DEVELOPMENT AND VALIDATION OF SEVEN PHENYL HYDRAZINE CHLORO ESTER ISOMERS (PGIs) BY RP-HPLC-UV METHOD IN ANTICOAGULANT DRUG SUBSTANCE; APIXABAN

SARADHI VENKATA RAMANA V.; DURGA RAJA K.; RAGHU BABU K.; PADMA M.; JAGADEESH KUMAR V.; PAVAN KUMAR K. S. R.; HEMANT KUMAR SHARMA

doi:10.22159/ijcpr.2020v12i2.37507

Authors

SARADHI VENKATA RAMANA V. Department of Engineering Chemistry, Andhra University, Vishakhapatnam, Andhra Pradesh, India 53003, Analyical Research Department, Aurobindo Pharma Limited Research Centre-II, Indrakaran (V), Kandi (M), Sangareddy Dist, Telangana, India 502329
DURGA RAJA K. Department of Engineering Chemistry, Andhra University, Vishakhapatnam, Andhra Pradesh, India 53003, Analyical Research Department, Aurobindo Pharma Limited Research Centre-II, Indrakaran (V), Kandi (M), Sangareddy Dist, Telangana, India 502329
RAGHU BABU K. Department of Engineering Chemistry, Andhra University, Vishakhapatnam, Andhra Pradesh, India 53003,
PADMA M. Department of Engineering Chemistry, Andhra University, Vishakhapatnam, Andhra Pradesh, India 53003
JAGADEESH KUMAR V. Analyical Research Department, Aurobindo Pharma Limited Research Centre-II, Indrakaran (V), Kandi (M), Sangareddy Dist, Telangana, India 502329
PAVAN KUMAR K. S. R. Analyical Research Department, Aurobindo Pharma Limited Research Centre-II, Indrakaran (V), Kandi (M), Sangareddy Dist, Telangana, India 502329
HEMANT KUMAR SHARMA Analyical Research Department, Aurobindo Pharma Limited Research Centre-II, Indrakaran (V), Kandi (M), Sangareddy Dist, Telangana, India 502329

DOI:

https://doi.org/10.22159/ijcpr.2020v12i2.37507

Keywords:

Seven potential genotoxic impurities, Reverse phase-HPLC-UV, Method development, Validation, Apixaban drug substance

Abstract

Objective: The objective of this work was to develop and validate a simple and sensitive reverse-phase high-pressure liquid chromatography method for the determination of seven potential genotoxic impurities in Apixaban drug substance.

Methods: The optimized separation was achieved by using ACE 3 C18 PFP (150 mm×4.6 mm, 3 µm) HPLC column. The mobile phase-A was a degassed mixture of 0.01M Ammonium acetate buffer(P^H adjusted 4.9±0.05 with diluted glacial acetic acid) and mobile phase-B was a degassed mixture of Acetonitrile, Isopropyl alcohol and Buffer P^H 4.9 in the ratio of 60:20:20 v/v/v. The gradient program was operated at a flow rate of 1.0 ml/min and UV detection was at 330 nm.

Results: The method was superior at linearity for seven impurities and correlation coefficient values were larger than 0.999, moreover, in the separation point of view, this method further achieved no matrix interference through chromatography by better resolution of the other impurities from the Apixaban drug substance and its related impurities for the accurate analysis of seven potential genotoxic impurities. The established limits of detection (LOD), limits of quantification (LOQ) values for the seven mutagenic impurities were each of 5 ppm (0.015µg/ml) and15 ppm (0.045µg/ml) respectively. The developed method was validated as per ICH guidelines and applied as a generic method to determine these seven potential genotoxic impurities for the pharmaceutical process control and drug material release.

Conclusion: Validation of this analytical method was carried out including stability, selectivity, linearity, accuracy, system precision, method precision and intermediate precision thus proving that the described RP-HPLC method could be employed for fast and simple analysis of sevenphenyl hydrazine chloro ester isomers in Apixaban drug substance.

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