Quantitative determination of trans-polydatin, a natural strong anti-oxidative compound, in rat plasma and cellular environment of a human colon adenocarcinoma cell line for pharmacokinetic studies.
A simple, accurate, precise, specific and reproducible high-performance liquid chromatography (HPLC) method was developed for determination of trans-polydatin, a natural strong anti-oxidative compound, in rat plasma and cell suspension. The assay procedure involved simple liquid-liquid extraction, the supernatant liquid was added an equal volume of water to avoid solvent effect. The detection of the analyte peak was achieved by monitoring the eluate using a UV detector set at 303 nm. The analysis used a Hypersil ODS2 C18 column (5 microm, 4.6 mm x 250 mm) and methanol/distilled water as the mobile phase (flow rate=1 mL/min). A total analytical run was achieved within 6.0 min and calibration curve was linear over a wide concentration range of 0.25-40 microg/mL for plasma sample and 1.0-500 microM for cell suspension, the coefficients of correlation were 0.9997 and 0.9999 or better, respectively. There was 80.7+/-7.86%, 96.8+/-3.20% and 102.7+/-9.72% recovery from 0.5, 10, and 40 microg/mL plasma samples, respectively. Intra- and inter-batch accuracy and precision were acceptable for the both matrices. The RSD of intra- and inter-day assay variations were all less than 10%. Both analyte and IS were stable in the battery of stability studies, freeze-thaw cycles. The described assay method was applied to pharmacokinetic studies in rats and a human colon adenocarcinoma cell line (Caco-2) successfully. The application of the assay to determine the pharmacokinetic is described.
He
Hui
H
Center of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing, Jiangsu, PR China. njhe@hotmail.com
Zhao
Ying
Y
Chen
Xijing
X
Zheng
Yuanting
Y
Wu
Xiaolan
X
Wang
Rui
R
Li
Tingting
T
Yu
Qiangling
Q
Jing
Jing
J
Ma
Le
L
Ren
Weichao
W
Han
Deen
D
Wang
Guangji
G
D000230:Adenocarcinoma; D000818:Animals; D000975:Antioxidants / Q000032:analysis* / Q000302:isolation & purification / Q000493:pharmacokinetics; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003110:Colonic Neoplasms; D005960:Glucosides / Q000097:blood* / Q000302:isolation & purification / Q000493:pharmacokinetics; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013267:Stilbenes / Q000097:blood* / Q000302:isolation & purification / Q000493:pharmacokinetics; D013997:Time Factors
D000975:Antioxidants; D005960:Glucosides; D013267:Stilbenes; C058229:polydatin