Simultaneous Determination of Amlodipine and Olmesartan in Human Plasma by Liquid Chromatography Tandem Mass Spectrometry and Its Application in Pharmacokinetic Study (original) (raw)
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Journal of Pharmaceutical Analysis, 2012
A rapid and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay method has been developed and fully validated for the simultaneous quantification of telmisartan and amlodipine in human plasma. Carbamazepine was used as an internal standard. Analytes and the internal standard were extracted from human plasma by solid-phase extraction technique using Waters Oasis s HLB 1 cm 3 (30 mg) extraction cartridge. The reconstituted samples were chromatographed on a Hypurity advance C 18 column (50 mm  4.6 mm, 5 mm) using a mixture of acetonitrile-5 mM ammonium acetate buffer (pH-4.0) (50:50, v/v) as the mobile phase at a flow rate of 0.8 mL/min. The calibration curve obtained was linear (rZ0.99) over the concentration range of 2.01-400.06 ng/mL for telmisartan and 0.05-10.01 ng/mL for amlodipine. Method validation was performed as per FDA guidelines and the results met the acceptance criteria. A run time of 2.5 min for each sample made it possible to analyze more than 400 human plasma samples per day. The proposed method was found to be applicable to clinical studies.
Pharmaceutical Sciences Asia, 2017
A sensitive liquid chromatography tandem mass spectrometry method was developed to quantify amlodipine in human plasma. Alkalinized plasma spiked with desipramine, an internal standard, was extracted by liquid-liquid extraction and evaporated an organic part to dryness. The residue was reconstituted and injected into an Acquity Ultra Performance LC TM , (Waters, Co., Ltd. USA) with C 18 column. The isocratic elution of mobile phase was performed by 90% of acetonitrile and 10% of 10 mM ammonium acetate pH 4 at flow rate of 0.20 mL/min with 5 minutes of total run time. Mass spectrometric analysis was performed using a Quattro Premier XE mass spectrometer, (Micromass Technologies, UK) coupled with an electrospray ionization (ESI) source in the positive ion mode. The MRM transitions of m/z 409.17>238.19 and 409.17>294.09 were selected for amlodipine and 267.09>208.06 for desipramine. The retention times were 1.63 and 1.69 minutes for amlodipine and desipramine, respectively. The linearity of the method revealed a correlation coefficient of >0.998 within the concentration range of 0.05-20 ng/mL. This work has been fully validated according to the Guidance for Industry: Bioanalytical Method Validation (USFDA CDER, 2001, BP) with high degree of accuracy and precision. This method was applied to quantify amlodipine concentrations in human plasma samples in a bioequivalence study.
2015
ANDREEA VARGA1, LENARD FARCZADI 2,3, LAURIAN VLASE2*, DANIELA P. PRIMEJDIE2, EMILIAN CARASCA4, IOAN TILEA1 1 University of Medicine and Pharmacy Tirgu Mures, Faculty of Medicine, Department of Family Medicine M3, 38 Gheorghe Marinescu Str., 540139, Tirgu Mures, Romania 2 University of Medicine and Pharmacy “Iuliu Hatieganu”, Department of Pharmaceutical Technology and Biopharmaceutics, Faculty of Pharmacy, 8 Victor Babes Str., 400012, Cluj-Napoca, Romania 3Vim Spectrum SRL, 409 Sighisoarei Str., 547367, Corunca, Tirgu-Mures, Romania 4 University of Medicine and Pharmacy Tirgu Mures, Faculty of Medicine, Department of Internal Medicine M3, 38 Gheorghe Marinescu Str., 540139, Tirgu Mures, Romania
Biomedical Chromatography, 2006
A simple, sensitive and rapid high-performance liquid chromatography/electrospray ionization tandem mass spectrometry method was developed and validated for the assay of amlodipine in human plasma. Following liquid-liquid extraction, the analytes were separated using an isocratic mobile phase on a reverse-phase C 18 column and analyzed by MS in the multiple reaction monitoring mode using the respective [M+H] + ions, m/z 409/238 for amlodipine and m/z 409/228 for the IS. The assay exhibited a linear dynamic range of 50-10,000 pg/mL for amlodipine in human plasma. The lower limit of quantification was 50 pg/mL with a relative standard deviation of less than 8%. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. The average absolute recoveries of amlodipine and the IS from spiked plasma samples were 74.7 ± 4.6 and 72.1 ± 2.0%, respectively. A run time of 1.5 min for each sample made it possible to analyze more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies. The observed maximum plasma concentration (C max) of amlodipine (2.5 mg oral dose) was 1425 pg/mL, time to observed maximum plasma concentration (T max) was 8.1 h and elimination half-life (T 1/2) was 50.
