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

• Type: Article in International Scientific Journal
• Date: 2010

Authors

Maria Joao Valente (Author) Other; Felix Carvalho (Author) FFUP; Lourdes L Bastos (Author) FFUP; Marcia Carvalho (Author) Other; de Pinho, PG (Author) FFUP

Journal

Title: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

Vol. 878

Pages: 3083-3088

ISSN: 1570-0232

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 15 Citations

Scopus - 18 Citations

Scientific classification

FOS: Natural sciences > Chemical sciences

Other information

Authenticus ID: P-003-100

DOI: 10.1016/j.jchromb.2010.09.010

Abstract (EN): 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 (r(2) > 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 6