Quantitative analysis of a novel HIV fusion inhibitor (sifuvirtide) in HIV infected human plasma using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry.
A sensitive method for measuring sifuvirtide, a novel HIV fusion inhibitor peptide drug in HIV-1(+) human plasma by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. The plasma samples were treated by solvent/detergent (S/D) method to inactivate viral activity before analysis. After protein precipitation sifuvirtide was determined by LC-MS/MS. A structure analog was used as internal standard (IS). The mass spectrometer was operated in positive ion and multiple reaction monitoring mode with transitions m/z 946.3-->159.0 for sifuvirtide and 951.7-->159.2 for IS. The intra-day precision ranged from 2.74% to 7.57% with accuracy from 91.63% to 102.53%. The inter-day precision ranged from 2.65% to 3.58% and the accuracy from 95.53% to 105.28%. Stability studies showed that sifuvirtide was stable both during the assay procedure and long-term storage. The lower limit of quantitation (LLOQ) was 9.75ngml(-1). The method was used for analyzing samples from phase IIa clinical study of sifuvirtide in China.
Che
Jinjing
J
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20, Dongda Street, Fengtai District, Beijing 100071, PR China.
Meng
Qingfang
Q
Chen
Zhihang
Z
Hou
Yunan
Y
Shan
Chengqi
C
Cheng
Yuanguo
Y
D011232:Chemical Precipitation; D002681:China; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D017322:Clinical Trials, Phase II as Topic; D004355:Drug Stability; D023581:HIV Fusion Inhibitors / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D015658:HIV Infections / Q000097:blood / Q000188:drug therapy* / Q000821:virology; D015497:HIV-1 / Q000472:pathogenicity*; D006801:Humans; D007279:Injections, Subcutaneous; D010455:Peptides / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D012015:Reference Standards; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000592:standards; D053719:Tandem Mass Spectrometry / Q000592:standards; D038301:Virus Inactivation
D023581:HIV Fusion Inhibitors; D010455:Peptides; C505768:sifuvirtide