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.
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.
Zheng
Yuanting
Y
Key Laboratory of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, 1 Shennong Road, Nanjing 210038, China.
Wang
Guangji
G
Wu
Xiaolan
X
Sun
Jianguo
J
Li
Xiaoyu
X
Gu
Yi
Y
Lv
Hua
H
Liang
Yan
Y
Yan
Bei
B
Ye
Wencai
W
D000230:Adenocarcinoma / Q000737:chemistry* / Q000188:drug therapy / Q000378:metabolism; D000970:Antineoplastic Agents / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D003110:Colonic Neoplasms / Q000737:chemistry* / Q000188:drug therapy / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000737:chemistry; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D014315:Triterpenes / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology
C468651:23-hydroxybetulinic acid; D000970:Antineoplastic Agents; D017077:Culture Media, Conditioned; D014315:Triterpenes