Determination of Sulfhydryl Groups in Milk Using Ellman's Procedure and Clarifying Reagent®

Abstract This work reports on a rapid method for determination of total sulfhydryl groups in raw and heated milks using 5,5'-dithio-bis(2-nitrobenzoic acid). The reaction medium is clarified after the chemical reaction by addition of a dissolving agent; thus, the spectrophotometric measurements are performed directly in milk without other sample treatment. This protocol correlated well with amperometric titrations (r=.994). Results on raw milks from individual cows showed seasonal variations in sulfhydryl groups. Sulfhydryl groups decreased in heated commercial milks. This method is simple, rapid, and easily applicable in routine measurements and may allow heated commercial milks to be distinguished.

[1]  G. Humbert,et al.  Measurement of proteolysis in milk and cheese using trinitrobenzene sulphonic acid and a new dissolving reagent , 1990, Journal of Dairy Research.

[2]  D. Irvine,et al.  Heat-Induced Protein Changes in Milk Processed by Vat and Continuous Heating Systems , 1988 .

[3]  A. Owen,et al.  A procedure for the complete clarification of milk of various species and its suitability for use with colorimetric measurements , 1984, Journal of Dairy Research.

[4]  H A Scheraga,et al.  Sensitive quantitative analysis of disulfide bonds in polypeptides and proteins. , 1984, Analytical biochemistry.

[5]  R L Blakeley,et al.  Ellman's reagent: 5,5'-dithiobis(2-nitrobenzoic acid)--a reexamination. , 1979, Analytical biochemistry.

[6]  H. Swaisgood,et al.  Sulfhydryl and Disulfide Groups in Skim Milk as Affected by Direct Ultra-High-Temperature Heating and Subsequent Storage , 1976 .

[7]  M. Kaláb Factors Affecting the Ellman Determination of Sulfhydryl Groups in Skimmilk Powder and Gels , 1970 .

[8]  D. G. Vakaleris,et al.  Effects of Heat on Sulfhydryl and Disulfide Groups of Milk Proteins as Measured by the Spectrofluorometric Method , 1968 .

[9]  I. A. Gould,et al.  Spectrophotometric Method for Determination of Heat-Activated Sulfhydryl Groups of Skimmilk , 1968 .

[10]  R. Lyster The free and masked sulphydryl groups of heated milk and milk powder and a new method for their determination , 1964, Journal of Dairy Research.

[11]  E. O. Herreid,et al.  Amperometric Titration of Sulfhydryl and Disulfide Groups in Milk Proteins , 1962 .

[12]  G. Ellman A colorimetric method for determining low concentrations of mercaptans. , 1958, Archives of biochemistry and biophysics.

[13]  S. Patton,et al.  The Origin of Sulfhydryl Groups in Milk Proteins and their Contributions to “Cooked” Flavor , 1952 .

[14]  J. Ackland,et al.  Effect of preheat treatment and storage on the properties of whole milk powder. Changes in physical and chemical properties , 1991 .

[15]  H. Kessler,et al.  Reaction kinetics evaluation of the oxidative changes in stored UHT milk , 1986 .

[16]  H. Kessler,et al.  The effect of head space volume on the quality of stored degassed UHT milk , 1986 .

[17]  H. Klostermeyer,et al.  Milcherhitzung und SH-Gruppenentwicklung. III , 1984 .

[18]  H. Klostermeyer,et al.  On the influence of SH-groups in UHT milk during storage , 1984 .

[19]  S. Nakai,et al.  DETERMINATION OF SH‐ AND SS‐GROUPS IN SOME FOOD PROTEINS USING ELLMAN'S REAGENT , 1974 .