Determination of paclitaxel in hyaluronic acid polymeric micelles in rat blood by protein precipitation-micelle breaking method: application to a pharmacokinetic study.
An efficient dissociation of paclitaxel (PTX) from the home-made hyaluronic acid-octadecyl (HA-C18) polymeric micelles formulation in rat blood could not be achieved using previously published PTX analytical methods. So, we intended to develop the micelle-breaking method to determine paclitaxel encapsulated in the HA-C18 polymeric micelles in blood. The pretreatment method of blood samples adopted a simple one-step protein precipitation-micelle breaking process with methanol as micelle-breaking and protein precipitant solvents for complete extraction of PTX from HA-C18 micelles in blood. The micelle breaking efficiency of methanol was as high as 97.7%. Separation was carried out by gradient elution on an Acquity UPLC BEH C18 column with a mobile phase consisting of water (containing 0.1% formic acid) and acetonitrile. A total single run time was as short as 3.0min. Detection was performed by triple-quadrupole mass spectrometry with positive electrospray ionization as source ionization in multiple-reaction monitoring mode at m/z 854.3→286.2 for PTX and m/z 808.5→527.3 for the internal standard, docetaxel. The method demonstrated good linearity at the concentrations ranging from 20 to 10,000ng/mL. The intra- and inter-day relative standard deviations were less than 9.9%. The mean extraction recoveries of PTX and IS were 94.7% and 87.5%, respectively. In summary, the methanol protein precipitation-micelle breaking method could extract PTX completely from the polymeric micelles. Finally, the method was successfully applied to a pharmacokinetic study of the home-made PTX-loaded HA-C18 polymeric micelles and Taxol solution after intravenous administration in rats.
Department of Biopharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, China.
Yanhua
Y
Liu
Jin
J
Sun
He
H
Lian
Xin
X
Li
Wen
W
Cao
Liming
L
Bai
Yongjun
Y
Wang
Zhonggui
Z
He
D000818:Animals; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004337:Drug Carriers / Q000737:chemistry*; D004355:Drug Stability; D005260:Female; D006820:Hyaluronic Acid / Q000737:chemistry*; D016014:Linear Models; D008823:Micelles; D017239:Paclitaxel / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
D004337:Drug Carriers; D008823:Micelles; D006820:Hyaluronic Acid; D017239:Paclitaxel
HA-C(18) polymeric micelles; Paclitaxel; Pharmacokinetic study; Protein precipitation-micelle breaking method; UPLC–MS/MS method