Simultaneous estimation of Azelnidipine and Metoprolol succinate with greenness assessment using HPLC and UV-spectrophotometric methods

Abstract

Azelnidipine is a novel dihydropyridine calcium channel antagonist and Metoprolol succinate is a beta blocker, this combination was approved by CDSCO for Phase III clinical trial in April 2021. This combination is utilized to achieve a better outcome in the treatment of stage 2 hypertension. The present research includes an HPLC and Absorbance Correction UV method for the determination in synthetic mixture.The HPLC chromatography was performed using a gradient technique on a reversed-phase C18 column with mobile phase based and optimized depending on the polarity of the molecules. The Unesphere C18 column Agela Tech., (250 mm x 4.6 mm i.d, 5 μm,) as stationary phase and Acetonitrile: Phosphate Buffer pH 3.5 (40:60%v/v) as mobile phase with UV detection at 275 nm at 1 ml/min for gradient elution was used to achieve chromatographic separation over the linearity 10.0 - 30.0 μg/mL and 50.0–150.0 μg/mL with r2 0.9995 and 0.9993 for Azelnidipine and Metoprolol succinate respectively. The retention time was found to be 6.367 and 2.308 min for Azelnidipine and Metoprolol succinate respectively.% RSD of intraday precision of AZL and MET was found to be 0.01 – 0.03% and 0.01 – 0.03%, respectively. Interday presion was found to be 0.02 – 0.06% and 0.02 – 0.05%, respectively which are indicated that the method is precise.% recovery for AZL and MET by this method was found in the range of 99.85 - 100.76% and 99.23 - 99.89%, respectively. The system suitability test parameters theoretical plates were found to be 7042.56 (AZL) and 2231.74 (MET) and tailing factor was found to be 1.47 (AZL) and 1.52 (MET).The Absorbance Correction UV method was developed validated at 313 nm (λmax of Azelnidipine) and 275.40 nm (λmax of Metoprolol succinate) in methanol: distilled water (20:80, v/v) for estimation of AZL and MET respectively and show linearity 5.0 - 25.0 μg/mL and 25.0 - 125.0 μg/mL with r2 0.9998 and 0.9991 for Azelnidipine and Metoprolol succinate respectively.% RSD of intraday precision of AZL and MET was found to be 0.46 – 0.64% and 0.42 – 0.71%, respectively and for interday precision was found to be 0.40 – 0.65% and 0.22 – 0.75%, respectively.% recovery for AZL and MET by this method was found in the range of 100.50 - 100.77% and 100.30 - 100.78%, respectively. The value of% RSD within the limit indicated that the method is accurate and% recovery shows that there is no interference from the excipients.Both methods were validated as per ICH Q2(R1) guidelines. The analytical method validation parameters were found to be as per acceptance criteria. In terms of accuracy and precision, statistical testing employing an analysis of variance test did not find any significant differences between the two methods. The ICH Q2R1 recommendations extensive statistical validation of the two established methodologies, proving their effective use in the simultaneous study of the two medications in bulk and tablets made within the laboratory.The Analytical eco scale (AES), National environmental method index (NEMI), Green analytical procedure index (GAPI), and software-based Analytical greenness metric (AGREE) tools were used to assess the ecological impact of the new UV spectrophotometric technique and HPLC method. This assessment confirmed the method's environmental friendliness in terms of solvent usage, chemical compounds, energy consumption, and trash creation. Both methods can be used to successfully analyze Azelnidipine and Metoprolol succinate in combination dose form without interference from excipients. There were no spectral or chromatographic interferences from formulation excipients discovered. Because the proposed procedures are simple, quick, precise, and accurate, they were effectively used to determine Azelnidipine and Metoprolol succinate in their dose forms. To evaluate the greenness and whiteness of an analytical method, a reliable assessment of the health and environmental risk associated with chemical use is essential. An objective and useful way of risk assessment should consider all pertinent characteristics of substances utilized, the actual quantities required for method implementation, and the result of the assessment should be quantified using a standard unit.