The Study Chemical Constituents Of The Aerial Part Of Gastrodia Elata Blume. And Quality Evaluation Of Gastrodiae Rhizoma

Objective:To study the chemical constituents of the aerial part of Gastrodia elata Bl. and to provide the theoretical basis for the further utilization of waste materials of GASTRODIAE RHIZOMA stalk. To establish HPLC fingerprint and to compare gastrodine content of GASTRODIAE RHIZOMA from different origins, to provides a method which can be used comprehensive evaluate of the quality of GASTRODIAE RHIZOMA.To set up a method for the determination of the content of heavy metals of GASTRODIAE RHIZOMA.To evaluate the safety of GASTRODIAE RHIZOMA and provide a reference for the formulation of limit standard ofharmful heavy metals in GASTRODIAE RHIZOMA.Methods:1.The aerial part of Gastrodia elata Bl. was extracted by heating reflux with 70% ethanol.The extraction was concentrated to obtain extractum.Followed by, the extractum was partitioned with ethyl acetate and n-butanol saturated with water.Then recycled reagent, to get two extraction parts. The above was separated and purified by variety of chromatographic techniques. The structures were identified by various methods that including thin layer chromatography, spectrum, mass spectrometry, literature comparison and so on.2.The separation was performed by HPLC gradient elution with the mobile phase composed of methanol(A)-0.05 % phosphoric acid water(B).The wavelength of detection was 220 nm and the flow rate was 0.8 ml.min-1.The temperature of column was 30℃.Elution procedure was:0 min~10 min(2% B), 10 min~20 min(2%~5% B),20 min~60 min(5 %~40 % B), 60 min~85 min(40%B).Gastrodine was taked as the reference compound. The HPLC chromatographic fingerprint of 39 samples of GASTRODIAE RHIZOMA were evaluated.3.The mobile phase of HPLC was acetonitrile-0.05 % phosphoric acid water(3:97), the wavelength of detection was 220 nm, the flow rate was 0.6 ml.min-1, And the temperature of column was 25℃.4.The sample pretreatment method was Microwave digestion. Then heavy metal Contents were determined by ICP-MS.Results:1.21 compounds were obtained from the aerial parts of the Gastrodia elata Bl. Among them, 20 structures were identified as 2-(4-hydroxybenzoyl)-3-hydroxyethyl indole(1), 4,4’-((4-hydroxy-2-(3-hydroxyphe-nethyl)-6-methoxy-1,3-phenylene) bis(methy lene)) diphenol(3), N- trans- feruloyltyramine(4), p-hydroxybenzoic acid(5), N-cisferuloyityramine(6), 3-(p-Hydroxy) benzoyl indole(7), p-hydroxybenzoic acid methylester(8), hydroquinone(9),4-(methylamino)-benzyl alcohol(10),beta-sitosterol(11), 2H-Pyran-2-one, tetrahydro-4-hydroxy-4,5-dimethyl-(9CI)(12), daucosterol(13), Botcinin F(14), Cytochalasin B(15), Botcinic acid methy ester(16), 2-(4-hydroxybe nzoyl)-3-(4-hydroxybenzyl) indole(17), E-cinnamicacid(18),(+)–neoechinulin A(19),gastrodin(20), 3β,5α,6β-tri–hydroxylated sterols(21).2. The HPLC fingerprint of main producing areas of GASTRODIAE RHIZOMA was established and the fingerprint consisted of 8 common peaks. Four common peaks were identified with the reference substance.3. The linear relationship of Gastrodin concentration which ranged from 20 μg.ml-1-320 μg.ml-1 was good. The average recovery was 100.87%, RSD was 2.89 %. The average content of 39 batchs of GASTRODIAE RHIZOMA was 0.42 %, the highest content from Anhui Jinzhai was 1.27 %, the lowest content from Yunnan Zhaotong was 0.08%, Most of the content of the samples ranged from 0.25 %-0.65 %.4.A method that is ICP with microwave digestion was established for determination of the contents of five kinds of heavy metals. The linear relationship of each element was good(r=0.991-0.999), and the average recovery ranged from 84.0 %-105.2 %. Comprehensived referenced two standards, five elements including lead,cadmium, shall,arsenic,copper exceed the standard number were 5, 8, 0,18, 3 in turn.Consults: 21 compounds were isolated from the aerial parts of the Gastrodia elata Bl. Among them, compound 1 and 17 were new compounds. For the first time,compounds 4,6,7,8, 9, 10, 12, 14, 15, 16, 19 were isolated from this plant. The HPLC fingerprint can be used to evaluate the quality of GASTRODIAE RHIZOMA from main producing areas. According to the limited standard of gastrodin, a total of 35 batches of samples conformed to the standard. Gastrodin content of samples from different regions existed differences.This method is convenient and quick.Arsenic and cadmium elements of GASTRODIAE RHIZOMA severely exceed the standard.