Development and Validation of RP-LC Method for Simultaneous Estimation of Amlodipine Besylate and Valsartan in Bulk and Its Pharmaceutical Formulations (original) (raw)
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Asian Journal of …
The present study describes development and validation of simple, rapid, sensitive and stability indicating highperformance liquid chromatographic assay method for simultaneous determination of Amlodipine besylate and Valsartan in bulk as well as in commercial formulation. Analytical separation was achieved with RP Waters Symmetry C18 Column [150×4.6 mm] using a combination of methanol and potassium dihydrogen phosphate buffer [0.01 M] pH 2.5, in ratio 60:40 v/v with flow rate of 1 mL min −1 . The retention times for Amlodipine and Valsartan were found to be 4.6 and 7.6 min respectively. Both the drugs were exposed to thermal, photolytic, hydrolytic, and oxidative stress conditions individually as well as in combination, subsequently samples were analyzed by the proposed method. Detection was done by PDA detector at 238 nm. Method was found to be very sensitive as LOD found to be 20 ng/mL and 44 ng/mL for Amlodipine besylate and Valsartan respectively. The method was found to be specific and stability indicating as no interfering peaks of degradation compounds and excipients were noticed. The developed method was validated with respect to linearity, accuracy, precision, specificity, robustness as per International Conference on Harmonization (ICH) guidelines. The proposed method hence useful for the application in quality control laboratories for quantitative analysis of both the drug individually or in combination, as it is simple and rapid method with excellent accuracy and precision.
Acta Chromatographica, 2012
A reverse-phase high-performance liquid chromatographic (RP-HPLC) method was developed for the simultaneous estimation of amlodipine besylate (AMB), valsartan (VAL), and hydrochlorothiazide (HCT) in pharmaceutical formulation using RP-C 18 column. The mobile phase (acetonitrile:methanol:50 mM phosphate buffer adjusted to pH 3 with orthophosphoric acid) was pumped at a flow rate of 1.0 mL min −1 in the ratio of 20:50:30% v/v and the eluents were monitored at 239 nm. Linearity was obtained in the concentration range of 0.5-5 μg mL −1 for AMB, 4-40 μg mL −1 for VAL, and 1-10 μg mL −1 for HCT. The method was validated as per International Conference on Harmonization (ICH) guidelines and statistically. The method was validated for accuracy and precision. For precision, the coefficient of variance (COV) was found to be 0.3794, 0.1703 and 0.0578, and for accuracy, it was found to be 0.6351, 0.7688 and 1.1305 for AMB, VAL, and HCT, respectively. The COV values for all the drugs were found to be less than 2%, indicating high degree of precision and accuracy of the proposed highperformance liquid chromatographic (HPLC) method. Owing to its simplicity, rapidness, high precision and accuracy, the proposed HPLC method can be applied for determining AMB, VAL, and HCT in bulk and in pharmaceutical dosage form.
2016
Title: Development and Validation of RP-LC Method for Simultaneous Estimation of Amlodipine Besylate and Valsartan in Bulk and Its Pharmaceutical Formulations K Geetha Bhavani , N Srinivasu, Nanduri Gayatri Devi and D. Ramachandran JMJ College for Women’s, Tenali, A.P, India 2 Department of Science and Humanities, Vignan University, Vadlamudi, A.P, India. 3 Department of Chemistry, Acharya Nagarjuna University, Nagarjuna Nagar, Guntur, A. P, India
Acta Chromatographica, 2015
A new ultra-performance liquid chromatography method for the simultaneous determination of valsartan, amlodipine besylate, and hydrochlorothiazide in pharmaceutical formulations has been developed. Chromatographic separation was achieved within 1 min on a sub-2 μm RP18 column using an isocratic mode mobile phase consisting of a mixture of methanol and phosphate buffer (0.05 M, pH 3 ± 0.05) in the ratio 70:30 (v/v). The flow rate was set at 0.3 mL min −1 with a detection wavelength of 239 nm. The method was validated over the concentration ranges from 6.5 to 15.0 μg mL −1 for hydrochlorothiazide, from 5.0 to 12 μg mL −1 for amlodipine besylate and from 76.5 to 178.5 μg mL −1 for valsartan. Calibration plots were linear for valsartan, amlodipine besylate, and hydrochlorothiazide with a correlation coefficient greater than 0.99. Relative variation coefficients for repeatability and reproducibility were less than 3.0%. Recoveries from standard addition essay of individual component in pharmaceutical formulations were varied from 97.0% to 102.0%. The proposed method was successfully applied for the determination of valsartan, amlodipine besylate, and hydrochlorothiazide in pharmaceutical formulations with recoveries with respect to label amount in pharmaceutical formulations that varied from 98.0% to 102.9%. The low flow rate, short analysis time, and simple mobile phase composition make the method cost effective, rapid, nontedious, and successfully employed for simultaneous determination of these drugs in commercial pharmaceutical products.
