International Science Community Association Research Journal of Pharmacutical Sciences 14 2 2025 12 30 Simultaneous Estimation of S-RRR Nebivolol and R-SSS Nebivolol in Human Plasma by Using Liquid Chromatography-tandem Mass Spectrometry 1 13 EN Zaware Sandeep 1 Bioanalytical Research Department, Synergen Bio pvt. Ltd. Pune, Maharashtra, India Bhoite Sangram 2 Bioanalytical Research Department, Synergen Bio pvt. Ltd. Pune, Maharashtra, India Kadam Akshay 3 Bioanalytical Research Department, Synergen Bio pvt. Ltd. Pune, Maharashtra, India Lahoti Neel 4 Bioanalytical Research Department, Synergen Bio pvt. Ltd. Pune, Maharashtra, India > Research Paper 2025 6 4 2025 12 30 The present study describes a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous estimation of S-RRR and R-SSS nebivolol in human plasma using solid phase extraction technique. Method of both S-RRR and R-SSS nebivolol has been developed, validated by using S-RRR nebivolol D4 and R-SSS nebivolol D4 as an internal standard. Analytes from human plasma were extracted by ion exchange cartridges and subsequently separated on chiral column using acetonitrile: 1.5 mM ammonium carbonate in water, 70:30% v/v as a mobile phase, at a flow rate of 1.0 ml min-1. Quantification of S- RRR and R-SSS nebivolol and nebivolol D4 was performed using multi-reaction monitoring mode (MRM) in positive mode. The calibration curve was linear (r2 > 0.99) over the concentration range of 20.0-20000.0 pg ml for S-RRR and R-SSS nebivolol. The intra-day and inter-day assay precision revealed within ±15% (at LLOQ level ±20%) with accuracy within 85-115% (at LLOQ level 80-120%). The LC-MS/MS method was fully validated for all the validation parameters as per M10 and current regulatory requirement such as selectivity, matrix effect, recovery and stability. Overall the present study revealed the selectivity and sensitivity of this method for the simultaneous estimation of S-RRR and R-SSS nebivolol in human plasma. Copyright@ International Science Community Association