Vol 14, Issue 2, 2021
Online - 2455-3891
Print - 0974-2441
STABILITY INDICATING METHOD DEVELOPMENT AND VALIDATION OF FIMASARTAN
BY REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY IN BULK AND
PHARMACEUTICAL DOSAGE FORM
SRUTHI A*, UTTAM PRASAD PANIGRAHY
Department of Pharmaceutical Analysis, CMR College of Pharmacy, Hyderabad, Telangana, India. Email: uttampanigrahy@gmail.com
Received: 23 October 2020, Revised and Accepted: 2 December 2020
ABSTRACT
Objective: A rapid, sensitive and specific reverse phase High performance liquid Chromatography (RP-HPLC) method was developed for the estimation
of Fimasartan in bulk and pharmaceutical dosage form.
Method: The RP-HPLC analysis was performed isocratically on a Primacel C
18
column (150 mm × 4.6 mm internal diameter, 5 μm particle size) using
mobile phase of composition Acetonitrile and 0.1% orthophosphoric Acid in 80:20, v/v proportions with a flow rate of 0.8 ml/min.
Results: The analyte was monitered with UV-detector at 265 nm. In the developed method Fimasartan elutes at a typical retention time of 2.4 min.
The proposed method is having linearity in the concentration ranging from 5-30 μg/ml of Fimasartan.
Conclusion: : The method was statistically validated and had been applied to analysis of the drug in bulk and pharmaceutical dosage form.
Keywords: Fimasartan, Reverse-phase high-performance liquid chromatography, C
18
column.
INTRODUCTION
Fimasartan is a typical anti-hypertensive agent. Fimasartan acts on the
kidney’s rennin-angiotensin cascade, which begins when renin release from
the kidney causes the breakdown of angiotensinogen into angiotensin I. In
blocking the AT
1
receptor, Fimasartan inhibits vasoconstriction favoring
vasodilation. Fimasartan is chemically known as 2-[2-butyl-4-methyl-6-
oxo-1-[[4-[2-(2H-tetrazol-5-yl) phenyl] phenyl] methyl] pyrimidin-5-yl]-N
and N-dimethylethanethioamide shown in Fig. 1.
Literature survey reveals the availability of few methods such as ultraviolet
(UV), high-performance liquid chromatography (HPLC), and LC-MS in
biological fluids and pharmaceutical dosage forms [1-16]. The present
method gives the accurate information regarding method development
and validation of Fimasartan in bulk and pharmaceutical dosage form
using UV detector at 265 nm which is simple, accurate, economic, rapid,
reproducible, and sensitive according to procedures and acceptance
criteria based on FDA guidelines and recommendations of ICH.
METHODS
Chemical and reagents
Fimasartan was procured as gift sample from Metrochem Pvt. Ltd.,
Hyderabad, India. Fimanta
®
120 mg (formulation) was manufactured
by Ajanta Pharma Limited. HPLC grade acetonitrile, HPLC grade water,
and orthophosphoric acid were obtained from Merck, Hyderabad, India.
All other chemical reagents used were of analytical grade.
Selection of wavelength
Wavelength was selected by dissolving Fimasartan in suitable solvent
(acetonitrile) and the resulting solution was scanned in UV region, that
is, 200–400 nm. Fimasartan has showed that maximum absorption at
265 nm was selected shown in Fig. 2.
Chromatographic conditions
A Shimadzu HPLC was used for the analysis of the entire method. The
method was carried out on Primacel C
18
(150 mm × 4.6 mm i.d., 5 μm)
column as a stationary phase and Acetonitrile:0.1% OPA (80:20,v/v) as
the mobile phase at a flow rate of 0.8 ml/min. Rheodyne injector with
25 μl loop was used for injecting the samples. Finally, the sample was
analyzed at 265 nm and the mobile phase was sonicated filtered and
degassed.
Preparation of mobile phase
Acetonitrile and 0.1% OPA in the ratio of 80:20 v/v were used as
a mobile phase for present study which is prepared by dissolving
80 ml of acetonitrile and 20 ml of 0.1% OPA in a reservoir. 0.1%
orthophosphoric acid was prepared by accurately measuring of 0.1 ml
of OPA and dissolving in 100 ml of HPLC grade water. The above mobile
phase prepared is sonicated and degassed.
Preparation of standard solution
Standard solution was prepared by accurately weighing 100 mg of
Fimasartan and transferring into 100 ml volumetric flask. To this add
few ml of acetonitrile and dissolve the drug properly. Finally make up
the solution to 100 ml with acetonitrile (1000 μg/ml).
Preparation of sample solution
Tablet powder equivalent to 100 mg of Fimasartan was accurately
weighed and transferred into 100 ml volumetric flask and make up
the volume to 100 ml with acetonitrile. Sonicate the solution for about
15 min and filter the solution through Whatman filter paper (No. 1)
into another 100 ml volumetric flask (1000 μg/ml). The solution was
further diluted with the diluent to the working concentration range of
calibration curve.
Calibration curve
Ten milliliters of standard solution were transferred into 100 ml
volumetric flask and the volume was made up to 100 ml using acetonitrile
(100 μg/ml). From this solution, further dilutions were made in the
concentration range of 5 μg/ml–30 μg/ml. Further these concentrations
were injected thrice into the system and the calibration curve was
constructed by plotting concentration on X-axis and peak area on Y-axis.
© 2021 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/
licenses/by/4.0/) DOI: http://dx.doi.org/10.22159/ajpcr.2021v14i2.39919. Journal homepage: https://innovareacademics.in/journals/index.php/ajpcr
Research Article