DEVELOPMENT AND VALIDATION OF STABILITY INDICATING, COST EFFECTIVE AND RAPID METHOD BY USING CONVENTIONAL COLUMN FOR QUANTIFICATION OF APIXABAN RELATED SUBSTANCE IN DRUG SUBSTANCE AND PHARMACEUTICAL PRODUCT

Abstract

Objective: The objective of this analytical research study is to establish new reliable, cost effective, rapid and stability-based indicating analysis support reverse phase liquid chromatographic (RP-HPLC) method by using conventional column for the estimation and identification of Apixaban related compound (impurity) in active pharmaceutical ingredient and pharmaceutical drug product.

Materials and methods: This method has fast eluted all impurities with good resolution by using laboratory conventional C18 with 5µm particle size column. Analytes were separated by simple buffer system and gradient elution program using RP-HPLC, where mobile phase A contains in a 99:1v/v ratio of orthophosphoric acid (OPA) with acetonitrile(ACN) and mobile phase B contains in a 40:60 v/v ratio of ACN and methanol (MeOH). The mobile phase was carried at a flow volume is 1.2mL/min, and injected 20 μL of sample through a C18 conventional column (25 × 0.46cm, 5 μm) at a 230 nm. The new established cost effective simple, fast HPLC method required to validated in accordance with ICH guidelines.

Results: The method validation is representing sensitivity, selectivity, specificity, precise, accurate and linear concentration range from 0.125- 5.0 ppm for known and from 0.05-2.0 ppm for unknown Impurities of Apixaban, robust with slight change in flow, column temperature and wavelength. The Solution was stable up to 72-hrs. All validation data found within pre-measurement acceptance criteria. The method was applied to drug product under various stress condition, major degradation observed in alkali condition.

Conclusion: The output come from method, its indicate that the method is simple, sensitive, robust, rugged, rapid, analysis cost and time saving. also reveals stability indicating for its intended application.