Budapest, September 4 - 11, 1963

Studies on the Color of Meat and Meat Products II. Effects of Polyphosphates on the Color of Cooked Sausage

Norihide A ndo, Yoshitaka Kako, Yukiharu Nagata, Tomio Ohashi,

Yoshihiko H irakata, Noriko Suematsu, and Eiko Katamoto

Laboratory of Chemistry and Technology of Aaimal Products, Kyushu University, Fukuoka, fapan

Reprinted from Bulletin of Meat and Meat Products

19 6 3 JAPANESE SOCIETY OF MEAT AND MEAT PRODUCTS Bull. Meat and Meat Prods., Vol. 2, p. 1 ~ 6 , (1963)

Studies on the Color of Meat and Meat Products II. Effects of Polyphosphates on the Color of Cooked Sausage

Norihide A ndo, Yoshitaka Kako, Yukiharu Nagata, Tomio Ohashi,

Yoshihiko Hirakata, Noriko Suematsu, and Eiko Katamoto

Laboratory of Chemistry and Technology of Animal Products,

Kyushu University, Fukuoka, fapan

(Received March 1, 1963)

As briefly reviewed in the previous trimmed to remove fat and connective tissue as paper15, according to the results of works far as possible, and the resulting lean meat was published previously2'85, the detailed action then ground three times with a meat grinder. of phosphates on the color of meat prod­ The ground meat was mixed thoroughly and ucts is still far from being explicitly was divided into five lots. Sausage samples were prepared in the same elucidated. manner as related in the previous paper, namely, It was found in our previous work each lot was treated with each of the mixture that the addition of acidic orthophosphates of curing ingredients given in Table 1, stuffed gave favorable effects on the color of into Krehalon (vinylidene chloride-vinyl chlo­ cooked sausage, but resulted in considable ride copolymer) casing, cured for 72 hours at decrease in water , holding capacity of 4°C, cooked at 75 °C for an hour, and then meat, whereas the supply of basic ortho­ cooled. phosphates developed a tendency to exert Determination of water holding capacity of undesirable influences on color and en­ m eat: couraging influences on water retention The total water content of ground fresh by meat. meat sample was determined in the usual way, The present study has been made to and the amount of cooking loss was determined investigate the behavior of polyphosphates in the same manner as described in the previous paper except for the modifications in the follow­ towards the formation and the protection ing two points. of color of cooked cured meat. First, cooking loss was separated at 7,000 METHODS rpm for 10 minutes instead of separating at 2,500 rpm for 10 minutes. Second, since usually In the present experiment, color formation a certain amount of many sorts of inorganic value (CFV), color retention value (CRV), pH and organic compounds are contained in cook­ value and TBA absorbance were determined in ing loss, the water content of cooking loss was the same ways as described in the previous determined in the usual way by drying at 100 paper, but determinations of water holding °C, and the water holding capacity ot meat was capacity of meat, reducing value, nitrite and calculated by the following formula: volatile basic nitrogen were made by modified Water holding capacity of meat methods given below. _% total water—% water of cooking loss Preparation of cooked sausage : % total water Beef chuck at five days after slaughter was By repeating experiments on centrifugal — 2 —

Table 1. Curing ingredients used for the preparation of cooked sausage

Curing ingredients

NaCl NaN02 Polyphosphate1*1 Neofurasukin

Control 2.5% 0.02% — -% 0.025 % 1 2.5 0.02 Na2H;P30 7 0.5 0.025 2 2.5 0.02 Na4P207 0.5 0.025 3 2.5 0.02 Na5P 3O10 0.5 0.025 4 2.5 0 02 Na6P60 18 0.5 0.025

