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Quantitative analysis for organic acids in biological samples: batch isolation followed by gas chromatographic-mass spectrometric analysis.

Clinical chemistry (1989-04-01)
G Hoffmann, S Aramaki, E Blum-Hoffmann, W L Nyhan, L Sweetman
ABSTRACT

This new method for qualitative and quantitative determination of organic acids, aldehydes, and ketones in biological samples is effective for use with urine, plasma, and amniotic fluid, and it requires no deproteinization. Isolation by batch-wise liquid partition chromatography on silicic acid follows formation of the O-(2,3,4,5,6-pentafluorobenzyl)oximes of oxoacids, aldehydes, and ketones. The total organic acid content of the sample provides a rapid screening test for metabolic abnormality. A wide-bore, bonded-phase capillary column was used for quantitative gas chromatographic-mass spectrometric analysis, followed by automated identification and quantification. Analytical recoveries were quantitative for a wide variety of metabolites. Gas-chromatographic retention indices, discriminating ions, and control ranges in amniotic fluid, plasma, and urine of adult subjects were determined for 61 biologically important compounds.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Citric acid, ACS reagent, ≥99.5%
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Oleic acid, ≥99% (GC)
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Oleic acid, technical grade, 90%
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Citric acid, 99%
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L-(+)-Lactic acid, ≥98%
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Glycolic acid, ReagentPlus®, 99%
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Stearic acid, Grade I, ≥98.5% (capillary GC)
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L-(+)-Tartaric acid, ACS reagent, ≥99.5%
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Fumaric acid, ≥99.0% (T)
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Adipic acid, 99%
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Oleic acid, suitable for cell culture, BioReagent
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α-Ketoglutaric acid, 99.0-101.0% (T)
Supelco
Benzoic acid, Pharmaceutical Secondary Standard; Certified Reference Material
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4-Hydroxybenzoic acid, ReagentPlus®, ≥99%
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Stearic acid, reagent grade, 95%
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Melting point standard 121-123°C, analytical standard
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Mettler-Toledo Calibration substance ME 18555, Benzoic acid, analytical standard, for the calibration of the thermosystem 900, traceable to primary standards (LGC)
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Supelco
Citric acid, Anhydrous, Pharmaceutical Secondary Standard; Certified Reference Material
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Palmitic acid, BioXtra, ≥99%
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Stearic acid, analytical standard
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Succinic acid, ACS reagent, ≥99.0%
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L-(−)-Malic acid, ≥95% (titration)
Sigma-Aldrich
Glutaric acid, 99%

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