title: Determination of telaprevir in plasma of HCV-infected patients by HPLC-UV.
abstract: Telaprevir is a direct acting antiviral agent, used with pegylated interferon and ribavirin for the management of chronic hepatitis C virus (HCV) genotype 1 infection, in patients not responding to therapy with pegylated interferon and ribavirin only. Although 75% of patients achieve a sustained virological response after treatment with telaprevir, adverse drug-drug interactions and undesirable events often occur. Therefore, telaprevir monitoring is pivotal to improve the management of HCV infection. Here, the first High-Performance Liquid Chromatography-Ultraviolet (HPLC-UV) method to quantify telaprevir in human plasma of HCV-genotype 1-infected patients is reported. The volume of the plasma sample was 700 μL. This method involved automated solid-phase extraction with Oasis HLB Cartridge 1 cc (divinylbenzene and N-vinylpyrrolidone). The extracted samples were reconstituted with 150 μL of 60/40 water/acetonitrile. Thirty microliters of these samples was injected into a HPLC-UV system, and the analytes were eluted on a X Terra(®) RP18 column (250 mm × 4.6 mm i.d.) with a particle size of 5 μm. The mobile phase (ammonium acetate buffer, 150 mM, pH 8.0, and methanol:acetonitrile 50:50) was delivered at 1.0 mL/min with linear gradient elution. The total run time for a single analysis was 16 min; telaprevir was detected by UV at 276 and 286 nm. The calibration curve was linear from 312.5 to 20,000 ng/mL (r(2) > 0.996). The absolute recovery of telaprevir ranged between 89 and 93% at concentrations of quality control samples of 800, 4,000, 8,000, and 16,000 ng/mL. Both precision and accuracy were always <15%. The HPLC-UV method reported here: (i) has been validated, (ii) is currently applied to monitor telaprevir in plasma of HCV-infected patients, and (iii) appears useful in a routine laboratory. ,
authors.affiliation: Istituto Nazionale per le Malattie Infettive IRCCS, Lazzaro Spallanzani, Roma, Italy. massimo.tempestilli@inmi.it
authors.forename: Massimo
authors.initials: M
authors.lastname: Tempestilli
authors.forename: Elisa
authors.initials: E
authors.lastname: Milano
authors.forename: Gianpiero
authors.initials: G
authors.lastname: D'Offizi
authors.forename: Marzia
authors.initials: M
authors.lastname: Montalbano
authors.forename: Antonio
authors.initials: A
authors.lastname: D'Avolio
authors.forename: Tecla
authors.initials: T
authors.lastname: Gasperi
authors.forename: Pasquale
authors.initials: P
authors.lastname: Narciso
authors.forename: Paolo
authors.initials: P
authors.lastname: Ascenzi
authors.forename: Leopoldo P
authors.initials: LP
authors.lastname: Pucillo
mesh_terms: D000368:Aged; D000998:Antiviral Agents / Q000097:blood* / Q000627:therapeutic use; D001774:Blood Chemical Analysis / Q000592:standards; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D005260:Female; D019698:Hepatitis C, Chronic / Q000097:blood* / Q000188:drug therapy; D006801:Humans; D057230:Limit of Detection; D008297:Male; D008875:Middle Aged; D009842:Oligopeptides / Q000097:blood* / Q000627:therapeutic use; D012015:Reference Standards; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000998:Antiviral Agents; D009842:Oligopeptides; C486464:telaprevir
keywords: HPLC-UV; anti-HCV therapy; telaprevir; therapeutic drug monitoring