By John N. Abelson, Melvin I. Simon, John R. Sokatch, Robert Adron Harris
This quantity comprises chapters at the most up-to-date thoughts for measuring metabolites of branched-chain amino acids, many new enzyme assays equivalent to the size of isozymes of acetohydroxacid synthase, and the purification of lately defined enzymes equivalent to isovaleryl-CoA dehydrogenase
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Extra info for Branched-Chain Amino Acids
273, 245 (1983). l0 G. Liveseyand W. T. E. Edwards, J. Chromatogr. 337, 98 (1985). METHODS IN ENZYMOLOGY, VOL. 166 Copyright © 1988 by Academic Press, Inc. All rights of reproduction in any form reserved.  QUANTITATIVE H P L C OF KETO ACIDS 21 Principles of the Method The proteins of blood and plasma are precipitated with acid and the solution is purified by retention of the keto acids on a hydrazide gel. The hydrazones are converted anaerobically to the quinoxalinols with ophenylenediamine which are then separated and quantified by HPLC using a reversed-phase column and a fluorescence detector.
Data were processed by a Pye Unicam Ltd. data control center. The optimum excitation and emission wavelengths given were obtained by scanning the sample in the detector cell under stopped-flow conditions. Procedure The dried quinoxalinols are dissolved in dimethylformamide (40 gl) and water (100 #l) and aliquots (20/zl) injected onto the column. A concave gradient (Perkin-Elmer Code 2) is used to change the mobile-phase composition from 20 to 80% solution A over 35 rain for plasma or whole blood extracts.
All rights of reproduction in any form reserved.  QUANTITATIVE H P L C OF KETO ACIDS 21 Principles of the Method The proteins of blood and plasma are precipitated with acid and the solution is purified by retention of the keto acids on a hydrazide gel. The hydrazones are converted anaerobically to the quinoxalinols with ophenylenediamine which are then separated and quantified by HPLC using a reversed-phase column and a fluorescence detector. Gel - CONH- NH 2 + R. 3 with NaOH 'The preparative procedure is based on the method of Hayashi et al.
Branched-Chain Amino Acids by John N. Abelson, Melvin I. Simon, John R. Sokatch, Robert Adron Harris