A Simple and Rugged Bioanalytical Method Development and Validation of Brivudine in Human Plasma by Using High-Performance Liquid Chromatography

Abstract

<jats:p>Objective: The current research work focus to simple and rugged bioanalytical method development and validation of brivudine in human plasma using high-performance liquid chromatography.&#x0D; Methods: The analyte (Brivudine) and internal standard (Sofosbuvir) were extracted using the Solid Phase Extraction (SPE) technique. The chromatographic separation was accomplished by using Zorbax eclipse XDB-C18 Column (150×4.6 mm, 5 μm) with a mobile phase consisted of Methanol: 0.5% Ortho-phosphoric acid (65:35%, v/v) respectively, at a flow rate of 0.7 mL/min. The developed method was validated by performing system suitability, carryover effect, linearity, selectivity, sensitivity, precision, accuracy, recovery, ruggedness, and stability studies. The method was validated as per USFDA guidelines.&#x0D; Results: The selected chromatographic condition was found to efficiently separated brivudine (RT-3.55 min) and ISTD (RT-7.87 min). The assay demonstrated a linear dynamic range of 85.205 to 4500.246 ng/ml for brivudine in human plasma with r2&gt;0.99. Demonstrated the lowest limit of detection at 85.205 ng/ml. This method established an intra-run and inter-run precision within the range of 2.99-6.31%CV and 3.67-5.80%CV, respectively. Additional intra-run and inter-run accuracy were within the range of 97.55-105.37% and 99.27-102.15%, respectively. The mean percentage recovery of brivudine and ISTD studies proved good extraction efficiency and the robustness was also evaluated.&#x0D; Conclusion: A simple, accurate, precise, linear and rugged RP-HPLC method was developed and validated for the estimation of brivudine in human plasma with K2EDTA anticoagulant and suitable for conducting BA/BE and TDM.</jats:p>

Bioanalytical Rp-HPLC Method Development and Validation of Clopidogrel Bisulfate in Wistar Rat Plasma and Its Application to Pharmacokinetic Study

“…Plasma sample's linearity was quantified using the peak-area ratio versus analyte nominal concentration [25]. For the establishment of the linearity range of the proposed method, calibration curves in plasma were prepared in the concentration range of 40-200 ng/ml.…”

“…The ruggedness of the method was studied by changing the experimental condition with precision and accurate batch [25]. Ruggedness was performed by using different analysts [29] in the same laboratory.…”

Bioanalysis Method Development and Validation of Small Molecules in Pharmaceutical Industry: A Bioanalyst Review Point

“…The system suitability test of the chromatographic system was performed to confirm the adequate suitability and reproducibility of the system before each day of a validation run. A system suitability test was performed by injecting six consecutive injections using a standard aqueous mixture equivalent to middle concentration of the calibration standard curve of analyte and working concentration of ISTD [23].…”

“…This denotes that a compound is getting carried over from one injection to the next. The chromatographic system should be tested to evaluate the possibility of carry over by injecting mobile phase followed by an extracted blank, extracted lower limit of quantification (LLOQ), extracted ULOQ, and then once again extracted blank [22,23].…”

A novel HPLC method for selexipag in human plasma and application to a prototype pharmacokinetic study

“…In addition, there are not enough methods that provide pharmacokinetic studies to be easily applicable for the determination of SLP in human plasma. Review of literature revealed that few methods were reported for the estimation of SLP including; spectrophotometric methods [5][6][7], gas chromatograhy [8] and chromatographic methods [9][10][11][12] in dosage form, in addition to, LC-MS/MS methods in plasma [13][14][15][16][17].…”