Validation of a method for ethanol analysis in biological and non-biological samples and its toxicological application Technical paper
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Abstract
A simple, cost-effective and fast headspace gas chromatography method coupled with flame ionization detection (HS-GC/FID) for determination of ethanol was developed and validated for clinical and forensic toxicology purposes. HS-GC/FID is often used for alcohol determination in different biological and non-biological samples. The calibration dependence of the method was linear in the range from 0.15 to 4.00 g dm-3 (r2=0.999) with adequate accuracy (99–106 %) and precision. The limit of detection (LOD) was 0.006 g dm-3. The method was quantitative (LOQ) above 0.020 g dm-3. The new method was successfully used for determination of ethanol in biological samples of intoxicated patients, car accidents participants, participants in criminal acts, and postmortem samples, non-biological samples such as alcoholic beverages, alcohol-based herbal preparations, cosmetic preparations, etc. This method is easy to perform, making it suitable not only for the routine applications in clinical biochemistry and forensic laboratories, but also in different fields of industry (e.g. for pharmaceutical preparations, cosmetics, dietary supplements, etc.). Some of the applications for ethanol determination in different samples related to various clinical-forensic cases are presented.
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References
Jones AW. Alcohol, its analysis in blood and breath for forensic purposes, impairment effects, and acute toxicity. WIREs: Forensic Science. 2019; 1(6): e1353. https://doi.org/10.1002/wfs2.1353
Tangerman A. Highly sensitive gas chromatographic analysis of ethanol in whole blood, serum, urine, and fecal supernatants by the direct injection method. Clin Chem. 1997; 43(6): 1003–1009. https://doi.org/10.1093/clinchem/43.6.1003
Zuba D, Parczewski A, Reichenbächer M. Optimization of solid-phase microextraction conditions for gas chromatographic determination of ethanol and other volatile compounds in blood. J Chromatogr B Analyt Technol Biomed Life Sci. 2002; 773(1): 75-82. https://doi.org/10.1016/s1570-0232(02)00143-5
Kolb B, Ettre LS. Static Headspace–Gas Chromatography. 2006. New York, NY:John Wiley & Sons. https://doi.org/10.1002/0471914584
Dubowski KM. Alcohol Determination in the Clinical Laboratory. Am J Clin Pathol. 1980; 74(5): 747-750. https://doi.org/10.1093/ajcp/74.5.747
ONeal CL, Wolf II CE, Levine B, Kunsman G, Poklis A. Gas chromatographic procedures for determination of ethanol in postmortem blood using t-butanol and methyl ethyl ketone as internal standards. Forensic Sci Int. 1996; 83(1): 31-38. https://doi.org/10.1016/0379-0738(96)02007-5
Snow NH, Slack GC. Head-space analysis in modern gas chromatography. TrAC Trends Anal Chem. 2002; 21(9–10): 608–617. https://doi.org/10.1016/s0165-9936(02)00802-6
Dorubet D, Moldoveanu S, Mircea C, Butnaru E, Astarastoae V. Development and validation of a quantitative determination method of blood ethanol by gas chromatography with headspace (GC-HS). Rom J Leg Med. 2009; 17(4): 303–308. https://doi.org/10.4323/rjlm.2009.303
De Martinis BS, Ruzzene MAM, Martin CCS. Determination of ethanol in human blood and urine by automated headspace solid-phase microextraction and capillary gas chromatography. Anal Chim Acta. 2004; 522(2): 163-168. https://doi.org/10.1016/j.aca.2004.07.007
Chun H-J, Poklis JL, Poklis A, Wolf CE. Development and Validation of a Method for Alcohol Analysis in Brain Tissue by Headspace Gas Chromatography with Flame Ionization Detector. J Anal Toxicol. 2016; 40(8): 653–658. https://doi.org/10.1093/jat/bkw075
