蒼耳子中酚酸類化合物及不同品種和居群蒼耳子中總酚酸含量的測(cè)定

1、蒼耳子中酚酸類化合物及不同品種和居群蒼耳子中總酚酸含量的測(cè)定         07-09-12 10:38:00     編輯：studa20          作者：韓婷，李慧梁，胡園，張巧艷，黃寶康，鄭漢臣，Khalid RAHMAN，秦路平 【關(guān)鍵詞】  蒼耳子    摘要  目的：研究中藥蒼耳子（Fructus Xanthii）中

2、的化學(xué)成分；測(cè)定不同居群和不同品種蒼耳屬植物果實(shí)中的總酚酸含量以評(píng)價(jià)其品質(zhì)。方法：運(yùn)用各種層析方法分離純化化合物，以綠原酸作對(duì)照，用紫外分光光度法測(cè)定總酚酸的含量。結(jié)果：（1）從蒼耳子中分離到6個(gè)咖啡?？鼘幩犷惢衔?，以及咖啡酸和阿魏酸；（2）從中國29個(gè)居群采集到的樣品，總酚酸含量為0.311.44%。在采集到2個(gè)品種及1個(gè)變種的蒼耳屬植物中，近無刺蒼耳X. sibiricum var. subinerme樣品的總酚酸含量較其他兩種都偏低。蒼耳X. sibiricum樣品中，從上海采集的3號(hào)蒼耳子樣品總酚酸含量最高，達(dá)1.44%，而從江西新建采集的12號(hào)樣品含量最低，僅0.38%。結(jié)論：5O

3、咖啡?？鼘幩?，1，4-二O咖啡?？鼘幩岷?，5二O咖啡?？鼘幩崾菑纳n耳屬植物中首次分離獲得。不同品種和不同居群蒼耳子藥材中，總酚酸含量的差異較為顯著。若以總酚酸含量作為指標(biāo)評(píng)價(jià)蒼耳子藥材的品質(zhì)，則認(rèn)為上海和福建三明產(chǎn)蒼耳子為優(yōu)質(zhì)種群。關(guān)鍵詞  蒼耳子; 酚酸; 咖啡?？鼘幩? 紫外分光光度法; 化學(xué)結(jié)構(gòu)鑒定; 居群; 含量測(cè)定; 質(zhì)量評(píng)價(jià)Phenolic acids in Fructus Xanthii and determination of contents of total phenolic acids in different species and populations

4、of Xanthium in ChinaABSTRACT  Objective: To study the chemical constituents of Fructus Xanthii and to determine the contents of total phenolic acids (TPA) in fruits of Xanthium from different populations for evaluating the quality of them. Methods: Components in Fructus Xanthii were isolated an

5、d purified by various column chromatographies and the contents of TPA were determined by ultraviolet spectrophotometry with chlorogenic acid (CHA) as reference substance. Results: Six caffeoylquinic acids along with caffeic acid and ferulic acid were isolated from Fructus Xanthii. The contents of TP

6、A of the samples collected from 29 populations in China varied from 0.31% to 1.44%. Among the samples originated from two species and 1 variety of Xanthium, the contents of TPA in X. sibiricum var. subinerme samples with an average of 0.36% were relatively lower than those in other 2 species. While

7、the content of TPA in Sample 3 collected from Shanghai was 1.44% and the highest among all the samples, and that in Sample 12 from Xinjian of Jiangxi Province was 0.38% and the lowest among the X. sibiricum samples. Conclusion: 5Ocaffeoylquinic acid, 1,4diOcaffeoylquinic acid and 4,5diOcaffeoylquini

8、c acid were isolated from Xanthium plant for the first time. The difference of contents of TPA in samples from different species and different populations in China was relatively significant. Fructus Xanthii in Shanghai and Sanming of Fujian Province were considered high quality if contents of TPA w