(a) Each amount of the polyphosphate added was calculated on the basis of anhydride. (b) Preservative permitted in Japan. Neofurasukin is a mixture of nitrofurazone and nitrofurylacryl amide. separation of cooking loss from cooked ground 8 ml of distilled water, 10 ml of 10% trichlor­ meat at 4,000, 5,000, 6,000, 7,000, 8,000, 9,000 acetic acid solution was added, mixed thorough­ 10,000 rpm for 10 minutes, respectively, it was ly, and allowed to stand for 10 minutes at room found that most successful results were ob­ temperature, then filtered through a filter paper. tained when samples were centrifuged at 7,000 The optical absorption of the filtrate was meas­ rpm for 10 minutes, because the variation in ured at 420 m/i using a 1cm cell, with a mixture, values for cooking loss separated from the very which underwent the same operation as before same samples was the slightest, viz., the coef­ except that 4 ml distilled water were added ficient of variation of the findings for cooking instead of 2g of meat sample and 2 ml of M/50 loss obtained from the same samples showed the K3Fe (CN)6 solution, as a blank. lowest value, and even a trifling difference in The mg % of cysteine was calculated by the cooking loss between two different samples could following formula, and the finding for mg % of be observed most explicitly. cysteine was expressed as RV (reducing value). Besides, through determining the water con­ RV (mg % of cysteine) =242.32 — 224.37 A tent of cooking loss, it was possible to get in­ A=Absorbance of sample observed. disputably more accurate figures for water Determination of nitrite : holding capacity of meat than before. Since there are several methods for the Determination of reducing value : determination of nitrite10-14', considerable efforts As a result of further experiments on the have been exerted to find out the most satisfacto­ determination of reducing value of meat and ry method for determining nitrite present in meat meat products, the following modification of and meat products. From the results given by the method of Kajita91 was found to be most repeated examinations, the following procedure, successful. a modified method of Shinn111, may confidently Two grams of a meat sample were homo­ be recommended. genized with 5 ml of distilled water for 3 mi­ Two grams of a meat sample were homoge­ nutes, cooling with ice water. The homogenate nized with 5 ml of distilled water in a Waring was transferred into a 20 ml graduated test tube blender for 3 minutes. The homogenate was with 10 ml of M/15 phosphate buffer of pH 7 transferred into a 150 ml graduated 300 ml beaker and with distilled water. After diluted to the with enough distilled water heated to 80 °C to 20 ml mark with distilled water, 2 ml of M/50 fill to the 150 ml mark. The beaker was placed K3Fe (CN)6 solution were added, the test tube on a boiling water bath and heated for an hour, being stopped with a well-fitted glass stopper, the content being stirred occasionally with mixed thoroughly, and the mixture was allowed a glass rod. The evaporated water during the to stand for 20 minutes at 0°C. Afterwards the heating process was supplied occasionally by mixture was removed into a 100 ml beaker with adding distilled water to the 150 ml mark s

throughout the heating operation. Two ml of supplied by disodium pyrophosphate for saturated HgCl2 solution were added, mixed, both CFV and CRV always showed the and cooled with running tap water to room highest figure among the five kinds of temperature, then transferred into a 200 ml cooked sausage samples. When treated volumetric flask, diluted to the mark with with sodium hexametaphosphate (Na6P6- distilled water, mixed thoroughly, filtered Oi8), no practically significant change in through a filter paper. pH value was effected and some favorable To 8 ml of the filtrate 2 ml of reagent (a mixture of 1.0 ml of 0.2% sulfanilamide effects were brought about on the color of aqueous solution, 0.2 ml of 6 NHC1 solution, 0.2 cooked sausage. Most considerable in­ ml of 0.1% N-(Vnaphthyl)-ethylenediamine crease of pH value and most undesirable dihydrochloride aqueous solution, and 0.6 ml of effects on color were observed in the sam distilled water) were added, mixed, and kept at pie treated with tetrasodium pyrophosphate 30 °C for 30 minutes, cooled with running tap (Na4P207), and the addition of sodium water for 10 minutes, transferred into a 1 cm tripolyphosphate (Na5P30lo) exhibited to cell and the absorbance was determined at 535 develop the same tendency as displayed mft with a mixture of 8 ml distilled water and by the supply of tetrasodium pyrophos­ 2 ml reagent as a blank. The content of nitrite phate, though the influence of sodium was expressed as ppm of nitrite group (N02~). tripolyphosphate was expressly less ef­ Determination of volatile basic nitrogen : In the present experiment, the content of fective than that of tetrasodium pyrophos­ volatile basic nitrogen was determined by phate. a modification of the micro-diffusion method of These results may be taken to in­ Conway15’. dicate, as previously related in cases of orthophosphates, that the decrease in pH RESULTS AND DISCUSSION value by supplying acidic polyphosphate, Color formation value (CFV) and color viz. disodium pyrophosphate, would mani­ retention value (CRV): fest desirable effects on color formation According to the results given in and protection from discoloration by light, Table 2, the addition of disodium pyro­ whereas the increase in pH by supplying phosphate (Na2H2P207) decreased the pH basic polyphosphate, i. e. tetrasodium value to some extent and produced most pyrophosphate and sodium tripolyphospate, A favorable effects on the color of cooked would produce undesirable effects on color sausage, i. e., each value of the sample of cooked sausage. ► Table 2. Effects of polyphosphates on pH, CFV, CRV and water holding capacity

