Aim To develop and validate a sensitive and specific liquid chromatography electrospray ionization ion-trap mass spectrometry （LC-ESI-MS/MS） method for the identification and concentration of rupatadine in human pla...Aim To develop and validate a sensitive and specific liquid chromatography electrospray ionization ion-trap mass spectrometry （LC-ESI-MS/MS） method for the identification and concentration of rupatadine in human plasma. Methods After the addition of the internal standard （IS, loratadine） and 0.01 mol·L^-1 sodium hydroxide solution, plasma samples were extracted with methylene chloride： ethyl acetate mixture （20：80, V/V）. The organic layer was evaporated under vacuum drying at 37 ℃. The residue was reconstituted with 200 μL mobile phase. Chromatography was performed on an Agilent Eclipse XDB-C18 （4.6 mm × 150 mm, 5 μm） column with a mobile phase consisting of acetonitrile （1% formic acid） -20 mmol·L^-1 ammonium acetate （76：24, V/V） at a flow-rate of 0.6 mL·min^-1. Detection was performed on Agilent MSD Trap XCT ion-trap mass spectrometry connected to a Agilent 1100 high performance liquid chromatography （HPLC） by selected reaction monitoring （SRM） mode via electrospray ionization （ESI） source. Rupatadine （MRM m/z 416 → 309） and loratadine （MRM m/z 383 → 337） were detected by Agilent MSD Trap XCT ion-trap mass analyser. Results The method was proved to be sensitive and specific by testing 20 different plasma batches. Linearity was established for the range of concentrations 0.05 - 14.0 ng·mL^ -1 with a coefficient of determination （r） of 0.998. The intra-and inter-day precision （RSD %） were lower than 15% and accuracy ranged from 85.1% to 114.0%. The lower limit of quantitation （LLOQ） was identifiable and reproducible at 0.05 ng·mL^-1 with a precision of 9.22% （n =5）. Conclusion The proposed method is sensitive and reproducible enough to be used in pharmacokinetic, bioavailability or bioequivalence studies of rupatadine.展开更多
A simple and sensitive high-performance liquid chromatography mass spectrometry（LC-MS）method for the simultaneous determination of irbesartan and hydrochlorothiazide in human plasma was developed and applied to a ph...A simple and sensitive high-performance liquid chromatography mass spectrometry（LC-MS）method for the simultaneous determination of irbesartan and hydrochlorothiazide in human plasma was developed and applied to a pharmacokinetic study. Acetaminophen was used as the internal standard（IS）.Sample pretreatment using liquid-liquid extraction with ethyl acetate was used.The analysis was carried out on an Elite SinoChrom ODS-BP C_（18）column with a mobile phase composed of acetonitrile-water （35：65,v/v）.Target ions were [M-H]^-m/z 427.25 for irbesartan,[M-H]^-m/z 295.95 for hydrochlorothiazide and [M-H]^- m/z 150.05 for the IS via an electrospray ionization（ESI）source.The intra-and inter-day precision（RSD%）was below 14.5% for irbesartan and hydrochlorothiazide,and the accuracy（RE%）was less than 1.9% and-2.0% for irbesartan and hydrochlorothiazide,respectively.The linear calibration curves were obtained in the concentration range of 10-5000 ng/mL （r0.99）for irbesartan and 1-200 ng/mL（r0.99）for hydrochlorothiazide with the lower limit of quantification（LLOQ）of 10 ng/mL and 1 ng/mL,respectively.The method was applied to a clinical pharmacokinetic study of a tablet containing irbesartan and hydrochlorothiazide in healthy Chinese volunteers after oral administration.展开更多
An efficient and sensitive ion-pair HPLC-UV method using atenolol as internal standard （IS） was developed and validated for the determination of metformin in the plasma of diabetic rats. Plasma samples were deprotei...An efficient and sensitive ion-pair HPLC-UV method using atenolol as internal standard （IS） was developed and validated for the determination of metformin in the plasma of diabetic rats. Plasma samples were deproteinated with 10% （v/v） perchloric acid. Separation was achieved on a UltimateTM AQ-C18 column （250 mm×4.6 mm, 5 μm） with a mobile phase （pH 5.05） composed of acetonitrile-water （31：69, v/v, containing 0.002 M sodium dodecyl sulfate, 0.0125 M potassium dihydrogen phosphate, 0.015 M triethylamine） at a flow rate of 1.0 mL/min. The calibration curve was linear （r〉0.994） between 7.5 and 4000 ng/mL. The lower limit of quantification （LLOQ） was 7.5 ng/mL. The precision was validated and the relative standard deviation was in the range of 1.87% to 15.70%; the accuracy was between 93.98%-106.89%. The mean recoveries were 95.40% and 95.31% for metformin and IS, respectively. The relative error （RE） of stability at different storage conditions was within ±9.00%. This method was used to determine the concentration-time profile of metformin in diabetic rat plasma following an oral administration of metformin at the dose of 10 mg/kg. Our results indicated that ion-pair HPLC-UV method using UltimateTM AQ-C18 column was effective for the pharmacokinetic studies of high polarity compounds like metformin.展开更多
