title: RP-HPLC separation and characterization of unknown impurities of a novel HIV-protease inhibitor Darunavir by ESI-MS and 2D NMR spectroscopy.
abstract: A selective RP-HPLC method for separation and determination of darunavir from its process related substances has been developed and validated. The separation was accomplished on a Hiber, LiChrospher 60, RP-select B, C₈ (250 mm × 4.6 mm, 5 μm) column connected to a photo diode array detector (PDA) using 10 mM phosphate buffer with 0.1% of triethylamine (pH: 4.0 adjusted with orthophosphoric acid)/acetonitrile as a mobile phase under gradient elution. Two unknown impurities of darunavir were isolated and characterized by ¹H, ¹³C, 2D-NMR and mass spectrometry. The method was validated in terms of accuracy, precision, linearity, robustness, LOD and LOQ.
authors.affiliation: D-215, Analytical Chemistry Division, Discovery Laboratory, Indian Institute of Chemical Technology, Tarnaka, Hyderabad 500 607, India. rnrao55@yahoo.com
authors.forename: Ramisetti
authors.initials: R
authors.lastname: Nageswara Rao
authors.forename: Bondigalla
authors.initials: B
authors.lastname: Ramachandra
authors.forename: Kondapalli
authors.initials: K
authors.lastname: Santhakumar
mesh_terms: D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D000069454:Darunavir; D004340:Drug Contamination; D004355:Drug Stability; D055664:Electrochemical Techniques; D017408:Guidelines as Topic; D017320:HIV Protease Inhibitors / Q000032:analysis / Q000737:chemistry*; D007194:India; D057230:Limit of Detection; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D008970:Molecular Weight; D011786:Quality Control; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D013449:Sulfonamides / Q000032:analysis / Q000737:chemistry*
chemical_list: D017320:HIV Protease Inhibitors; D013449:Sulfonamides; D000069454:Darunavir