Gram-scale purification of dehydroevodiamine from Evodia rutaecarpa fruits, and a procedure for selective removal of quaternary indoloquinazoline alkaloids from Evodia extracts.

Dehydroevodiamine (DHE) is a major bioactive constituent in the traditional Chinese herbal drug Evodiae fructus (Wu zhu yu). The compound has been shown to possess pronounced cardiovascular and neuropharmacological activities in vitro and in vivo. For quality control purposes and follow-up studies assessing potential safety risks of DHE, we developed a simple and efficient two-step protocol for gram-scale purification of DHE. An alkaloidal fraction was obtained by cation-exchange solid phase extraction, and DHE and the minor alkaloid hortiamine were purified by isocratic preparative RP-HPLC. The DHE content in different commercial batches of Evodiae fructus, and in a series of commercially available Evodia-containing TCM products was assessed. A daily intake of up to mg amounts of DHE was calculated from recommended doses of these products. A method for the selective removal of quaternary indoloquinazoline alkaloids from Evodia extracts was developed.

Schramm A ，Hamburger M 《-》

Quantitative analyses of indoloquinazoline alkaloids in Fructus Evodiae by high-performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry.

Fructus Evodiae (Wuzhuyu), the fruits of Evodia rutaecarpa and related varieties, is widely used in traditional Chinese medicine. The bioactive constituents include the indoloquinazoline alkaloids rutaecarpine, evodiamine and dehydroevodiamine. A new assay based on high-performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry (HPLC/UV/APCI-MS/MS) was developed for the measurement of the indoloquinazoline alkaloids in commercial Fructus Evodiae products. Initially, the MS/MS fragmentation pathways of indoloquinazoline alkaloids were investigated to identify fragment ions that might be useful for the sensitive and selective detection of trace indoloquinazoline alkaloids during LC/MS/MS. Then, quantitative MS analysis of five indoloquinazoline alkaloids in 12 commercial Fructus Evodiae products from different geographical sources was performed. Analyte recovery was in the range of 97.5-105.3% for all with relative standard deviations (RSDs) below 6%, the intra-assay and inter-assay RSDs were less than 7%, and good linear relationships were shown with correlation coefficients for the analytes exceeding 0.999. Therefore, this LC/MS/MS assay facilitated the rapid quantitative analysis of rutaecarpine, evodiamine, evodiamide, 14-formyldihydrorutaecarpine and dehydroevodiamine in 12 commercial Fructus Evodiae products with excellent recovery, repeatability, accuracy and sensitivity. This method is simple and specific and can be used for identification and quality control of this traditional Chinese remedy.

Zhou Y ，Li SH ，Jiang RW ，Cai M ，Liu X ，Ding LS ，Xu HX ，But PP ，Shaw PC ... -  《RAPID COMMUNICATIONS IN MASS SPECTROMETRY》

Dehydroevodiamine and hortiamine, alkaloids from the traditional Chinese herbal drug Evodia rutaecarpa, are I(Kr) blockers with proarrhythmic effects in vitro and in vivo.

Evodiae fructus is a widely used herbal drug in traditional Chinese medicine. Evodia extract was found to inhibit hERG channels. The aim of the current study was to identify hERG inhibitors in Evodia extract and to investigate their potential proarrhythmic effects. Dehydroevodiamine (DHE) and hortiamine were identified as IKr (rapid delayed rectifier current) inhibitors in Evodia extract by HPLC-microfractionation and subsequent patch clamp studies on human embryonic kidney cells. DHE and hortiamine inhibited IKr with IC50s of 253.2±26.3nM and 144.8±35.1nM, respectively. In dog ventricular cardiomyocytes, DHE dose-dependently prolonged the action potential duration (APD). Early afterdepolarizations (EADs) were seen in 14, 67, 100, and 67% of cells after 0.01, 0.1, 1 and 10μM DHE, respectively. The proarrhythmic potential of DHE was evaluated in 8 anesthetized rabbits and in 8 chronic atrioventricular block (cAVB) dogs. In rabbits, DHE increased the QT interval significantly by 12±10% (0.05mg/kg/5min) and 60±26% (0.5mg/kg/5min), and induced Torsade de Pointes arrhythmias (TdP, 0.5mg/kg/5min) in 2 rabbits. In cAVB dogs, 0.33mg/kg/5min DHE increased QT duration by 48±10% (P<0.05*) and induced TdP in 2/4 dogs. A higher dose did not induce TdP. In human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), methanolic extracts of Evodia, DHE and hortiamine dose-dependently prolonged APD. At 3μM DHE and hortiamine induced EADs. hERG inhibition at submicromolar concentrations, APD prolongation and EADs in hiPSC-CMs and dose-dependent proarrhythmic effects of DHE at micromolar plasma concentrations in cAVB dogs should increase awareness regarding proarrhythmic effects of widely used Evodia extracts.

Baburin I ，Varkevisser R ，Schramm A ，Saxena P ，Beyl S ，Szkokan P ，Linder T ，Stary-Weinzinger A ，van der Heyden MAG ，Houtman M ，Takanari H ，Jonsson M ，Beekman JHD ，Hamburger M ，Vos MA ，Hering S ... -  《-》

New glycosidic alkaloid from the nearly ripe fruits of Euodia rutaecarpa.

Hu CQ ，Li KK ，Yang XW 《-》

Development and validation of HPLC methods for the analysis of phenethylamine and indoloquinazoline alkaloids in Evodia species.

Pellati F ，Benvenuti S ，Yoshizaki F ，Melegari M ... -  《-》