A sensitive LC-ESI-MS/MS method for determination of isochlorogenic acid B in rat plasma was developed and validated in the present study. Plasma samples were prepared by a simple protein precipitation with methanol c...A sensitive LC-ESI-MS/MS method for determination of isochlorogenic acid B in rat plasma was developed and validated in the present study. Plasma samples were prepared by a simple protein precipitation with methanol containing resveratrol as internal standard (IS). The chromatographic separation was performed on a Zorbax SB-Cjg column (3.5 pm, 2.1 mmx 100 mm, Agilent, USA) at a flow rate of 0.2 mL/min using methanol/water containing 0.1% formic acid (v/v) as mobile phase. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with Electronic Spray Ion by selected reaction monitoring (SRM) of the transitions at m/z 515.3->352.9 for isochlorogenic acid B and m/z 227.1-143.1 for IS, respectively. The calibration curve of the method was linear over the range of 5-2500 ng/mL (r^2= 0.9982). The intra- and inter-day precisions (R.S.D.%) were less than 12.46%, and the accuracy (R.E.%) was within ±5.80%. Isochlorogenic acid B was sufficiently stable under all relevant analytical conditions. The validated method was successfully applied to the plasma pharmacokinetic studies of isochlorogenic acid B in rats. It was found that isochlorogenic acid B had non-linear pharmacokinetic characteristics in rats within the dosage ranges from 5 to 20 mg/kg.展开更多
In the present study, we developed and validated a selective, specific and sensitive liquid chromatography-electrospray ionization mass spectrometry （LC-ESI-MS/MS） method for the determination of tenofovir in human ...In the present study, we developed and validated a selective, specific and sensitive liquid chromatography-electrospray ionization mass spectrometry （LC-ESI-MS/MS） method for the determination of tenofovir in human plasma. Entecavir was used as an internal standard, and plasma samples were prepared by solid-phase extraction performed on Phenomenex Strata cartridges （30 mg）. The mobile phase consisted of 10 mM ammonium acetate in water and methanol （60：40, v/v）. The chromatographic separation was performed isocratically on a Phenomenex C18 （4.6 mm×150 mm, 5 μm）, and analytes were analyzed in multiple reaction monitoring （MRM） mode with positive electrospray ionization （ESI） interface using the respective [M＋H] ＋ ions, m/z 288.2→m/z 176.1 for tenofovir and m/z 278.1→m/z 152 for entecavir. The calibration curve （r2 = 0.9962） of tenofovir was established within the range of 4.096-1000 μg/L. The intra- and inter-day precisions were less than 10%. This validated method was successfully applied to a pharmacokinetic study in 12 healthy Chinese volunteers after the oral administration of tenofovir disonroxil fumarate.展开更多
A rapid, sensitive and selective UPLC-MS/MS method was developed to determine paeoniflorin and astragaloside IV, This method was validated via a pharmacokinetic study using rat plasma. The internal standard was clarit...A rapid, sensitive and selective UPLC-MS/MS method was developed to determine paeoniflorin and astragaloside IV, This method was validated via a pharmacokinetic study using rat plasma. The internal standard was clarithromycin. A simple one-step deproteinization procedure was used to prepare plasma samples. Separation was achieved on a CAPCELL CORE ADME CI8 column with a gradient mobile phase consisting of solution A （water containing 0.1% formic acid） and solution B （acetonitrile） at a flow rate of 0.3 mL/min. Multiple reaction monitoring （MRM） was used with an electrospray ionization source （ESI） in positive mode. A good linear response was observed within the ranges of 0.01 to 5.00 ~g/mL for paeoniflorin and 0.000l to 0.05 ~tg/mL for astragaloside IV. The accuracy （RE） was within the range of-3.5% to 6.3%, and the intra- and inter-day precisions （RSD） were within 14.2%. The extraction recoveries were all above 78.9%. The pharmacokinetic study of the two analytes in rats after oral administration of Huangqi Guizhi Wuwu Decoction （HGWD） was successfully completed through this method. The method develooed in this studv will fill a gap in oharmacokinetic studies of HGWD.展开更多
