mmum .wcfiasaq Ne— mYlllllllAU Easum> .umma smouw .wcfiae:e Nun nYuinfunuAu see .wseu wceanszh one emu co ow~ o- ow . . . 1.- . - 4 z<=m a: 53:: 3 l l 14 cu Nd ed ed w~ om N :eououm 50 Fat content in the tissue and exudate after tumbling and in the final product is shown in Appendix B-2. Fat con- tent in the exudate was significantly affected (P50.01) by pressure during tumbling and by tumbling time (P=0.013) as shown in ANOVA table in Appendix CS. The interactions among the three factors are shown in Figure 4. It is important to note that fat level in the exudate increased soon after 60 minutes tumbling without vacuum. There seems to be a rapid release of fat from the muscles to the exudate after a short period of tumbling. This may be due to the fact that most of the fat in the ham muscles is superficial rather than intramuscular fat. Figure 5a shows the effects of tum- bling time and pumping level on fat content in the exudate. Although both effects, tumbling time and pumping level, significantly affected fat percent in the exudate (Appendix C-4), the direction of the interactions does not show a trend of variation of fat content in the exudate and in the finished product (Figures 5a and 5b). Moisture content in tissue and exudate after tumbling and in the final product is shown in Appendix B-3. Moisture content in the exudate was found to be significantly affected by tumbling time and pressure during tumbling (P50.01), as shown in the statistical analysis (Appendix c-5). However, the direction of the interactions (Figures 6a and 6b) indi- cates no clear effect of tumbling time, pressure during tum- bling and condition of the meat on moisture content in the exudate. A decrease in moisture level in the exudate should 51 :cd w .5 283:6 hegemoemficwem mum muwuuma uefiuomuoesm .Anv ume :mnouw 1cm Amv anus zmwuu Eouu wbnwsxo men :a ucoucoo any so mafia wcefibezu mo uoouuw one I a muswfim Essom>lcoz I'll-'0... acoumumwv sufi3 o>u=o mama may ca mucfiom "wuoz Essom> 1IIIIIIIIJ :HE .wseu wcfiunase owe o- co cw— ON~ Cc . . w q _ _ o w. I N o 1 x .x 4 e \\om I N x . 2e \ x s \ \ n O‘I-Illln'cllucllu0\ e A 1 w m9w~ mcuesse weeps can seem wcefime:u >2 emuowumw me Any mew: mocmficfiu czm Amv wummsxw use cm ucwucou use I m muawwm :cd w t 286:2. >_u:m0wuwcwmm ohm muobuoa uefihuwuoesm Essum> .umms :mwum .wcfie53m NNm mw I.|.IIlmv mcwhmwwww :uwB m>u=o mEcw msu co mucfiom ”muoz Essom> .ummE smouw .m:«eE=e N©~ mVlllllllmU :wE .mEHm mewfiaszw ow_ o- Co one o- Ce _ .III ..J.. _ . . _ _ o . a .9 m J N 7. 5 I. -.V I N Hu=o mama one :o mucfiom "wuoz :fiE .wswu wcwamESH ow— o- cc and Enzom>Icoz E:=om> o- oo .II“.“ I - q H _ eema mafiaaae maeun vcm mafia wcfifiaazu he nwuomuum mm Amy new; uw:me:ee can Amv wumvsxm msu cw unmucoo musumwoz I N ouswfim 293.: 28623 . Nuucmowwficwmm mum muouuofi uefiuomumezm E::om> have :mwuu .wcwdE3e Nwm mWiII-IEIAU "muoz Ezzom> .ume smoum .w:eda:e No. lellllllAU bcmumuuwm :uw3 w>uso mEmm ecu :o museom :AE .wsfiu wcefissse owl o~_ co ow. . o~_ co . q . . d . Q s a 4 oh X N K W -6 ..l. \\ m 2 ...Q\ .. .. 2m nV.\I\ a my ya u B./, I I // / IIIIIG Q: m. l ow m z<= Ase ma2 nouooumm mm odomse xhoe uo eumnsxo use . Cod w .3 accumumuv Neucmouuecwqm mum muuuuofi uamuomumesm uceumeevv :ue3 o>u=o mama 6:» :o mango; "muoz umws cmuoum "ll'"‘ u mos Lumen Illa c«S .wsqu mCN‘nEDH ow— ON“ co q _ q “A \r \m I x . x xx \ \ x xx... \ o \ x x \ \ O a usoueo ousmmmue ue>o nmwmum>m mm3~a> <99 Aev .oswb mcu~253u us: news ecu mo co_uLucoo :« Aoaesmw w ooc_\¢c>;ou#m:o_ms may conE:c 353 I_ew=w_m one :oeuancou name mean use Cob uouuoa newuomuoasm mcuumuemu and: when u Esnom>lcoz Ezaum> uses mzu we cowuwvcoo :mN0um umwa :mmum uo>o nwwwum>w mH wceesne new mafia wcwanssu as vmuomuum mm Amy new: mesmecfiu use Any oumwsxm no“ AmHeEmm w ocod\mv%:mnfimcoame hcmv Mensa: .umos :mmuu .wceease NNm me.I.IIIAO ucouomwfiu :uws u>uao ween men so museom "muoz Essom> .umws :mmum .wree55a No~ lellllllAmv :qE .mEHu wceanE:H S: 8. 8 2: oi 8 II a . H . . . 1 oo.c x G I 85 Q a I 25 I 26 z<= Aav _ me2 wouowuum mm yams Anv :mN0um mzm Amv :mwuu mo wumcsxm w:u cfi ucmucou uHmm I o— muswem Sod .v. .5 298:? maucmoeuwcwfim mum mumuuma uefiuomuoezm acmpmmuem sues m>u=o mEmm ecu :o muceom "euoz Easom>Icoz o. I I I I I 1 :«E .meeu mafianssh ow— Owe co ow~ o- Go a _ a H d - o.~ ..I x x m d n / III 0 [Ir x N x II . uamm I. III. ...V/ «W... // . ION / / x / l. m -.m.~ mhwa wceesza wan mafia wceaneau Nb nmuooeum me any man; nwLmHCwu new Amy mummsxm ecu .ucmuume .ucmucoo uamm I _— muswfim .A_o.cmwmv ucmumueen heucmowuecwwm mum mumuume uawuomumeam usmuwuuwc :ufi3 m>u=o wEmm mzu co mucfiom "muoz Eamon? .umma :mmuu .wcwease NNm @IIIIG Essom> .umos smouw .wcweazn Nod lell. mw cue .mEHu wcdfinE:h ow— o- co om— cum co . . a . . . o.~ E: as 3333. A3 c.m 63 presence of ascorbate in the brines, the color is converted to the rather dark red of nitrosylmyoglobin. During cooking, this pigment is converted into the stable nitrosylhemochrome, which is pink. Nitrite can also react with non-heme protein (Kubberod gt gt., 1974) binding to the sulfhydryl groups. According to Goutefongea gt gt (1977) nitrite reacts with adipose tissue when conditions are similar to those for meat curing. Nitrite can also be converted into nitrate (Lee gt gt., 1978), especially in the presence of the reductant sodium ascorbate, as is the case 'tn this study (Newmark gt gt. 1974). Finally, nitrite can be converted into NO and N2 gases during the mixing stage of cured meat manufacture (Sebranek gt gt. 1973). It was observed in this study that during tumbling ofthe meat,nitrite finds great chances of reaction inside the tumbler, not only with the meat components discussed above but with ascorbate present in the cure brine. It is important to note that during tumbling nitrite can also interact with some components of the connective tissue frac- tion of the meat, such as proline, which may lead to the formation of nitrosamines in the final product (Gray and Dugan, 1975). However, there are some factors during the processing steps followed in this study which tend to decrease the possibility of nitrosamine formation: (1) The source of meat used in this experiment was quite low in both fat and connective tissue (muscles were trimmed to less than 1 milli- meter fat thickness) which decreases the proline content in the meat. (2) The formation of various N-nitroso compounds 64 can be blocked by the presence of ascorbate in the system. According to Mirvish gt gt. (1972) the inhibitory effect of ascorbate on formation of nitrosamines is that nitrite is "used" so that it is unavailable for N-nitrosationbecause the rate of reaction of nitrite with the reductant is greater than it is with given amines in the system. (3) During cook- ing of the meat internal temperature was raised to 68°C which is below the critical temperature (BO-100°C) at which N-nitro- samine formation is accelerated (Gray and Dugan, 1975). Tumbling time and pressure during tumbling significant- ly affected (P<0.01) the content of nitrite in the exudate (Appendix C-8). The significance of the interactions is shown in Figure 12. Tumbling fresh meat under vacuum or without vacuum resulted in significantly higher (P<0.01) nitrite levels in the exudate after 180 min. tumbling. The opposite pattern was observed for frozen meat (Figures 12a and 12b). Assuming that the initial concentration of nitrite in the exudate is higher than in the tissue (some injected brine comes out of the tissue-right after stitch pumping), then the recapture of nitrite by frozen meat seemed to