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A procedure for the complete clarification of milk of various species and its suitability for use with colorimetric measurements

Published online by Cambridge University Press:  01 June 2009

A. Jane Owen
Affiliation:
*National Institute for Research in Dairying (University of Reading), Shinfield, Reading RG2 9 AT, UK
Anthony T. Andrews
Affiliation:
*National Institute for Research in Dairying (University of Reading), Shinfield, Reading RG2 9 AT, UK

Summary

Bovine, caprine, ovine and human milks have been clarified by addition of Triton X-100 and EDTA followed by a short incubation. Generally, addition of 4·2% Triton X–100 and 5·4% EDTA was sufficient and with skim-milk samples no dilution was necessary. With whole milks some dilution and greater amounts of Triton X-100 or longer incubation times or higher temperatures were required. These factors were interrelated to some extent and the optimum conditions will be determined by convenience and any particular requirements of the experiment. The attainment of optically transparent milk samples greatly facilitates colorimetric measurements; as examples we have shown that total protein may be estimated by the biuret reaction, NH2 groups measured with 2,4,6-trinitrobenzene sulphonic acid and SH groups by reaction with 5, 5′-dithiobis(2-nitrobenzoic acid). Acid phosphatase activity in bovine milk was followed colorimetrically by reaction with p-nitrophenyl phosphate to demonstrate the suitability of the clarification procedure in the measurement of enzymic reactions.

Type
Original Articles
Copyright
Copyright © Proprietors of Journal of Dairy Research 1984

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References

REFERENCES

Andrews, A. T. 1976 Bovine milk acid phosphatase. III. Purification and characterisation of the enzyme. Biochimica et Biophysica Acta 434 345353CrossRefGoogle ScholarPubMed
Folch, J., Lees, M. & Sloane Stanley, G. H. 1957 A simple method for the isolation and purification of total lipides from animal tissues. Journal of Biological Chemistry 226 497509CrossRefGoogle ScholarPubMed
Gornall, A. G., Bardawill, C. J. & David, M. M. 1949 Determination of serum proteins by means of the biuret reaction. Journal of Biological Chemistry 177 751766CrossRefGoogle ScholarPubMed
Linden, G., Humbert, G., Desnouveaux, R. & Picard, J. 1982 [Applications of total dissolution of milk to the determination of enzyme activities]. Lait 62 209219CrossRefGoogle Scholar
Linden, G. & Paquet, D. 1981 Alkaline phosphase activity in transparent milk and cream. Journal of Dair Science 64 568571CrossRefGoogle Scholar
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. 1951 Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193 265275CrossRefGoogle ScholarPubMed
Nakai, S. & Le, A. C. 1970 Spectrophotometric determination of protein and fat in milk simultaneously. Journal of Dairy Science 53 276278CrossRefGoogle Scholar
Nakai, S., Wilson, H. K. & Herreid, O. 1964 Spectrophotometric determination of protein in milk. Journal of Dairy Science 47 356358CrossRefGoogle Scholar