文摘Aim To develop and validate a sensitive and specific liquid chromatography electrospray ionization ion-trap mass spectrometry （LC-ESI-MS/MS） method for the identification and concentration of rupatadine in human plasma. Methods After the addition of the internal standard （IS, loratadine） and 0.01 mol·L^-1 sodium hydroxide solution, plasma samples were extracted with methylene chloride： ethyl acetate mixture （20：80, V/V）. The organic layer was evaporated under vacuum drying at 37 ℃. The residue was reconstituted with 200 μL mobile phase. Chromatography was performed on an Agilent Eclipse XDB-C18 （4.6 mm × 150 mm, 5 μm） column with a mobile phase consisting of acetonitrile （1% formic acid） -20 mmol·L^-1 ammonium acetate （76：24, V/V） at a flow-rate of 0.6 mL·min^-1. Detection was performed on Agilent MSD Trap XCT ion-trap mass spectrometry connected to a Agilent 1100 high performance liquid chromatography （HPLC） by selected reaction monitoring （SRM） mode via electrospray ionization （ESI） source. Rupatadine （MRM m/z 416 → 309） and loratadine （MRM m/z 383 → 337） were detected by Agilent MSD Trap XCT ion-trap mass analyser. Results The method was proved to be sensitive and specific by testing 20 different plasma batches. Linearity was established for the range of concentrations 0.05 - 14.0 ng·mL^ -1 with a coefficient of determination （r） of 0.998. The intra-and inter-day precision （RSD %） were lower than 15% and accuracy ranged from 85.1% to 114.0%. The lower limit of quantitation （LLOQ） was identifiable and reproducible at 0.05 ng·mL^-1 with a precision of 9.22% （n =5）. Conclusion The proposed method is sensitive and reproducible enough to be used in pharmacokinetic, bioavailability or bioequivalence studies of rupatadine.
文摘A simple and sensitive high-performance liquid chromatography mass spectrometry（LC-MS）method for the simultaneous determination of irbesartan and hydrochlorothiazide in human plasma was developed and applied to a pharmacokinetic study. Acetaminophen was used as the internal standard（IS）.Sample pretreatment using liquid-liquid extraction with ethyl acetate was used.The analysis was carried out on an Elite SinoChrom ODS-BP C_（18）column with a mobile phase composed of acetonitrile-water （35：65,v/v）.Target ions were [M-H]^-m/z 427.25 for irbesartan,[M-H]^-m/z 295.95 for hydrochlorothiazide and [M-H]^- m/z 150.05 for the IS via an electrospray ionization（ESI）source.The intra-and inter-day precision（RSD%）was below 14.5% for irbesartan and hydrochlorothiazide,and the accuracy（RE%）was less than 1.9% and-2.0% for irbesartan and hydrochlorothiazide,respectively.The linear calibration curves were obtained in the concentration range of 10-5000 ng/mL （r0.99）for irbesartan and 1-200 ng/mL（r0.99）for hydrochlorothiazide with the lower limit of quantification（LLOQ）of 10 ng/mL and 1 ng/mL,respectively.The method was applied to a clinical pharmacokinetic study of a tablet containing irbesartan and hydrochlorothiazide in healthy Chinese volunteers after oral administration.
基金National Integrity Innovational Technology Platform of New Drug and Research and Development (Grant No.2009ZX09201-010)Innovation Team of Ministry of Education(Grant No. BMU20110263)
文摘An efficient and sensitive ion-pair HPLC-UV method using atenolol as internal standard （IS） was developed and validated for the determination of metformin in the plasma of diabetic rats. Plasma samples were deproteinated with 10% （v/v） perchloric acid. Separation was achieved on a UltimateTM AQ-C18 column （250 mm×4.6 mm, 5 μm） with a mobile phase （pH 5.05） composed of acetonitrile-water （31：69, v/v, containing 0.002 M sodium dodecyl sulfate, 0.0125 M potassium dihydrogen phosphate, 0.015 M triethylamine） at a flow rate of 1.0 mL/min. The calibration curve was linear （r〉0.994） between 7.5 and 4000 ng/mL. The lower limit of quantification （LLOQ） was 7.5 ng/mL. The precision was validated and the relative standard deviation was in the range of 1.87% to 15.70%; the accuracy was between 93.98%-106.89%. The mean recoveries were 95.40% and 95.31% for metformin and IS, respectively. The relative error （RE） of stability at different storage conditions was within ±9.00%. This method was used to determine the concentration-time profile of metformin in diabetic rat plasma following an oral administration of metformin at the dose of 10 mg/kg. Our results indicated that ion-pair HPLC-UV method using UltimateTM AQ-C18 column was effective for the pharmacokinetic studies of high polarity compounds like metformin.
