By P. Alexander, H. P. Lundgren
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Extra info for A Laboratory Manual of Analytical Methods of Protein Chemistry. Volume 4
This should not be necessary more frequently than every two months. MeHgl is made by exposing metallic mercury (10 g) and methyl iodide (20 g) in a spring-loaded glass-stoppered Pyrexflaskto sunlight for 3-4 weeks. After a variable induction period, the mixture solidifies to a yellow mass which is ground up*, then extracted from a Soxhlet thimble, using boiling MeOH. On cooling each consecutive extract, pale yellow needles are deposited Attempts to speed up the synthesis by use of ultraviolet lamps and quartz reaction vessels have not been successful in the hands of the reviewer.
Mercuribenzoate (Section IV, A. 3) which reacts further and more rapidly with protein —SH groups at pH 4-6 than at 7. If the mercurial absorbs strongly in the ultraviolet or visible, or gives a well-defined current plateau in a polarograph, the reaction is best carried out in a spectrophotometer cuvette or a polarographic cell, so that the rate of reaction may be measured and recorded automatically. This is a more sound procedure than the usual titration method described above since the results are then obtained in the form of a continuous spectrum of —SH reactivities and can be used for calculating rate constants for each —SH group involved.
28 ANALYTICAL METHODS OF PROTEIN CHEMISTRY When complete amino acid analysis is not to be carried out and the hydrolysate is not therefore fractionated by column chromatography, there are two sound methods available for cystine analysis of the amino acid mixture, viz. 3). The former is the method of choice where coloured hydrolysates are to be analysed. It also has the advantage that all of the —SS— groups become available for reaction even after short times of acid hydrolysis (2 hr) at 105° so that extensive destruction due to prolonged hydrolysis may be avoided.