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. 2024 Oct 15;16(10):5311-5325.
doi: 10.62347/VYDX5901. eCollection 2024.

Comprehensive analysis of Aspirin and Apixaban: thedevelopment, validation, and forced degradation studies of bulk drugs and in-house capsule formulations using the RP-HPLC method

Tarang Patel  1 , Mehul Patel  1 , Umang Shah  1 , Ashish Patel  1 , Swayamprakash Patel  2 , Nilay Solanki  3 , Siddharth Shah  4
Affiliations

Comprehensive analysis of Aspirin and Apixaban: thedevelopment, validation, and forced degradation studies of bulk drugs and in-house capsule formulations using the RP-HPLC method

Tarang Patel et al. Am J Transl Res. .

Abstract

Objectives: This study aimed to develop a robust Reverse Phase-High-Performance Liquid Chromatography (RP-HPLC) method for simultaneous determination of Aspirin (ASP) and Apixaban (API) in bulk and in-house capsule formulations.

Methods: The separation was conducted on a Phenomenex Luna C18 column using a Shimadzu LC20AT High-performance liquid chromatography (HPLC) system. The mobile phase consisted of 40:60 Acetonitrile (ACN): phosphate buffer (pH 4) modified by O-Phosphoric Acid (OPA). The parameters included a flow rate of 1 ml/min, a column temperature of 30°C, and Ultra-Violet (UV) detection at 227 nm. Method validation encompassed linearity, precision (Intraday and Interday), accuracy (% Recovery), and sensitivity (Limit of Detection (LOD) and Limit of Quantification (LOQ)). Stability testing followed The International Council for Harmonization (ICH) guidelines.

Results: The developed method demonstrated reliable separation of Aspirin and Apixaban with retention times of 5.37 min and 7.10 min, respectively. It exhibited linearity over the concentration ranges of 50-300 μg/mL for Aspirin and 5-15 μg/mL for Apixaban. The recovery percentage ranged from 90.02% to 101% for Aspirin and 98.18% to 101.18% for Apixaban. LOD and LOQ were determined as 0.84 μg/mL and 2.55 μg/mL for Aspirin, and 0.41 μg/mL and 1.24 μg/mL for Apixaban, respectively. Stability testing confirmed the method's robustness under various stress conditions.

Conclusions: The validated RP-HPLC method offers a reliable tool for routine analysis of Aspirin and Apixaban in pharmaceutical formulations, highlighting its potential for combined dosage applications and routine quality control.

Keywords: Apixaban; Aspirin; ICH; RP-HPLC; forced degradation studies; method development and validation.

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Conflict of interest statement

None.

Figures

Figure 1
Figure 1
Chemical structures (A) Chemical structure of aspirin (acetylsalicylic acid), (B) Chemical structure apixaban.
Figure 2
Figure 2
Chromatogram analysis under various conditions. A. Blank chromatogram. B. Optimized chromatogram of Aspirin and Apixaban in mixture. C. Peak purity curve of aspirin. D. Peak purity curve of apixaban.
Figure 3
Figure 3
Chromatogram analysis of placebo and formulation samples under optimized conditions. A. Placebo. B. Chromatogram of In-house capsule formulation. C. Peak purity curve of aspirin (In-house formulation). D. Peak purity curve of apixaban (In-house formulation).
Figure 4
Figure 4
Chromatographic profiles illustrating peak degradation under acidic conditions in (A) bulk and (B) formulation samples.
Figure 5
Figure 5
Chromatographic analysis of peak degradation under alkaline conditions in (A) bulk and (B) formulation samples.
Figure 6
Figure 6
Chromatographic profiles illustrating peak degradation under oxidative conditions in (A) bulk and (B) formulation samples.
Figure 7
Figure 7
Chromatographic profiles illustrating peak degradation under photolytic stress conditions in (A) bulk and (B) formulation samples.
Figure 8
Figure 8
Chromatographic profiles illustrating peak degradation under thermal stress conditions in (A) bulk and (B) formulation samples.
Figure 9
Figure 9
Chromatographic profiles illustrating peak degradation under refrigerated temperature conditions in (A) bulk and (B) formulation samples.
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