Development and validation of an LC–MS/MS method for quantification of NC-8 in rat plasma and its application to pharmacokinetic studies

Authors

B.H. Kachingwe
Y.-S. Uang
T.-J. Huang
L.-H. Wang
S.-J. Lin

Abstract

ent-16-Oxobeyeran-19-N-methylureido (NC-8) is a recently synthesized derivative of isosteviol that showed anti-hepatitis B virus (HBV) activity by disturbing replication and gene expression of the HBV and by inhibiting the host toll-like receptor 2/nuclear factor-κB signaling pathway. To study its pharmacokinetics as a part of the drug development process, a highly sensitive, rapid, and reliable liquid chromatography tandem mass spectrometry (LC–MS/MS) method was developed and validated for determining NC-8 in rat plasma. After protein precipitation extraction, the chromatographic separation of the analyte and internal standard (IS; diclofenac sodium) was performed on a reverse-phase Luna C18 column coupled with a Quattro Ultima triple quadruple mass spectrometer in the multiple-reaction monitoring mode using the transitions, m/z 347.31 → 75.09 for NC-8 and m/z 295.89 → 214.06 for the IS. The lower limit of quantitation was 0.5 ng/mL. The linear scope of the standard curve was between 0.5 and 500 ng/mL. Both the precision (coefficient of variation; %) and accuracy (relative error; %) were within acceptable criteria of < 15%. Recoveries ranged from 104% to 113.4%, and the matrix effects (absolute) were non-significant (CV ≤ 6%). The validated method was successfully applied to investigate the pharmacokinetics of NC-8 in male Sprague–Dawley rats. The present methodology provides an analytical means to better understand the preliminary pharmacokinetics of NC-8 for investigations on further drug development.

Abstract Image

ScienceDirect Link

10.1016/j.jfda.2017.09.003

Recommended Citation

Kachingwe, B.H.; Uang, Y.-S.; Huang, T.-J.; Wang, L.-H.; and Lin, S.-J. (2018) "Development and validation of an LC–MS/MS method for quantification of NC-8 in rat plasma and its application to pharmacokinetic studies," Journal of Food and Drug Analysis: Vol. 26 : Iss. 1 , Article 3.
Available at: https://doi.org/10.1016/j.jfda.2017.09.003

Creative Commons License

This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Fulltext URL

https://www.sciencedirect.com/science/article/pii/S1021949817301825/pdfft?md5=4b3fad2e61cf427e01ff02caf3d52fff&pid=1-s2.0-S1021949817301825-main.pdf