A rapid, sensitive, selective and validated reverse phase high-performance liquid chromatography （RP-HPLC） method for the estimation of paclitaxel in micro-sample of rat plasma and in culture of cancer cells was per...A rapid, sensitive, selective and validated reverse phase high-performance liquid chromatography （RP-HPLC） method for the estimation of paclitaxel in micro-sample of rat plasma and in culture of cancer cells was per- formed in this study. The mobile phase consisted of an （80： 20： 0.1, v/v/v）. Column elution at a flow rate of 1 optimized mixture of methanol：water： trifluroacetic acid mL/minute with UV detection at 225 nm at room tern- perature was used. The RP-HPLC method was successfully applied for the determination of paclitaxel in plasma samples and in culture of cancer cells with nano-quantity of estimation. The validation studies were performed in accordance with the International Conference on Harmonization （ICH） guidelines. The intra- and inter-day pre- cision showed that the coefficients of variation ranged from 1.07% to 4.27% at different levels of concentrations. To the best of our knowledge, this study also reported for the first time the optimization of different solvents for effective extraction of paclitaxel wherein tert.-butyl methyl ether （TBME）： diethyl ether （DEE） in 50：50 v/ v composition was found most efficient with extraction efficiency ranging between 77.99% and 91.74% and be- tween 76.14 and 93.66% in the plasma and cell culture, respectively. This proposed method was successfully ap- plied to study the pharmacokinetics of paclitaxel and the influence of verapamil and all-trans retinoic acid （atRA） on paclitaxel pharmacokinetics in rat models. This proposed method might emerge as a valuable aid in the labo- ratory monitoring of paclitaxel in a variety of in vitro as well as in vivo scenarios.展开更多
The purpose of the present research was to develop a suitable, simple, precise, accurate,robust, and reproducible RP-HPLC method for a reliable simultaneous quantification of docetaxel(DTX) and curcumin(CCM) in rat pl...The purpose of the present research was to develop a suitable, simple, precise, accurate,robust, and reproducible RP-HPLC method for a reliable simultaneous quantification of docetaxel(DTX) and curcumin(CCM) in rat plasma samples using paclitaxel(PTX) as an internal standard. The samples were assayed by the Agilent 1260 Infinity HPLC instrument using a Capcell Pak C8 column(4.6 mm × 150 mm, 5 μm) under isocratic conditions. The mobile phase consisted of acetonitrile and triple distilled water(40/60, v/v) with a flow rate of 1.0 ml/min. The eluent was monitored at 230 nm for simultaneous measurement of curcumin and docetaxel. The method was validated by determining system suitability, selectivity, sensitivity, linearity, inter-day and intra-day precision, accuracy, robustness, and stability in accordance with the guidelines of the United States Food and Drug Administration(FDA).The developed chromatographic method proved to be simple, precise, accurate, robust and reproducible. Moreover, the samples showed stability at room temperature over a period of 48 h. Thus, this method would be employed for routine simultaneous quantification of docetaxel and curcumin in rat plasma samples.展开更多
A novel isocratic reverse phase high performance liquid chromatography （RP-HPLC） with photo diode array （PDA） detection method for the determination of dobutamine （DBT） in rat plasma was developed and validated ...A novel isocratic reverse phase high performance liquid chromatography （RP-HPLC） with photo diode array （PDA） detection method for the determination of dobutamine （DBT） in rat plasma was developed and validated after optimization of various chromatographic conditions and other experimental parameters. Homoveratrylamiue was used as an internal standard. Methanol was used as the extracting solvent for the preparation of plasma samples. Samples were separated on a Symmetry C18 （250ram x4.6mm i.d., 5 pro） analytical column. Acetonitrile and 15raM potassium dihydrogen phosphate （pH 5.0 with 0.3% TEA） （20：80, v/v） was used. The column oven temperature was optimized at 35 ~C and the flow rate was 0.8 mL/min. The detection wavelength was fixed at 230 nm for entire analysis. The calibration curve was found to be linear over the concentration range of 50-2000 ng/mL （ra=0.9992）. The limit of quantification （LOQ） of the method was 50 ng/mL. The % RSD values of accuracy and precision values for intra and inter days were 〈 15% at quality control （QC） concentrations. Recovery, stability and robustness were studied within the acceptable range according to ICH guidelines. The method was efficiently applied to a pharmacokinetic study in healthy Wistar rats.展开更多
