Liquid Chromatography–Mass Spectrometry Method for the Determination of Amlodipine in Human Plasma and its Application in a Bioequivalence Study

Abstract

A sensitive and selective liquid chromatographic-mass spectrometric (LC-MS) method for the determination of amlodipine (CAS 88150-42-9) in human plasma has been developed. Sample preparation was based on liquid-liquid extraction using NaOH and cyclohexane. Analytical determination was carried out on a C18 column interfaced with a single quadrupole mass spectrometer. Positive electrospray ionization was employed as the ionization source. The mobile phase was 10 mmol/L ammonium acetate solution-methanol (30:70, v/v) at the flow rate of 0.2 mL/min. The method was linear in the concentration range of 0.2-20 ng/mL. The lower limit of quantification was 0.2 ng/mL. Amlodipine was sensitive to UV light. The method was validated in terms of accuracy, precision, absolute recovery, and stability. The intra- and interday relative standard deviation across three validation runs over the entire concentration range was less than 8.17%. The accuracy determined at three concentrations (0.4, 2.0 and 10 ng/mL for amlodipine maleate) was within +/- 3.17% in terms of relative error. The method herein described was successfully applied for the evaluation of pharmacokinetic profiles of amlodipine maleate tablets in 20 healthy volunteers. The results showed that AUC, Tmax, Cmax and T 1/2 between the test and reference formulation have no significant difference (P > 0.05). The relative bioavailability was 103.7 +/- 12.3%.

HPLC–MS–MS for the Simultaneous Determination of Atorvastatin and Amlodipine in Plasma of Hypertensive Patients

“…According to the reported data, our method appears sensitive enough for determination of atorvastatin and amlodipine in human plasma samples collected up to 24 or 96 h after administration. In addition, the sensitivity of the present assay was comparable to the reported studies for the individual determination of atorvastatin or amlodipine [9][10][11][12][13]. However, the sample pretreatment method employed in this study was much simpler than those of reported studies.…”

“…There were many studies available for the determination of amlodipine or atorvastatin in human plasma [9][10][11][12][13]. However, the study for the simultaneous determination of amlodipine and atorvastatin in human plasma was limited.…”

Simultaneous Estimation of Amlodipine and Atenolol in Human Plasma: A Sensitive LC–MS/MS Method Validation and its Application to A Clinical PK Study

“…A thorough literature review revealed that several LC-MS/MS methods have been published for the determination of AMD and ATL individually (for AMD [9][10][11][12][13][14][15][16] and for ATL [17]), or along with other drug(s) (ATL with other drugs [18][19][20][21][22] and AMD with other drugs [23][24][25][26][27]) in various biological matrices. It is important to develop a sensitive, rapid and reliable bioanalytical method for simultaneous quantitation of two or more drugs in combination therapy in human plasma.…”

Multivariate Versus Classical Univariate Calibration Methods for Spectrofluorimetric Data: Application to Simultaneous Determination of Olmesartan Medoxamil and Amlodipine Besylate in their Combined Dosage Form

“…Several analytical methods have been reported for the determination of AML in pharmaceutical formulations and/or biological fluids. These methods include spectrophotometry [19][20][21], spectrofluorimetry [22,23], anodic stripping voltammetry [24,25], HPLC [26][27][28][29][30][31], HPTLC [32], capillary electrophoresis [33] and micellar electrokinetic chromatography [34].…”