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Licensed Unlicensed Requires Authentication Published by De Gruyter September 21, 2011

Analysis of γ-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

  • Paul Van hee , Hugo Neels , Mireille De Doncker , Nicolas Vrydags , Katinka Schatteman , Wim Uyttenbroeck , Nicole Hamers , Dirk Himpe and Willy Lambert


Analysis of blood of severely intoxicated patients always requires prompt investigation. Diagnosis of intoxication with ethylene glycol, γ-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, γ-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 γ-hydroxybutyric acid, while the limit of quantification ranged from 0.4 mg/l for ethylene glycol to 3.15 mg/l for γ-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 γ-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.

Corresponding author: Dr. Hugo Neels, Laboratory of Biochemistry and Toxicology, Ziekenhuis Netwerk Antwerpen Stuivenberg, Lange Beeldekensstraat 267, 2060 Antwerp, Belgium. Phone: +32-3-217-7804, Fax: +32-3-217-7358, E-mail:


1 Porter WH, Auansakul A. Gas-chromatographic determination of ethylene glycol in serum. Clin Chem 1982; 28:75–8.10.1093/clinchem/28.1.75Search in Google Scholar

2 Robinson DW, Reeve DS. A gas chromatographic procedure for quantitation of ethylene glycol in postmortem blood. J Anal Toxicol 1981; 5:69–72.10.1093/jat/5.2.69Search in Google Scholar

3 Horton H, Solomons ET. An improved procedure for the determination of ethylene glycol in blood. J Anal Toxicol 1982; 6:253–4.10.1093/jat/6.5.253Search in Google Scholar

4 Fraser AF. Clinical toxicologic implications of ethylene glycol and glycolic acid poisoning. Ther Drug Monit 2002; 24:232–8.10.1097/00007691-200204000-00005Search in Google Scholar

5 Porter WH, Rutter PW, Bush BA, Pappas AA, Dunnington JE. Ethylene glycol toxicity: the role of serum glycolic acid in hemodialysis. J Toxicol Clin Toxicol 2001; 39:607–15.10.1081/CLT-100108493Search in Google Scholar

6 Uribarri J, Oh MS, Carroll HJ. D-Lactic acidosis. A review of clinical presentation, biochemical features, and pathophysiologic mechanisms. Medicine 1998; 77:73–82.10.1097/00005792-199803000-00001Search in Google Scholar

7 Pan YM, Gill GN, Wall CH, McCurdy HH. Improved procedure for the analysis of gamma-hydroxybutyrate and ethylene glycol in whole blood. J Anal Toxicol 2001; 25:328–32.10.1093/jat/25.5.328Search in Google Scholar

8 Elian AA. A novel method for GHB detection in urine and its application in drug-facilitated sexual assaults. Forensic Sci Int 2000; 109:183–7.10.1016/S0379-0738(99)00221-2Search in Google Scholar

9 Elian AA. GC-MS determination of gamma-hydroxybutyric acid (GHB) in blood. Forensic Sci Int 2001; 122:43–7.10.1016/S0379-0738(01)00441-8Search in Google Scholar

10 Gembus V, Goullé JP, Lacroix C. Determination of glycols in biological specimens by gas chromatography-mass spectrometry. J Anal Toxicol 2002; 26:280–5.10.1093/jat/26.5.280Search in Google Scholar PubMed

11 Goullé JP, Gembus V, Lacroix C. Dosage simultané de huit glycols dans les milieux biologiques par chromatographie en phase gazeuse couple´e a` la spectrometrie de masse. Ann Toxicol Anal 2001; 13:247–52.10.1051/ata/2001022Search in Google Scholar

12 Van hee P, Dedoncker M, Vrydags N, Schatteman K, Uyttenbroeck W, Lambert W, et al. Comment on determination of glycols in biological specimens by gas chromatography-mass spectrometry. J Anal Toxicol 2003; 27:173–4.10.1093/jat/27.3.173Search in Google Scholar

13 Maurer HH, Peters FT, Paul LD, Kraemer T. Validated gas chromatographic-mass spectrometric assay for determination of the antifreezes ethylene glycol and diethylene glycol in human plasma after microwave-assisted pivalylation. J Chromatogr B Biomed Sci Appl 2001; 754:401–9.10.1016/S0378-4347(01)00022-6Search in Google Scholar

14 Baffi P, Elsener S, Baffi M, De Melin M. Quantitative determination of diethylene glycol contamination in pharmaceutical products. J AOAC Int 2000; 83:793–801.10.1093/jaoac/83.4.793Search in Google Scholar

15 Williams RH, Shah SM, Maggiore JA, Erickson TB. Simultaneous detection and quantitation of diethylene glycol, ethylene glycol, and the toxic alcohols in serum using capillary column gas chromatography. J Anal Toxicol 2000; 24:621–6.10.1093/jat/24.7.621Search in Google Scholar

16 Lora-Tamayo C, Tena T, Rodriguez A, Sancho JR, Molina E. Intoxication due to 1,4–butanediol. Forensic Sci Int 2003; 133:256–9.10.1016/S0379-0738(02)00424-3Search in Google Scholar

17 Elliott SP. Gamma hydroxybutyric acid (GHB) concentrations in humans and factors affecting endogenous production. Forensic Sci Int 2003; 133:9–16.10.1016/S0379-0738(03)00043-4Search in Google Scholar

Published Online: 2011-9-21
Published in Print: 2004-11-1

© Walter de Gruyter

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