be more efficient than that by the fresh meat. Fresh meat pro- bably retains the injected brine better than frozen meat in that during extensive tumbling, the nitrite tends to leave the tissue rather than to diffuse into it. The similarity of the pattern of nitrite and salt diffusion from the exudate to the tissue (Figures 10b and 12b) indicates that the fro- zen meat tended to absorb much of the curing salts during tumbling. .wEHu mafianssu mo :ofiuocse m mm Any ummE cououu new Amy ume cmmuw Eouw wumnsxw w:u :« acmucoo mueuuwz I - wuswwm .Soé w .c 238:3 I oulllll... Naucmowuficwwm mum muwuuma uneuomumesm Eszom> :oz ucmuouufiv :ue3 w>u=o mEmm emu :« muceom "muoz Easom> 1 11 see .mEHu magenE3H 65 ow— o~— cc owe ON~ cc 4 q a _ 1 . o J on "N I. 3 1 o I I. ./l IIIIIIIIIII. AE— m \II\ n m \‘|IV4I|I|III~N o\\\ M h I I D o\nw m Ito! X a I!!! x o: .m w I o8 me2 venomuum we have Aev concuu can Amy smoum Eouw amaze ofinafiom us» :« acmucou :«ououm I m— muswqm .Coéwt ucmummwwv Naucmofiwacwqm mum muwuumd uefiuom Imogen acmumMan sues mafia m macaw mucfiom "muoz E:=om> n n Essum>I=oz o. I I. I I I no see .mENU weeHAEDH Im/Sm ‘sseqd annIos u: UIBJOld 2: o2 8 c2 o2 8 _ _ a _ _ _ . o .. o. .. ON I 9.. l S m \\m m a Ion \\ 14 colllllilcl'll m. \ \ m 0 II. lulu. an a m m «loo ... I E 1 cm 5333. Em: zeuome 3v Egan.— .Se: smug... A3 I ca a O 2: 72 60 x x 4 x 55 _ Protein in ‘50 ’ soluble phase a a ”’9 mg/ml ’0." 45 _ .II [I / a I O’ 40 1 1 L 60 120 180 H 16% pumping, fresh meat, vacuum 9.---..9 32% pumping, fresh meat, vacuum Note: Points on the same curve with different superscript letters are significantly different (P S 0.01) Figure 16 - Protein content (mg/ml) in the exudate soluble phase as effected by tumbling time and pumping level. 73 Composition of the Soluble Phase Figure 17 shows the relative mobility of several pro- teins as obtained in sodium dodecyl sulfate-polyacrylamide gel electrOphoresis (SDS-PAGE) as a function of molecular weight. The regression line shown in this figure was calcu- lated and built on the basis of four standard proteins: myosin heavy chain, MHC (Approximate MW = 200,000); bovine serum albumin, BSA (Approximate MW = 60,000); ovalbumin (Approximate MW = 45,000) and lysozyme (Approximate MW = 14,000). Next, the most common myofibrillar proteins were marked on the standard line according to their molecular weight and assigned a gel relative mobility value. According to this procedure eight major myofibrillar protein bands were identified in our samples as shown in Table 8. Figure 18 illustrates the scanning of a gel with the major protein bands showing different peaks. Relative concentration of these proteins in the soluble phase were calculated from the area under the band peaks in Figure 18. These values are shown in Figures 19 and 20 as a function of tumbling time. It is important to note that after 60 minutes of tum- bling the relative concentration of the high molecular weight proteins (myosin heavy chain; M-line and C-protein; a-actinin; and tropomyosin ) are higher in the soluble phase of the meat tumbled without vacuum than in the meat tumbled under vacuum. The case of the myosin band is particularly important since this major myofibrillar protein is primarily responsible for 74 5:32: 03.32 use 2225 83020:. .0 3:25. a «a 32?QO c. 5:. 520:. 22:35 2.9... eueaweoz m>eumamm 3 Be 2.: we... 8.: 2...: mad «2 8... _ . A _ _ a _ 3% 6 ~83 52... Eu: 5822 52.0 22. 53»: Ease Eu: 5ng L x 8a.. . ...:Q n» so. «3865:. Emanono; b-5530; Emmy—.333 Eamm ossom Emoafioeo; Scone... 55.8- 1 5205-0 529:. 25.: 8:2. 58.: ® 2:205 5:29.022 + 3.22.. Eaves-m ® 68.:— Sada Sada 865v Geode 898. ecodow 898m 898v JqBIaM letnoarow 75 (OOO‘QL - OOO‘BI.) ugeuo m6" ugsko} :5 (000%) ugsokwodou} >- (OOO‘LS) l UIUOOOJJ (ooo‘ge) Jowouow ugskoodo: 1} (000‘sz uuov - s} m (ooo‘oflugsoflwodou 1 (00009) mode: 11 -* (OOO‘OOIWIunae-n 3. m (000‘0vI) UIaImd - 0, (000990 umwd sun - w1 W ‘— (000sz amp bum; ugsko} .. Figum No. 18 Scanning oI a SOS-PAGE gel mm the major bands assigned I0 8 o! Iho mosI common myofibrillar meoIns. 76 Hm.0 00.0 No.0 on H0.0 em. 0e. am. ca. 00000 0H. mama was mo sunaunos 0>Humuom 000.0H ou 000.0H 000.0m 000.Nm ou 000.0m 000.00 000.05 000.00H 000.mmH ou 000.00H 000.00N Amaousmec ufimeB Hmasomfioz Amaze manage “swam sumo»: A000.¢mv HmEocoE Canoefiomoufi A000.0mV Hmaocoa CHmo%EOQOHH mam HIEHGOQOHH econ Cwuom I 0 meeEoo cho%Eoeoue cwaHuom I s :flmuoumuo mam Cwmuoue mCHHIE AumZv Geese h>mm£ ch0%Z mcwououe HmHHHunHmoxz .m0HumHmm I m maeme 40 d C'- Q) U H 0 G- .30 Q) 0) CU ..C Q. 3 .920 3 H 0 CD Q ...".3 U :10 n... fi 0 '14 U ('6 H U 5 at. C O U 2 “3 U m H OJ H g2 H G) U 0 E 1 0 77 (a) Myosin (b) M line protein Z-protein N 4 \ 6I. \ <3 - \ I / \ x 4__ \ //’,.43 a- '_ '. 2. " 0 I L 1 60 120 180 (d) Tropomyosin - 15. 10_ I- 5 .. I— “ l l j 0 l L L 60 120 180 60 120 180 Tumbling time, min G>----49 Frozen meat, vacuum 0 - .. .. .. Q Fresh meat , vacuum E¥----4§ Frozen meat, non-vacuum I- .. .. .. a , Fresh meat , non-vacuum Figure 19 - Relative concentration of Myosin (a); M-line protein and Z-protein (b); a-actinin (c); and Tropomyosin complex (d) in the exudate soluble phase as a function of tumbling time. .30 U I: Q U B Q) C. .25 Q) 0) a a 0 €20 .4 O U} (D .2 U =15 1.1 I: O ...; 3 h 5 212 811 0,10 .39 58 87 =6 CH 35 El. 3 2 1 o 78 (b) Troponin T and (a) G‘ACtin high MW Tropomyosin monomer " 30 I- Q‘ "' 20 I- I _I I 15 I 60 120 180 60 120 180 (c) Low MW TrOpomysin (d) Myosin light chains monomer - 121. _ 11_, I- 9 - b 8 - 7 - 6 - 5 _ 4 g 3 t 2 P 1 I 4 I 0, 60 120 180 Tumbling time, min e; 0 Frozen meat, vacuum G... .. .... .. ..g Fresh meat, vacuum Ep, 43 Frozen meat, non-vacuum Q. .. .. - .. .. a Fresh meat, non-vacuum Figure 20 - Relative concentration of G-actin (a); Troponin-T and high MW Tropomyosin monomer (b); Low MW Tropomyosin monomer (c); and Myosin light chains (d), in the exudate soluble phase as a function of tumbling time. 79 the binding properties in this type of sectioned and formed product (Hegarty, 1963). Statistical analysis showed a significant effect of the condition of the meat and pressure during tumbling on relative myosin concentration in the solu- ble phase (Appendix C-ll). After 60 minutes tumbling myosin relative concentration was significantly higher (P50.01) in both the fresh and frozen meats tumbled without vacuum than in the same meats tumbled under vacuum (Figure 19a). In the system.with fresh meat tumbled under vacuum only after 180 minutes tumbling did the level of myosin reach a value com- parable to that for the fresh meat tumbled 60 minutes with- out vacuum (Figure l9a). This situation with myosin is quite similar to the other three high molecular weight proteins (Figures 19b, 19c, and 19d), which probably indicates that the use of vacuum with short periods of tumbling did not contribute to the extraction of myosin and the other high molecular weight myofibrillar proteins. Low molecular weight myofibrillar proteins (G-actin; troponin-T and trOpomyosin nonomer [36,000]; tropomyosin monomer [34,000]; and myosin light chains) showed an Opposite pattern of extraction. After 60 minutes tumbling under vacuum fresh and frozen meat tended to show higher relative concentrations of these low molecular weight myofibrillar proteins than in the system tumbled without vacuum.