Clinical Chemistry, Vol 34, 1077-1083, Copyright © 1988 by American Association for Clinical Chemistry

Solid-phase extraction with strong anion-exchange columns for selective isolation and concentration of urinary organic acids

BJ Verhaeghe, MF Lefevere and AP De Leenheer
Laboratoria voor Medische Biochemie en voor Klinische Analyse, Gent, Belgium.

This is a new method for isolating and concentrating urinary organic acids before gas chromatography--mass spectrometry. Sulfate and phosphate anions are removed by precipitation with Ba(OH)2, and the urine pH is adjusted to 8-8.5. The sample is then applied onto small, disposable, strong-anion-exchange columns. Neutral and basic compounds are washed out with water, which is then removed by centrifugation and by rinsing the columns with methanol and diethyl ether. The organic acids are eluted with a 4/1 (by vol) mixture of an organic solvent-- either n-butanol or ethyl acetate--and formic acid containing HSO4- (0.1 mol/L) as a highly selective counter-ion, and finally with methanol alone. Sulfate ions are retained, and the eluate is evaporated before trimethylsilyl derivatization. Analytical recoveries (about 100%) of organic acids compare favorably with those obtained with solvent extraction. This convenient procedure is selective and reproducible and is a suitable alternative to the more cumbersome diethylaminoethyl-Sephadex extraction method.

The following articles in journals at HighWire Press have cited this article:

				J. M. Halket, D. Waterman, A. M. Przyborowska, R. K. P. Patel, P. D. Fraser, and P. M. Bramley Chemical derivatization and mass spectral libraries in metabolic profiling by GC/MS and LC/MS/MS J. Exp. Bot., January 1, 2005; 56(410): 219 - 243. [Abstract] [Full Text] [PDF]

				J. W. Suh, S. H. Lee, and B. C. Chung GC-MS determination of organic acids with solvent extraction after cation-exchange chromatography Clin. Chem., December 1, 1997; 43(12): 2256 - 2261. [Abstract] [Full Text] [PDF]