Development of a Sustainable High-Performance Liquid Chromatography (HPLC) Method for Quantifying Metformin, Sitagliptin, and Empagliflozin in Type 2 Diabetes Treatment

Abstract

Objective: To develop and validate a green, environmentally sustainable high-performance liquid chromatography (HPLC) method for the simultaneous determination of three commonly used antidiabetic agents: Metformin Hydrochloride (MET), Sitagliptin Phosphate (STG), and Empagliflozin (EMP). 

Study Design: Experimental analytical study.

Place and Duration of Study:  This study was conducted at the State Company for Drugs Industry and Medical 
Appliances Samarra (SDI) Iraq from April 2024 to May 2024. 

Methods: The chromatographic separation was achieved using an isocratic elution on a C10 column (4.6 × 250 mm, 5 µm) with a mobile phase consisting of phosphate buffer (20 mM), methanol, and acetonitrile in the ratio of  65:30:5 (v/v/v), adjusted to pH 2.9. The flow rate was maintained at 1.0 mL/min, with a column temperature of  30°C and a total run time of 15 minutes. Detection was performed using UV spectroscopy at 208 nm. Linearity was established for MET (0.08–0.13 mg/mL), STG (0.035–0.065 mg/mL), and EMP (0.014–0.026 mg/mL), with correlation coefficients exceeding 0.998 and RSD% not exceeding 1.06%. The method’s greenness was evaluated using AGREE and GAPI tools. Additionally, the Blue Applicability Grade Index (BAGI) was employed to assess operational and environmental suitability. 

Results: The method exhibited high accuracy, reproducibility, and compliance with green analytical chemistry standards. The AGREE and GAPI assessments confirmed the method's minimal environmental impact. BAGI scoring yielded a value of 82.5, indicating strong sustainability and applicability. The method was successfully applied to the analysis of 13 commercial antidiabetic products, including branded and generic formulations available in the Iraqi market. 

Conclusion: The developed HPLC method provides a reliable, accurate, and environmentally sustainable approach for the simultaneous analysis of MET, STG, and EMP. It demonstrates strong analytical performance and practical utility, contributing to green pharmaceutical quality control and sustainable laboratory practices.