Analytical HPLC used LiChrospher 60 RP column and eluted with potassium phosphate

<p>. (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.</p

Scheme of all necessary steps in obtaining Epiisopiloturine with >98% purity from Jaborandi leaves.

<p>Scheme of all necessary steps in obtaining Epiisopiloturine with >98% purity from Jaborandi leaves.</p

Mass spectrum obtained from ESI+/Ion Trap.

<p>(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.</p

Infrared (IR) and Raman wavenumbers (cm⁻¹) of solid state EPI.

<p>Calculated vibrational wavenumbers (cm-1) for the isolated EPI molecule. A tentative assignment of the observed vibrational modes is also shown. See text for theoretical details. ν =  stretching, δ =  bending, β =  bending in plane, γ =  bending out of plane, r =  rocking, τ =  twist, sc =  scissoring, ω =  wagging, νs =  symmetric stretching, νa =  antisymmetric stretching, sh  =  shoulder.</p

EPI bond distances obtained through x-ray diffraction (Experimental) and DFT results (Calculated).

<p>Atom labels accordingly to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066702#pone-0066702-g004" target="_blank">Figure 4</a>.</p

Epiisopiloturine FT-IR spectra: A) experimental and B) calculated.

<p>Epiisopiloturine FT-IR spectra: A) experimental and B) calculated.</p

Isolated Epiisopiloturine molecular structure.

<p>Isolated Epiisopiloturine molecular structure.</p

EPI bond angles obtained through x-ray diffraction (Experimental) and DFT results (Calculated).

<p>Atom labels accordingly to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066702#pone-0066702-g004" target="_blank">Figure 4</a>.</p

Epiisopiloturine FT-Raman spectra: A) experimental and B) calculated.

<p>Epiisopiloturine FT-Raman spectra: A) experimental and B) calculated.</p

Epiisopiloturine TGA-DSC (A) and DTG-MS (B) curves under air atmosphere.

<p>Epiisopiloturine TGA-DSC (A) and DTG-MS (B) curves under air atmosphere.</p