LC-MS/MS Bioanalytical Method Development and Validation for the Quantification of (2S, 3S)-sec-butylpropylacetamide (SPD) in Rat and Mouse Plasma
Abstract number :
3.402
Submission category :
2. Translational Research / 2E. Other
Year :
2021
Submission ID :
1886487
Source :
www.aesnet.org
Presentation date :
12/6/2021 12:00:00 PM
Published date :
Nov 22, 2021, 06:56 AM
Authors :
Chun-Yi Wu, PhD - University of California, Davis; Michael Rogawski, MD, PhD. - Professor, Neurology, University of California, Davis
Rationale: SPD is a novel valproic acid amide derivative with broad spectrum activity in diverse animal seizure models, including models of status epilepticus. SPD is particularly effective in terminating status epilepticus induced by chemical nerve agents. SPD has two asymmetric carbons so that there are four stereoisomers, each of which possesses antiseizure activity. The (2S,3S) isomer has modestly improved pharmacokinetic properties in relation to the other stereoisomers and is non-teratogenic in highly inbred SWV/Fnn mice known to be susceptible to valproic acid-induced neural tube defects. (2S,3S)-SPD has been selected for evaluation as a potential treatment for nerve agent-induced status epilepticus. Formulations for administration by various routes of administration are being developed. To assess the pharmacokinetic characteristics of the formulations in rodent models, we developed and validated a rapid LC-MS/MS bioanalysis method for the quantification of (2S,3S)-SPD in rat and mouse plasma with K2EDTA as the anticoagulant.
Methods: The plasma samples were processed with all-in-one protein precipitation: Twenty microliter of the sample was spiked with 80 µL of the internal standard, valnoctamide, dissolved in acetonitrile (100 ng/mL). (2S,3S)-SPD was extracted to the supernatant fraction, resolved in the UPLC column and eluted into the mass spectrometer (MS) with a 5-minute LC gradient.
Results: The linear quantification range of the assay was 1 to 1000 ng/mL in both rat and mouse matrices. The extraction yields were above 80% and no MS signal inhibition was observed due to matrix effect. The inter and intra-batch precision (%CV) and accuracy (%deviation) across low (5 ng/mL), medium (50 ng/mL), and high (500 ng/mL) QC sample concentration levels were all below 7%. The inter and intra-batch precision and accuracy at the lower level of quantification (LLOQ, 1 ng/mL) were between 4 and 13%.
Conclusions: The method, which provides a rapid, sensitive assay of (2S,3S)-SPD in plasma across a wide range of concentrations, is suitable for pharmacokinetic studies to assess the performance of novel formulations for delivery by diverse routes of administration.
Funding: Please list any funding that was received in support of this abstract.: Supported by NINDS grant 5U01NS112102.
Translational Research