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DEVELOPMENT AND VALIDATION OF BIOANALYTICAL METHOD FOR SIMULTANEOUS ESTIMATION OF ETAMSYLATE AND MEFENAMIC ACID IN HUMAN PLASMA BY RP-HPLC METHOD
Mona Karia*1, Bhargav Gohel2, Bina Thanki3, Darshan Madiya4, Shital Faldu5
1,3M.Pharm, Smt. R.D.Gardi B.Pharmacy College. Rajkot, Gujarat, India
2Q.A. Officer, Intas Pharmaceutical Ltd., Ahemdabad.
4Assistant Professor of Smt. R.D.Gardi B.Pharmacy College, Rajkot
5Principal of Smt. R.D.Gardi B.Pharmacy College, Rajkot
An accurate, simple, precise and sensitive HPLC method with UV detection was developed and validated for simultaneous estimation of Etamsylate and Mefenamic acid in human plasma as per USFDA guidelines. Etamsylate and Mefenamic acid were spiked with human plasma and 5 % w/v Trichloro acetic acid used for protein precipitation in ration of 1:9 %v/v of human plasma. The column used was SUSEIDO C18 column (250mm x 25 mm, i.d, 5 µ). Analysis was carried out with Methanol: ACN (70:30 % v/v): Buffer (0.05M Ammonium acetate buffer) pH 6.0 (85:15,% v/v) as mobile phase at flow rate of 1.5 ml/min at 298 nm with UV detection. The retention time of Mefenamic acid observed was 1.98 min and for Etamsylate 3.85 min respectively. The method was validated over the range of 0.5-30 μg/ml for Etamsylate and 0.2-18 µg/ml for Mefenamic acid with coefficients of regression 0.9939 and 0.9914 for Etamsylate and Mefenamic acid respectively. LOD and LOQ were 0.17 ng/ml and 0.51 ng/ml for Etamsylate and 2.62 ng/ml and 7.95 ng/ml for Mefenamic acid.The mean % recovery for MEF and ETM were found to be 89.07% and 88.98%with % C.V. of 1.87% and 1.91% respectively. Inter-day as well intra-day replicate, gave % RSD below 4.44 and 2.63 for Etamsylate and 3.20 and 3.75 for Mefenamic acid respectively. This RP-HPLC method is sensitive, selective, accurate, precise and rapid for the simultaneous estimation of Etamsylate and Mefenamic acid in human plasma with limit of Quantification in nano-gram level was developed and validated as per USFDA guideline and can be applied for a bioequivalence study to compare relative bioavailability of drug.
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About the University:
Nirma University, Ahmedabad was established in the year 2003 as a Statutory University under the Gujarat State Act at the initiative of the Nirma Education & Research Foundation(NERF). The University is also recognized by the University Grants Commission (UGC) under section 2(f) of the UGC Act. Institute of Pharmacy, Institute of Technology, Institute of Management, Institute of Diploma Studies, Institute of Science & Institute of Law are constituents of this University. The university is also accrediated by National Assessment and Accreditation Council (NAAC). Nirma University is a provisional memeber of the Association of Indian Universities(AIU) and the Assoication of Commonwealth Universities (ACU).
This Institute started as “Occupational Health Research Institute” (OHRI) in the year 1966 and was re-christened as “National Institute of Occupational Health” (NIOH) in 1970 presently located in the Eastern part of Ahmedabad. Two Regional Occupational Health Centers (ROHCs) were started at Bangalore in 1977 and at Kolkata in 1980.
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