Studies On The Effects Of The Main Chemical Components In Japanese Atractylodes Rhizome By Processing

Objective:Studies on the effects of the main chemical components in Japanese Atractylodes Rhizome by determining the chemical composition in Japanese Atractylodes Rhizome samples by GP-MSE-GC-MS and HPLC before and after process and determining the contents of water-soluble extract, alcohol-soluble extract and volatile ether extract in Japanese Atractylodes Rhizome samples. To provide the experimental basis for the processing of Japanese Atractylodes Rhizome.Methods:1. The comparative study of Japanese Atractylodes Rhizome by GP-MSE-GC-MS before and after processDB-5MS capillary column (30mx0.25mm x0.25mm) was used. Helium of high-purity was used as the carrier gas at a flowrate of0.98mL/min. The injector temperature was280℃. The GC oven temperature was programmed as follows:the initial oven temperature was45℃and held for1min, and then to250℃at a rate of4℃/min and finally to280℃at a rate of6℃/min and held for5min. Splitless injection was conducted and the injection volume is2μL.2. The comparative study of Japanese Atractylodes Rhizome by HPLC before and after processHPLC chromatographic behavior analysis:Kroasil C18column(4.6mm x250mm,5μm), using acetontrile-water solution as mobile phase gradient elution. The flow rate was1.0mL.min-1with detective wavelength of220nm. Column temperature was28℃. The injection volume is20μL.Content determination of Atractylenolide â… å'Œâ…¢:C18column, using methanol-acetontrile-water solution (59.5:0.5:40) as mobile phase. The flow rate was1.0mL.min-1with detective wavelength of220nm. Column temperature was28℃.3. The comparative study of extracts content in Japanese Atractylodes Rhizome before and after processIn accordance with the Appendix XA in "Chinese Pharmacopoeia" of2010edition, determine the contents of water-soluble extract, alcohol-soluble extract and volatile ether extract in Japanese Atractylodes Rhizome samples, compare the contents of water-soluble extract, alcohol-soluble extract and volatile ether extract in Japanese Atractylodes Rhizome samples.Results:1. The comparative study of Japanese Atractylodes Rhizome by GP-MSE-GC-MS before and after process42components were identified in rhizome of Atractylodes japonica Koidz.et Kitam and46components were identified in root tuber of Atractylodes japonica Koidz.et Kitam.41same components were identified in rhizome and root tuber of Atractylodes japonica Koidz.et Kitam. There is one specific component in rhizome, it is Cyclotene; there are five specific components in root tuber of Atractylodes japonica Koidz.et Kitam, they are1,2,3,6-tetramethyl-bicycle [2,2,2] octa-2,5-diene, alpha.-bulnesene, alpha.-caryophyllene, gamma.-selinene and alpha.-ionone.Detected91peaks and identified46components in the samples of Japanese Atractylodes Rhizome. After soaking, there are five same components in the components which the relative content is reduced in the rhizome and root tuber of Atractylodes japonica Koidz.et Kitam, three of them are Furyl alcohol,5-methyl-2-Furaldehyde and Elemol. the other two are uncertain components. There are seven same components in the components which the relative content is increased in the rhizome and root tuber of Atractylodes japonica Koidz.et Kitam, six of them are1,2,3,6-tetramethyl-bicycle [2,2,2] octa-2,5-diene, alpha.-Bulnesene, beta.-Caryophyllene, gamma.-Elemene, beta.-Eudesmene and Valencene, the other one is uncertain component.Analysis the samples before and after process by the method of principal component anlysis. The samples of Atractylodes japonica Koidz. at Kitam could be clustered into four kinds, they are rhizome original medicine, root tuber original medicine, rhizome processed samples and root tuber processed samples. The samples of rhizome of Atractylodes japonica Koidz. at Kitam could be clustered into three kinds, they are rhizome original medicine, rhizome soaked samples and rhizome heated afer soaking samples. The samples of root tuber of Atractylodes japonica Koidz. at Kitam could be clustered into three kinds, they are root tuber original medicine, root tuber soaked samples and root tuber heated afer soaking samples.2. The comparative study of Japanese Atractylodes Rhizome by HPLC before and after processHPLC chromatographic behavior analysis:Compare the chromatogram similarity of Japanese Atractylodes Rhizome samples before and after process by correlation coefficient method. In the rhizome samples of Japanese Atractylodes Rhizome, the similarities of soaked in water sample or soaked in washing water of rice and the original medicine were0.647and0.799. The similarities of soaked in water sample afer heating under80℃or heating under100℃or heating under120℃and soaked in water sample were0.965,0.897and0.878. The similarities of soaked in washing water of rice sample afer heating under80℃or heating under100℃or heating under120℃and soaked in washing water of rice sample were0.778,0.744and0.731.In the root tuber samples of Japanese Atractylodes Rhizome, the similarities of soaked in water sample or soaked in washing water of rice and the original medicine were0.623and0.745. The similarities of soaked in water sample afer heating under80℃or heating under100℃or heating under120℃and soaked in water sample were0.916,0.868and0.854. The similarities of soaked in washing water of rice sample afer heating under80℃or heating under100℃or heating under120℃and soaked in washing water of rice sample were0.733,0.721and0.709.Content determination of Atractylenolideâ… å'Œâ…¢:The Atractylenolide I was linear within the range of70～560ng (r=0.9997),the average recovery of Atractylenolide I was100.79%, RSD was4.87%. The Atractylenolide â…¢ was linear within the range of39～390ng (r=0.9999),the average recovery of Atractylenolide â…¢ was101.83%, RSD was5.43%. Compared with the original herbs, the contents of Atractylenolide â…  and Atractylenolide â…¢in soaking with water and washing water of rice samples were all decreased; After heating at80℃～120℃for20min, the contents of Atractylenolide â…  in soaked samples showed a increasing trend, the contents of Atractylenolide â…¢ in soaking with water sample of rhizome showed a decreasing trend, the contents of Atractylenolide â…¢in washing water of rice sample showed a increasing trend; The contents of Atractylenolide III in soaked samples of root tuber showed a increasing trend.3. The comparative study of extracts content in Japanese Atractylodes Rhizome before and after processCompared with the original herbs, the contents of extracts in soaking with water and washing water of rice samples were all decreased; After heating at80℃-120℃for20min, the contents of water-soluble extract and alcohol-soluble extract in soaked samples did not have significantly change, the content of volatile ether extract showed a decreasing trend.Conclusion:1. The category and content of volatile components identified in rhizome and root tuber of Atractylodes japonica Koidz.et Kitam have no significant difference.2. Water soaking or washing wanter of rice soaking can affect the volatile components and polar components in Atractylodes japonica Koidz.et Kitam.3. Heating at80℃-120℃for20min can affect the volatile components and polar components in soaking samples.The effect has a certain relationship with temperature. on the contents of extracts in soaked samples; The impact of soaking in water and washing water of rice on the extracts is different.