(Figures 20a, 20b, 20c and 20d). According to these results, after a short period of tumbling (60 minutes) high molecular weight myofibrillar pro- teins are higher in the exudate of the meat tumbled without 80 vacuum than in that of the meat tumbled under vacuum (Figure 19). At the same time low molecular weight myofibrillar pro- teins are higher in the exudate of the meat tumbled under vacuum than in that of the meat tumbled under no vacuum (Fig- ure 20). This pattern of protein extraction was not apparent after 120 minutes and 180 minutes tumbling. These observa- tions suggest that the use of vacuum during tumbling does not necessarily contribute to the extraction of myofibrillar proteins. Furthermore, the use of vacuum at the end of a tumbling Operation rather than throughout the whole process of tumbling may be more advantageous for myosin extraction. Parameters Related to the Final Product Figure 21 shows processing steps affecting final pro- cessing yield of hams. Pork meat injected 16% and 32% with cure brine showed average processing yields of 100.5% and 112.3%, respectively. Figure 22 shows the effect of tumbling time on actual ham yields. Statistical analysis for yields showed a significant effect (P<0.01) of condition of the meat (Appendix C-l2). Although there is a trend for better per- formance of fresh meat over frozen meat in terms of yield (Figure 22), at only 120 minutes level of tumbling time this effect was statistically significant (P50.01). This differ- ence in yield may occur because the water-holding capacity of fresh meat appears to be better than that of frozen meat. According to Kramlich gt gt. (1973) federal meat inspection regulations recognize three ham categories depending on the FRESH [FROZEN MEAT, MEAT If _- v— ~ '— L f STUFFING L '— ‘— COOKING FINISHED HAM A STORAGE Note: Final yields calculated based on actual processing 7' 81 THAWING LOSSES BONES SKIN AND FAT FINES CURING BRINES TUMBLING LOSSES COOKING LOSSES FINAL YIELDS 19 13. 16 32 10 100 112 .29% .86% .66% 61% .00% or .00% .93% .34% or .11% .50% or .30% losses. Figure 21 - Processing factors affecting final yields in the process of'manufacturing boneless hams. 105 104 Yield 103 % 102 101 100 82 ' CL a .- a P I I L 60 120 180 Tumbling time, min G——€ Frozen meat, vacuum 9.... -0 Fresh meat, vacuum Ep_____4a Frozen meat, non-vacuum 3..---.9 Fresh meat, non-vacuum Note: Pairs of points belonging to the same meat condition with different letters within the same tumbling time are signifi- cantly different (P S 0.01) . Figure 22 - Percent conversion of pork meat into boneless ham as a function of tumbling time. Yields calculated from actual processing losses. 83 amount of added substance remaining in hams after processing. Added substance refers to water and salt present in the cured product in excess of the normal amount occurring in the un- cured product. This control is exercised through calculation based on chemical analysis. The following formula is used for yields: estimated yield = % moisture + % salt - k x % protein-+3“M);(Kramlich gt gt. 1973). The protein multiplier or k factor is an average figure representing the approxi- mate ratio of moisture to protein. For smoked hams this factor is 3.79. Table 9 shows the yield of the hams obtained in this study as calculated according to the procedure fol- lowed by federal inspection. Table 9a - Estimated yields of hams as calculated by Federal inspection procedures using 3.79 as k factor. Estimated_yiélds according to Fed. ingpection Tumbling 16% pumping 32% pumping time (min.) Vacuum Non Vacuum Vacuum ~ Fresh Frozen Fresh Frozen Fresh 60 107.82 99.61 93.41 98.21 _ 116.13 120 101.82 103.52 97.81 101.52 113.83 180 97.71 100.42 102.92 95.41 120.73 1Fully cooked hams with no label restrictions 2Fully cooked "water-added” hams (According to Federal inspection) 3Hams not eligible for sale. 84 No labeling restrictions are imposed for those hams with estimated yields equal or lower than 100%. Those hams with added substance up to 10% are labeled "water added" hams and those with added substance over 10% are ineligible for sale. according to Federal labeling restrictions (USDA, 1979b). Our results show that 50% of the hams pumped 16% brine should be classified as regular hams (no labeling restrictions) and 50% should be classified as water added hams. No appreciable effect of processing treatments on estimated yields, as cal- culated according to Federal inSpection, was observed in this study (Table 9). Hams pumped 32% with brine showed esti- mated yields between 113.8% and 120.7% and they fall in the category of not legal hams. Cooking losses during thermal processing averaged 10.7% with .a range from 10.0% to 12.4%. NO significant differences due to main effects and/or interactions were observed (Appen— dix C-13). As expected, cooking losses for the meat injected 32% with brine were significantly higher (P<0.01) than those for the meat injected 16% (Figure 23). According to Federal inspection regulations the hams processed in this study are fully cooked or ready-to-eat hams because they were cooked to an internal temperature over 64 5°C (68°C actually). Figures 24a and 24b show nitric oxide pigments content and percent conversion (the fraction of the total pigments converted into nitric oxide pigments), respectively, as functions of tumbling time. From this figures it can be noted that processing treatment did not drastically affect 85 13 ”’0‘ 12 .. 9'” \\ 0 Cooking 11 ' losses % 10 .. 9 I J_ I 60 120 180 Tumbling time, min 3 0 16% pumping, fresh meat, vacuum 0"-"G 32% pumping, fresh meat, vacuum Figure 23 - Cooking losses (%) of the meat as a function of tumbling time and pumping level. 86 the content of nitric oxide pigments content and pigment conversion in the meat. This is probably due to the multiple needle stitch pumping system used in this study to inject the muscles, which gives a relatively high initial concentration of the cure inside the meat. Tumbling time produced a slight increase in nitric oxide pigments and pigment conversion in the meat tumbled without vacuum. This observation agrees with results reported by Krause gtht. (1978b). These authors found a significant improvement in internal color of hams tumbled for 18 hours over hams tumbled for 3 hours. The effect of tumbling on the rate and uniformity of diffu- sion of curing ingredients probably accounts for the color development. The use of vacuum during tumbling, however, did not improve nitric oxide pigment levels and/or pigment conversion. Furthermore, the meat tumbled without vacuum showed higher levels of both nitric oxide pigments and pigment conversion than that tumbled under vacuum after 180 minutes (Figures 24a and 24b). Although no statistical analysis was possible for these parameters the results tend to indicate that vacuum during tumbling does not have a beneficial effect on pigment conversion and nitric oxide pigments in the meat. NO noticeable effect of brine pumping level on nitric oxide pigments or pigment conversion was observed (Figure 25). Figure 26 shows L, a and b color parameters for ham slices as measured by the Hunter color meter. Although a significant effect (P50.01) of tumbling time on L, a and b color parameters was shown in the statistical study Curing pigments, PPm Pigment conversion, percent 87 100 _ (a) Nitric oxide pigments 75 .. 50 .. 25 L I I 100 .. (b) Pigment conversion 90 _ 80 .. 70 - 60 _ 50 L I I 60 120 180 Tumbling time, min cg. Frozen, vacuum Q- - .. __ :0 Fresh, vacuum B——-E Frozen , non-vacuum a... - - .3 Fresh, non-vacuum Figure 24 - Nitric oxide pigment content (a) and percent pig- ment conversion (b) in hams as a function of tumbling time. 88 (a) Nitric oxide pigments 100 ‘ 75 ’ Nitric oxide pigumts PF“1 50 b 25 A L L (b) Pigment conversion 100 " 90 " Pigment conversion go .