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Quantitative analysis of cocaine and its metabolites in whole blood and urine by high-performance liquid chromatography coupled with tandem mass spectrometry.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jun 01; 852(1-2):338-44.JC

Abstract

AIM

In forensic toxicology it is important to have specific and sensitive analysis for quantification of illicit drugs in biological matrices. This paper describes a quantitative method for determination of cocaine and its major metabolites (ecgonine methyl ester, benzoylecgonine, norcocaine and ethylene cocaine) in whole blood and urine by liquid chromatography coupled with tandem mass spectrometry LC/MS/MS.

METHOD

The sample pre-treatment (0.20 g) consisted of acid precipitation, followed by centrifugation and solid phase extraction of supernatant using mixed mode sorbent columns (SPEC MP1 Ansys Diag. Inc.). Chromatographic separation was performed at 30 degrees C on a reverse phase Zorbax C18 column with a gradient system consisting of formic acid, water and acetonitrile. The analysis was performed by positive electrospray ionisation with a triple quadropole mass spectrometer operating in multiple reaction monitoring (MRM) mode. Two MRM transitions of each analyte were established and identification criteria were set up based on the retention time and the ion ratio. The quantification was performed using deuterated internal analytes of cocaine, benzoylecgonine and ecgonine methyl ester. The calibration curves of extracted standards were linear over a working range of 0.001-2.00 mg/kg whole blood for all analytes. The limit of quantification was 0.008 mg/kg; the interday precision (measured by relative standard deviation-%RSD) was less than 10% and the accuracy (BIAS) less than 12% for all analytes in whole blood. Urine samples were estimated semi-quantitatively at a cut-off level of 0.15 mg/kg with an interday precision of 15%.

CONCLUSION

A liquid chromatography mass spectrometric (LC/MS/MS) method has been developed for confirmation and quantification of cocaine and its metabolites (ecgonine methyl ester, benzoylecgonine, norcocaine and ethylene cocaine) in whole blood and semi-quantitative in urine. The method is specific and sensitive and offers thereby an excellent alternative to other methods such as GC-MS that involves derivatisation.

Authors+Show Affiliations

Department of Forensic Chemistry, Institute of Forensic Medicine, University of Copenhagen, Frederik V's vej 11, DK-2100 Copenhagen OE, Denmark. SSJ@forensic.ku.dkNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17301000

Citation

Johansen, Sys Stybe, and Helle Merete Bhatia. "Quantitative Analysis of Cocaine and Its Metabolites in Whole Blood and Urine By High-performance Liquid Chromatography Coupled With Tandem Mass Spectrometry." Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, vol. 852, no. 1-2, 2007, pp. 338-44.
Johansen SS, Bhatia HM. Quantitative analysis of cocaine and its metabolites in whole blood and urine by high-performance liquid chromatography coupled with tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;852(1-2):338-44.
Johansen, S. S., & Bhatia, H. M. (2007). Quantitative analysis of cocaine and its metabolites in whole blood and urine by high-performance liquid chromatography coupled with tandem mass spectrometry. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 852(1-2), 338-44.
Johansen SS, Bhatia HM. Quantitative Analysis of Cocaine and Its Metabolites in Whole Blood and Urine By High-performance Liquid Chromatography Coupled With Tandem Mass Spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jun 1;852(1-2):338-44. PubMed PMID: 17301000.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Quantitative analysis of cocaine and its metabolites in whole blood and urine by high-performance liquid chromatography coupled with tandem mass spectrometry. AU - Johansen,Sys Stybe, AU - Bhatia,Helle Merete, Y1 - 2007/02/01/ PY - 2006/07/13/received PY - 2006/12/19/revised PY - 2007/01/22/accepted PY - 2007/2/16/pubmed PY - 2007/8/2/medline PY - 2007/2/16/entrez SP - 338 EP - 44 JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences JO - J Chromatogr B Analyt Technol Biomed Life Sci VL - 852 IS - 1-2 N2 - AIM: In forensic toxicology it is important to have specific and sensitive analysis for quantification of illicit drugs in biological matrices. This paper describes a quantitative method for determination of cocaine and its major metabolites (ecgonine methyl ester, benzoylecgonine, norcocaine and ethylene cocaine) in whole blood and urine by liquid chromatography coupled with tandem mass spectrometry LC/MS/MS. METHOD: The sample pre-treatment (0.20 g) consisted of acid precipitation, followed by centrifugation and solid phase extraction of supernatant using mixed mode sorbent columns (SPEC MP1 Ansys Diag. Inc.). Chromatographic separation was performed at 30 degrees C on a reverse phase Zorbax C18 column with a gradient system consisting of formic acid, water and acetonitrile. The analysis was performed by positive electrospray ionisation with a triple quadropole mass spectrometer operating in multiple reaction monitoring (MRM) mode. Two MRM transitions of each analyte were established and identification criteria were set up based on the retention time and the ion ratio. The quantification was performed using deuterated internal analytes of cocaine, benzoylecgonine and ecgonine methyl ester. The calibration curves of extracted standards were linear over a working range of 0.001-2.00 mg/kg whole blood for all analytes. The limit of quantification was 0.008 mg/kg; the interday precision (measured by relative standard deviation-%RSD) was less than 10% and the accuracy (BIAS) less than 12% for all analytes in whole blood. Urine samples were estimated semi-quantitatively at a cut-off level of 0.15 mg/kg with an interday precision of 15%. CONCLUSION: A liquid chromatography mass spectrometric (LC/MS/MS) method has been developed for confirmation and quantification of cocaine and its metabolites (ecgonine methyl ester, benzoylecgonine, norcocaine and ethylene cocaine) in whole blood and semi-quantitative in urine. The method is specific and sensitive and offers thereby an excellent alternative to other methods such as GC-MS that involves derivatisation. SN - 1570-0232 UR - https://www.unboundmedicine.com/medline/citation/17301000/Quantitative_analysis_of_cocaine_and_its_metabolites_in_whole_blood_and_urine_by_high_performance_liquid_chromatography_coupled_with_tandem_mass_spectrometry_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1570-0232(07)00078-5 DB - PRIME DP - Unbound Medicine ER -