Addition of Clarase or pig mucosa to soybean meal slurries digested in ivater (pH 6.5) caused loss of added thiamine. No loss was observed by addition of Lvheat germ phosphatase, or by autolysis of the raw meal. The cause of losses by Clarase and pig mucosa was not apparent. Likeivise. the good recovery with kvheat germ phosphatase is not understood. About 60% of the thiamine in soybeans is present in bound form presumed to be cocarboxylase. Raw meal contains a phosphatase with optimum activity at pH 6.0 which liberates free thiamine from cocarboxylase on autolysis. In the Johnson analysis. soybean meal is digested a t pH 4.5 with Clarase to liberate thiamine from its phosphate form. and Ivith papain presumably to liberate protein-bound thiamine. No additional thiamine was liberated by papain. trypsin? or pepsin; hence, their presence in the digestion is apparently unnecessar!.. The use of Clarase or some other phosphatase is advisable because cooking so)-bean meal for feeds inactivates the natural phosphatase; also the natural phosphatase is not very active at p H 4..5. Destruction of meal thiamine ivith sulfite and .\.it11 alkali, folloived by addition of ferric!-anide. failed to yield fluoresing materials i n escess of those in the blank. .Also. fluorescent materials obtained from the meal acted like thio-
chrome. in that they were entirely destroyed by ultraviolet irradiation. Hence, there \vas no evidence that fluorescence was not a measure of thiamine in the meal. Literature Cited
(1) Benesch. R., Benesch, R . E., Gutcho, M., Laufer, L.? Science 123, 981-2 (1956). (2) Bhagvat, K., Indian J . M e d . Research 31, 145-52 (1943). (3) Bhagvat, K., Devi, P., Zbid., 32, 1317. 138-44 (1944). (4) Block. R. J.? Durrum, E. L., Zweig: G., “Manual of Paper Chromarography and Paper Electrophoresis,” p. 295, Academic Press: Neiv York, 1955. (5) Cordy, D. R.. Cornell Vet. 42, 108-17 (19 52). (6) Goldzieher, J. \V.) Rawls, W. B.! Goldzieher, XI. A , . J . Biol. Chem. 203, 519-26 (1953). ( 7 ) Hasegawa, E., lVitamins ( & y o t o ) 8, 415-21 (1955). (8) Hasegaiva, E.: Tamaki: T., Tanaka, T., Fujita. A,, J . Vitaminol. (Osaka) 3, 30-8 (1957). (9) Hennessy. D. J.. Bonvicino. G., Abstracts, p. 63C, 116th Meeting, ACS, Atlantic City, S . J., Sept. 18-23, 1949. (IO) Johnson, B. C.: “Methods of Vitamin Determination,’‘ pp. 52-6. Burgess Publ. Co.. Minneapolis, 1948. (11) McKinney. L. L.. Weakley, F. B., Campbell. R . E.: Eldridge. .4. C., Cowan. J. C.? Picken. J. C.. Jr.,
Jacobson, N. L., J . A m . Oil Chemists’ SOC.34, 461-6 (1957). (12) Mason, H. L., Williams, R . D., J . Biol. Chem. 146, 589-94 (1942). (13) Matsukawa, D., Kawakami, T., J . Vitaminol. ( O s a k a ) 1,208-16 (1955). (14) Matsukawa, T., Yurugi, S., Science 118, 109-11 (1953). (1 5) Naftalin, J. M., Cushnie. G . H., J . Comp. Pathol. 64, 54-74. 75-86 (1954). (16) Sose, Y . , J . Japan. Biochem. Soc. 24, 25-9 (1952-3). (17) Roberts. E. -4.H.? Chem. & Ind. (London) 1959, 995. (18) Roberis, H. E., Evans. E. T. R., Evans, CV. C.: I’et. Record 61, 549 (1949). (19) Rosenberg, H. R . . ”Chemistry and Physiology of the Vitamins,“ pp. 99150. Interscience, New York, 1942. (20) Rubin, S. H.. DeRitter. E., Febbraro, E., Jahns, F. b’,> Cereal Chem. 25, 52-60 (1948). (21) Thomas, .4. .J.. \Vatkin. J. E., Evans. I. A , , Evans, LV. C.. Biochem. J . 61, viii (1955). (22) Walter, E. D., J . A m . Chem. SOL.63, 3273-6 (1941). Receiued f o r reisisre’ June 20, 7960. Acceptea March 6. 7967. Diaision of Agricultural and Food Chemistry, Symposium on Deleierious Compounds in Foods and Feeds. 734th Meeting, ,dCLy.Chicqo, Ill., Spptember 7958. RFferFncF to a commercial product in this publication is not intended to be a recommendation of that product by the C:. S. Department of dgriculture ouer others not mentioned. The AVorthern Laboratory is part of the Xorthern L‘tiliration Research and Deuelopment Dirision, Agicultziral Rpsearch Seri,ice, c‘.S. Department of Ag,iculture.