Just after preparation i After storage for 4 weeks at 4°C Poly- Before After Before After Water Sample phosphate exposing to exposing to exposing to exposing to light light light light holding number added, PH pH 0.5% capacity Absor­ Absor­ CFV CRV Absor- ppy AbSOr* pnrr bance bance bance v bance CRV

Control None 6.05 1028 100 0.468 46 6.05 0.935 91 0.314 31 \ 55.48 1 Na2H.,P207 5.95 1.076 105 0.518 50 5.95 0.978 95 0.382 37 53.23 2 Na4P207 6.35 0.941 92 0.341 33 6.36 0.885 86 0.257 25 72.98 3 Na5P3O10 6.25 0.984 96 0.371 36 6.24 0.921 90 0.295 29 69.70 4 Na6P60,B 6.04 1.046 102 0.487 47 6.03 0.979 95 0.358 35 56.91 — 4 —

It seems to be conceivable, however, Since the main purpose of the present that changes in some factors other than experiment is to investigate the effects pH resulted from the supply of poly­ of polyphosphates on the color of cooked phosphates might exert some particular sausage, nothing further should be here influences upon the color of cooked sau­ concerned with the problem of water sage. holding capacity of meat. As shown in Table 2, no significant Nevertheless, as pointed out in the changes occurred in pH values during previous paper, since the supply of poly­ the storage period of 4 weeks at 4 °C, phosphates produces quite an opposite but a considerable decrease took place effect on color to those on the water in values for CFV and CRV during the holding capacity of meat, in cases of storage. orthophosphates as well, it should be Water holding capacity of meat: forcibly emphasized here again that a pru­ In almost the same manner as mani­ dent attitude must be taken when phos­ fested in the previous experiment on phates are put to practical use. orthophosphates, the water holding capa­ Reducing value CRV) and N 02“: city of meat was most remarkably im­ The supply of disodium pyrophosphate, proved by the addition of most basic the most acidic polyphosphate used for polyphosphate, i. e. tetrasodium pyrophos­ the present experiment, resulted in most phate, and the effect of sodium tripoly­ considerable increase of RV and most phosphate on water holding capacity rank­ remarkable decrease of N 02~ content, ed next. whereas the addition of tetrasodium pyro­ The addition of disodium pyrophos­ phosphate, the most basic polyphosphate phate, an acidic polyphosphate, resulted employed in the present experiment, re­ in a slight decrease in pH value and a dis­ sulted in most marked decrease of RV couraging decline in the water holding and most striking increase of N 02” con­ capacity of meat, while the supply of tent, as compared with the findings for sodium hexametaphosphate brought about control. an insignificant change in pH value and Since the high reducing power and a just faint increase in the water holding the low pH value of meat promote the capacity of meat. decomposition of nitrite into nitric oxide, According to the theories on the and in consequence yields favorable results mechanism of the action of alkaline phos­ on the formation and retention of cooked phates, increases in pH value, ionic cured meat color, it would be conceivably strength and solubility of fibrous myofi­ reasonable that the content of N 02' in brillar proteins caused by the addition of cooked sausage may increase with decreas­ phosphates, abilities of phosphates to re­ ing reducing power and increasing pH move bivalent cations, such as calcium value of cooked sausage, and that the and magnesium, from peptide chains by higher remaining content of N(V denotes complexing with those cations and to split the lesser formation of nitric oxide heme actomyosin into its components myosin pigment and accordingly exerts infallibly and actin, are considered to be most con­ discouraging influences on the color of ceivably contributable to the favorable cooked sausage. effects of phosphates on the water holding But the reasons why the RV of the capacity of meat. cooked sausage has remarkably increased — 5 —

