Research Article

Assay comparison of rivaroxaban by new HPLC method with an existing method in tablet dosage form

Suraj Sahoo1*, Suman Kumar Mekap2

1School of Pharmaceutical Education and Research, Berhampur University, Berhampur, Odisha, India

2Roland Institute of Pharmaceutical Sciences, Berhampur, Odisha, India

*For correspondence

Dr. Suraj Sahoo,

School of Pharmaceutical Education and Research, Berhampur University, Berhampur, Odisha, India.

Email: montesus@gmail.com

 

 

 

 

 

 

Received: 02 April 2017

Accepted: 14 May 2017

ABSTRACT

Objective: In the present work, RP-HPLC procedure is optimized to finalize a different approach for the estimation of rivaroxaban in tablet dosage form. A novel drug rivaroxaban used as anti-coagulant in the patients for the prevention of thromboembolism.

Methods: The molecule is identified with the molar mass of 435.882 g/mol and molecular formula C19H18ClN3O5S. The determination was executed by C18 column (Phenomenex 250 x 4.6 mm, 5 μm maintained at 35°C) at 251 nm with a mobile phase (ACN: Water, 55:45 v/v) and flow of 1.2 ml/min.

Results: The retention time found to be about 3.8minutes.The validation parameters performed as per ICH guidelines and found to be within acceptance criteria. Linearity of the method is found to be accepted across five concentration level i.e. being studied by calibration curve. Accuracy was executed at three different concentrations, the amount being recovered are close to 100%. The % RSD values obtained for repeatability, intermediate and reproducibility under precision are within acceptance criteria.

Conclusions: The method was accurate, precise, robust and rapid for quantitative determination of rivaroxaban by High Performance Liquid Chromatography.

Keywords: Rivaroxaban, Validation, RP HPLC, ICH

Introduction

Anticoagulants are administered to prevent the clots from getting larger or blood from clotting. Clot can develop serious conditions like heart attack or stroke in cases where clots block blood flow to heart or brain.

Rivaroxaban, an oxazolidinone based oral anticoagulant is characterized with molecular mass of 435.882 g/mol and formula of C19H18ClN3O5S i.e. sparingly soluble in water.1

Rivaroxaban is used in the prevention of venous thromboembolism in adult patients after total hip replacement or total knee replacement surgery and first available, potent, selective direct inhibitor of factor Xa.2 Rivaroxaban is also used in two more indications: prevention of stroke and systemic embolism in adult patients with nonvalvular atrial fibrillation and treatment of deep vein thrombosis and pulmonary embolism.3 The drug is invented and manufactured by a pharmaceutical company, Bayer. The brand Xarelto is marketed in a number of countries.

Literature survey express colorimetric method, spectrphotometric methods, HPLC method and bio analytical method for determination of Rivaroxaban.4,5 The present study was aimed to develop a suitable assay for the quantitative estimation of Rivaroxaban in its dosage form, validate (as per ICH guidelines) the developed assay method by using HPLC then comparing the assay result with one existing method.

Figure 1: Chemical structure of rivaroxaban.

Materials and Methods

Chemicals and reagents

Rivaroxaban API was procured from Nerus lifecare Pvt. Ltd., New Delhi, marketed pharmaceutical formulation XARELTO 10 mg Tablet, Acetonitrile and water of HPLC grade were purchased from local seller.

Apparatus and chromatographic conditions

Analysis was performed over Shimadzu Prominence Modular HPLC. The chromato-graphic parameters set in the HPLC were phenomenex 250 mm × 4.6 mm, 5 µm C 18 column (maintained at 35°C), 20 µl injection volume, 251nm λ max for UV detection, 1.2 ml/min flow rate (isocratic mode) with mobile phase (ACN: Water, 55:45 v/v). Weighing balance make was metler Toledo, sonicator (make) were used to degass the mobile phase. Column was equilibrated at least for 30 minutes to minimize noise and other possible interference from previous injections.

Preparation of standard solution

Rivaroxaban standard stock solution was prepared by using diluent (ACN: Water, 75:25). 10 mg of API was dissolved in about 60 ml of diluent, shaken well and volume was make up to 100 ml. above stock solution was further diluted serially to get working standard of 20 µg/ml and for linearity study. The standard solution with concentration of 20 µg/ml was injected into HPLC system to get the chromatogram.

Preparation of sample solution

Ten tablets were weighed to calculate average weight, triturated to fine powder, weight equivalent to 10 mg sample fine powder was taken in 100 ml volumetric flask, added about 80 ml of diluent, shaken well and kept under sonication for 30 minutes under controlled temperature of about 25°C, then volume was make up to 100 ml again shaken well. 20 ml of filtered (through 0.45 µm membrane filter paper) solution were diluted in 100 ml diluent in a volumetric flask. The sample solution with concentration of 20 µg/ml was injected into HPLC system to get the chromatogram.

Results and Discussion

For better HPLC results the vital aspect to be considered is optimization of mobile phase keeping the physiochemical properties of drug of interest in view. Various trials with different considerations were tried to optimize the final chromatographic conditions. A well-defined peak when obtained along with acceptance criteria of analytical validation under ICH guidelines were important aspects for development of assay procedure. The important parameters of validation were performed by using newly developed method, simultaneously compared with an established method, the results obtained from both the methods are found to be within acceptance range.

Table 1: Results of validation parameters from proposed and established method.

Sl. No Parameter Proposed method Established method
1 System Suitability 3.8 5.4

Retention time (min)

Theoretical plates (N)

2100 2000
2 Specificity Peak purity pass (without any interference) at 3.8 min Peak purity pass (without any interference) at 5.4 min
3 Linearity Range (μg mL-1) 5.0-40.0 5.0-40.0
4 Accuracy (% Recovery) 99.5 99.0

50%

100%

99.0 99.8

150%

100.0 101.0
5 Precision (%RSD) 1.05 1.49

Intermediate

Ruggedness

0.89 1.0
6 Robustness No Significant changes in retention time No Significant changes in retention time

Mobile Phase

λ Max

Flow Rate

Conclusions

In the above study, it is tried to optimize an analytical method which is precise, accurate and robust From above, it can be concluded that the proposed method can be advantageous for the estimation of Rivaroxaban in routine laboratory work.

Acknowledgements

I, on behalf of all the concerned express my deep sense of gratitude who are directly or indirectly involved in the work.

Funding: No funding sources

Conflict of interest: None declared

References

  1. Roehrig S, Straub A, Pohlmann J. Discovery of the novel antithrombotic agent (BAY 59-7939): an oral, direct factor Xa inhibitor. J Med Chem. 2005;48(19):5900–8.
  2. Kakkar AK, Brenner B, Dahl OE. Extended duration rivaroxaban versus short term enoxaparin for the prevention of venous thromboembolism after total hip arthroplasty: a double-blind, randomised controlled trial. Lancet. 2008;372:31–9.
  3. Eriksson BI, Borris LC, Friedman RJ. Rivaroxaban versus enoxaparin for thromboprophylaxis after hip arthroplasty. New England J Med. 2008;358(26):2765–75.
  4. Rohde G. Determination of rivaroxaban - a novel, oral, direct Factor Xa inhibitor - in human plasma by high-performance liquid chromatography-tandem mass spectrometry. J Chromatogr B. 2008;872:43-50.
  5. Satyanarayana PVV, Madhavi AS. Department of Chemistry. New Spectrophotometric methods for the quantitative estimation of Rivaroxaban in formulations, Int J Res Rev Pharm Applied Sci. 2010:611-20.




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