Abstract
Mice are the most frequently used animals in pharmacokinetic studies; however, collecting series of blood samples from mice is difficult because of their small sizes and tiny vessels. In addition, due to the small sample size, it is problematic to perform high required quantification. Thus, present work aims to find an effective strategy for overcoming these challenges using trans-resveratrol as a tool drug. Based on the idea of a joint technology, the capillary microsampling (CMS) was chosen for blood sample collection from mice after delivery of trans-resveratrol (150 mg/kg) by gavage, and a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for the determination of trans-resveratrol and its main metabolites. All the mouse blood samples were exactly collected by CMS without obvious deviation. This provided credible samples for subsequent quantitative analysis. The HPLC-MS/MS method was found to be sensitive, accurate, and repeatable, and the pharmacokinetic parameters for all analytes were comparable with those reported in previous studies. However, the present joint technology offers the advantages of less animal damage, easy for sample preparation, and improved reliability. It has overcome some of the major limitations revealed in previous pharmacokinetic studies in mice and therefore provides a more effective option for future studies.
Highlights
Preclinical pharmacokinetic studies, including single- or multiple-dose toxicology pharmacokinetic studies, conducted in animals have been considered a key approach for obtaining toxicological data for a new compound
capillary microsampling (CMS) could solve the problem of insufficient sample quantities collected from a little mouse, and the high sensitivity of HPLC-MS/MS could compensate for the disadvantages of CMS for collecting very small sample volumes per time point. erefore, a strategy that combines the advantages of CMS with the high sensitivity of HPLCMS/MS might solve the problems in mice pharmacokinetic studies. e present study aimed to determine the feasibility of this joint technology for overcoming those challenges using trans-resveratrol just as a tool compound
Linearity was validated over three consecutive days through the use of the calibration-standard samples (2.24–223.5 ng/ mL for the trans-resveratrol and 1–200 ng/mL for the R3G and resveratrol-3-sulfate salt (R3S)), which were analyzed by the present method after sample preparation. e calibration curves were drawn by calibrating the peak area ratio (y) of the trans-resveratrol or its metabolites to the internal standard (IS) with the concentration (x) of the standard sample. e regression parameters were calculated from the linear least squares regression (1⁄ x2). e lower limit of quantification (LLOQ) was the lowest concentration, with a coefficient of variation (CV) precision of no more than 20%, accuracy 80–120%, and signal-to-noise ratio (S⁄ N) no less than 10
Summary
Preclinical pharmacokinetic studies, including single- or multiple-dose toxicology pharmacokinetic studies, conducted in animals have been considered a key approach for obtaining toxicological data for a new compound. A pharmacokinetic study usually requires several biological samples, such as blood, serum, and plasma, from the experimental animals at different series of time points [3, 4]. DBS is the collection of whole blood with filter paper, and dried biological samples can be conveniently kept and shipped It seems more suitable for the qualitative examination of diseases but offers no more advantages over CMS for quantitative determination [10]. One of the major reasons might be the stringent requirements for the sensitivity of the quantitation instruments in pharmacokinetic studies because of the very small sample volumes obtained from mice until the rapid progress of mass spectrometry. HPLC-MS/MS could significantly reduce the consumption of sample volume during the detection process It provides a good detection technology platform for the study of pharmacokinetics in mice. CMS could solve the problem of insufficient sample quantities collected from a little mouse, and the high sensitivity of HPLC-MS/MS could compensate for the disadvantages of CMS for collecting very small sample volumes per time point. erefore, a strategy that combines the advantages of CMS with the high sensitivity of HPLCMS/MS might solve the problems in mice pharmacokinetic studies. e present study aimed to determine the feasibility of this joint technology for overcoming those challenges using trans-resveratrol just as a tool compound
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