Quantifying pomalidomide in human plasma employing UPLC-MS/MS Alekhya K.*, Nataraj K. S.**, Pawar A. K. M.***, Vaheeda**** Shri Vishnu College of Pharmacy, Vishnupur, Bhimavaram - 534202West Godavari District AU College of Pharmaceutical Sciences, Andhra University, Visakhapatnam - 530017 *Corresponding Author E-mail: alekhya@svcp.edu.in
**kalakondasrinataraj@gmail.com
***akmpawarau@rediffmail.com
****vaheedarehmans@gmail.com
Online Published on 13 October, 2023. Abstract The current work aims to create a simple, fast, clear and accurate chromatography-tandem mass spectrophotometric (UPLC-MS/MS). 50μl ISTD (250ng/ml concentration) and 250μl of 5mM ammonium formate buffer (pH - 2.5) were put into the samples just before liquid-liquid extraction (LLE) using 2.5ml of tertiary butyl methyl ether (TBME). Chromatographic fraction was done using Hypersil gold column (50 mmX4.65mm, 5μ) with use of 5mM ammonium formate buffer (pH - 2.5) and acetone in 20: 80 v/v (mobile phase). The rate of flow was around 0.50ml/min. Pomalidomide and Celecoxib were ionized into ions with polarity (positive) and were recognized using MRM. Quantitation was made by transitions m/z 274.43 precursor ion to m/z 201.15 for drug Pomalidomide and m/z 382.12 precursor ion to m/z 362.0 of celecoxib. The concentrations of eight standards exhibited linearity between 1.9900 to 199.8380ng/ml (r2 ≥ 0.9968). Separation of chromatogram was obtained in two minutes. The recovery values in an average for three QC's was 51.49% of Pomalidomide which was in concordance with acceptance. The Coefficient of variation was ≤15% in intra and inter batch assay values. The %CV ruggedness ranges from 1.26 - 2.25. Short and long term stability percentages of stock solutions were found acceptable like 99.76 and 98.69. Results show that all the validation parameters were in acceptable limits. Top Keywords Pomalidomide, Human plasma, LC-MS/MS, Liquid-liquid extraction. Top |