Development and validation of high-performance liquid chromatography (HPLC) method for acetylsalicylic acid, ramipril and indapamide simultaneous analysis

Abstract

Introduction: It is established that 1.13 billion people worldwide have been diagnosed with hypertension [1]. To achieve effective blood pressure reduction most patients have to use different drug combinations [2]. However, increased drug consumption is associated with an increased risk of poisoning [3]. Therefore, it is useful to develop a method for simultaneous analysis of different drugs. The aim of this research is to develop a HPLC method suitable for qualitative and quantitative analysis of acetylsalicylic acid, ramipril and indapamide mixture. Materials and methods: For HPLC method optimization 0.1 mg/ml concentration ethanolic standard solutions of acetylsalicylic acid and indapamide, methanolic standard solution of ramipril and their mixture were analysed. The analysis was carried out on Waters HPLC system equipped with PDA detector and ACE C18 (250 mm × 4.6 mm, 5 μm) column. Samples were chromatographed in a gradient mode (eluent A (aqueous 0.1 % trifluoracetic acid), eluent B (acetonitrile)), eluent flow rate was 1 ml/min, injection volume - 10 μl. Method validation was performed by evaluating specificity, precision, linearity, limit of detection (LOD) and limit of quantification (LOQ). Results: Method specificity was approved when retention times (acetylsalicylic acid – 11.565 min., ramipril – 13.601 min., indapamide – 14.164 min.) and UV spectra of substances derived from medicinal products matched with the standards. Method precision was measured as the relative standard deviation (RSD, %). Repeatability and intermediate precision RSD value for retention time was 0.1 % for all compounds. Linearity was obtained in the concentration range of 0.03125 - 1.0 mg/ml for acetylsalicylic acid, 0.03125 - 0.5 mg/ml for ramipril and indapamide. LOD for acetylsalicylic acid, ramipril and indapamide were 1.399 μg/ml, 7.966 μg/ml, 0.075 μg/ml and LOQ were 4.666 μg/ml, 20.332 μg/ml, 0.251 μg/ml r[...].