Wasfi IA, Al-Awadhi AH, Al-Hatali ZN, Al-Rayami FJ, Al Katheeri NA. Rapid and sensitive static headspace gas chromatography–mass spectrometry method for the analysis of ethanol and abused inhalants in blood. J Chromatogr B Analyt Technol Biomed Life Sci. 2004; 799(2): 331-336. https://doi.org/10.1016/j.jchromb.2003.11.003
Lachenmeier DW. Rapid quality control of spirit drinks and beer using multivariate data analysis of Fourier transform infrared spectra. Food Chem. 2007; 101(2): 825-832. https://doi.org/10.1016/j.foodchem.2005.12.032
Betz JM, Nikelly JG. Determination of Ethanol In Alcoholic Beverages by Liquid Chromatography Using the UV Detector. J Chromatogr Sci. 1987; 25(9): 391-394. https://doi.org/10.1093/chromsci/25.9.391
Yarita T, Nakajima R, Otsuka S, Ihara T, Takatsu A, Shibukawa M. Determination of ethanol in alcoholic beverages by high-performance liquid chromatography–flame ionization detection using pure water as mobile phase. J Chromatogr A. 2002; 976(1-2): 387-391. https://doi.org/10.1016/s0021-9673(02)00942-1
Westland JL, Dorman FL. Comparison of SPME and static headspace analysis of blood alcohol concentration utilizing two novel chromatographic stationary phases. Forensic Sci Int. 2013; 231(1-3): e50-e56. https://doi.org/10.1016/j.forsciint.2013.05.007
Corrêa CL, Pedroso RC. Headspace gas chromatography with capillary column for urine alcohol determination. J Chromatogr B Biomed Sci Appl. 1997; 704(1-2): 365-368. https://doi.org/10.1016/s0378-4347(97)00445-3
Morris-Kukoski CL, Jagerdeo E, Schaff JE, LeBeau MA. Ethanol analysis from biological samples by dual rail robotic autosampler. J Chromatogr B. 2007; 850(1-2): 230-235. https://doi.org/10.1016/j.jchromb.2006.11.034
Bursová M, Hložek T, Čabala R. Simultaneous Determination of Methanol, Ethanol and Formic Acid in Serum and Urine by Headspace GC-FID. J Anal Toxicol. 2015; 39(9): 741-745. https://doi.org/10.1093/jat/bkv075
Scientific Working Group for Forensic Toxicology (SWGTOX) Standard Practices for Method Validation in Forensic Toxicology. J Anal Toxicol. 2013; 37(7): 452–474. https://doi.org/10.1093/jat/bkt054
Monteiro C, Franco JM, Proença P, Castañera A, Claro A, Vieira DN, Corte-Real F. Qualitative and quantitative analysis of a group of volatile organic compounds in biological samples by HS-GC/FID: application in practical cases. Forensic Sci Int. 2014; 243:137-43. https://doi.org/10.1016/j.forsciint.2014.07.016.
Tiscione N, Alford I, Yeatman DT, Xiaoqin Shan X. Ethanol Analysis by Headspace Gas Chromatography with Simultaneous Flame-Ionization and Mass Spectrometry Detection, J Anal Toxicol. 2017; 35(7):501–511. https://doi.org/10.1093/anatox/35.7.501
Borman P, Elder D. Q2(R1) Validation of Analytical Procedures. ICH Quality Guidelines. 2017; 127–166. https://doi.org/10.1002/9781118971147.ch5
Mihretu LM, Gebru AG, Mekonnen KN, Asgedom AG, Desta YH. Determination of ethanol in blood using headspace gas chromatography with flameionization detector (HS-GC-FID): Validation of a method, Cogent Chem. 2020; 6:1 https://doi.org/10.1080/23312009.2020.1760187
Bursová M, Hložek T, Čabala R. Simultaneous Determination of Methanol, Ethanol and Formic Acid in Serum and Urine by Headspace GC-FID. J Anal Toxicol. 2015; 39(9):741-5. https://doi.org/10.1093/jat/bkv075
Schlatter J, Chiadmi F, Gandon V, Chariot P. Simultaneous determination of methanol, acetaldehyde, acetone, and ethanol in human blood by gas chromatography with flame ionization detection. Hum Exp Toxicol. 2014;33(1):74-80. https://doi.org/10.1177/0960327113482845
Neo MS, Gupta SM, Khan TM, Gupta M. Quantification of Ethanol Content in Traditional Herbal Cough Syrups. Pharmacogn J. 2017; 9(6): 821-827. https://doi.org/10.5530/pj.2017.6.128