Quantitative Assessment of the Influence of Rhizoma Zingiberis on the Level of Aconitine in Rat Gut Sacs and Qualitative Analysis of the Major Influencing Components of Rhizoma Zingiberis on Aconitine Using UPLC/MS

Abstract

This study attempted to clarify the material basis for the detoxification of Rhizoma Zingiberis (RZ) on aconitine, an analgesic drug, by quantitatively assessing the influence of RZ on the in vitro intestinal concentration of aconitine using an everted gut sac model and by qualitatively identifying the components in the RZ extract. To quantify aconitine in rat everted gut sacs, both an accurate processing method and a sensitive detection method were required. We developed a three-step sample processing method to protect the components from decomposition and applied ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC/TQMS) to quantify aconitine, glucose and digoxin. In addition, ultra-performance liquid chromatography coupled with linear ion trap mass spectrometry (UPLC/ITMS) was applied to detect the potential antidotal components in the RZ extract. Finally, the RZ extract reduced the level of aconitine in everted gut sacs, and eleven gingerols were successfully identified, which could be considered potential antidotal components for aconitine. This study demonstrated the application of two UPLC/MS methods for analyzing the material basis for the reciprocity between Chinese medicine components in everted gut sacs.

Fig 1. Multiple reaction monitoring (MRM) ion chromatogram of glucose.

Fig 2. MRM ion chromatogram of aconitine.

Fig 3. MRM ion chromatogram of digoxin.

Fig 4. MRM ion chromatogram of aconitine(a) and digoxin(b) in blank intestinal nutritious solution.

Fig 5. Ratios of glucose content in the serosal side to that in the mucosal side in different sacs.

Fig 6. Effect of verapamil on the absorptive profile of aconitine.

Fig 7. Effect of the Rhizoma Zingiberis extract on the absorptive profile of digoxin.

Fig 8. Base peak ion chromatogram of the extract of Rhizoma Zingiberis: (A) blank solvent, (B) 6-gingerol standard, and (C) the Rhizoma Zingiberis extract.

Fig 9. Effect of 6-gingerol on the absorptive profile of digoxin.