Development of a highly sensitive and specific ELISA method for the determination of l-corydalmine in SD rats with monoclonal antibody

J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Jan 15:1073:163-169. doi: 10.1016/j.jchromb.2017.09.022. Epub 2017 Sep 14.

Abstract

l-Corydalmine (l-CDL) is a potent analgesic constituent of the traditional Chinese medicine, Rhizoma Corydalis. However, the pharmacokinetic process and tissue distribution of l-CDL in vivo are still unknown. Therefore, it is necessary to establish a simple and sensitive method to detect l-CDL, which will be helpful to study its distribution and pharmacokinetic process. To determine this compound in biological samples, a monoclonal antibody (mAb) against l-CDL was produced and a fast and highly sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed in this study. The icELISA was applied to determine l-CDL in biological samples. The limit of detection (LOD) of the method was 0.015 ng/mL with a liner range of 1-1000 ng/mL (R2 = 0.9912). The intra- and inter-day precision were below 15% and the recoveries were within 80-117%. Finally, the developed immunoassay was successfully applied to the analysis of the distribution of l-CDL in SD rats. In conclusion, the icELISA based on the anti-l-CDL mAb could be considered as a highly sensitive and rapid method for the determination of l-CDL in biological samples. The ELISA approach may provide a valuable tool for the analysis of small molecules in biological samples.

Keywords: Enzyme-linked immunosorbent assay; Monoclonal antibody; Tissue distribution; l-Corydalmine.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / metabolism*
  • Berberine / analogs & derivatives*
  • Berberine / analysis
  • Berberine / chemistry
  • Berberine / pharmacokinetics
  • Chromatography, High Pressure Liquid
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Limit of Detection
  • Linear Models
  • Liver / chemistry
  • Liver / metabolism
  • Myocardium / chemistry
  • Myocardium / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Reproducibility of Results
  • Tissue Distribution

Substances

  • Antibodies, Monoclonal
  • Berberine
  • corydalmine