- percent I '70 ' 60 " 50 I I I 60 120 180 g I; 16% pumping, fresh meat, vacuum. @.....a 32% pumping, fresh meat, vacuum. Figure 25 - Nitric oxide pigments (a) and percent pigment conversion (b) as a function of tumbling time and pumping level. 89 53 (1) Color parameter L Color parameter L Color parameter a 9.0 , 80 L L I ° '(III) Color parameter b 7.5 _ Color 7.0 _ parameter b 6.5 , 6.0 .L I it I 60 120 180 o e e----o e————-a sun-Ia Tumbling time, min Frozen meat, vacuum Fresh meat, vacuum Frozen meat, non-vacuum Fresh meat, non-vacuum Figure 26 - Color parameter L (I); a (II) and b (III) in the slices as a function of tumbling time. 90 (Appendices C-14; C-15; and C-l6), no trends were evident in the study of the interactions shown in Figures 26(I); 26(II), and 26(III). No relationship between this color determina- tion and the level of nitric oxide pigments discussed above was found either. The main limitation of the assessment of color in the ham by this technique is, of course, the rela- tively large variability in color intensity from one type of muscle to another within the same ham piece. Results from the microscopy study are shown in the next series of microphotographs. Figures 27 to 29 show the effect of tumbling on the muscular fiber arrangement in a transversal cut through the tissue. This effect goes from a state in which the fibers are quite ordered, showing cir- cular sections characteristic of intact fresh muscle, and ‘with very little exudate material around them after 60 min. tumbling (Figure 27); to a state in which considerable amount of soluble protein can be seen around the fibers, with increased spacing between fibers and some degree of cell disrupture, after 120 min. tumbling (Figure 28); and to a state in which the fibers have lost their circular shape to the transversal cut, with large spaces filled by protein exudate, air bubbles and/or fat globules (Figure 29). These pictures are quite similar to those reported by Rejt gt gt. (1978), for massaged porcine bicep femoris muscles. However the presence of exudate material among the fibers is much more evident in the pictures shown in this study than those by Rejt gt gt (1978). A similar pattern of fiber 91 FIGURE 27. Microphotograph of the cross section of bicep femoris fibers in ham from fresh meat, vacuum, 60 minutes tumb- ling (X80). f g',..‘ - ::' . f,§;4l( ,’ ‘ :‘ ”um-’1 «V .\ \L, , ’ - ' l- I ; x m . F FIGURE 28. Microphotograph of the cross section of bicep femoris fibers in ham from fresh meat, vacuum, 120 minutes tumb- ling (X80). %,7, ,l, 92 FIGURE 29. Microphotograph of the cross section of bicep femoris fibers in ham from fresh meat, vacuum, 180 minutes tumb- ling (X80). I H.731. ’l"ll Ifl’rll. M f‘ ) I‘M! ' «K 1"“ FIGURE 30. Microphotograph of the longitudinal cut of bicep femoris fibers in ham from fresh meat, non vacuum, 60 minu- tes tumbling (X80). 93 damage with tumbling time could be observed when the muscle was cut along the direction of the fibers (Figures 30, 31 and 32). It was also evident that tissue fibers damage was greater on the periphery of the muscle chunks than in the interior part of the meat. These observations indicate that the pattern of tissue disruption with tumbling observed in this study can be found in a single chunk muscle which has been tumbled for a relatively short period of time by sampling at different locations from the interior to the periphery of the meat piece. The pattern can also be found in a muscle which is sampled at about the same location but at different times during the tumbling Operation. Figure 33 shows a typi- cal seam area in which two chunks of meat bind together. The cross sections of the fibers from one of the meat pieces can be seen in the left side of the picture, separated from the exudate material by some connective tissue layer. Some fat droplets or air bubbles can be seen in the exudate in the lower right corner of the picture. Tumbling seems to have considerably damaged the fibers near the edge of the tissue as the large spacing among them and their irregular shape at the transversal cut demonstrate. Yet, this damage resulted from a processing treatment which used an intermediate tum- bling length (120 min.). The effect of vacuum during tumbling can be seen in Figures 34 and 35. In the former picture the exudate soluble material appears to be quite homogeneous on the edge of the tissue with a few small fat drOplets. In the latter picture 94 FIGURE 31. Microphotograph of the longitudinal cut of bicep femoris fibers in ham from fresh meat, non vacuum, 120 min— utes tumbling (X64). \ _ v"; “NJ-(‘1); .974. fig' ‘ . {“JS 2“ .5\ (\C‘f3 FIGURE 32. Microphotograph of the longitudinal cut of bicep femoris fibers in ham from fresh meat, non vacuum, 180 min- utes tumbling (X80). ¥¥ 95 51% $1 .. 1 v I3; . ’ ufllfifé‘ ' _4 x". ., ~44 "'2' P" I I FIGURE 33. Microphotograph of a seam or binding junction area in ham from fresh meat, vacuum and 120 minutes tumbling (X64). 96 ~' 'I,;.;:.\.:. g‘i IN ,, 4 , ‘; ~ ,‘I';:‘ . -..”, .- .QIqwmmgfig \ . “‘1‘: , 4 '4 ..,\. . x . 9 . . ‘ ‘K‘vIlI’IgI- ‘23- I ' '-1:4;;‘5Ix\,\r( 32' ”I ‘ . QIII’IIII .I 3.44.4‘4444C4 7' “"Z .297 'i‘inI )5. N ' 4i , I’,4 4‘ VWII % If I #49. a. ‘ .“ 4.\.-4 4.4‘ ' ' .‘y II . ‘gfi-II‘JIUIIZ‘I' ' g ' ‘ ‘ r 27.4" '.'\.. 7.14mi '*=II\'4II.I;£'~ 44444.43: 7.4,“ . iufi w- :- FIGURE 34. Microphotograph of a seam or binding junction area in ham from fresh meat, vacuum, 120 minutes tumbling (X64). ;~.'. . . 2“- 'I: Meafl FIGURE 35. Microphotograph of a seam or binding junction area in ham from fresh meat, non vacuum, 120 minutes tumbling (X64). 97 the exudated soluble material on the left side of the picture shows some air bubbles. Picture 8 belongs to a ham tumbled under vacuum for 120 min. and picture 9 to a ham tumbled without vacuum for 120 min. The use of vacuum during tum- bling seems to eliminate much of the air entrapped in the tissue and exudate. This effect will produce hams with a more uniform surface texture and better binding. A foamy exudate interfers with binding (Anonymous, 1981). The effect of the condition of the meat used in this study can be observed in Figures 36 and 37. The former one shows a transversal cut across the fibers of a ham piece from fro- zen and thawed muscle, and the latter one of a fresh muscle. Both preparations belong to treatments tumbled under vacuum for 120 minutes. The damage produced by the tumbling Oper- ation is much more evident in the tissue from frozen and thawed meat than in that from fresh meat. Fibers from frozen and.thawed muscle seemed to be more fragile than those from fresh muscle, the reason probably being the physical stress <3n the fibers during freezing and thawing. No clear differ- einces due to pumping level were observed in the tissue under 'the light microsc0pe. Binding strength in ham pieces is shown in Table 10 ill the form of tensile strength parameters (g/cmz) obtained Withthe Instron universal testing machine. These results indicated that meat tumbled without vacuum binds signifi- cantlybetter (PSO.Ol) than the meat tumbled under vacuum, forprocessing conditions which included fresh meat tumbled 98 FIGURE 36. Microphotograph of a cross section of bicep femoris fibers in ham from frozen meat, vacuum, 120 minutes tumb- ling (X80). 'oI " v .. mm: If 9 4 f ’ 4‘ I. I /« (7"..- .-" -I‘&‘ // ' HI” “awn?" fix“ FIGURE 37. Microphotograph of a cross section of bicep femoris fibers in ham from fresh meat, vacuum, 120 minutes tumb- ling (X80). 