基金Chinese Academy of Medical Sciences(Grant No.CAMS-2017-I2M-1-011)
文摘A sensitive LC-ESI-MS/MS method for determination of isochlorogenic acid B in rat plasma was developed and validated in the present study. Plasma samples were prepared by a simple protein precipitation with methanol containing resveratrol as internal standard (IS). The chromatographic separation was performed on a Zorbax SB-Cjg column (3.5 pm, 2.1 mmx 100 mm, Agilent, USA) at a flow rate of 0.2 mL/min using methanol/water containing 0.1% formic acid (v/v) as mobile phase. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with Electronic Spray Ion by selected reaction monitoring (SRM) of the transitions at m/z 515.3->352.9 for isochlorogenic acid B and m/z 227.1-143.1 for IS, respectively. The calibration curve of the method was linear over the range of 5-2500 ng/mL (r^2= 0.9982). The intra- and inter-day precisions (R.S.D.%) were less than 12.46%, and the accuracy (R.E.%) was within ±5.80%. Isochlorogenic acid B was sufficiently stable under all relevant analytical conditions. The validated method was successfully applied to the plasma pharmacokinetic studies of isochlorogenic acid B in rats. It was found that isochlorogenic acid B had non-linear pharmacokinetic characteristics in rats within the dosage ranges from 5 to 20 mg/kg.
基金The Science and Technology Development Plan Project of Wujin District,Changzhou City,Jiangsu Province(Grant No.WS201413)the Research Funds for the Teachers of Central South University(Grant No.2014JSJJ028)
文摘In the present study, we developed and validated a selective, specific and sensitive liquid chromatography-electrospray ionization mass spectrometry （LC-ESI-MS/MS） method for the determination of tenofovir in human plasma. Entecavir was used as an internal standard, and plasma samples were prepared by solid-phase extraction performed on Phenomenex Strata cartridges （30 mg）. The mobile phase consisted of 10 mM ammonium acetate in water and methanol （60：40, v/v）. The chromatographic separation was performed isocratically on a Phenomenex C18 （4.6 mm×150 mm, 5 μm）, and analytes were analyzed in multiple reaction monitoring （MRM） mode with positive electrospray ionization （ESI） interface using the respective [M＋H] ＋ ions, m/z 288.2→m/z 176.1 for tenofovir and m/z 278.1→m/z 152 for entecavir. The calibration curve （r2 = 0.9962） of tenofovir was established within the range of 4.096-1000 μg/L. The intra- and inter-day precisions were less than 10%. This validated method was successfully applied to a pharmacokinetic study in 12 healthy Chinese volunteers after the oral administration of tenofovir disonroxil fumarate.
文摘A rapid, sensitive and selective UPLC-MS/MS method was developed to determine paeoniflorin and astragaloside IV, This method was validated via a pharmacokinetic study using rat plasma. The internal standard was clarithromycin. A simple one-step deproteinization procedure was used to prepare plasma samples. Separation was achieved on a CAPCELL CORE ADME CI8 column with a gradient mobile phase consisting of solution A （water containing 0.1% formic acid） and solution B （acetonitrile） at a flow rate of 0.3 mL/min. Multiple reaction monitoring （MRM） was used with an electrospray ionization source （ESI） in positive mode. A good linear response was observed within the ranges of 0.01 to 5.00 ~g/mL for paeoniflorin and 0.000l to 0.05 ~tg/mL for astragaloside IV. The accuracy （RE） was within the range of-3.5% to 6.3%, and the intra- and inter-day precisions （RSD） were within 14.2%. The extraction recoveries were all above 78.9%. The pharmacokinetic study of the two analytes in rats after oral administration of Huangqi Guizhi Wuwu Decoction （HGWD） was successfully completed through this method. The method develooed in this studv will fill a gap in oharmacokinetic studies of HGWD.
