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UPLC method Validation for bioanalysis of Gemcitabine in rat PK study using VAMS methodology (Volumetric Absorptive Microsampling) | Abstract

Der Pharma Chemica
Journal for Medicinal Chemistry, Pharmaceutical Chemistry, Pharmaceutical Sciences and Computational Chemistry

ISSN: 0975-413X
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Abstract

UPLC method Validation for bioanalysis of Gemcitabine in rat PK study using VAMS methodology (Volumetric Absorptive Microsampling)

Author(s): Raja Rajeswari K*, Hafis Jamal Ayyil and Kathirvel S

Volumetric absorptive microsampling (VAMS) is a simple intuitive technique for collecting and quantitative analysis of dried blood samples. It enables the collection of an accurate blood volume regardless of blood hematocrit. A bioanalytical method for the determination of gemcitabine in dried blood supported on VAMS samplers has been validated and used to support a pharmacokinetic study in rat. The calculated PK parameters were comparable to those obtained from blood–water (1:1, v/v) samples. VAMS is demonstrated to be a robust method that simplifies both the blood sample collection and bioanalytical laboratory procedures and generates high quality quantitative data. Waters Acquity UPLC system using an Acquity BEH C18 column (100x2.1 mm, 1.7 μm) was used for chromatographic separation by isocratic elution using acetonitrile-water (35-65) as the mobile phase at a flow rate of 0.5 mL/min. Gemcitabine was administered to rat orally at 3 mg/kg for conducting the PK study and the blood was collected at various time intervals using VAMS sampler which consists of a hydrophilic polymeric tip, absorbs an accurate sample volume within 2–4 s by wicking, attached to a molded plastic handle. The tip is white before use and turns completely red when filled with blood, and the blood samples were processed after collection and analyzed by UPLC. The intra-day and inter-day accuracy of gemcitabine were 93.6–108.7% and 94.2–111.5% respectively, and the precision (RSD, %) was less than 15% for both intra-day and inter-day measurements. Gemcitabine has a good linear relationship in the range of 50-500 ng/mL with r2 value of 0.997. A robust and reliable UPLC method was fully optimized and developed to detect the blood concentration of gemcitabine in rats and the samples were analyzed by Empower software.


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