99 Table 10 - Tensile strength values (g/cm?) measured tO separate pieces Of hams by the seam or bind- ing area. TREATMENT IDENTIFICATION PROCESSING FACTOR TENSILE STRENGTH NUMBER TESTED g/cm2 2 Vacuum 513.43: 11.3 VS b 4 Non Vacuum 922.2 1 84.9 11 Frozen meat 86.93: 20.3 VS . 12 Fresh meat 155.0b: 12.3 3 Long tumbling 193.33: 27.5 VS 11 Short tumbling 86.9b: 20.3 10 16% pumping 331.7a: 42.2 15 32% pumping 256.33: 31.2 “......— 1Means (within the same processing factor tested) with different superscript letters are Significantly different '(PS0.0l). N=3 100 for 180 minutes. Although a better binding was expected with vacuum.tumbling, the values Of binding for both treat- ments under comparison are far above those needed for good slicing prOperties in the ham. According to Theno gt El- (1978c), lOO g/cm2 binding strength is necessary for ham rolls to exhibit acceptable slicing characteristics. Anon— ymous (1981) reported that with vacuum tumbling a bind Of 200 g/cm2 between muscle sections was considered good for sliced product and was achieved in about four hours. Results in Table lO.also sflunv, as expected, that fresh meat bound significantly better than frozen meat, longer tumbling bound significantly better than short tumbling and 16% pumping better than 32% pumping. It is important to note, ‘however, that this physical determination involves a series of factors which may lead to erroneous measurements or inter- pretation of the results. One such factor is the difficulty to take the sample from the finished hams. It is hard tO localize seams or junction areas in highly trimmed pieces Of ImUScles like the ones used in this study. Sometimes the direction Of the fibers on one side Of the seam runs parallel ‘33 the seam line which makes it very easy tO tear apart the meat rather than separate apart the chunks Of meat, when the I118tron is used. Another factor which may lead to erroneous restflxs is the fact that there are natural seams between 1lesales which can be mistaken in the final product for pro- teiJl seams between two individual chunks Of meat. In this CaSe the values Of tensile strength will be misleadingly lOl lower or higher depending on the density Of the connective tissue between muscles. Finally, there are several muscle components in a ham piece with different intrinsic strength or tenderness. Very tender muscles like the semi membranous can be easily ruptured during the tensile strength determina- tion Of the bonding Of meat pieces. Table 11 shows the overall appearance Of ham slices by visual inspection. According tO these results the color distribution in ham slices from meat tumbled without vacuum ‘was significantly more uniform than that in those from meat tumbled under vacuum. This unexpected result might be due to the great variability in color within some ham slices. Since various types Of muscles may be present in a same ham slice the rate Of cure penetration and/or the rate Of color development is probably different from one muscle type to another. Some characteristics Of muscle type such as firm- ness, fat content and connective tissue content may effect the rate Of cure penetration. Moreover, some muscles in Pork.ham present different proportions Of white and red fibers with the consequent difference in pigment level avail- able to react with the cure. Red muscle fibers have a higher myoglobin content, more lipid and higher activity Of OXidative enzymes than do white fibers. Lee gE_al, (I976) fol-Ind lower residual nitrite in cured meat made from white musCle than that in meat from red muscle. These authors reported that the cause Of this phenomena was the low pH Of white muscle. NO significant differences were Observed for 102 Table 11 - Evaluation Of the overall appearance Of ham slices by a visual inspection panel. Characteristic evaluated, expressed as a preference ratio Treatment Processing COLOR SURFACE TEXTURE ‘EXTRANEOUS MATERIAL ID factor Uniform/ NO defects/ Absence/ number tested Non Uniform defects Presence 1 Vacuum 0/12* 2/10 9/3 vs 3 Non vacuum 6/6 2/10 5/7 5 Frozen 5/7 8/4 3/9 vs 6 Fresh 3/9 2/10 4/8 2 Long tumb. 6/6 7/5 0/12* vs 10 Short tumb. 4/8 4/8 7/5 6 167. pump. 3/9 * 2/10 4/8 vs 14 327. pump. 11/1 3/9 5/7 1 jRatios with an asterisk mark within the same processing factor tested are significantly different (P<0.01). 103 surface texture and presence Of non-lean material in the ham slices from meat tumbled with or without vacuum (Table 11). Condition Of the meat did not significantly affect color, texture or presence Of non-lean material in the ham slices. Moreover, according to the panelists, tumbling time did not affect color and surface texture Of ham slices. Short tum- bling time (60 minutes) however, produced a significantly higher (P<0.05) presence Of non-lean material, such as fat and connective tissue. The meat pumped to 32% showed a color uniformity that was significantly higher (P<0.05) than that pumped tO 16%. NO significant differences were detected for surface texture and presence Of non-lean material in the meat tumbled 16% and 32%. It should be noticed, however, that the panel failed to detect surface texture differences between the meat tumbled with and without vacuum. Probably, the most evident organoleptic characteristic Of the hams Obtained in this study was the different surface texture Of the hams tumbled under vacuum.and no vacuum. The use Of 'Vacumm.produced hams with a very uniform.and homogeneous $1lrface texture; seams or binding joints between chunks Of meat were very hard to localize in these products and the WhOle piece Of ham had the appearance Of an intact muscle PifiDduct. When the meat was tumbled without vacuum hams Stkmwed a fine porosity at the seam areas and in some muscle °I7‘tissue area. In other words, the effect Of air bubbles eljlnination by vacuum was usually apparent on the final pro- duct upon slicing the hams. 104 Table 12 shows the binding strength Of ham pieces as evaluated by the taste panel. NO significant differences were detected at this pOint as consequence Of tumbling time and pressure during tumbling. Frozen meat bound significantly stronger than fresh meat, a situation Opposite tO that found with the use Of the Instron, above. Sixteen percent pumping produced hams which bound significantly better than those pumped 32% brine, which agree with the results Obtained by the Objective evaluation Of binding using the Instron (Table 10). It should be noted that values presented in Table 12 represent 24 Observations per factor tested (a twelve persons panel judging the same treatment samples in two sessions). Although the group Of people participating in this ham quality evaluation was supposed to be a semi-trained panel high Variability in the scoring was Observed. Scores from the first session did not correlate well with those from the Second session. This fact indicates that taste panel eval- uation Of binding strength in ham pieces is a quite subjec- tlive estimation Of the force necessary to separate pieces (If meat at the binding junction or seam line. Among the 3facrnrs involved in this problem are the difficulty in SEEIecting and preparing samples, the difference, in binding all different points in the same seam or junction line, the IMay the panelist pulls the pieces Of meat apart, the judg- umfilt by the panelist Of the binding strength, etc. Table 13 indicates the values Of tenderness Of ham 105 Table 12 - Evaluation Of binding strength between pieces Of meat in a ham slice by semi-trained panelists. Panelist preference for binding strength Coding Of Factor A stronger B stronger Cannot tell treatments tested than B than A the difference A = treatment 1 Vacuum vs 103' 11a 3 B = treatment 3 Non Vacuum A = treatment 5 Frozen vs 1491 4b 6 B = treatment 6 Fresh A = treatment 2 Long tumbling vs 78' 11a 6 B = treatment 10 Short tumbling A = treatment 6 16% pumping vs 16a 5b 3 B = treatment 14 32% pumping 1”Values in the same row with different superscript letters iire significantly different (P50.05). 