Chiral LC method development: Stereo-selective separation, characterization, and determination of cabotegravir and related RS, RR, and SS isomeric impurities on coated cellulose-based chiral stationary phase by HILIC-LC and LC-MS

• Published in: Journal of pharmaceutical and biomedical analysis 2023-01, Vol.222, p.115062-115062, Article 115062
• Main Authors: Krishna Murthy Kasa, S.R., Venkatanarayana, Muvvala, Chennuru, Lakshmi Narayana, Chandra Sekhara Rao, B., Vemparala, Manohar, Chaman, Abdul Fareed, Talluri, M.V.N. Kumar
• Format: Article
• Language: English
• Subjects: Cabotegravir; Characterization; CHIRALCEL OX-3R column; HPLC and LC-MS; RS, RR, and SS-isomers separations
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2022.115062

The first, sensitive, and rapid chiral method was developed for enatioseperations and determination of cabotegravir and its enantiomeric impurities by using HPLC and LC-MS. Cabotegravir is an antiretroviral medication used for the treatment of HIV/AIDS approved by the food and drugs administration (FDA) in the year 2021. The cabotegravir chiral separation was achieved on the coated cellulose-tris (4-chloro-3-methyl phenyl carbamate) (CHIRALCEL OX-3R) column in HILIC mode and the total run time is less than 15 min. The effects of mobile phase composition, elution mode, and percentage of organic modifier as well as the effect of mobile phase-additives and column temperature were investigated on selectivity, resolution, and peak symmetry. The mobile phase consisted of acetonitrile and water with 0.1% (v/v) addition of formic acid additive with the flow rate of 1 mLmin−1. UV detection was carried out at 220 nm. The calibration curves of cabotegravir and its enantiomers were linear over the concentration range of 0.04–1.125 µgmL−1. The limits of detection and quantification for cabotegravir and its enantiomer (RS isomer) were ≤ 0.02 and ≤ 0.06, and the RR and SS-isomers limits were ≤ 0.02 and 0.03 µgmL−1 respectively. It was demonstrated that the proposed method is selective, precise, and robust. Finally, the validated method was applied for the determination and identification of cabotegravir and its chiral enantiomers in the bulk drugs by using HPLC and LC-MS techniques. •First analytical method for the separation of cabotegravir and its isomers.•Synthesis and characterization of cabotegravir and its RS, RR, and SS-isomers.•Method development approach by using different analytical modes (NP/PO and RP Mode) on polysaccharide-based CSPs.•Method optimization: Analytical mode, mobile phase composition, additive effects, and column temperatures.•Method validation and application of developed LC and LC-MS methods for bulk drug analysis.