Intermedine n oxide

Lasiocarpine (Las) and lasiocarpine N-oxide were ordered from Cfm Oskar Tropitzsch (Marktredwitz, Germany). Lycopsamine (Lyc), Lycopsamine N-oxide, europine (Eur), europine N-oxide, senecionine (Sen), senecionine N-oxide, echimidine (Ech), echimidine N-oxide, intermedine, intermedine N-oxide, jacobine, jacobine N-oxide, heliosupine, heliosupine N-oxide, retrorsine N-oxide, retronecine, and senkirkine were obtained from Phytolab (Vestenbergsgreuth, Germany). Retrorsine (Ret) was delivered by AppliChem (Darmstadt, Germany). Methanol (MeOH, LC–MS grade) and water (H₂O, LC–MS grade) were purchased from Merck KGA (Darmstadt, Germany). 7,9-Diglutathionyl-6,7-dihydro-1-hydroymethyl-5H-pyrrolizidine was obtained from ASCA (Berlin, Germany). Potassium cyanide (KCN), isotopically labeled potassium cyanide (KCX, K¹³C¹⁵N), glutathione and isotopically labeled glutathione (GSX, ¹³C₂¹⁵N-GSH), and all other chemicals and co-factors were purchased from Sigma-Aldrich (Steinheim, Germany). All chemicals were obtained in analytical grade, if available. Human liver microsomes (mixed gender from 50 donors; protein concentration 20 mg/mL; cytochrome P450 content 310 pmol/mg protein) and rat liver microsomes (male Sprague–Dawley rats; protein concentration 20 mg/mL; cytochrome P450 content 700 pmol/mg protein) were obtained from Corning (Corning, USA).

A mixture of 7-acetyl-intermedine/lycopsamine was from PlantaAnalytica (Danbury, US), retrorsine from Cfm O.Tropitzsch (Marktredwitz, Germany), and echimidine, intermedine, intermedine N-oxide, and senecionine were from PhytoLab (Vestenbergsgreuth, Germany). Echivulgarine was isolated in-house [38 (link)] and echimidine N-oxide and retrorsine N-oxide were N-oxidized in-house, starting from the free bases according to [39 (link)], except that dichloromethane was used instead of trichloromethane.
D3-atropine was from CDN isotopes (Quebec, Canada), methanol was from Honeywell (LC-MS grade, Grogg AG, Stettlen, Switzerland). Sulfuric acid, formic acid, and aqueous ammonia solution 25%, all of analytical grade, were from Merck (Grogg AG, Stettlen, Switzerland). Ultrapure water was used (Elga purelab ultra, Labtec, Villmergen, Switzerland).

PA standards used for analysis were 7-acetylintermedine, 7-acetylintermedine-N-oxide, 7-acetyllycopsamine, 7-acetyllycopsamine-N-oxide, echinatine, echinatine-N-oxide, heliosupine, heliosupine-N-oxide, heliotrine, heliotrine-N-oxide, indicine hydrochloride, indicine-N-oxide, integerrimine, integerrimine-N-oxide, intermedine, lycopsamine, lycopsamine-N-oxide, retrorsine, riddelliine, riddelliine-N-oxide, rinderine, rinderine-N-oxide, senecionine, senecionine-N-oxide, seneciphylline, and senkirkine, from PhytoPlan (Heidelberg, Germany). Echimidine, Echimidine-N-oxide, erucifoline, erucifoline-N-oxide, europine hydrochloride, europine-N-oxide, intermedine-N-oxide, jacobine, jacobine-N-oxide, lasiocarpine, Monocrotaline-N-oxide, retrorsine-N-oxide, seneciphylline-N-oxide, senecivernine, and senecivernine-N-oxide were from PhytoLab (Vestenbergsgreuth, Germany). lasiocarpine-N-oxide was from Cfm Oskar Tropitzsch (Marktredwitz, Germany). Monocrotaline was from Carl Roth (Karlsruhe, Germany) and trichodesmine from Latoxan (Valence, France). For the palynological analysis and sample preparation, the following reagents were used: demineralized water, ultrapure water, dish soap, Kaiser’s glycerol gelatin, dry ice, 0.05 M sulfuric acid, ammonia solution, and methanol. For sample analysis, 5 mM ammonium formate and 0.1% (v/v) formic acid in methanol were used.