106 Table 13 - Evaluation Of meat tenderness for ham slices by tast panel.1 Panelist preference for tenderness Coding of Factor A B treatments tested more tender more tender Cannot tell . than B than A the difference A = treatment 2 Vacuum vs 15a 6b 3 B = treatment 4 Non Vacuum A = treatment 7 Frozen vs 16a 1b 7 B = treatment 8 Fresh A = treatment 1 Long Tumbling vs 11a 9a 4 B = treatment 9 Short Tumbling A = treatment 6 16% pumping vs 10a 9a 5 B = treatment14 32% pumping 1Values in the same line with different superscript letters are Significantly different (PS0.05) 107 slices as judged by a taste panel. According tO these results the meat tumbled under vacuum produced hams signifi- cantly more tender (PS0.05) than that tumbled without vacuum. These results agree with those found by Rejt g5 §l(l978). These authors reported that vacuum massaged meat showed higher tenderness and lesser cooking loss than non-massaged meat. In this study, frozen meat produced hams significantly more tender than fresh meat. Overall rating Of frozen meat by taste panel was equal or better than fresh meat. These Observations are substantiated by the results Of estimated yields (Table 9) and cooking losses (Page 39) which Show no significant effect Of condition Of the meat. According tO the taste panel tumbling time and pumping level did not sig- nificantly effect the tenderness Of ham slices. Although tenderness Of the meat is a quality attribute relatively easy to evaluate by mouth feeling, the intrinsic difference in tenderness from one muscle type to another within a same piece Of ham may produce some variation in the response by panelists. SUMMARY AND CONCLUSIONS The primary Objective Of this study was to determine the effects Of tumbling time, pressure during tumbling, condition Of the meat and brine pumping level on the nature Of the exudate after tumbling and the quality parameters Of sectioned and formed meats. Fully cooked boneless hams were manufactured as a model system for the experiment. Four sources Of variation were considered in this study: (1) tumbling time (60, 120 and 180 minutes); (2) pressure during tumbling (vacuum and non vacuum); (3) condition Of the meat (fresh meat and frozen and thawed meat); and (4) brine injection level (16% and 32% pumping). The meat system was analyzed at four differ- ent stages during the process Of ham manufacture: (a) the raw meat sample was collected from the trimmed pork muscles just before brine injection and it was analyzed for moisture, fat, protein and lipid oxidation; (b) the tissue sample was collected from the core Of the meat chunks, after tumbling, and analyzed for moisture, fat, protein, lipid oxidation, salt and nitrite; (c) the exudate sample, also collected after tumbling, was anlayzed for moisture, fat, protein, lipid oxidation, salt, nitrite, soluble phase volume, pro- tein in the soluble phase and protein composition Of the 108 109 soluble phase; (d) the ham sample was collected from the finished product and analyzed for moisture, fat, protein, lipid oxidation, salt, nitrite, color distribution, tensile strength, taste panel and microscopic structure Of the meat. Results indicated that protein and fat are extracted with tumbling at different rates, with protein being extracted gradually with tumbling time and fat being extracted mainly at the beginning Of tumbling. Protein from fresh meat is extracted with more difficulty than from frozen meat. The use Of vacuum does not affect protein and fat extraction. Although the meat system remained relatively free Of lipid oxidation throughout the process, the results indicated that tumbling time and absence Of vacuum during tumbling increase lipid oxidation, with the effect being more evident with frozen meat. After pumping Of the brine into the meat nitrite and salt are retained better by fresh meat than frozen meat. Frozen meat tends tO absorb much Of the cure during tumbling. NO effect Of vacuum on cure distribution was Observed. The soluble phase extracted from the exudate varied in viscosity with protein content. Small volumes Of soluble phase were collected from exudates with high protein content, the amount Of total protein being similar for all the treat- ments under study. Since high myosin contents in the exudate after tumbling are associated with gOOd binding characteristics in the final product the effect Of processing treatments is particularly important. The results Of this study suggest 110 that the use Of vacuum during tumbling does not contribute to the myosin extraction. Furthermore, with short tumbling (60 minutes) myosin is extracted more easily by tumbling without vacuum. The results also show that the effect Of tumbling time on myosin extraction is not conclusive and further research in this area is suggested. Results related tO the final product show that hams pumped 16% with brine are not affected, in terms of yield, by tumbling time and the use Of vacuum during tumbling. Fresh meat shows slightly better yields than frozen meat. According to Federal regulations about 50% Of the hams pumped 16% in this experiment should be labeled "water added hams". Meat pumped 32% with brine produces hams "not eligible for sale" since they contain more than 10% added substance. Nitric oxide pigment content and pigment conversion in the hams tumbled without vacuum were higher than in those tumbled under vacuum. However, this difference was not evident when the product was assessed by Hunter color para- meters. Results from the micrOSCOpic study show a pattern Of increased cell disrupture in the muscle tissue with tumbling time. However, the same pattern Of fiber damage can also be Observed going from the interior parts to the peripheral parts Of the tissue in a single muscle chunk tumbled for a short time. Fibers from frozen meat showed more damage after tumbling than those from fresh meat. The use Of vacuum during tumbling eliminated presence Of small air bubbles 111 in the exudate. Binding strength determinations by the Instron instru- ment show that treatments without vacuum produced hams which bind better than those from treatments with vacuum. However, binding values for both treatments were highly acceptable for slice-ability characteristics in the ham. NO differences in binding due tO vacuum effect were detected by the taste panel. Tumbling time does not affect binding strength, according tO both the Objective and subjective evaluations used in this experiment. Frozen meat binds better than fresh meat, according to the taste panel, but not according tO the Objective evaluation with the Instron. Although the results show some discrepancies between the subjective and Objective evaluation Of binding strength as affected by tumbling time, pressure during tumbling and condition Of the meat the effect Of pumping level is the most evident one. Hams with 16% pumped brine bind significantly better than those pumped 32%. - Color distribution in hams from meat tumbled without vacuum is more uniform than that from meat tumbled under vacuum. However, tenderness