by the addition of disodium pyrophosphate went during the storage, but consistent and why the RV of the cooked sausage and evident decreases in the figures for significantly decreased by the treatment NCV content were observed during the with tetrasodium pyrophosphate still re­ storage. main unknown. As to the influence of polyphosphates From the findings for volatile basic on changes in the figures for RV during nitrogen and TBA absorbance given in storage, many further experiments should Table 3, it would be conceivably suggest­ be required before a definite conclusion ed that faintly slight changes in protein can be safely drawn. and no significant changes in fat under­

Table 3. Effects of polyphosphates on reducing value and N02~ content

Sample Polyphos­ Jus t after preparation After storage for 4 weeks at 4°C phate number added, Reducing xjp, _ Volatile TBA Reducing n o 2- Volatile TBA 0.5% value - basic N absorbance value basic N absorbance 1 Cysteine Cysteine mg% ppm m g% mg% ppm mg% Control None 64.2 46.0 18.84 0.274 45.0 30.8 21.10 0.188 1 Na2H2P,07 77.8 36.8 19.92 0.268 50.2 22.8 21.97 0.184 2 Na4P20- 35.1 85.4 22.01 0.253 38.7 58.7 26.35 0.206 3 aO10 Na5P 41.4 73.4 19.91 0.258 42.9 46.2 24.17 0.222 4 Na6P60 I8 67.8 44.5 20.78 0.270 50.5 28.1 24.06 0.209

aging effects on the color of cooked cured SUMMARY meat were manifested by the supply of A study on the effects of four kinds sodium hexametaphosphate. of polyphosphates -----disodium pyrophos­ As a markedly considerable increase phate (Na2H2P207), tetrasodium pyrophos­ in pH value and a definite decrease in phate (Na4P207), sodium tripolyphosphate reducing power of meat resulted from the (Na5P3O10), and sodium hexametaphos- addition of tetrasodium pyrophosphate, phate (Na6P60 18)-----on the color of cooked a fairly large amount of nitrite was al­ sausage were performed in the present lowed to remain unchanged, and this experiment. deservedly brought about the most unde­ The addition of disodium pyrophos­ sirable effects on producing and protecting phate resulted in a significant decrease in cooked cured meat color of all the sausage pH value and a remarkable increase in samples used for the present experiment. reducing power of meat, and accordingly Quite similar tendencies, as displayed promoted the decomposition of nitrite into in the case of tetrasodium pyrophosphate, nitric oxide, thus brought about the most were observed when the meat sample was favorable effects on color formation and treated with sodium triphoshate, but every protection from fading by light of all the effect on pH value, nitrite content, reduc­ cooked sausage samples employed in the ing power of meat, development and present experiment. retention of cooked cured meat color was Although no sinificant shift in pH consistently and explicitly less intensive value was effected, a slight increase in than that achieved in the case of tetraso­ reducing power of meat and some encour­ dium pyrophosphate. — 6 —