QUALITY A N D FLAVOR O F D A I R Y PRODUCTS
Review of Biochemical Properties of Milk and the Lipide Deterioration in Milk and Milk Products as Influenced by Natural Varietal Factors
T
F L ~ V O R S O F MILK and milk products and their behavior in normal use seriously affect their consumption and ma)- make them totally unfit for human food. T h e psychic effect of smell and taste cannot be lightly dismissed ( 4 0 ) . and oxidized fats in the diet may IoLver the utilization of vitamin A and actually be injurious to health (38, 4 4 ) . Consequently. i t is very desirable to make inilk products both palatable and relatively resistant to the causes of bad flavors and losses of nutritive value. The prevailing tendency, however, is still to emphasize quantity production of feed and milk to reduce production costs. to promote consumption of milk by stressing its unique nutritive properties. and to lean largely on processing methods as a source of aid, rather than to improve production methods on the farm. Yet.
HE
fat-soluble vitamin content of milk and certain biochemical properties which control palatability and thus consumption are related not to quantity production of feed and milk. but to the type and quality of roughages and supplements fed. the physiological response of cows of a given breed to the feed consumed, and the handling of milk after withdra~val from the mammary gland. Major Causes of Undesirable Flavors
Major causes of undesirable flavors in milk and milk products include: odorous principles or their metabolites which pass from the feed via the colv‘s blood to the milk. giving it a “feedy flavor”; splitting of fat by the milk enzyme lipase, products of which give milk a rancid odor and bitter taste and
VLADlMlR
N.
KRUKOVSKY
Deportment of Dairy and Food Science, N e w York State College of Agriculture, Cornell University, Ithaca, N. Y.
increase the acid degree of fat; and certain other chemical reactions, enhanced by specific substances or by exposure of milk and milk products to light, \vhich result in metallic-to-fishy. oily, and chalky-to-soapy-tallo\\l (cardboardlike) flavors. Lipolytic Rancidity in Raw Milk
Shortly before \\’orld War 11: lipolytic rancidity caused by by-products of fat splitting was one of the greatest problems of the dairy industry. It \vas solved here at Cornell University by a careful study of the methods used for handling raw milk on the farm and in the milk plant. The milk enzyme lipase was found to be activated by slight Lvarming of cold milk and subsequent recooling; activity increased as the temperature \vas lowered
V O L . 9, N O . 6, N 0 V . - D E C .
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439
The purpose of this paper is to provide a perspective and better understanding of biochemical processes which control palatability and nutritive value of milk and milk products. Causes of loss in palatability of prime interest are: splitting of fat in raw milk by lipase, which gives rancid odor and bitter taste; and other chemical reactions, enhanced by specific substances or by exposure of milk to light, which result in metallic-to-fishy, oily, and cardboard-like flavors. The importance of each varies with the method of handling raw milk, the type of feed the cow eats and her physiological response to the feed consumed, the type of product held, and processing and storage conditions. Although we are still in the first stage of solving the problem, several methods are available for producing palatable and nutritious milk and milk products.
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I I I I I I C O O L E D R A P 1 D L Y O V E R T H E S U R F A C E C O O L E R T O 0-5'C. C O O L E D S L O W L Y I N W A T E R A T 0-5OC
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Figure 1.
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Activation of milk lipase b y temperature changes Holding time 24 hours
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Figure 2. Effects of temperature of storage on rates of lipolysis in milk with natural-surfaced and resurfaced milk f a t globules Holding time 2 4 hours
(28, 29). hIaximum activation occurred when rau milk originally below 10' C. was \varmed to 30" C. and then slowly recooled to below 10' C. (Figure 1). Milk so treated became unfit to drink. sometimes shortly after milking.
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AGRICULTURALAND
This could be prevented by cooling both the fresh and the temperature-activated milk rapidly to below IO' C . and holding it there until pasteurization; this caused the enzyme to be reversibly inactivated. Lipase can he inactivated irreversibly FOOD CHEMISTRY
and completely by heating milk at 58' to 63' C. for 30 minutes in the presence of occluded oxygen (35);by rennet and acids a t the isoelectric point of casein (20): and by preformed H?Ou added in amounts needed to oxidize all of the ascorbic acid in the milk to dehydroascorbic acid (27). Splitting of milk fat occurred also when the raw milk \vas allo\i-ed to churn and the fat to resurface on shaking or foaming (20, 33). T h e lipolysis of the milk-fat globules resurfaced by churning of cream and the re-emulsification of pure fat in ra\v skim milk, as \vel1 as of triolein and trielaidin substrates, proceeded faster, hojvever: as the incubation temperature increased (Figure 2), shoiving a "positive temperature coefficient" ( 3 4 : \\hereas the lipolysis of natural-surfaced milk-fat globules in temperature-activated milk accelerated as the temperature decreased, showing a "negative temperature coefficient." Triolein and trielaidin \
