| Cimicifuga, Latin scientific name is Rhizoma cimicifugae, which is contained in the "Shen Nong's Herbal Classic", mainly produced in Sichuan, Chongqing, Qinghai, Shanxi, Henan and Northeast of China. The plants of Cimicifuga belong to Ranunculaceae which are the main members of the Cimicifuga plants. There are three original plants are received in "Chinese Pharmacopoeia" (2005) which are used as Cimicifuga for the common drugs:Cimicifuga heracleifolia Kom. Cimicifuga dahurica (Turcz.)Maxim. and Cimicifuga foetida L. They are widely distributed in China and have large resources. Cimicifuga rhizome is mainly composed of triterpenoid saponin, organic acids, and color of the original ketones. Organic acid is generally considered as the active ingredients for the effect of reducing pathogenic fire and detoxication, meanwhile the ferulic acid and isoferulic acid are considered as the indexs to control their quality. Recent studies showed that the triterpenoid saponin contained in Cimicifuga had anti-tumor, inhibition of nucleoside operation, regulation of endocrine and neurological function, was calcium antagonist, and could inhibit inflammatory bone loss and pain or other physical activity, so it was a class of highly developed prospects for natural products.Nanchuan Cimicifuga (Cimicifuga nanchuanensis) is a specific variety to China. Its roots is used as medicine and also known as "Cimicifuga green bean". It is well-known herbs in Chongqing. Because of their narrow distribution, meanwhile their ecological environment was being destroyed serious in recent years; it is endangered and is classified as National Plant Protection II. To know the causes and mechanisms of its endangered, some scholars studied its reproductive biology and photosynthetic characteristics systematically. But the ingredients for the determination of Cimicifuga nanchuanensis in the literature are few. The current reports focusing on the researches and studies of chemical constituents of Cimicifuga plant are mainly concentrated in analyzing the chemical composition of its roots, but just few studies for its aerial parts. This is contrary to the sustainable development of resources for the Cimicifuga, especially for the Study of Cimicifuga nanchuanensis which is in more endangerous status desirably. "Chinese Pharmacopoeia" edited 2005 adopts to TLC method using silica gel G with isoferulic acid as the standard for identification tests and controlling the quality of the medicinal Cimicifuga. by measuring the content of isoferulic acid. With the development of chromatography, especially high performance liquid chromatography technology getting more sophisticated, an increasing number of Chinese herbal medicines have established their own quality control standards. So this study used gel G thin-layer chromatography method to identify Cimicifuga nanchuanensis, while used high performance liquid chromatography (RP-HPLC) measure the content of ferulic acid and isoferulic acid in its sub-sites. On this basis, we studied variety in different parts of the Cimicifuga nanchuanensis. And also studied the content differences of other varieties in the same part between ferulic acid and isoferulic acid. Meanwhile we also compared the differences between ferulic acid and isoferulic acid aboveground and underground parts of the Cimicifuga nanchuanensis. and provided some references to save the endangered state of Cimicifuga nanchuanensis. The main findings were as follows:1 Through the thin layer chromatographic method, could visually see in various parts of Cimicifuga nanchuanensis contained ferulic acid and isoferulic acid, which was consistent with other Cimicifuga species.2 Extraction conditioning 10% ethanol,30% ethanol,60% ethanol,80% ethanol, were refluxed and ultrasonicly treated, to determine the same material, the results showed that 60% ethanol refluxed 180min was better.3 Chromatographic conditions:This study tested varieties of mobile phase, detection wavelength and column and finalized naive conditions:mobile phase:methanol: 0.1% formic acid (35:65); UV detection wavelength:310nm; column temperature: 35℃; column:Waters C18 (4.6mm×150mm 5um); Sensitivity:2.0 AUFS; flow rate: lml/min; injection volume:10ul.4 The determination of the standard curve:In this study, according to the established chromatographic conditions, injected the concentration profile of the reference standard into the liquid chromatograph, and recorded the peak area., then used the injection volume of the ferulic acid and isoferulic acid as the horizontal respectively, the peak area as the vertical coordinates, plot the standard curve. The result showed that ferulic acid were in the range of 0.000390625～0.0125mg, isoferulic acid were in the range of 0.000390625～0.0125mg with good linear relationship in the peak area, the ferulic acid standard curves was:y= 3551.4x+0.848 R2= 0.9976; the isoferulic acid standard curves was:y= 2604.9x+0.6102 R2= 0.9987.5 RP-HPLC determination results:This study measured three kinds of Cimicifuga species that were collected on the same region, the results showed that the content of ferulic acid and isoferulic acid of the Cimicifuga nanchuanensis in aboveground and underground parts were higher than the other two species. The contents of ferulic acid and isoferulic acid in various different parts in Cimicifuga nanchuanensis were lower:The content of ferulic acid in leaves was 0.021414%, isoferulic acid was 0.110615%; the content of ferulic acid in stems was 0.011188%, isoferulic acid was 0.148814%; the content of ferulic acid in inflorescences was 0.011993%, isoferulic acid was 0.216306%; the content of ferulic acid in roots was 0.019105%, isoferulic acid was 0.855491%; the content of ferulic acid in fibrous roots was 0.019052%, isoferulic acid was 0.494472%. Thus the following conclusions were:The ferulic acid and isoferulic acid of Cimicifuga nanchuanensis were mainly concentrated in the roots and fibrous roots of the underground parts; The ferulic acid and isoferulic acid were mainly concentrated in the stems, leaves and inflorescences of Cimicifuga nanchuanensis aboveground were in the low level relatively.
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