Of the meat was better in the hams from meat tumbled with vacuum than that from meat tum- bled without vacuum. From the results summarized above it can be concluded that: (l) Tumbling allowed for more economical usage Of added cure substances producing hams Of generally 112 gOOd acceptance by consumers and panelists. (2) Either fresh or frozen and thawed pork showed tO be suitable for this type Of processing. (3) Although protein increased with tumbling time in the exudate, hams with highly acceptable character- istics could be produced with tumbling times as short as 60 minutes (four hours in the tumbler). (4) The use Of vacuum during tumbling improved the overall appearance Of the final product primarily by elimination Of air bubbles from the exudate. Vacuum did not contribute tO extraction Of myosin and nitric oxide pigment development in the pro- duct. Vacuum should be used in the later stages Of the tumbling cycle tO improve surface texture Of the meat. (5) When tumbling procedures are used, percent pumping showed to be a critical factor on quality charac- teristics Of the final product. Sixteen percent pumping produced hams Of good acceptance character- istics. However, 32% pumping produced hams ineli- gible for sale,due tO the excess moisture retained with poorer slicing and binding properties, although the finished product exhibited acceptable color, flavor and texture. APPENDIX A Taste Panel Score Sheets 113 APPENDIX A APPENDIX A-l: Score sheet for the evaluation Of ham Slices by visual inspection. INSTRUCTIONS In this part Of the panel, you are requested to evaluate the overall appearance Of 15 ham slices just by visual inspection. You should concentrated on three types of defects: color uniformity (not color intensity or color differences from one piece of muscle to another); surface texture (presence Of holes, air pockets or brine pockets); and presence of non-muscle material (connective tissue lines or fat accumu- lation). You are asked tO stop by each sample displayed on the table and evalu— ate the three characteristics before going On the the next sample. Mark your decision on the logo sheet with a / COLOR UNIFORMITY SURFACE TEXTURE NON-MUSCLE MATERIAL Presence Of Sample Good Non NO Presence Of NO white NO. distribution uniform defects defects appreciable material 114 APPENDIX A-Z: Score sheet for the evaluation Of binding strength and ham tenderness. HAM TASTE PANEL Date 1. Evaluation Of the binding strength. In this part Of the test compare the binding strength Of the two pieces Of meat inside each plate separately. Pull apart the meat piece by using either your fingers or by using the two forks. Mark with a / the corresponding square. Plate 1 stronger than [ I stronger than [ I is not different than I I Plate 2 stronger than [ I stronger than [ I is not different than I I Plate 3 stronger than [ I stronger than I l is not different than [ I Plate 4 stronger than [ I stronger than [ I is not different than I I 2. Evaluation Of ham tenderness. In this part Of the test compare the tenderness Of ham pieces separately in each plate. Chew both ham samples in a plate before making your decision. Mark with a / the corresponding square. llS APPENDIX A-Z: (Continued) Plate 1 more tender than [ l more tender than [ ] not different than [ ] Plate 2 more tender than [ 1 more tender than [ ] not different than [ ] Plate 3 more tender than [ ] more tender than [ 1 not different than [ 1 Plate 4 more tender than [ ] more tender than [ 1 not different than [ ] APPENDIX B Chemical Analysis 116 Aamzv mm.o H H~.mfi Am.aa NA.HH Hm.¢H oA.¢H mo.w~ omH Aoumbsxmv N~.o H mm.ma ¢©.mH mm.oa ON.mH om.ma H¢.wH ONH Amsmmwev om.o H mm.na mH.mH cm.m m~.HH Nu.ma m¢.wH om uouum nasalwwM mesa Noa mEDm Nun mend flea mafia Nmm mafia Rea NCHEM wumwcmum Em: muonsxm mammwh oEwu wmaoom wcwaneah N .szeomm N~.ma oH.mH mm.mH mm.ma owH Em: NN.o H mm.wa mo.o~ mm.aa «o.ma oNH Em: om.om wm.o~ HH.o~ mH.wH oo Em: mo.ma mo.¢a ¢¢.ma Hm.qH owH mumwsxm HH.o H om.ma ¢©.NH mm.NH oN.MH oNH wuwbsxm wN.NH mm.HH mm.ma m~.HH ow muonsxm mm.aH mo.oa H©.©H mo.wa owH mommwe om.o w mw.mH m©.wa oo.ma H¢.wa ONH osmmfie No.mH mm.ma mm.ma m¢.wa ow oomme Hound cowoum meum cwmoub amoub ACMEV mama rumwcmum Essom>ncoz E::om> wEHu mamamm wmfioom meanness x .szaomm 9W2v.mucoaumwuu wcfimmmUOHm >3 nmuoomwm mm Ewummm ume map CH ucmucoo Camuoum .Hum xHozmmm< ll7 Az<=v oo.o H wm.m NH.N os.~ oH.~ Hm.o cm.o omH Ameucoz Ensom> mafia oHQEmm emHoom wcHHnEDH N .Hucoz Essom> mEHu deEmm Hence: wsHHnssH N .H4H Amuzv .mquEummHu wcflmmmooum kn noncommm mm Emummm ummE m5» CH Damucoo musumwoz .mn: xHszmm< 119 Aamzv Ho. H oHH. NHH. woo. «mo. NoH. mmm. om: Amumcsxmv Nc. H cmc. ccc. mmc. ch. ccc. Ncc. cNH AmnmmHHv mc. H HcH. ccc. cca. Ncc. mcH. mma. cc Houum QED: Nmm aEsmNcH NEsm.NRm QEDQNMH mEnm Nwm mE:m NbH. NCHEV cumccmum L. Em: mumcdxm mammwe mEHu cmaoom wGHHQEDB .AmHmEmm w cccH\mc%£mUHmconE wEv HmnEnz «:9 mmH. wad. mwc. NHH. cca Em: ac. H mma. mmc. mma. ccc. cNH Em: ccH. mmc. mac. cmc. cc Em: ccH. Nca. «Ha. cmc. cwH mumcsx: Hc. H ccc. mmc. mac. ch. cNH mumcsxm mca. mad. I Ncc. cc mumcsxm «ca. HcH. mNH. mmm. cwH mammfie Nc. H mcc. ch. ccc. Ncc. cm: mammHH mma. cma. ccc. mma. cc msmmwe Houum :mmonh :mmum cmuoum :wmu: NSHEV mmwu cumccmom Essom>usoz Essom> mEHu chEmm cmfiooe wcHHnese .AchEmm w cccH\mcwsmchconE.wEv HmnEsz scoz EDDom> mEHu mHmEmm cmHoo: wcHHAEDH Amuzv.mucmEummHu wcwmmmooua mp wmuomwwm mm Emumhm ummE msu CH quucoo uHmm .mum :Hmzmmm< AEm:V Hm.H H HN.¢¢ mm.ca cm.cHH cc.wmfi mc.mNH NH.HNH cwa Amumcoxmv Hc.H H Nc.Hm c¢.NN cm.mNH cc.cma Hm.HHH ac.ccH cm: AmSmmHHv NN.N H cc.mN mm.mm Nc.mHH Hm.cNH cm.cc cN.cc cc Houum QED: Nwm mam: NMH QED: Nmm mEDm NcH QED: Nwm NED: NcH ACHEV cnmccmom Em: mumcsx: msmeH mEHu cmaoo: wcHHnEDH Mammy EHHEHHZ 121 mH.HN cm.mm c¢.Nm mm.cH cwH Em: mm.H H cm.cm mo.NN cw.Nc m¢.mm cNH Em: cm.cN cw.¢a m¢.~¢ mm.mm cc Em: ¢N.NcH N©.@HH wc.NNH «w.wca cwH mumcsx: cc.~ H cc.cMH um.ccH cm.cHH mw.cma ONH mumcsx: qc.HmH cm.mcH ww.c¢a Hm.cNH cc mumcnx: mw.HcH cm.¢c cH.m¢H NH.HNH cwH mSmmHH mc.q H Hw.c¢H cc.ccH wH.ccH Hc.ccH cNH mammwe Hm.cmH cc.cc ¢¢.¢¢H cN.wc cc mzmmwfi Moupm cmmouh :mmu: cmmou: meum NEHEV mmNu oumccmum E:=om>|:oz Endom> mEHu m Emm _ . Ha cmaoo: wCHHQEDH Aaeev EHHEHHZ Amie.mucmEummHu wCHmmmooua >c cmuomwmm mm Emumxm ummE m:u CH ucmucoo muHHqu .cI: chzmmm< APPENDIX C Analysis of Variance Tables APPENDIX C-l. ANOVA table for protein content in the exudate as affected by tumbling time, pressure during tumbling and condition of the meat. 05/28/81 05/28/01 1 h :nm .0 No: NF. A03 ONS ACTI A02 A01 EXPLAINED RESIDUAL 3'UAY INTER TOTAL 11 13.838 34.528 .001 152.217 .401 4.624 24 9.618 161.835 35 MISSING. APPENDIX C-6. ANOVA table for TBA number in the exudate as affected by tumbling time, pressure during tumbling and condition of the meat. 05/28/81 05/28/81 1 V A R I A N C E A A A A A ZlHOa 127 ILLI— ZLL (DO ZLLJ (I U< 2: OF SUM OF SQUARES SOURCE OF VARIATION «II—Iifi—t 666° GONO o o o o tho—n econ-t "MONCD O O O O nub—nu NN N O‘o-o—td‘ cdco GOOD 0 o o o can-4N \Dv-Ov-Im ”dc“ COO: o o o o FFECTS E 1 2 3 COO N A A A MAI NOD—I OG'VOQ 9906 o o o o CG’NO ammnw (Uh—ND I o o 0 10¢ U‘ NNDG' COCO coco o o o o [Du-INN HNOQ «106° COCO . o o o .005 12.399 .001 .000 .002 .047 EXPLAINED RESIDUAL TOTAL 23 .010 .057 32 MISSING. E 1X9 UPPRESSED. APPENDIX C—7. ANOVA table for salt content in the exudate as affected by tumbling time, pressure during tumbling and condition of the meat. 05/28/81 05/28/81 1 V V LEVEN tag: 32": l—O-LLW V A R I A N C E O F AAAA A 128 LLLI. ZU- (DO ZLl-I <¢ LU< 23 OF SUM OF SQUARES SOURCE OF VARIATION «Oh—0 acne coco O O 0 O ccnm soNN—c ..nmm o o o o rmmm: v-1 N \DNOMr dam-r: “DO—1 o o o . #F'N-‘N Q'NO‘Q \OG’QHO {COW o o o o FFECTS E 1 2 3 666 A A A MAIN anN @950 ON'OO O O O O Gino-1N hutch—o FOG-Ow o o o . Ind—ltd NHO‘N G'v-IIOO COO—1 o o o o 1.0—ANN chow:- n—o—oc NOON . o o . NF- ccc <=c 0. NF- hfi- Inn 0. hp- ~0c CO 0. POM V310 pow-I o 0 A03 IONS 2 ACT A0 A01 EXPLAINED RESIDUAL 3-UAY INTER TOTAL 8.607 .001 .076 .009 .041 11 12 23 .833 .106 .939 RE MISSING. D E U) Y , pressure during tumbling and condition of the meat -3, ANOVA table for nitrite content in the exudate as affected b tumbling time 05/28/81 05/28/81 1 A A A A A A A A 129 LLLL 21L LDC DF SOURCE OF VARIATION _t.