Enantiomeric Resolution And Quantitative Estimation Of Pirtobrutinib In Tablet Formulation: Method Development And Validation By NP-HPLC

Abstract

Pirtobrutinib, a next-generation non-covalent BTK inhibitor sold under the brand name Jaypirca, requires exact enantiomeric separation and measurement in order to maintain regulatory compliance and quality control.This study set out to create and validate a robust, accurate, and dependable NP-HPLC method for identifying Pirtobrutinib's R- and S-isomers in pharmaceutical formulations and bulk.The need for a validated analytical method in compliance with ICH Q2 (R1) guidelines and the therapeutic relevance of the active S-enantiomer served as the driving forces behind the reasoning. A YMC Chiral ART Cellulose-SC column (250 × 4.6 mm, 5 μm) was used in the procedure, and a mobile phase made up of acetonitrile and diethylamine (100:0.1% v/v) was supplied at a flow rate of 1.5 mL/min. Acetonitrile was used as the diluent, the column oven ran for 15 minutes, and the temperature was kept at 35 °C.With a resolution of 5.5, the results showed distinct, symmetrical peaks at retention periods of 4.42 minutes (R-isomer) and 6.21 minutes (S-isomer).With a tailing ratio of 1.4 and theoretical plates more than 4000, the system's applicability satisfied acceptance standards. For both enantiomers, linearity was established over the range of 0.5–3.5 µg/mL, with correlation values (R²) greater than 0.9999.The limits of quantitation (LOQ) were 1.0 µg/mL (S/N ratios: 18.05 and 13.54, respectively), while the limits of detection (LOD) were 0.5 µg/mL (S/N ratios: 6.34 for R-isomer and 4.71 for S-isomer).Recovery values within 98.1–101.8% were confirmed by accuracy studies to meet ICH requirements.For retention duration, peak area, tailing factor, and plate count, precision experiments produced %RSD values <2%, indicating repeatability and intermediate precision.Resolution was routinely greater than 5, and robustness was validated despite small intentional changes in flow rate (±0.2 mL/min) and column oven temperature (±5 °C).  Results from tests for ruggedness by various analysts were repeatable.  The active S-isomer was recovered 99.7% of the time in the Jaypirca tablet assay. For the enantiomeric study of Pirtobrutinib, the validated NP-HPLC method demonstrated specificity, sensitivity, precision, accuracy, and robustness. Its appropriateness for regular quality monitoring of pharmaceutical dosage forms and bulk drugs was validated by the simplified validation values.