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Development and validation of a gas chromatography/ion trap-mass spectrometry method for simultaneous quantification of cocaine and its metabolites benzoylecgonine and norcocaine: application to the study of cocaine metabolism in human primary cultured renal cells.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Nov 15; 878(30):3083-8.JC

Abstract

Acute renal failure is a common finding in cocaine abusers. While cocaine metabolism may contribute to its nephrotoxic mechanisms, its pharmacokinetics in kidney cells is hitherto to be clarified. Primary cultures of human proximal tubular cells (HPTCs) provide a well-characterized in vitro model, phenotypically representative of HPTCs in vivo. Thus, the present work describes the first sensitive gas chromatography/ion trap-mass spectrometry (GC/IT-MS) method for measurement of cocaine and its metabolites benzoylecgonine (BE) and norcocaine (NCOC) using a primary culture of HPTCs as cellular matrix, following solid phase extraction (SPE) and derivatization with N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA). The application of this methodology also enables the identification of two other cocaine metabolites: ecgonine methyl ester (EME) and anhydroecgonine methyl ester (AEME). The validation of the method was performed through the evaluation of selectivity, linearity, precision and accuracy, limit of detection (LOD), and limit of quantification (LOQ). Its applicability was demonstrated through the quantification of cocaine, BE and NCOC in primary cultured HPTCs after incubation, at physiological conditions, with 1 mM cocaine for 72 h. The developed GC/IT-MS method was found to be linear (r² > 0.99). The intra-day precision varied between 3.6% and 13.5% and the values of accuracy between 92.7% and 111.9%. The LOD values for cocaine, BE and NCOC were 0.97±0.09, 0.40±0.04 and 20.89±1.81 ng/mL, respectively, and 3.24±0.30, 1.34±0.14 and 69.62±6.05 ng/mL as LOQ values.

Authors+Show Affiliations

REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha 164, 4050-047 Porto, Portugal. mjoao.pcv@gmail.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Validation Study

Language

eng

PubMed ID

20965795

Citation

Valente, Maria João, et al. "Development and Validation of a Gas Chromatography/ion Trap-mass Spectrometry Method for Simultaneous Quantification of Cocaine and Its Metabolites Benzoylecgonine and Norcocaine: Application to the Study of Cocaine Metabolism in Human Primary Cultured Renal Cells." Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, vol. 878, no. 30, 2010, pp. 3083-8.
Valente MJ, Carvalho F, Bastos ML, et al. Development and validation of a gas chromatography/ion trap-mass spectrometry method for simultaneous quantification of cocaine and its metabolites benzoylecgonine and norcocaine: application to the study of cocaine metabolism in human primary cultured renal cells. J Chromatogr B Analyt Technol Biomed Life Sci. 2010;878(30):3083-8.
Valente, M. J., Carvalho, F., Bastos, M. L., Carvalho, M., & de Pinho, P. G. (2010). Development and validation of a gas chromatography/ion trap-mass spectrometry method for simultaneous quantification of cocaine and its metabolites benzoylecgonine and norcocaine: application to the study of cocaine metabolism in human primary cultured renal cells. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 878(30), 3083-8. https://doi.org/10.1016/j.jchromb.2010.09.010
Valente MJ, et al. Development and Validation of a Gas Chromatography/ion Trap-mass Spectrometry Method for Simultaneous Quantification of Cocaine and Its Metabolites Benzoylecgonine and Norcocaine: Application to the Study of Cocaine Metabolism in Human Primary Cultured Renal Cells. J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Nov 15;878(30):3083-8. PubMed PMID: 20965795.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and validation of a gas chromatography/ion trap-mass spectrometry method for simultaneous quantification of cocaine and its metabolites benzoylecgonine and norcocaine: application to the study of cocaine metabolism in human primary cultured renal cells. AU - Valente,Maria João, AU - Carvalho,Félix, AU - Bastos,M Lourdes, AU - Carvalho,Márcia, AU - de Pinho,Paula Guedes, Y1 - 2010/10/19/ PY - 2010/07/12/received PY - 2010/09/14/revised PY - 2010/09/15/accepted PY - 2010/10/23/entrez PY - 2010/10/23/pubmed PY - 2011/1/29/medline SP - 3083 EP - 8 JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences JO - J Chromatogr B Analyt Technol Biomed Life Sci VL - 878 IS - 30 N2 - Acute renal failure is a common finding in cocaine abusers. While cocaine metabolism may contribute to its nephrotoxic mechanisms, its pharmacokinetics in kidney cells is hitherto to be clarified. Primary cultures of human proximal tubular cells (HPTCs) provide a well-characterized in vitro model, phenotypically representative of HPTCs in vivo. Thus, the present work describes the first sensitive gas chromatography/ion trap-mass spectrometry (GC/IT-MS) method for measurement of cocaine and its metabolites benzoylecgonine (BE) and norcocaine (NCOC) using a primary culture of HPTCs as cellular matrix, following solid phase extraction (SPE) and derivatization with N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA). The application of this methodology also enables the identification of two other cocaine metabolites: ecgonine methyl ester (EME) and anhydroecgonine methyl ester (AEME). The validation of the method was performed through the evaluation of selectivity, linearity, precision and accuracy, limit of detection (LOD), and limit of quantification (LOQ). Its applicability was demonstrated through the quantification of cocaine, BE and NCOC in primary cultured HPTCs after incubation, at physiological conditions, with 1 mM cocaine for 72 h. The developed GC/IT-MS method was found to be linear (r² > 0.99). The intra-day precision varied between 3.6% and 13.5% and the values of accuracy between 92.7% and 111.9%. The LOD values for cocaine, BE and NCOC were 0.97±0.09, 0.40±0.04 and 20.89±1.81 ng/mL, respectively, and 3.24±0.30, 1.34±0.14 and 69.62±6.05 ng/mL as LOQ values. SN - 1873-376X UR - https://www.unboundmedicine.com/medline/citation/20965795/Development_and_validation_of_a_gas_chromatography/ion_trap_mass_spectrometry_method_for_simultaneous_quantification_of_cocaine_and_its_metabolites_benzoylecgonine_and_norcocaine:_application_to_the_study_of_cocaine_metabolism_in_human_primary_cultured_renal_cells_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1570-0232(10)00586-6 DB - PRIME DP - Unbound Medicine ER -