-new! ccmc DOOO o o o o €31.05 U‘FMD? QU‘TDO‘ o o o o cue-0‘ U‘U-i \O DQNV’) mmurd ID'ODO‘ also. MFMMO ammonia «no In «rod-IN IOQNTD c-IG'Q'N Nncm o o o o crmnn 01.0705 he c Nan c-d FFECTS E 1 2 3 COO <<< MAIN fink-I'd COCO @066 o o o 0 Nd’h—i Odom can-c O O O 0 N00? Qh—Hn Q'fl’NLO NmCON VO‘LD‘ hnmcc o o o o meom coco—«n OOQN ran—oa- Inc-0N0! \Dlnf‘e' WON-IN FNO‘O o o o 0 come: NQNF‘ d’xDNlfi 1.0an NN F070 GO 0 o .-u-o NN \OVD O O -¢¢ No: NH- '3’? 0. mm: nva or, 1¢¢ cos 0. «cc FN- .A03 ACTIONS A02 A01 EXPLAINED RESIDUAL 3-UAY INTER TOTAL .001 1660.536 216.330 11 12 23 18265.892 7.676 798.174 92.111 18358.004 E MISSING. APPENDIX C-9. ANOVA table for VOIUWG 0f the soluble phase in the exudate as affected by tumbling time, pressure during tumbling and condition of the meat. 05/28/81 05/28/81 1 A A A A A A A 130 SIGNIF 0F F F 21...! <¢ 2:3 DF LII-(fl CU Z< (00 SOURCE OF VARIATION pun—4.4 QQOQ o—ncc o o o o IDQNPO O‘Q—‘N \D—‘eN o o o o NNOLD N Inn cecm Nd’d'ln \D¢N—i o o o o In Oh ¢~~N N¢°¢ ccec mewn C... N cc N ~~ FFECTS E 1 2 3 65° N A A A MAI «“0630 Na'Nc-I c—iO‘O O O O 0 QNLI'MD O‘c-ONG OVQND o o o o G’N In NCDIIW~ DOM-4v QG'Qv-I o o o o 1.0—00164 NOOQ‘ \OO‘IOO‘ ufi'aN o o o o G' N 9 3.186 15.686 .001 28.676 EXPLAINED RESIDUAL TOTAL .203 1.482 2.438 31.113 21 fit MISSING. D E U: ATRIXI N SUPPRESSED. 4 EE SINGULAR l S HAVE 8 A ION OR CT Y CELLS R INTERA y tumbling -10, ANOVA table for protein in the soluble phase as affected b time, pressure during tumbling and condition of the meat APPENDIX C 05/28/81 05/28/81 1 131 LLLL. 21.1. 66 DF SUM OF SQUARES SOURCE OF VARIATION MAI c-u-u-o—o 6666 66’66 . . o 0 66¢? Iowa-s \OG'IDN . o o 0 666.1 n—INQ’ cured comma @6h13 o o O o mmhm «mun: a-NNLD VHfiN NOG'N NhnN 0‘6h—a . . o . @656 _uncnh hNN—t .4 u-n FFECTS E 1 2 3 666 N A A A v-tO‘N—o 6—‘66 6666 O o O 0 66¢“ 60‘6N C!‘N~.D(7~ O O O O ch—uo N ...¢ hem—I "30“?10 hoxo—I I Q 0 O QNMN c6xom POM—HO LOU-1N0) heuw cccc ccmn O... nth eoNd hdfin 0" 1'4 u-n—dN >666 <<<< 2-U NH- ccc 13° .0 ccc NF- 00 O. NH- A03 ONS CTI A02 A01 EXPLAINED RESIDUAL 3-UAY INTERA TOTAL 334.042 23.851 .001 11 12 23 3674.461 14.005 167.066 168.065 3842.526 MISSING. HERE PROCESSED. I 33.3 PCT) HERE CASES CASES 36 12 -11. ANOVA table for myosin relative content in the soluble phase as affected by tumbling time, pressure during tumbling and condition of the meat. APPENDIx C 05/28/81 05/28/81 1 V A111 AAIC E F A A A A LLLI. 21.5 66 ZLIJ <0! LIJ< 26 DF SUM OF SQUARES SOURCE OF VARIATION 132 “Go-I6 6661" 666M 0 o o o chw cmcc nmcc .... cmmd H w cNLnN schn ncnc I O 0 O \DN'O6 G‘O‘hu— cod—0N 6N6” 6¢f~m €666 o o o 0 106166 QO‘BN W N FFECTS 6.!de o-AO‘v-Am 6'06N 666m 0 o o o comm-n N506 Nn—NO O O O O cmcv .-o N dNNm now—I 066'?) 0 O O O eraunc Chet: mdNN 60166 L066?) dQQQ O O O O d'a‘c-iN her—.— 4:- N6c 2 Oct) ZCZFi-I < OLLII Znnc 0— ”N N 24: x i-o-n U261! > n N ..z—u t/Hl (ALAN. u. .30: 6.11.524: 6 >11:— luv-ix. .J Z 264: m~¢o~ rave—a ..J C < C 2 .r «(tn—scum coca.- <<<< «I a )- a m c u c a: .- A. c 4: 133 LL11. 21L 66 21.1.1 <6 U< 26 DF SOURCE OF VARIATION MAI VDo-QNO‘ '“m6h 6'466 . . . . 610601 107036 NNd'c-i o o O o 6N6” QNN‘O 666x13 {\Ofl'N o o o o ”FINN {F4601 \ONNPO —mwnn ccc-In O O O O I'm-45¢ ... N EFFECTS A01 A02 A03 {HOLD'O O‘N66 {Oh—m . o . . N666 P0666 @6106 O O O O QNQC 66106 \06Nd' o o o o 1.0—.0101 chcc 106—dun 70666 000 0 IO N <2: Ice TOLD NN POM \DVD 0 o '0") LDLD d’fi' o 0 06 66 60‘ O 0 A03 IONS 2 ACT A0 A01 EXPLAINED RESIDUAL 3-UAY INTER TOTAL 11 1.624 2.266 .088 17.860 .717 1.150 12 23 8.600 26.460 56 MISSING. o" E APPENDIX C-l3. ANOVA table for ham cooking losses as affected by tumbling time, pressure during tumbling and condition of the meat. 05/28/81 05/28/81 1 a 4: c 4r 4- «I a a c c c a c c .— t d A Lu 22} L4 u—u 023:4: U (U >N6a 2 cc zap-H: < OLLII 2"")! H ”N N 243 6 I-H U<£¢ < «11...: >26; > H N Fez—w mu wUNc u. .36 1..)ch 6 >11— two-42¢ <4 I 264! m MONO exact—HG moat-d _Imu-o c woman: UUZZC >- 16" 60.00-¢ «I 4 a Z «a (Ft—INK) moccqx <<<< A A >- 6 CD 4: a c c «I c a c c 134 LLLL. 211.. 66 ZLIJ €01 U< 26 DF SUM OF SQUARES SOURCE OF VARIATION Nnm—I 6666 6‘66 0 O O O «ac-cc €051.00 ~066h O C O O mNnco INN—06' (bu-INN LDNVOO" O O A O Q'FIN-‘N \DQF‘F- QdNV ”N106 0 O O O N w. EFFECTS 01 02 03 N A A A MAI ennw 6661.0 6d66 o o 0 o ccdm Nhnm sown .... emhn ~066N U‘v-imtfi Q'Nhf’) o o o . LD—‘NN NQv-‘Q' 6'166 {NI-ON o o O o N H 66 NF NN o 0 NW Lmn P0” 0 o '40-! No: Pee '41-! 0. e: In: No: 0. A03 ACTIONS A02 A01 EXPLAINED RESIDUAL 3-NAY INTER TOTAL .468 4.461 .001 .105 .219 11 24 5.152 2.520 35 7.672 A6 NISSING. 6U 135 time, pressure during tumbling and condition of the meat. APPENDIX C-14. ANOVA table for L color parameter in hams as affected by tumbling ‘ 6 \ 6 N \ In 6 4: «I u.u. NNU‘d' ”ONO NN .-4 H 6‘66 6¢36l'~ mm 6 c c 21L 66006 6666 66 6 66 o o o o o o o o o o o c a u— (I) an c V066") hv-"Oln dv-I r~ 4! i OMNO‘ Nina‘h 8"? 6 LL 6066 O‘HG‘Q 'OI'O 0‘ «I: c o o . o . o o o o o o mh rs enhN ran er c p— < c L4 26.: Noe-c (In-nah e-e- In '5 r~ :2. <0: Ken-”MO 6\ON\O SONG '0 o h Lu .— LIJ< 0‘60‘6 #1066 min N 6 PO 622. Z: 00.. .000 .0 o o 0 U <6 C3 \Oh h 10'0va 10") In .-0 N >N6c VI 2 66 Zen-NO < on." ZH'Oa u-o IIN LL e-s-o—oN InflNN NN .-n e- In N Z. 6 .-o N n C! t—u—n U26c .... > II N :0 div-HI u..m soceN the Nh- : c c \ mu 6“ 6¢nn 0'6er NN 6 6 6 Q ”UNA 6 660—4 “3'06—0 «H LO \o .-a g N LL ..ch Z< o o a o o o o o o o o o o o \ 141.52.: 2: nr~ In cnhc hh '5 m n 2 L0 6 >H~ WC! N .-. N w-t In N 6 ~— 6 two-oz. m m (.1 m I 26: .... III 0‘) U66 5: < cram-nu ¢ 3:11.: Honk-d U 6 )- 0:6!- (0 0321..) .JQLM-d m.- ~6..i _J P0 Lg.— Zr-U c 6 60 ~P: < 4 on 15¢ i x U 2 (La 0:0: ‘I 2 CU -> > >- r-o b666 1-6 66 Lu 4: an «I I: Q<<< u< mm 62 ..J < (I) < < MU h—<)-LIJ c c > I- a: a: mm (A < 6 U Lu << >1... 4 c u. 1...: I— II- 6 UL.) 6(2 6 u. 2 ? Lu .1 23 U i A LL .— H 2 < ~06 H ...l U LEI—4N” pun-IN .-o u- : n ..JuJ .... t C U ccc >ccc >c .1 cs _; muum m z<<< <<<< << 4 u < 26.16 4' 4' D '— 3 3 0.. m '- :IHD c < I I x L.) c I-i-uJfl & 4! (h I N '0 LA at I— APPENDIX C-lS. ANOVA table for a color parameter in hams as affected by tumbling time, pressure during tumbling and condition of the meat. 05/28/81 I 05/28/81 i t i t i t t .— £ < c to 12¢ Lg :— 622' 6 <6 >N6C Z 66 26“; < 6U." ZIIN‘MI n—o "N N 2‘ 6 I—u-n 6424- < <6 >Z6‘ > II N HI—tfi (DH (AUN‘I u. .16 ULLZ‘I 6 >lh-1 IMF-42¢ <.J I 26* (I) 666 66—1". Mosh—l _Jm— C (06666 UUZIC >- 66" <0.60—« _1 a < a Z q <6—9Nn U‘r066.‘ <<<< ¢ 4' ). a 6 ‘t t fl t t i i 136 LLLI. ZU- 66 DF LL03 CLIJ =< DD (06 SOURCE OF VARIATION nhhn N666 ONNO‘ o o o 0 #606 666K) \DNN6 o o o o .-o—a IOIOIOG' \ONN6 adv-16 o o o o VFW-NV N666 IONN6 {VFW-‘6 O O C O FFECTS E l 2 3 666 N A A A MAI NNa-uf) 6N6~O 666m 0 O O O 6666 {N66 N666 o o o o ION 60466 6666 V6—t6 o o o o UNI-INN U‘NLDC‘J 66".“ 06nd o o o o N N _t.—N >666 -> > 66 La 62 _J l-(i-LLJ (I) 4 >6 6