Development and qualification of an LC–MS/MS method for investigating the biological implications of micelle entrapped paclitaxel in cell culture and rats

Abstract

Paclitaxel is a front-line antineoplastic drug used in chemotherapeutic modalities for treatment of various types of malignancies. However, its efficacy is limited by dose-related toxicities. In this study, we have explored two important biological aspects of entrapping paclitaxel in PEG -DSPE micelles. First, we evaluated the impact of this micellar delivery system on P-glycoprotein (P-gp)-paclitaxel interaction, and we investigated differences in plasma pharmacokinetics of free and micelle-entrapped paclitaxel. For quantification of paclitaxel, an LC-MS/MS method was developed. Paclitaxel was extracted from samples using a simple one-step protein precipitation. Chromatographic conditions included a C column with a mobile phase consisting of 0.1% formic acid in acetonitrile-water (60:40, v/v) pumped at 1 mL/min. The lower limit of quantitation in both plasma and cell lysate was 1.0 ng/mL. The quantitative linear range was 1-1000 ng/mL. In addition, P-gp efflux studies on free and micellar paclitaxel showed the proficiency of PEG -DSPE micelles in evading P-gp-mediated efflux, thus increasing paclitaxel uptake. Furthermore, the micellar paclitaxel levels were maintained in the body for longer time as compared with taxol, which is desirable for increasing the efficacy of paclitaxel in cancer treatment.

Preparation and characterization of paclitaxel palmitate albumin nanoparticles with high loading efficacy: an in vitro and in vivo anti-tumor study in mouse models

“…The supernatant was vortically mixed for 1 min and centrifuged at 13000 r/min for 10 min, from which 150 lL supernatant was mixed with 300 lL 30% acetonitrile and prepared into a 5 lL sample injection, analyzed by HPLC-MS to determine the concentration of PTX-PA and PTX. (Kaddoumi et al, 2017).…”

Preparation and Characterization of Paclitaxel Palmitate Albumin Nanoparticles With High Loading Efficacy: an in Vitro and in Vivo Anti-tumor Study in Mouse Models

“…100 µL tissue homogenate was transferred into a 1.5 mL centrifuge tube, and protein was precipitated with 300 µL acetonitrile containing internal standard liquid [21,22]. The supernatant was vortically mixed for 1 min and centrifuged at 13000 r/min for 10 min, from which 150 µL supernatant was mixed with 300 µL 30% acetonitrile and prepared into a 5 µL sample injection [23].…”

Simultaneous LC‐MS/MS bioanalysis of etoposide and paclitaxel in mouse tissues and plasma after oral administration of self‐microemulsifying drug‐delivery systems

“…To investigate the effect of SMEDDS on the bioavailability and tissue distribution of ETP and PTX, an analytical method for simultaneous quantification of ETP and PTX in tissues and plasma is required. Until now, several studies have been carried out to assay PTX in plasma or tissues by HPLC-UV or HPLC-MS/MS method (Guo, Ma, et al, 2003Green, Vretenbrant, et al, 2006;Vainchtein, Thijssen, et al, 2006;Rezazadeh, Emami, et al, 2015;Malhi, Stesco, et al, 2017) and the most recent study displayed a robust and accurate LC-MS/MS method to assay the PTX concentration in rat plasma (Kaddoumi, Gill, et al, 2017). As for ETP, a number of LC-UV and LC-MS/MS methods have been reported for the determination in plasma (Carcel-Trullols, Torres-Molina, et al, 2004;Gong, Yang, et al, 2017;Harivardhan Reddy, Sharma, et al, 2005;Patlolla & Vobalaboina, 2005;Saadati & Dadashzadeh, 2011).…”