ABSTRACT:
Dolutegravir is an antiviral agent, a second-generation HIV integrase strand transfer inhibitor (INSTI). A precise liquid chromatography-tandem mass spectrometry method has been developed and validated for the detection of potential degradants in Dolutegravir. The goal of this study was to detect major and minor potential degradants that can develop during the various stages of the drug molecule, start from acquiring of raw material for the synthesis, manufacturing, formulation and storage of the final product. With this objective in mind preparative scale HPLC was used to detect possible oxidative degradants in Dolutegravir. The oxidative degradation sample was isolated using Prominence preparative high performance liquid chromatograph equipped with a reverse phase C18 Inertsil ODS (300mm×19mm ×7µ) column. Mobile phase was 0.1% trifluoro acetic acid in water (A) and acetonitrile (B) (50:50). Ethyl acetate was used for the extraction of aqueous layer and the same was distilled and the collected fractions were evaporated to dryness for LCMS studies. To detect the degradation products mass spectra of the drug was established by direct infusion of degradant into mass spectrometry systems. The LC-MS spectrum of Dolutegravir was recorded by Shimadzu LCMS spectrometer in the electron spray ionization mode. The outcome was intense protonated molecular ion peak at m/z 304.27 and 249.06 in the positive ionization mode, scan range 120–560 amu and the other possible degradant structures with m/z 320.32, m/z 207.18, m/z 185.17, m/z 385.41, m/z 155.14. The information obtained from degradation behaviour of dolutegravir by stress degradation studies, under peroxide oxidation and the degradation products formed were identified by HPLC, LCMS analytical techniques are useful information regarding manufacturing, formulation and storage of the drug.