Development of a stability-indicating CE assay for the determination of amlodipine enantiomers in commercial tablets (original) (raw)
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Biomedical Chromatography, 2013
A rapid, simple, specific and sensitive LC-MS/MS method has been developed and validated for the enantiomeric quantification of amlodipine (AML) isomers [R-amlodipine (R-AML) and S-amlodipine (S-AML)] with 200 mL of human plasma using R-AML-d 4 and S-AML-d 4 as corresponding internal standards as per regulatory guidelines. A simple liquid-liquid extraction process was used to extract these analytes from human plasma. The total run time was 3.5 min and the elution of R-AML, S-AML, R-AML-d 4 and S-AML-d 4 occurred at 1.62, 2.51, 1.63 and 2.53 min, respectively. This was achieved with a mobile phase consisting of 0.2% ammonia-acetonitrile (20:80, v/v) at a flow rate of 1 mL/min on a Chiralcel OJ RH column. A linear response function was established for the range of concentrations 0.1-10 ng/mL (r >0.998) for each enantiomer. The intra-and inter-day precision values for both enantiomers met the acceptance criteria. Both enantiomers were stable in a set of stability studies, viz. bench-top, auto-sampler, freeze-thaw cycles and long-term. The current assay was successfully applied to a pharmacokinetic study to quantitate AML enantiomers following oral administration of 10 mg AML tablet to humans.
Journal of Separation Science, 2011
An LC method was developed and prevalidated for the enantiomeric purity determination of S-amlodipine in polar organic solvent chromatography using a chlorine-containing cellulose-based chiral stationary phase (CSP). The concentration of formic acid (FA) (0.01-0.2%) in the mobile phase containing acetonitrile as the main solvent was found to influence the elution order of amlodipine enantiomers as well as the enantioresolution. A reversal of the enantiomer elution order of amlodipine was only observed with chiral stationary phases with both electron-withdrawing (chloro) and electron-donating groups (methyl) on the phenyl moieties of the chiral selector, namely cellulose tris(3-chloro-4-methylphenylcarbamate) and cellulose tris(4-chloro-3-methylphenylcarbamate). The highest enantioresolution (R s : 4.1) value was obtained at the lowest FA concentration (0.01%) using cellulose tris(4chloro-3-methylphenylcarbamate) as the chiral selector and the enantiomeric impurity, Ramlodipine, eluted first under these conditions. Therefore, the mobile phase selected for the prevalidation of the method consisted of ACN/0.1% DEA/0.01% FA and the temperature was set at 251C. The method was prevalidated by means of the strategy based on the total measurement error and the accuracy profile. The method was found to be selective and the limit of quantification was found to be about 0.05% for R-amlodipine, while the limit of detection was close to 0.02%.
Comparative Enantioseparation of Amlodipine by HPLC and Capillary Electrophoresis
Acta Medica Marisiensis
Objective: The purpose of this study was to separate the enantiomers of amlodipine by High Performance Liquid Chromatography (HPLC) using ovomucoid (OVM) as chiral selector, respectively by Capillary Electrophoresis (CE) using cyclodextrines and to evaluate the analytical performance of the both proposed methods. Material and methods: HPLC enantioseparation of amlodipine was performed on an HPLC Agilent Technologies 1100 series using as chiral stationary phase an Ultron ES OVM, 150x4.6 mm column with ovomucoid as chiral selector. The stereoselective CE analysis of amlodipine was achieved on Agilent Technologies 7100 CE using uncoated fused-silica capillaries 48 cm x 50 mm and different type of cyclodextrins as chiral selectors. Results: A mobile phase consisting of 80% Na2HPO4 10 mM at a pH level of 5.0 and 20% ACN, isocratic elution at a flow of 1 ml/min turned to be the optimal experimental conditions for HPLC analysis (R=5.51; α=1.71) with retention times shorter than 10 minutes ...
