A validated HPLC-MS method for quantification of the CCR5 inhibitor maraviroc in HIV+ human plasma

• A very easy and simple method to quantify maraviroc in human plasma using only 100 μL of sample.
• Run time was only of 20 min and no matrix effect was observed.
• Use of a cheaper instrument (HPLC-MS vs. HPLC-MS/MS), but more sensitive than a UV detector.
• Validation of the assay following FDA guideline.
• Eligibility of this method for routine dosage of maraviroc in plasma samples.

Maraviroc is a CCR5 inhibitor approved in 2007 for treatment of therapy experienced adult patients infected with CCR5-tropic HIV-1 virus. International guidelines for HIV therapy indicate a plasma concentration cutoff of maraviroc for response. We developed and validated a new HPLC-MS method to quantify maraviroc concentrations in human plasma. 6,7-Dimethyl-2,3-di(2-pyridyl)quinoxaline was used as internal standard and added to 100 μL of plasma. Samples were then treated with 500 μL of acetonitrile for the protein precipitation procedure. An analytical T3 Atlantis column (150 mm × 4.6 mm i.d.) with a particle size of 5 μm was used to separate the compounds and ions were detected at m/z 257.5 and 313.3 for maraviroc and quinoxaline respectively. The calibration curve was linear up to 2500 ng/mL. The mean recovery of maraviroc was 89.1%. All validation data results were in accordance to Food and Drug Administration and European Medicines Agency requirements. The HPLC-MS method reported here could be used routinely to monitor plasma concentrations of maraviroc in HIV-infected patients.

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