High Throughput Lc-Ms/Ms Method for the Quantitation of Emtricitabine in Human Plasma by Solid Phase Extraction Using 96 Well Plate Format

Aim: To develop and validate sensitive, high-throughput Liquid chromatography–mass spectrometry method for the quantification of Emtricitabine in human plasma using Lamivudine as an internal standard (ISTD) using 96 well plate format solid phase extraction (SPE) technique.

Methodology: The samples were extracted from plasma using solid phase extraction (SPE) in 96 well plate format. After extraction the analyte and internal standard were analyzed on Inertsil ODS column with dimensions 4.6 X 100 mm, 5 µm using a mobile phase consisting of Methanol: 0.2% formic acid in water 85:15, v/v. The precursor and product ions of the analytes were monitored on a triple quadrupole instrument operated in the positive ionization mode.

Results: The method was validated with concentration range of 5.024 to 5023.725 ng/mL. The relative recoveries were ranging from 60.7 to 65.9%. Three validation batches were performed with five QC levels (LLOQ, LQC, MQC II, MQC and HQC). Across three validation runs the inter batch precision (%CV) was ≤ 9.2% and the accuracy was between 98.9–106.0%. All the stability experiments found satisfactory.

Conclusion: According to the method validation results, the current method was found to be specific, accurate, sensitive, precise and high throughput method. This method can be used for the estimation of Emtricitabine in human plasma during routine analysis.