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Bioanaltical Method Development And Validation Of Primidone In Rat Plasma By K3edta Using Lc-Ms/Ms
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| Author:
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KHEDKAR NAVNATH, S. SATHESH KUMAR
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| Abstract:
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Bio-analytical methods are widely used to quantitate drugs and their metabolites in physiological matrices. Primidone is an anticonvulsant of the barbiturate class. It is a structural analog of phenobarbital and related to barbiturate-derivative anticonvulsants. Identification of drug and internal standard was done by using Mass spectrometer. Parent ion and product ion of drug and ISTD (Primidone D5) tuning for drug and ISTD polarity were found in positive method, ion source as Turbo Ion Spray (TIS) was used. Trials were taken at different combination of solvents and final mobile phase selected as 0.1% Formic Acid in water, v/v: Organic Mixture (Acetonitrile: Methanol, 50:50, v/v), 20:80, v/v) Column: Gemini C18 (250 mm). Spiked concentration ranges lies between 3000.000 to 10.000 ng/mL for STD1 to STD8 respectively. 2400.000 to 10.000 ng/mL for HQC to LLOQ QC samples respectively. Final extraction procedure LLE was selected on the basis of Chromatography and response i.e.100 µL Sample + 50 µL ISTD Dilution + 2% OPA in water + 1.5 mL Ethyl acetate + extract on extractor at 50 rpm for 20 min + Centrifuge 10 min 4000RPM at 5 ± 3 ºC + Collect 1ml supernant + Drying + 300 µL RS. All the parameters for validation i.e. specificity, ruggedness, precision and accuracy and recovery were found satisfactory and within the acceptance criteria. The methods of estimation of Primidone in rat plasma successfully developed and validated as per protocols.
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| Keyword:
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Primidone, LLE, Specificity, Precision and Accuracy, Validation
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2020.12.04.679
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| Download:
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