Multiplex Liquid Chromatography-Tandem Mass Spectrometry Assay for Simultaneous Therapeutic Drug Monitoring of Ribavirin, Boceprevir, and Telaprevir

Abstract

g New directly acting antivirals (DAAs) that inhibit hepatitis C virus (HCV) replication are increasingly used for the treatment of chronic hepatitis C. A marked pharmacokinetic variability and a high potential for drug-drug interactions between DAAs and numerous drug classes have been identified. In addition, ribavirin (RBV), commonly associated with hemolytic anemia, often requires dose adjustment, advocating for therapeutic drug monitoring (TDM) in patients under combined antiviral therapy. However, an assay for the simultaneous analysis of RBV and DAAs constitutes an analytical challenge because of the large differences in polarity among these drugs, ranging from hydrophilic (RBV) to highly lipophilic (telaprevir [TVR]). Moreover, TVR is characterized by erratic behavior on standard octadecyl-based reversed-phase column chromatography and must be separated from VRT-127394, its inactive C-21 epimer metabolite. We have developed a convenient assay employing simple plasma protein precipitation, followed by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) for the simultaneous determination of levels of RBV, boceprevir, and TVR, as well as its metabolite VRT-127394, in plasma. This new, simple, rapid, and robust HPLC-MS/MS assay offers an efficient method of real-time TDM aimed at maximizing efficacy while minimizing the toxicity of antiviral therapy.

Simultaneous determination of newly developed antiviral agents in pharmaceutical formulations by HPLC-DAD

“…A review of the literature revealed that CE [15, 16], HPLC [17–21], UPLC–MS/MS [22–24], LC–MS/MS [25, 26] and HPTLC [27, 28] methods have been reported for the analysis of RIT, individually or in combination with other drugs. However, a method for the simultaneous determination of OMB, PAR, RIT and DAS has not yet been reported.…”

The emerging role of multiplex tandem mass spectrometry analysis for therapeutic drug monitoring and personalized medicine

“…This approach has allowed the setting-up of numerous multiplex assays for the quantification of drugs from the same therapeutic class in a single chromatographic run. This multiplex approach has been applied notably for the simultaneous analysis of antiretroviral agents (in plasma and cells) [40][41][42][43][44], anti-HCV drugs [45], targeted anticancer agents [46], antimalarials [47], antifungals [48], new generation antiepileptics [49], (Figure 2), antibiotics such as several beta-lactams (carbapenems, penicillins, cephalosporins) altogether [50][51][52][53] or combined with other antibiotics classes including linezolid, ciprofloxacin, daptomycin and rifampicin [54][55][56][57][58]. Overall, the main advantages of this approach are: i) a unique sample extraction procedure and short analytical run; ii) time saving through the establishment of simultaneous calibration curves; iii) easy application to blood samples from patients receiving either single-drugs or combination regimens.…”

Sofosbuvir and ribavirin before liver re-transplantation for graft failure due to recurrent hepatitis C: a case report

“…Details of the methodology developed to measure plasma concentrations of SOF, based on liquid chromatography coupled with tandem mass spectrometry, will be reported elsewhere (LAD, unpublished data). RBV plasma concentrations were determined using a previously described analytical method [ 14 ]. Cyclosporine trough levels were determined by enzyme multiplied immunoassay on a Cobas Integra® 400 plus system using reagents from Roche Diagnostics (Rotkreuz, Switzerland).…”