Development and Validation of HPLC Method for quantification of Zonisamide in Spiked Human Plasma
Journal Title: Jordan Journal of Pharmaceutical Sciences - Year 2022, Vol 15, Issue 1
Abstract
In present work a simple, economic, accurate and precise HPLC method with UV detection was developed for the quantification of zonisamide (ZON) in spiked human plasma using trimethoprim (TRI) as an internal standard. Both ZON and TRI were well separated and resolved from each other on C18 column using mobile phase blend of methanol: acetonitrile: 20 mM phosphate buffer (pH 3.0) in an isocratic mode at flow rate of 1 mL/min with proportion of 25:5:70 %, v/v/v. The detector wavelength was set at 240 nm. Maximum recovery of ZON and TRI from plasma were obtained with dichloromethane (DCM) as extracting solvent. The calibration curve was found to be linear in the range of 3-60 mg/mL with regression coefficient (r2) = 0.997. The LLOQ of the method was 3 mg/mL. Although, with acceptable r2, heteroscedasticity of the calibration data was observed which further was reduced with use of weighted linear regression with weighting factor 1/x. Finally, the method was validated with respect to sensitivity, accuracy, precision, recovery, stability and for carry-over as per US-FDA guidance for Bioanalytical Method Validation, May 2018.
Authors and Affiliations
Sandeep S. Sonawane, Gayatri J. Joshi, Sanjay J. Kshirsagar
Keywords
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