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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 850; no. 1; pp. 575 - 580
• Main Authors: Hu, Ze-Ping, Yang, Xiao-Xia, Chen, Xiao, Chan, Eli, Duan, Wei, Zhou, Shu-Feng
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.05.2007; Elsevier Science
• Subjects: Accumulation; Analysis; Analytical, structural and metabolic biochemistry; Animals; Antineoplastic Agents, Phytogenic - metabolism; Biological and medical sciences; Camptothecin - analogs & derivatives; Camptothecin - metabolism; Cell Line, Tumor; Chromatography, High Pressure Liquid - methods; CPT-11; Culture Media; Fundamental and applied biological sciences. Psychology; General pharmacology; HPLC; Medical sciences; Metabolism; Pharmacology. Drug treatments; Rats; SN-38; Spectrometry, Fluorescence; SN-38; CPT-11; Metabolism; Accumulation; HPLC; Antineoplastic agent; Culture medium; Chemical analysis; Enzyme; CPT-11; SN-38; Metabolism; Accumulation; HPLC; DNA topoisomerase; Enzyme inhibitor; HPLC chromatography; Tissue culture; In vitro; Irinotecan; Alkaloid; Isomerases; Simultaneous measurement; Pharmacokinetics; Tumor cell
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2006.12.056

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.