9、ere used as reference for quality evaluating.KEY WORDS  Fructus Xanthii; phenolic acid; caffeoylquinic acid; ultraviolet spectrophotometry; chemical structure identification; population; content determination; quality evaluationThe genus Xanthium (family Compositae) is represented by 25 species

10、 in the world and 3 species and 1 variety in China1, which are Xanthium sibiricum Patr. (Xanthium strumarium L.), X. mongolicum Kitag., X. inaequilaterum DC. and X. sibiricum var. subinerme (Winkl.) Widder. Xanthium species have been used as traditional herb medicines for a long history in oriental

11、countries. Xanthium sibiricum is the principle species abundantly found throughout China, and its fruits (Fructus Xanthii) are used in China for the treatment of nasal sinusitis, headache caused by windcold, urticaria and arthritis2. In continuation of research on biologically active compounds, we c

12、onducted the study on X. sibiricum and obtained 8 phenolic acids from nbutanol fraction. Pharmacological studies revealed that phenolic acids were the main active components and the reports from some publications showed that these naturally occurring phenolic acids had various pharmacological proper

13、ties and could be used to act as cholagogues, stomach stimulants, and immunostimulants, as well as antiinflammatory, antibacterial, and antifungal agents35. In order to utilize this crude drug more reasonably and scientifically, we determined the contents of total phenolic acids (TPA) in 2 species a

14、nd 1 variety Xanthium samples collected from 29 populations in China by ultraviolet (UV) spectrophotometry6.1  MATERIALS1.1  Apparatus  UV analysis was operated on Shimadzu UV2550 spectrophotometer. Nuclear magnetic resonance (NMR) spectra were operated on a Bruker DRX500 spectrometer

15、 at 500 MHz for 1HNMR and 125 MHz for 13CNMR. Chemical shift was expressed in values with reference to tetramethylsilane (TMS) as internal standard, and coupling constants (J) were given in Hz; Electron ionization mass spectrometry (EIMS) was recorded on a Varian MAT212 mass spectrometer and HRESI o

16、n a QTOF micro mass spectrometer; Melting point was measured on a RY2 melting point apparatus that was uncorrected; Infrared (IR) spectrum was recorded on a Bruker Vector22 spectrometer with KBr pellet.1.2  Chemicals and drugs  Column chromatography was performed on silica gel (200300 mesh

17、, Yantai, China), silica gel H (1040 m, Yantai, China), and sephadex LH20 (Pharmacia); Thinlayer chromatography (TLC) analysis was run on HSGF254 precoated silica gel plates (1040 m, Yantai, China).  Chemicals are all ARgrade and purchased from Shanghai Chemical Reagent Co., Ltd (Shanghai, Chin

18、a); Distilled deionized water was produced by a MilliQ Reagent Water System (Millipore, MA, USA); Chlorogenic acid (CHA) was prepared in our lab and its purity was determined to be over 98% by HPLC analysis.1.3  Plant materials  The ripe fruits of Xanthium sibiricum were collected from a l

19、ocal research farm in Sunqiao town, Shanghai, China in November 2003 and authenticated by Prof. HanChen ZHENG, Second Military Medical University. A total of 29 samples of the wild herbs of Xanthium were collected from 19 provinces of China in 2003 and 2004 (Table 1). The voucher specimens of these

20、plants were deposited at the Herbarium of Department of Pharmacognosy, Second Military Medical University, Shanghai, China.2  METHODS AND RESULTS2.1  Extraction, isolation and identification  Dried fruits of X. sibiricum (20 kg) were ground and extracted with 75% aqueous ethanol by re

21、flux. The solvent was evaporated under vacuum to afford 1 100 g crude extract (yield, 5.5%). Then the extract was suspended in water and partitioned with petroleum ether, chloroform, ethyl acetateandaquasaturatednbutanolsuccessively.Table 1  TPA in different species and different populations of Xanthium in China（略）Each fraction was evaporated under vacuum to yield the residues of petroleum ether fraction 55 g (5.0%), chloroform fraction 60 g (5.4%), ethyl acetat