A simple and validated liquid chromatographic-mass spectrometric method (LC-MS) for amlodipine in human plasma was quantifi ed using LC-MS (ESI). Chromatography was performed on a C 18 analytical column, the mobile phase used was Acetonitrile-10mM Ammonium acetate in the ratio of 90:10%v/v and the retention times were 0.829 and 1.281 min for azithromycin (Internal standard) and amlodipine respectively. The ionization was optimized using ESI (+) and enhanced selectivity was achieved. The method is validated as per FDA guidelines. The analyte was shown to be stable over the timescale of the whole procedure. The pharmacokinetic parameters such as peak plasma concentration (C max ), Time to peak Concentration (t max ), Area under the plasma concentration-time curve (AUC 0-t & AUC 0-∞ ), elimination rate constant (K eli ), Elimination half-life (t ½ ) were calculated. Log transferred values were compared by Analysis of Variance (ANOVA) followed by classical 90% confi dence interval for C max AUC 0-t .and AUC 0-∞ and was found to be within the range. These results indicated that the Test and Reference formulation is bioequivalent.
Pharmaceutical Methods, 2012
Introduction: A simple, rapid and sensitive liquid chromatography-tandem mass spectrometric assay method has been developed and fully validated for simultaneous quantification of losartan and its active metabolite, losartan carboxylic acid, and amlodipine in human plasma. Irbesartan was used as an internal standard. Materials and Methods: The analytes were extracted from human plasma samples by solidphase extraction technique using Oasis HLB cartridges, (Waters Corporation, Mumbai, India). The reconstituted samples were chromatographed on a C 18 column by using an 85:15, v/v mixture of methanol and 0.1% v/v formic acid as the mobile phase at a flow rate of 1.0 mL/min. A detailed validation of the method was performed as per the FDA guidelines. Results: The calibration curves obtained were linear (r ≥ 0.99) over the concentration range of 0.5-1000 ng/mL for losartan and for its active metabolite losartan acid and 0.05-10.1 ng/mL for amlodipine. The results of the intra-and inter-day precision and accuracy studies were well within the acceptable limits. Conclusions: A run time of 2.5 min for each sample made it possible to analyze more than 300 plasma samples per day. The proposed method was found to be applicable to clinical studies.
Journal of chromatographic science, 2009
An accurate, sensitive, and reproducible high-performance liquid chromatographic method for the quantitation of amlodipine besylate in human plasma has been developed and validated. The drug, internal standard, and major metabolite were eluted from a C 18 hypersil HyPurity column (3 µm, 3.9 mm i.d. × × 150 mm) at room temperature with a mobile phase consisting of acetonitrile-potassium dihydrogen phosphate buffer (0.05 M) and acetic acid (62:38:0.1) with the pH adjusted to 3.5 using phosphoric acid. The flow-rate was 1.8 mL/min. The limit of detection was 1.0 ng/mL, and the limit of quantification of amlodipine besylate in plasma was 10 ng/mL. The intra-and inter-day precisions showed coefficients of variation ranging from 5.98-11.4% and from 5.60-11.74%, respectively at three different levels of concentration. The averages of the absolute and relative recoveries were found to be 96.74-98.51% and 95.96-100.71%, respectively. Stability studies showed that amlodipine besylate is stable for at least 2 months in plasma after freezing at -20°C. The method was successfully applied for a pharmacokinetic study and for the determination of commercial amlodipine tablet content. . Chemical structure of amlodipine.
Journal of Young Pharmacists, 2012
Simultaneous quantitative analysis of olmesartan, amlodipine and hydrochlorothiazide in their combined dosage form utilizing classical and alternating least squares based chemometric methods Simultaneous spectrophotometric analysis of a multi-component dosage form of olmesartan, amlodipine and hydrochlorothiazide used for the treatment of hypertension has been carried out using various chemometric methods. Multivariate calibration methods include classical least squares (CLS) executed by net analyte processing (NAP-CLS), orthogonal signal correction (OSC-CLS) and direct orthogonal signal correction (DOSC-CLS) in addition to multivariate curve resolution-alternating least squares (MCR-ALS). Results demonstrated the effi ciency of the proposed methods as quantitative tools of analysis as well as their qualitative capability. The three analytes were determined precisely using the aforementioned methods in an external data set and in a dosage form aft er optimization of experimental conditions. Finally, the effi ciency of the models was validated via comparison with the partial least squares (PLS) method in terms of accuracy and precision.
Il Farmaco, 2005
A rapid, simple and sensitive high-performance liquid chromatography (HPLC) method has been developed for quantification of amlodipine in plasma. The assay enables the measurement of amlodipine for therapeutic drug monitoring with a minimum detectable limit of 0.2 ng ml −1 . The method involves simple, one-step extraction procedure and analytical recovery was about 97%. The separation was performed on an analytical 125 × 4.6 mm i.d. Nucleosil C 8 column. The wavelength was set at 239 nm. The mobile phase was a mixture of 0.01 M sodium dihydrogen phosphate buffer and acetonitrile (63:37, v/v) adjusted to pH 3.5 at a flow rate of 1.5 ml min -1 . The calibration curve was linear over the concentration range 0.5-16 ng ml −1 . The coefficients of variation for inter-day and intra-day assay were found to be less than 10%.