Development and validation of RP-HPLC and UV-spectrophotometric methods for rapid simultaneous estimation of amlodipine and benazepril in pure and fixed dose combination

• Main Authors: Abhi Kavathia, Manju Misra
• Format: Article
• Language: English
• Published: Elsevier 2017-05-01
• Series: Arabian Journal of Chemistry
• Subjects: Amlodipine besylate (AM); Benazepril hydrochloride (BZ); Correlation equation; RP-HPLC
• Online Access: http://www.sciencedirect.com/science/article/pii/S1878535213004139

High-performance liquid chromatographic (HPLC) and UV spectrophotometric methods were developed and validated for the quantitative determination of amlodipine besylate (AM) and benazepril hydrochloride (BZ). Different analytical performance parameters such as linearity, precision, accuracy, specificity, limit of detection (LOD) and limit of quantification (LOQ) were determined according to International Conference on Harmonization ICH Q2B guidelines. The RP-HPLC method was developed by the isocratic technique on a reversed-phase Shodex C-18 5e column. The retention time for AM and BZ was 4.43 min and 5.70 min respectively. The UV spectrophotometric determinations were performed at 237 nm and 366 nm for AM and at 237 nm for BZ. Correlation between absorbance of AM at 237 nm and 366 nm was established and based on developed correlation equation estimation of BZ at 237 nm was carried out. The linearity of the calibration curves for each analyte in the desired concentration range was good (r2 > 0.999) by both the HPLC and UV methods. The method showed good reproducibility and recovery with percent relative standard deviation less than 5%. Moreover, the accuracy and precision obtained with HPLC co-related well with the UV method which implied that UV spectroscopy can be a cheap, reliable and less time consuming alternative for chromatographic analysis. The proposed methods are highly sensitive, precise and accurate and hence successfully applied for determining the assay and in vitro dissolution of a marketed formulation.