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Gas Chromatographic Resolution of Pyruvic, Lactic, Glyoxylic, Oxalic, Malonic, Fumaric, Malic, Alphaketoglutaric, and Citric Acids in Forage and Rumen Fluid^{1, 2,}

Departments of Animal Sciences and Agronomy Purdue University, Lafayette, Indiana

ABSTRACT

A number of procedures (1–6) have been reported for the resolution of esters of some TCA cycle and other related organic acids by gas-liquid chromatography (GLC). However, difficulties in their isolation and separation have been encountered. Keto acids were found to be difficult to extract and methylate because of their instability (4), while others decomposed when chromatographed (1).

This paper presents a procedure for the qualitative analyses of the methyl esters of pyruvic, lactic, glyoxylic, oxalic, malonic, fumaric, succinic, malic, -ketoglutaric, and citric acids. A. procedure is also presented for preparation of forage and rumen fluid samples for GLC analyses.

APPARATUS AND PROCEDURE

An Aerograph Hy-Fi gas chromatograph³ equipped with a gold-plated hydrogen flame ionization detector and a Speedomax H Leeds and Northrup recorder⁴ were used. Hydrogen gas for the detector was supplied from a model A-650 hydrogen generator³ and a commercial source of high-purity nitrogen was used as the carrier gas.

Column preparation.

¹ Published with the approval of the Director of the Indiana Agricultural Experiment Station as Journal Series Paper No. 2310.

² This research was financed in part from a trust agreement between Purdue University and Normandy Farm, New Augusta, Indiana, Mr. and Mrs. Herman C. Krannert, owners.