Ephedrine

HPLC-based fingerprinting was accomplished using two-dimensional HPLC (Agilent HP1200, Agilent Technologies, San Jose, CA, USA). Ephedrine (Sigma-Aldrich, St. Louis, MO, USA), as a principal component of Es, was used as a standard. Briefly, 100 μg of water extract of Es, 10 μg of Ephedrine and 200 mg of rat fecal samples from the end of microbiota preparation experiment were dissolved in 1 mL 50% of methanol separately. After sonication and centrifuging, the supernatant of samples were filtrated by 0.45 μm membrane filter (Sartorius AG, Goettingen, Germany) and prepared for HPLC analysis. The Agilent HPLC system consisted of a quaternary pump, on-line solvent degasser, Agilent sample manager/column heater module, and Agilent UV detector. An Agilent Eclipse XDB-C18 (5 mm, 4.6 mm × 250 mm) column was used and the compounds were eluted with solvents composed of acetonitrile: water: phosphoric acid (40:60:0.1 v/v/v) containing sodium lauryl sulfate at a concentration of 5 g/L as an ion-pairing agent. Peak identity of Ephedrine in the samples was confirmed by comparison of spectrum and retention time. HP1200 ChemStation software (Agilent Technologies, Santa Clara, CA, USA) was applied and all chromatograms were obtained using a wavelength of 215 nm.

Bupropion hydrochloride and 6-hydroxybupropion were obtained from Toronto Research Chemicals (Toronto, Canada), ibogaine from Ibogaworld, cocaine from Medisca (Montreal, Canada), β-PEA, pinoline, indole, tryptamine, ephedrine, frovatriptan, alprenolol, isoproterenol, evodiamine, and tyramine from Sigma (Oakville, Canada), fluoxetine and amphetamine were from Tocris Bioscience (Bristol, United Kingdom). PAL analogs, RTI analogs, and bicifadine were synthesized at RTI in the Blough laboratory as described previously (Carroll et al., 2009 (link), 2011 (link); Blough et al., 2011 , 2014 (link)). Noribogaine (Batch No. 606950002) was from Deborah Mash, Ph.D. (Obach et al., 1998 ). Ibogamine was obtained from Specs (Zoetermeer, The Netherlands). Sodium phenylbutyrate was purchased from Enzo Life Sciences Inc. (Farmingdale, NY). Ibogaminalog, ibogainealog, noribogainalog, fluorogainalog, and tabernanthalog were synthesized in the lab of David E. Olson (Cameron et al., 2020 (link)).

Indigo, Aliso B acetate, gentiopicroside, peimine, perillaldehyde, wedelolactone, bergapten, picroside I, berberine, cinnamaldehyde, ephedrine, p-synephrine, 6-shogao, and 6-gingerol were purchased from Sigma-Aldrich. Phellodendrine was purchased from Abcam. Baicalein, baicalin, wogonin, wogonoside, and Ephedra sinica were kindly provided by Y.L. Leu (Chang Gung University, Taiwan). Scutellaria baicalensis (precursor of Baicalein, baicalin, wogonin, wogonoside) and Ephedra sinica were supplied and authenticated by Department of Pharmacy Services, Chang Gung Memorial Hospital at Taoyuan, Taiwan. See Supplementary Information for more details on compound preparation.