Chromatographia, 2009
This paper describes a new, simple, precise, and accurate TLC method for simultaneous quantitation of amlodipine (AML) besylate and valsartan (VAL) as the bulk drug and in tablet dosage forms. Chromatographic separation of the drugs was performed on aluminum plates precoated with silica gel 60 F 254 as the stationary phase and the solvent system consisted of toluene:methanol:acetic acid 7:3:0.1 (v/v/v). Densitometric evaluation of the separated zones was performed at 244 nm. The two drugs were satisfactorily resolved with R F values of 0.41 ± 0.02 and 0.54 ± 0.02 for AML and VAL, respectively. The accuracy and reliability of the method was assessed by evaluation of linearity (100-600 ng spot-1 for AML and 1,600-9,600 ng spot-1 for VAL), precision (intra-day RSD 1.51-1.83% and inter-day RSD 1.24-1.95% for AML, and intra-day RSD 0.14-0.39% and inter-day RSD 0.19-0.52% for VAL), accuracy (98.29 ± 1.16% for AML and 98.72 ± 0.50% for VAL), and specificity, in accordance with ICH guidelines.
Simple and rapid assay method is primary prerequisite in quality assurance department for any pharmaceutical dosage form. When two drugs are combined in a single dosage form, it becomes difficult to estimate them simultaneously with required accuracy and precision. Here we have proposed a simple and rapid UV spectrophotometric assay method for simultaneous determination of amlodipine besylate and valsartan in both bulk drug and commercial dosage forms. Method employed was Zero absorbance method where amlodipine and valsartan were determined at 363 nm and 290 nm respectively. At 363 nm valsartan don't have absorption and at 290 nm amlodipine has zero absorption. Linearity was established at 8 µg/mL -80 µg/mL for amlodipine (y= 0.012X + 0.002 R 2 = 0.999) and 4 µg/mL -300 µg/ml for Vasartan (y= 0.002X + 0.017 R 2 = 0.999). Method was found to be accurate and precise with %RSD<2 for all the validation parameters employed. Method was found to be very sensitive with LOD 2.4 µg/mL and 3.6 µg/mL whereas LOQ was 7.5 µg/mL and 11 µg/mL for Amlodipine besylate and Valsartan respectively. As method uses methanol as a solvent to dissolve, it is cost effective for routine analysis in quality assurance department for determination of these drugs individually or simultaneously.
2011
Three methods were developed for simultaneous determination of amlodipine and valsartan without previous separation. The first method depends on first derivative of the ratios spectra (DD1) by measurements of the amplitudes at 234.5 and 247 nm for amlodipine using 30μg/mL of valsartan as a divisor and at 282 and 292 nm for valsartan using 80μg/mL of amlodipine as a divisor. Calibration graphs were established in the range of 10-100 μg/mL and 5-40 μg/mL for amlodipine and valsartan, respectively. The second method describes the use of multivariate spectophotometric calibration for the simultaneous determination of the analyzed binary mixture, where the resolution is accomplished by using partial least squares (PLS) regression analysis. In the third method, High performance liquid chromatography (HPLC), separation was performed by using reversed phase column and a mobile phase composed of acetonitrile: KH 2 PO 4 (50:50v/v) adjusted to pH 3.5 by phosphoric acid. The proposed methods were validated and results obtained by adopting the three methods were statistically analyzed and compared with those obtained from compendial method.