Nevertheless, since the addition of 265 (1956) polyphosphates which manifestly increases ( 4 ) Henrickson, R. L., Sleeth, R. B. and Brady. the water holding capacity of meat has D. E .: Food Technol., 10, 500 (1956) ( 5 ) Pohja, M. S. and Niinivaara, F. P .: Dtsch. always produced an indisputably discourag­ Lebensmitt. Rdsch., 53, 64 (1957) ing effects on color of cooked sausage, and (6 ) Swift, C. E. and Ellis, R.: Food Technol., vice versa, some efficacious devices, which 11, 450 (1957) not only produce favorable effects on water (7 ) Wismer-Pedersen, J .: Fleischwirtschaft., holding capacity of meat but also promote 10, 830 (1959) explicitly desirable effects on color of cook­ (8 ) Kamstra, L. D. and Saffle, R. L.: Food ed cured meat, seem to be indispensable Technol., 13, 652 (1959) when polyphosphates are put to practical (9) Kajita, A.: /. Japanese Biochem. Soc., 26, use in processing meat products. 547 (1954) (10) Tannaeff, N. A. and Schapowalenko, A.M .: Z. Anal. Chem., 100, 351 (1935) LITERATURE CITED (11) Shinn, M. B .: Ind. Eng. Chem., Anal. Ed., 13, 33 (1941) (1) Ando, N., Kako, Y., and Nagata, Y.: (12) Rider, B. F .: Ind. Eng. Chem., Anal. Ed., Bulletin of Meat and Meat Products., 1, 1 18, 96 (1946) (1961) (13) Novak, R. and Wilson, P. W .: J. Bade- (2 ) Janicki, J., Niearowicz, A. and Bury, S .: riol., 55, 517 (1948) Przem. Spozyw., 9, 414 (1955) (14) Method of Analysis, A. O. A. C., 389 (1955) (3 ) Mahon, P., Hogue, D., Leeking, P., Lim, (15) Hatano, H. and Kirita, T .: Kagaku no E. and Fenton, F .: Food Technol., 10, Ryoiki, Extra Edition., 34, 46 (1960)

Studies on the Color of Meat and Meat Products II. Effects of Polyphosphates on the Color of Cooked Sausage

Norihide A ndo , Yoshitaka Kako , Yukiharu N agata , Tomio Ohashi , Yoshihiko H irakata , Noriko Suematsu , and Eiko Katamoto

Laboratory of Chemistry and Technology of Animal Products,

Kyushu University, Fukuoka, Japan

SUMMARY

A study on the effects of four kinds a fairly large amount of nitrite was al­ of polyphosphates -----disodium pyrophos­ lowed to remain unchanged, and this phate (Na2H2P207), tetrasodium pyrophos­ deservedly brought about the most unde­ phate (Na4P207), sodium tripolyphosphate sirable effects on producing and protecting (Na5P3O10), and sodium hexametaphos- cooked cured meat color of all the sausage phate (Na6P60i8) ---- on the color of cooked samples used for the present experiment. sausage were performed in the present Quite similar tendencies, as displayed experiment. in the case of tetrasodium pyrophosphate, The addition of disodium pyrophos­ were observed when the meat sample was phate resulted in a significant decrease in treated with sodium triphoshate, but every pH value and a remarkable increase in effect on pH value, nitrite content, reduc­ reducing power of meat, and accordingly ing power of meat, development and promoted the decomposition of nitrite into retention of cooked cured meat color was nitric oxide, thus brought about the most consistently and explicitly less intensive favorable effects on color formation and than that achieved in the case of tetraso­ protection from fading by light of all the dium pyrophosphate. cooked sausage samples employed in the Nevertheless, since the addition of present experiment. polyphosphates which manifestly increases Although no sinificant shift in pH the water holding capacity of meat has value was effected, a slight increase in always produced an indisputably discourag­ reducing power of meat and some encour­ ing effects on color of cooked sausage, and aging effects on the color of cooked cured vice versa, some efficacious devices, which meat were manifested by the supply of not only produce favorable effects on water sodium hexametaphosphate. holding capacity of meat but also promote As a markedly considerable increase explicitly desirable effects on color of cook­ in pH value and a definite decrease in ed cured meat, seem to be indispensable reducing power of meat resulted from the when polyphosphates are put to practical addition of tetrasodium pyrophosphate, use in processing meat products. Table Effects of polyphosphates on pH, CFV (color formation value), reducing value, N02~ content and water holding capacity of cooked sausage just after preparation.

Polyphosphate Reducing Water holding Sample No. pH CFV n o 2- added, 0.5% value capacity

Cysteine mg% ppm Control None 6.05 100 64.2 46.0 55.48 1 Na2H2P2Ov 5.95 105 77.8 36.8 53.23 2 Na4P207 6.35 92 35.1 85.4 72.98 3 Na5P 3O10 6.25 96 41.4 73.4 69.70 4 Na6P c0la 6.04 102 67.8 44.5 56.91

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