title: Validation of simultaneous quantitative method of HIV protease inhibitors atazanavir, darunavir and ritonavir in human plasma by UPLC-MS/MS.
abstract: OBJECTIVE
  HIV protease inhibitors are used in the treatment of patients suffering from AIDS and they act at the final stage of viral replication by interfering with the HIV protease enzyme. The paper describes a selective, sensitive, and robust method for simultaneous determination of three protease inhibitors atazanavir, darunavir and ritonavir in human plasma by ultra performance liquid chromatography-tandem mass spectrometry.
  
  
  METHODS
  The sample pretreatment consisted of solid phase extraction of analytes and their deuterated analogs as internal standards from 50 μL human plasma. Chromatographic separation of analytes was performed on Waters Acquity UPLC C18 (50 × 2.1 mm, 1.7 μm) column under gradient conditions using 10 mM ammonium formate, pH 4.0, and acetonitrile as the mobile phase.
  
  
  RESULTS
  The method was established over a concentration range of 5.0-6000 ng/mL for atazanavir, 5.0-5000 ng/mL for darunavir and 1.0-500 ng/mL for ritonavir. Accuracy, precision, matrix effect, recovery, and stability of the analytes were evaluated as per US FDA guidelines.
  
  
  CONCLUSIONS
  The efficiency of sample preparation, short analysis time, and high selectivity permit simultaneous estimation of these inhibitors. The validated method can be useful in determining plasma concentration of these protease inhibitors for therapeutic drug monitoring and in high throughput clinical studies.
authors.affiliation: Chemistry Department, Kadi Sarva Vishwavidyalaya, Gandhinagar 382015, India.
authors.forename: Tulsidas
authors.initials: T
authors.lastname: Mishra
authors.affiliation: Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad 380009, India.
authors.forename: Pranav S
authors.initials: PS
authors.lastname: Shrivastav
mesh_terms: D000069446:Atazanavir Sulfate; D000069454:Darunavir; D017320:HIV Protease Inhibitors / Q000032:analysis* / Q000493:pharmacokinetics; D006801:Humans; D009842:Oligopeptides / Q000032:analysis* / Q000493:pharmacokinetics; D010949:Plasma / Q000737:chemistry* / Q000378:metabolism; D011725:Pyridines / Q000032:analysis* / Q000493:pharmacokinetics; D019438:Ritonavir / Q000032:analysis* / Q000493:pharmacokinetics; D013449:Sulfonamides / Q000032:analysis* / Q000493:pharmacokinetics
chemical_list: D017320:HIV Protease Inhibitors; D009842:Oligopeptides; D011725:Pyridines; D013449:Sulfonamides; D000069446:Atazanavir Sulfate; D019438:Ritonavir; D000069454:Darunavir