Pharmaceutica Analytica Acta, 2014
Amlodipine besylate is a calcium channel blocker which is used in treatment of hypertension alone or in combination with other antihypertensive drugs like angiotensin-II-receptor antagonists (ARA II) group (Losartan potassium and Valsartan) or in combination with anti hyperlipidemic agent like Atorvastatin calcium. RP-HPLC method was developed for the assay of these drugs. The method was performed by reversed phase high performance liquid chromatography using a mobile phase 0.01 M ammonium acetate buffer (pH 5.5): acetonitrile with detection at 240 nm on a spherical monomeric C18 column (250 mm × 4.6 mm, 5 μm) at flow rate of 1.5 ml/min. The proposed method was validated in terms of linearity ranged between [(2-12, 10-60, 16-96, 4-24 µg/ml) corresponding levels of 20-120% w/w of the nominal analytical concentration] with linear regression equations were [{y=64.627x-3.6383 (r= 0.9998), y=75.385x-8.3856 (r= 0.9997), y=64.492x-25.981 (r= 0.9998), y=70.964x-28.505 (r= 0.9998}], accuracy [100.18 ± 1.38, 100.79 ± 0.59, 100.45 ± 0.58 and 100.8 ± 1.69%], precision [99.29, 99.33, 99.30 and 99.30%], limits of detection [0.03, 0.18, 0.15, 0.007 µg/ml] and limits of quantitation [0.1, 0.54, 0.45, 0.024 µg/ml] for Amlodipine besylate, Losartan potassium, Valsartan and Atorvastatin calcium respectively. Method validation was developed following the recommendations for analytical method validation of International Conference on Harmonization (ICH) and Food and Drug Administration (FDA) organizations.
Journal of Chromatographic Science, 2013
Accurate, sensitive and reproducible reversed-phase high-performance liquid chromatography (RP-HPLC), high-performance thin-layer chromatography (HPTLC) and ultraviolet (UV) spectrophopometric methods were developed for the concurrent estimation of amlodipine besylate (AMLO), hydrochlorothiazide (HCTZ) and valsartan (VALS) in bulk and combined tablet dosage forms. For the RP-HPLC method, separation was achieved on a C18 column using potassium dihydrogen orthophosphate buffer (50 mM, pH 3.7) with 0.2% triethylamine as the modifier and acetonitrile in the ratio of 56:44 (v/v) as the mobile phase. Quantification was achieved using a photodiode array detector at 232 nm over a concentration range of 2-25 mg/mL for AMLO, 5-45 mg/mL for HCTZ and 20-150 mg/mL for VALS. For the HPTLC method, the drugs were separated by using ethyl acetatemethanol-toluene-ammonia (7.5:3:2:0.8, v/v/v/v) as the mobile phase. Quantification was achieved using UV detection at 242 nm over a concentration range of 100-600 ng/spot for AMLO, 150-900 ng/spot for HCTZ and 1,200-3,200 ng/spot for VALS. The UVspectrophotometric simultaneous equation method was based on the measurement of absorbance at three wavelengths; i.e., at 237.6 nm (l max of AMLO), 270.2 nm (l max of HCTZ) and 249.2 nm (l max of VALS) in methanol. Quantification was achieved over the concentration range of 2-20 mg/mL for AMLO, 5-25 mg/mL HCTZ and 10-50 mg/mL for VALS. All methods were validated according to International Conference on Harmonization guidelines and successfully applied to marketed pharmaceutical formulations. Additionally, the three methods were compared statistically by an analysis of variance test, which revealed no significant difference between the proposed methods with respect to accuracy and precision. Experimental Chemicals and reagents Standard drug samples of AMLO, VALS and HCTZ were gifted by Torrent Research Centre (Ahmedabad, India). A marketed
Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 2009
A rapid, precise, specific, and accurate ultra performance liquid chromatography (UPLC) method has been developed and subsequently validated for simultaneous determination of amlodipine present as amlodipine basylate (AB) and benazepril hydrochloride (BH) in capsule dosage form. The chromatographic separation was achieved on an Acquity UPLC, BEH C8 (100 mm x 2.1 mm, 1.7 microm) column using a photodiode array detector. The mobile phase used consisted of a mixture of phosphate buffer pH 3.0 (0.01 M aqueous potassium dihydrogen phosphate, pH 3.0 adjusted with orthophosphoric acid) and solvent mixture (equal mixture of acetonitrile and methanol) in the ratio of 45:55 (v/v) at a flow rate of 0.3 mL/min. The described method was linear over ranges of 5.21-15.63 microg/mL for AB and 20.24-60.72 microg/mL for BH. The mean recoveries were 100.47 and 99.97% for AB and BH, respectively. The lower limit of quantification was determined on the basis of signal-to-noise ratio method; it is 0.01 m...