Simultaneous determination of irinotecan (CPT-11) and SN-38 in tissue culture media and cancer cells by high performance liquid chromatography: application to cellular metabolism and accumulation studies

Hu, Z, Yang, X, Chen, X, Chan, E, Duan, W and Zhou, S 2007, 'Simultaneous determination of irinotecan (CPT-11) and SN-38 in tissue culture media and cancer cells by high performance liquid chromatography: application to cellular metabolism and accumulation studies', J Chromatogr B Analyt Technol Biomed Life Sci 2007, vol. 850, no. 1, pp. 575-580.


Document type: Journal Article
Collection: Journal Articles

Title Simultaneous determination of irinotecan (CPT-11) and SN-38 in tissue culture media and cancer cells by high performance liquid chromatography: application to cellular metabolism and accumulation studies
Author(s) Hu, Z
Yang, X
Chen, X
Chan, E
Duan, W
Zhou, S
Year 2007
Journal name J Chromatogr B Analyt Technol Biomed Life Sci 2007
Volume number 850
Issue number 1
Start page 575
End page 580
Total pages 6
Publisher Elsevier Science, Amsterdam, PAYS-BAS
Abstract A simple and sensitive HPLC method was developed to simultaneously determine CPT-11 and its major metabolite SN-38 in culture media and cell lysates. Camptothecin (CPT) was used as internal standard (I.S.). Compounds were eluted with acetonitrile-50 mM disodium hydrogen phosphate buffer containing 10 mM sodium 1-heptane-sulfonate, with the pH adjusted to 3.0 using 85% (w/v) orthophosphoric acid (27/73, v/v) by a Hyperclon ODS (C18) column (200 mm × 4.6 mm i.d.), with detection at excitation and emission wavelengths of 380 and 540 nm, respectively. The average extraction efficiencies were 96.9-108.3% for CPT-11 in culture media and 94.3-107.2% for CPT-11 in cell lysates; and 87.7-106.8% for SN-38 in culture media and 90.1-105.6% for SN-38 in cell lysates. Within- and between-day precision and accuracy varied from 0.1 to 10.3%. The limit of quantitation (precision and accuracy <20%) was 5.0 and 2.0 ng/ml for CPT-11 and 1.0 and 0.5 ng/ml for SN-38 in culture media and cell lysates, respectively. This method was successfully applied to quantitate the cellular accumulation and metabolism of CPT-11 and SN-38 in H4-II-E, a rat hepatoma cell line.
Subject Basic Pharmacology
DOI - identifier 10.1016/j.jchromb.2006.12.056
ISSN 1570-0232
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