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Spectrophotometric Method for Determination of Heat-Activated Sulfhydryl Groups of Skimmilk¹

Department of Dairy Technology, The Ohio State University, Columbus

ABSTRACT

By suitable modification, adaptation was made of the 5,5'-dithiobis (2-nitrobenzoic acid) reagent method for the spectrophotometric determination of activated sulfhydryl (-SH) groups of heated skimmilk. The modifications consisted of using a nitrogen-purged system, controlling pH, adding ammonium sulfate, filtering, and determining absorbance of the filtrate at 412 mµ. A standard cysteine-HCl curve was prepared in a similar manner. The modified procedure yielded results consistent with other currently used methods. The activation of -SH groups in heated skimmilk, as determined by the modified procedure, followed the kinetics of a first order reaction with reaction rate constants (k_min-1) of 0.078, 0.142, 0.384, and 0.805 for 75, 80, 85, and 90 C, respectively.

¹ Article 5:67. Department of Dairy Technology, The Ohio State University. This investigation was supported by Public Health Services Research Grant EF-00180 from the Division of Environmental Engineering and Food Protection.

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