Green RP-HPLC method for impurity profile of amlodipine in tablets
Arhiv za farmaciju, 2024
Increased awareness of nature preservation has encouraged the introduction of the green analytical chemistry (GAC) practice concepts concerning several important aspects, including sustainable development, environmental impact, and minimum waste. The aim of this research was to contribute to the implementation of this approach for the pharmaceutical industry while retaining the crucial aspects and strict requirements of quality control of medicines. Therefore, an ethanolbased, green and robust high-performance liquid chromatography (HPLC) method for the determination of related substances of amlodipine (AML) in film-coated tablets was developed and optimized using the Design of Experiments (DoE). The chromatographic separation was performed on an RP-select B column (250 x 4.0 mm, 5 μm), using a mixture of 0.04 M sodium dihydrogen phosphate monohydrate (pH 4.0) and ethanol (60:40 % v/v) as a mobile phase. The optimized conditions provided the separation of two specified impurities (impurity D and impurity F). The selectivity of the method was confirmed using forced degradation studies. The Analytical Eco-scale approach and AGREE metrics confirmed that the method conforms to the GAC principles. The validated method was successfully applied for the determination of related substances in three samples from the market, demonstrating the applicability of the method in routine analysis.
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.
American Journal of Analytical Chemistry, 2013
A stability indicating LC method was developed for the simultaneous determination of Amlodipine and Benazepril capsules in pharmaceutical dosage form. Efficient chromatographic separation was achieved on Symmetry C 18 stationary phase with simple combination of amobile phase containing 750 mL of DI Water, 250 mL of Acetonitrile and 2 mL of Octylamine into suitable container with adjusted pH to 2.50 ± 0.05 with the aid of Ortho phosphoric acid delivered in an isocratic mode and quantification was carried out using UV detection at 240 nm at a flow rate of 1.0 mL•min −1 with an injection volume of 20 μl and ambient column temperature. This method is capable to detect both the drug components of Amlodipine and Benazepril in presence of their degradation products (Amlodipine Imp-A and Benazepril Impurity-C) with a detection level of 0.05%. Amlodipine/Benazepril in their combination drug product were exposed to thermal, photolytic, hydrolytic and oxidative stress conditions, and the samples were analysed. Peak homogeneity data of Amlodipine and Benazeprilis were obtained using PDA detector, in the stressed sample chromatograms, demonstrating the specificity. The method shows excellent linearity over a range of 0.05%-2.0% for Amlodipine, Amlodipine Impurity-A and 0.05%-5.0% for Benazepril and Benazepril Impurity-C. The correlation coefficient for Amlodipine and Benazepril is 1. The relative standard deviation was always less than 2%. The proposed method was found to be suitable and accurate for quantitative determination and the stability study of Amlodipine and Benazepril in pharmaceutical preparations. The developed HPLC method was validated with respect to linearity & range, accuracy, precision and robustness.
Journal of Separation Science, 2010
Electro membrane extraction as a new microextraction method was applied for the extraction of amlodipine (AM) enantiomers from biological samples. During the extraction time of 15 min, AM enantiomers migrated from a 3 mL sample solution, through a supported liquid membrane into a 20 mL acceptor solution presented inside the lumen of the hollow fiber. The driving force of the extraction was 200 V potential, with the negative electrode in the acceptor solution and the positive electrode in the sample solution. 2-Nitro phenyl octylether was used as the supported liquid membrane. Using 10 mM HCl as background electrolyte in the sample and acceptor solution, enrichment up to 124 times was achieved. Then, the extract was analyzed using CD modified CE method for separation of AM enantiomers. Best results were achieved using a phosphate running buffer (100 mM, pH 2.0) containing 5 mM hydroxypropyl-a-CD. The range of quantitation for both enantiomers was 10-500 ng/mL. Intra-and interday RSD (n 5 6) were less than 14%. The limits of quantitation and detection for both enantiomers were 10 and 3 ng/mL respectively. Finally, this procedure was applied to determine the concentration of AM enantiomers in plasma and urine samples.
International journal of health sciences
Objective: A simple, Accurate, precise method was developed for the simultaneous estimation of the Amlodipine besylate and Enalapril maleate in pharmaceutical dosage form. Methods: Chromatogram was run through BEH C18 column (2.1 × 50mm i.d., 1.7 μm particle size). Mobile phase containing methanol: 1N HCl (1:1) was pumped through column at a flow rate of 1ml/min. Temperature was maintained at Ambient. Optimized wavelength for Amlodipine besylate and Enalapril maleate was 272 nm. Results: Retention time of Amlodipine besylate and Enalapril maleate were found to be 1.04 min and 0.59 min. The % purity of Amlodipine besylate and Enalapril maleate was found to be 100.03 % and 99.75 % respectively. The system suitability parameters for Amlodipine besylate and Enalapril maleate such as theoretical plates were found to be 5659.11 and 3214.07. the resolution was found to be 4.58,2.91. The linearity study for Amlodipine besylate and Enalapril maleate was found in concentration range of 10μg-...