Abstract
A simple, sensitive and cost-effective HPLC-UV bioanalytical method for determination of lopinavir (LPV) in rat and human plasma was developed and validated. The plasma sample preparation procedure includes a combination of protein precipitation using cold acetonitrile and liquid-liquid extraction with n-hexane-ethyl acetate (7:3, v/v). A good chromatographic separation was achieved with a Phenomenex Gemini column (C18 , 150 mm × 2.0 mm, 5 μm) at 40°C with gradient elution, at 211 nm. Calibration curves were linear in the range 10-10,000 ng/mL, with a lower limit of quantification of 10 ng/mL using 100 μL of plasma. The accuracy and precision in all validation experiments were within the criteria range set by the guidelines of the Food and Drug Administration. This method was successfully applied to a preliminary pharmacokinetic study in rats following an intravenous bolus administration of LPV. Moreover, the method was subsequently fully validated for human plasma, allowing its use in therapeutic drug monitoring (TDM). In conclusion, this novel, simple and cost-efficient bioanalytical method for determination of LPV is useful for pharmacokinetic and drug delivery studies in rats, as well as TDM in human patients.
Highlights
| INTRODUCTIONLopinavir (LPV) is an HIV (human immunodeficiency virus) protease inhibitor (PI) coadministered with a low dose of ritonavir (RTV) under the brand name Kaletra (LPV/r) as part of antiretroviral treatment (ART) in people affected by HIV
Lopinavir (LPV) is an HIV protease inhibitor (PI) coadministered with a low dose of ritonavir (RTV) under the brand name Kaletra (LPV/r) as part of antiretroviral treatment (ART) in people affected by HIV
A volume of 10 μL of working standard solutions was spiked into 100 μL rat plasma to obtain LLOQ, low quality control (LQC), medium quality control (MQC) and high quality control (HQC) samples at concentrations of 10, 25, 400 and 8000 ng/mL, respectively
Summary
Lopinavir (LPV) is an HIV (human immunodeficiency virus) protease inhibitor (PI) coadministered with a low dose of ritonavir (RTV) under the brand name Kaletra (LPV/r) as part of antiretroviral treatment (ART) in people affected by HIV. Cost-effective and sensitive bioanalytical methods for therapeutic drug monitoring (TDM) of LPV are needed, mostly in developing countries. To be relevant for TDM, these methods need to cover the range of clinically relevant plasma concentrations of LPV in HIV-infected individuals receiving LPV/r regimen (Eron et al, 2004; Ribera et al, 2004). In this work, a simple, sensitive, cost-efficient and low sample volume bioanalytical method for determination of LPV in rat plasma was developed and fully validated using HPLC with UV detection. This method was successfully implemented in a pharmacokinetic study following intravenous administration of LPV in rats.
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