Figure 1.
Scheme of all necessary steps in obtaining Epiisopiloturine with >98% purity from Jaborandi leaves.
Figure 2.
Analytical HPLC used LiChrospher 60 RP column and eluted with potassium phosphate
. (A) Standard EPI (20 µg/mL), (B) Standard pilocarpine (50 µg/mL), (C) “cultivated jaborandi leaves” solution, resulted from first extraction step, (D) “cultivated jaborandi acid” solution, obtained EPI under salt form, (E) Solution of “crude EPI” with some impurities as pilocarpine and other alkaloids, (F) last step of isolation showing EPI >98% purity.
Figure 3.
Mass spectrum obtained from ESI+/Ion Trap.
(A) free EPI with a pseudo molecular ion m/z 287.1 Da [M+H]+, (B) MS2 with characteristic fragment at m/z 269.1 Da [M – H2O + H]+, (C) MS3 with fragments at m/z 251.0 Da [M – 2H2O + H+] and 168.06 Da with proposed chemical structure.
Figure 4.
Isolated Epiisopiloturine molecular structure.
Figure 5.
Epiisopiloturine Crystalline form with the molecules represented in stick format.
Color code: carbon (gray), hydrogen (white), nitrogen (blue) and oxygen (red). Cyan lines are only guide lines to illustrate hydrogen bonds between the hydroxyl group and the imidazole ring of neighboring molecules in the solid.
Table 1.
EPI bond distances obtained through x-ray diffraction (Experimental) and DFT results (Calculated).
Table 2.
EPI bond angles obtained through x-ray diffraction (Experimental) and DFT results (Calculated).
Figure 6.
Epiisopiloturine FT-IR spectra: A) experimental and B) calculated.
Figure 7.
Epiisopiloturine FT-Raman spectra: A) experimental and B) calculated.
Table 3.
Infrared (IR) and Raman wavenumbers (cm−1) of solid state EPI.
Figure 8.
Epiisopiloturine TGA-DSC (A) and DTG-MS (B) curves under air atmosphere.