Quantitative analysis of 23-hydroxybetulinic acid, a novel anticancer substance, in the cellular environment of a human colon adenocarcinoma cell line for cellular pharmacokinetic studies.

Abstract

In order to study the cellular distribution and kinetics of a new anticancer substance, 23-hydroxybetulinic acid, a simple, reproductive and sensitive high-performance liquid chromatography/mass spectrometric (HPLC/MS) method was developed to quantify its trace concentration in cell suspension and cell culture medium. This method involved a liquid-liquid extraction with diethyl ether and a subsequent analysis performed on a Shimadzu LCMS 2010A system which contained an electrospray ionization interface. Separation was achieved by HPLC on a Zorbax Extend-C18 column with gradient elution using a mix of acetonitrile and water containing triethylamine and acetate-triethylamine as the mobile phase. A total analytical run was achieved within 6.5 min and the calibration curve was linear over a wide concentration range of 1.0-1000.0 nM for both cell suspension and culture medium. Intra- and inter-batch accuracy and precision were acceptable for both matrices. The described assay method was successfully applied to cellular pharmacokinetic studies in a human colon adenocarcinoma cell line (Caco-2) and its application of measuring the cellular concentrations of 23-hydroxybetunilic acid could be extended to different cultured cell lines.

• Authors: Yuanting Zheng, Guangji Wang, Xiaolan Wu, Jianguo Sun, Xiaoyu Li, Yi Gu, Hua Lv, Yan Liang, Bei Yan, Wencai Ye
• Journal: Rapid communications in mass spectrometry : RCM (Rapid Commun Mass Spectrom) Vol. 21 Issue 5 Pg. 779-86 ( 2007) ISSN: 0951-4198 [Print] England
• PMID: 17279599 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Validation Study)
• Copyright: Copyright (c) 2007 John Wiley & Sons, Ltd.

Chemical References

• 23-hydroxybetulinic acid
• Antineoplastic Agents
• Culture Media, Conditioned
• Triterpenes

Topics

• Adenocarcinoma (chemistry, drug therapy, metabolism)
• Antineoplastic Agents (analysis, metabolism, pharmacology)
• Cell Line, Tumor
• Chromatography, High Pressure Liquid
• Colonic Neoplasms (chemistry, drug therapy, metabolism)
• Culture Media, Conditioned (chemistry)
• Humans
• Reproducibility of Results
• Sensitivity and Specificity
• Spectrometry, Mass, Electrospray Ionization
• Triterpenes (analysis, metabolism, pharmacology)