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HomePharmacology & Drug Discovery ResearchAssay and Organic Impurity Profiling of Apixaban Using an Ascentis® Express C18 Column and UV Detection

Assay and Organic Impurity Profiling of Apixaban Using an Ascentis® Express C18 Column and UV Detection

Sonal Shinde Application Scientist

Introduction

Apixaban is a selective, orally bioavailable, and reversible inhibitor of free and clot-bound factor Xa (which catalyzes the conversion of prothrombin to thrombin). Apixaban is sold under the trade name Eliquis®, and is typically prescribed to lower the risk of stroke and embolism in people with nonvalvular atrial fibrillation. It is also used to prevent deep vein thrombosis (DVT) which may lead to pulmonary embolism (PE) in knee or hip replacement surgery patients. Apixaban was developed in a joint venture by Pfizer and Bristol-Myers Squibb, and it was approved in 2012 in Europe, and in 2014 in the U.S.

At present, there are no pharmacopoeial monographs available with either an assay and/or organic impurity profiling method for Apixaban. We have therefore developed and validated a new HPLC method for said purposes using a column with C18 Fused-Core® particles and UV detection at 235 nm. The limit of detection (LOD) is 0.33 ppm and the limit of quantitation (LOQ) is 1.0 ppm at 235 nm for Apixaban using HPLC-UV detection.

Apixaban
Chromatographic data - blank & standard solution

Figure 1. Chromatographic data - blank & standard solution.

Chromatographic data - system suitability test (SST) solution

Figure 2.Chromatographic data - system suitability test (SST) solution.

Chromatographic data - sample solution

Figure 3. Chromatographic data - sample solution,

Validation and Verification

1. Specificity

Inject standard solution and determine the retention time of desired analyte in the presence of other components like impurities and excipient.

2. Standard Repeatability (25 ppm)

3. Linearity, LOD & LOQ

Apixaban calibration curve data (0-37.5 ppm).

Figure 4. Apixaban calibration curve data (0-37.5 ppm).

Materials
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