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Paul Van hee, Hugo Neels, Mireille De Doncker, Nicolas Vrydags, Katinka Schatteman, Wim Uyttenbroeck, Nicole Hamers, Dirk Himpe, Willy Lambert
Analysis of ?-hydroxybutyric acid, DL-lactic acid, glycolic acid, ethylene glycol and other glycols in body fluids by a direct injection gas chromatography-mass spectrometry assay for wide use
Keywords: bis-N,O-trimethylsilyl trifluoroacetamide (BSTFA) derivatization, gas chromatography-mass spectrometry, glycols, hydroxy acids
Analysis of blood of severely intoxicated patients always requires prompt investigation. Diagnosis of intoxication with ethylene glycol, ?-hydroxybutyric acid or D-lactic acid takes hours, since several different procedures are required. Rapid derivatization of the common hydroxyl function may resolve this analytical problem.
Here we describe a fast method for the simultaneous measurement of ethylene glycol, glycolic acid, ?-hydroxybutyric acid and racemic lactic acid. Only 20 µl of serum, plasma or urine are required for immediate derivatization at 70°C with 750 µl of bis-N,O-trimethylsilyl trifluoroacetamide after adding 20 µl of internal standard solution (1,3-propylene glycol) and 20 µl of the catalyst dimethylformamide. After centrifugation an aliquot is transferred to a gas chromatographic system and analyzed with electron-impact mass spectrometry in selective ion monitoring mode.
The derivatized acids and ethylene glycol are well separated and detected with a limit of detection ranging from 0.12 mg/l for ethylene glycol to 0.95 mg/l for ?-hydroxybutyric acid, while the limit of quantification ranged from 0.4 mg/l for ethylene glycol to 3.15 mg/l for ?-hydroxybutyric acid. The method is linear from 0.5 to 1800 mg/l blood for ethylene glycol, from 0.7 to 1200 mg/l for lactic acid, from 1.2 to 1800 mg/l for glycolic acid, and from 3.2 to 200 mg/l for ?-hydroxybutyric acid, with analytical recoveries, accuracy, day-to-day and within-day precision well within the required limits. Total analysis time with one calibrator was 30 min, derivatization time included.
This method is very suitable for emergency toxicology, since several toxic substances can be quantified simultaneously in a fast and sensitive manner.
Clinical Chemical Laboratory Medicine, Walter de Gruyter
Print ISSN: 1434-6621
Volume: 42, 11/2004
Pages: 1341 - 1345