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Development of a Validated RP-HPLC Method for Estimation of Ethionamide in Spiked Human Plasma with UV Detection


Affiliations
1 Department of Pharmaceutical Analysis, MET’s Institute of Pharmacy, Bhujbal Knowledge City, Adgaon, Nashik – 422 003, India
     

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A simple, rapid and economic RP-HPLC method was developed and validated for estimation of Ethionamide in spiked human plasma. The drug along with Guaifenesin (used as internal standard) was extracted from plasma by liquid-liquid extraction (LLE) procedure using ethyl acetate as organic solvent. The drug was well resolved from the plasma interference and internal standard in a reversed phase mode on C 18 (250 × 4.6 mm, 5 μ) column with methanol: water (40: 60 %, v/v) as mobile phase, at a flow rate of 1 mL/min. The detection was performed at 275 nm. The developed method was validated as per the US-FDA guidelines, where the weighted linear regression was used in calibration experiments to obtain the homoscedasticity. In accuracy and precision studies, intra-day and inter-day, % relative error was found between ± 15 and % RSD was less than 15 %. Stability experiments indicated that the drug remained stable after three freeze-thaw cycles.

Keywords

Ethionamide, Bioanalytical, Liquid-Liquid Extraction, Weighted Regression, RP-HPLC.
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  • Development of a Validated RP-HPLC Method for Estimation of Ethionamide in Spiked Human Plasma with UV Detection

Abstract Views: 19  |  PDF Views: 1

Authors

Priya Rahade
Department of Pharmaceutical Analysis, MET’s Institute of Pharmacy, Bhujbal Knowledge City, Adgaon, Nashik – 422 003, India
Sandeep Sonawane
Department of Pharmaceutical Analysis, MET’s Institute of Pharmacy, Bhujbal Knowledge City, Adgaon, Nashik – 422 003, India
Atharva Bhalerao
Department of Pharmaceutical Analysis, MET’s Institute of Pharmacy, Bhujbal Knowledge City, Adgaon, Nashik – 422 003, India
Sanjay Kshirsagar
Department of Pharmaceutical Analysis, MET’s Institute of Pharmacy, Bhujbal Knowledge City, Adgaon, Nashik – 422 003, India

Abstract


A simple, rapid and economic RP-HPLC method was developed and validated for estimation of Ethionamide in spiked human plasma. The drug along with Guaifenesin (used as internal standard) was extracted from plasma by liquid-liquid extraction (LLE) procedure using ethyl acetate as organic solvent. The drug was well resolved from the plasma interference and internal standard in a reversed phase mode on C 18 (250 × 4.6 mm, 5 μ) column with methanol: water (40: 60 %, v/v) as mobile phase, at a flow rate of 1 mL/min. The detection was performed at 275 nm. The developed method was validated as per the US-FDA guidelines, where the weighted linear regression was used in calibration experiments to obtain the homoscedasticity. In accuracy and precision studies, intra-day and inter-day, % relative error was found between ± 15 and % RSD was less than 15 %. Stability experiments indicated that the drug remained stable after three freeze-thaw cycles.

Keywords


Ethionamide, Bioanalytical, Liquid-Liquid Extraction, Weighted Regression, RP-HPLC.