基金supported by Senior Research Fellowship and Commonwealth Split Site Fellowship awards from the Council for Scientific Industrial Research(CSIR)New Delhi(India) and the Association of Commonwealth Universities+1 种基金UKrespectively
文摘A rapid, sensitive, selective and validated reverse phase high-performance liquid chromatography （RP-HPLC） method for the estimation of paclitaxel in micro-sample of rat plasma and in culture of cancer cells was per- formed in this study. The mobile phase consisted of an （80： 20： 0.1, v/v/v）. Column elution at a flow rate of 1 optimized mixture of methanol：water： trifluroacetic acid mL/minute with UV detection at 225 nm at room tern- perature was used. The RP-HPLC method was successfully applied for the determination of paclitaxel in plasma samples and in culture of cancer cells with nano-quantity of estimation. The validation studies were performed in accordance with the International Conference on Harmonization （ICH） guidelines. The intra- and inter-day pre- cision showed that the coefficients of variation ranged from 1.07% to 4.27% at different levels of concentrations. To the best of our knowledge, this study also reported for the first time the optimization of different solvents for effective extraction of paclitaxel wherein tert.-butyl methyl ether （TBME）： diethyl ether （DEE） in 50：50 v/ v composition was found most efficient with extraction efficiency ranging between 77.99% and 91.74% and be- tween 76.14 and 93.66% in the plasma and cell culture, respectively. This proposed method was successfully ap- plied to study the pharmacokinetics of paclitaxel and the influence of verapamil and all-trans retinoic acid （atRA） on paclitaxel pharmacokinetics in rat models. This proposed method might emerge as a valuable aid in the labo- ratory monitoring of paclitaxel in a variety of in vitro as well as in vivo scenarios.
基金supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (NRF-2015R1C1A1A01051698)
文摘The purpose of the present research was to develop a suitable, simple, precise, accurate,robust, and reproducible RP-HPLC method for a reliable simultaneous quantification of docetaxel(DTX) and curcumin(CCM) in rat plasma samples using paclitaxel(PTX) as an internal standard. The samples were assayed by the Agilent 1260 Infinity HPLC instrument using a Capcell Pak C8 column(4.6 mm × 150 mm, 5 μm) under isocratic conditions. The mobile phase consisted of acetonitrile and triple distilled water(40/60, v/v) with a flow rate of 1.0 ml/min. The eluent was monitored at 230 nm for simultaneous measurement of curcumin and docetaxel. The method was validated by determining system suitability, selectivity, sensitivity, linearity, inter-day and intra-day precision, accuracy, robustness, and stability in accordance with the guidelines of the United States Food and Drug Administration(FDA).The developed chromatographic method proved to be simple, precise, accurate, robust and reproducible. Moreover, the samples showed stability at room temperature over a period of 48 h. Thus, this method would be employed for routine simultaneous quantification of docetaxel and curcumin in rat plasma samples.
基金Mr.Thippani Ramesh thanks MHRD,Government of India for providing financial assistance
文摘A novel isocratic reverse phase high performance liquid chromatography （RP-HPLC） with photo diode array （PDA） detection method for the determination of dobutamine （DBT） in rat plasma was developed and validated after optimization of various chromatographic conditions and other experimental parameters. Homoveratrylamiue was used as an internal standard. Methanol was used as the extracting solvent for the preparation of plasma samples. Samples were separated on a Symmetry C18 （250ram x4.6mm i.d., 5 pro） analytical column. Acetonitrile and 15raM potassium dihydrogen phosphate （pH 5.0 with 0.3% TEA） （20：80, v/v） was used. The column oven temperature was optimized at 35 ~C and the flow rate was 0.8 mL/min. The detection wavelength was fixed at 230 nm for entire analysis. The calibration curve was found to be linear over the concentration range of 50-2000 ng/mL （ra=0.9992）. The limit of quantification （LOQ） of the method was 50 ng/mL. The % RSD values of accuracy and precision values for intra and inter days were 〈 15% at quality control （QC） concentrations. Recovery, stability and robustness were studied within the acceptable range according to ICH guidelines. The method was efficiently applied to a pharmacokinetic study in healthy Wistar rats.
