【作者】 陶燕；

【导师】 巢建国；

【作者基本信息】 南京中医药大学 ， 生药学， 2007， 硕士

【摘要】 为保护江苏省的野生茅苍术资源，进一步分析组培茅苍术药材的质量，探索控制苍术类药材内在质量的新方法，在参考相关文献的基础上，本论文进行了以下研究：采挖新鲜茅苍术根茎制作石蜡切片，比较组培茅苍术、栽培茅苍术、野生茅苍术三者的显微结构，发现三者在显微结构上没有显著性差异，组培品的油室数量相对较多。总体而言，随着苍术生长年限的增加，木栓层层数和油室数量也不断增加。在茅苍术的花期对其实行人工辅助授粉，结果显示人工辅助授粉可以辅助茅苍术的雌蕊完成受粉过程，有利于茅苍术在野外顺利繁殖，能够显著提高茅苍术的结实率。同时摸索了采用人工补种方式增加茅苍术种群数量的具体条件，发现选择已发芽，且芽高在2～7cm的根茎进行切段，栽种于只有少量灌木遮阴而多草本的环境（遮阴度30％左右），苍术的成活率较高，生长情况较好。初步表明人工辅助授粉及人工补种可以作为增加茅苍术种群数量及种群内个体数的新措施。收集了十二个苍术样品并对其进行薄层鉴别，结果表明，组培茅苍术、人工补种茅苍术所含成分与野生和栽培的茅苍术相似。茅苍术、罗田苍术、北苍术在成分上有明显区别，可通过薄层层析进行鉴别。初步建立了茅苍术及苍术类药材的顶空气相色谱指纹图谱质量控制模式。首先确定茅苍术HSGC最佳分析条件：HP-INNOWAX毛细管柱；30℃，保持5min，以8℃·min^-1的速率升至120℃，保持10min，再以6℃·min^-1的速率升至180℃，保持5min，然后以5℃·min^-1的速率升至200℃，保持2min的程序升温条件；2cm³样品粉末（40目）充填10mL顶空进样瓶，以100℃平衡80 min，进样0.7mL顶空气体的样品制备条件。方法学考察显示所确定的HSGC分析方法精密度和重现性均良好。将所得图谱导入中药色谱指纹图谱相似度评价系统中进行相似度分析，逐个比较各个样品的差异，得到了苍术药材和茅苍术药材的共有模式，发现12个苍术样品与标准图谱相似度的差异很大，可初步分为五类：第一类为江苏茅山地区、江苏镇江地区野生茅苍术，江苏南京地区人工补种茅苍术及安徽产苍术；第二类为江苏镇江栽培品和江苏宜兴栽培的组培品；第三类为湖北英山产茅苍术、湖北产罗田苍术、山西盂县产野生北苍术，另一个山西收集的北苍术药材和内蒙1号药材归为第四类；内蒙2号药材单独归为第五类。总体而言，苍术药材的成分差异较大，受地域及生长环境的影响较为显著。更多还原

【Abstract】 By consulting the correlational data and literature, researches were performed in theseaspects listed below in order to protect the wild resource of Atractylodes lancea (Thunb.)DC. in Jiangsu Province, and to analysis the quality of tissue cultured Atractylodes lancea(Thunb.) DC., and to search for a new method for the purpose of controlling the qualitativeof Rhizoma Atractylodis:Atractylodes lancea’s fresh rhizomes were dug and made into paraffin section for thepurpose of comparing tissue cultured rhizomes’s microstructure to cultured ones’ and wildones’. There are no significant differences in the microstructure of tissue culturedAtractylodes lancea, wild Atractylodes lancea and cultured Atractylodes lancea. Tissuecultured Atractylodes lancea has more oil sacs than the others. Follow the increasing ofgrowing years, the numbers of cork layer and oil sac are raising unceasingly.Atractylodes lancea was pollinated artificially at its florescence. It was showed thatartificial pollination can help gynoecia to accomplish the process of fecundation. Artificialpollination is good for for Atractylodes lancea to propagate prosperously in field, and it canraise seed set propagation coefficient significantly.At the same time, specific conditions ofadditional seeding that should be noticed were groped when using this method to increasethe population quantity of Atractylodes lancea. It was found that Atractylodes lancea cangrow better when choosing the rhizome with a bud which height is 2～7cm and whenplanting it to a place with little bush and many weeds. Artificial pollination and additionalseeding can be new measures to increase the population quantity of Atractylodes lancea.Twelve Rhizoma Atractylodis samples were collected. They were treated with themethod of TLC (thin layer chromatography). The chemical compositions of tissue culturedrhizomes and additional seeding rhizomes are similar to cultured ones and wild ones. It wasfound that there are significant differences in the chemical constituents of Atractylodeslancea (Thunb.) DC., A. lancea (Thunb.) DC. subsp. Luotianensis S. L. Hu et X. F. Fengand A. chinesis (DC.) Koidz.. These species can be identifid by TLC.The HSGC (headspace gas chromatography) fingerprint identification pattern wasinitially established to control the quality of Rhizoma Atractylodis. The best HSGC analysis conditions of Rhizoma Atractylodis were firstly determined as listed below.HP-INNOWAX capillary column is suitable. Temperature programming is that keep30℃for 5 minutes, then rise to 120℃at the rate of 8℃per minute, keep 120℃for 10minutes, then rise to 180℃at the rate of 6℃per minute, keep 180℃for 5 minutes, andthen rise to 200℃at the rate of 5℃per minute, finally keep 200℃for 2 minutes.Theway of sample preparation is that get 2cm~3 powder (40 mesh) into a headspace samplebottle of 10 millilitres, then heat the bottle in a baking oven at 100℃for 80 minutes,inject the headspace gas of 0.7 millilitres into the gas chromatography instrument. It wasshowed that the precision and reproducibility of this method was good. Then put thechromatograms into a fingerprint chromatograms analysis system of Chinese medicinalmaterials in order to calculate the similarity and pertinence of all samples. By comparingall the samples to each other, common chromatograms of Rhizoma Atractylodis andAtractylodes lancea were obtained.There are many differences between twelve sampleswhen comparing to the common chromagrams. These samples can be divided into fivecategories. The first category includes Rhizoma Atractylodises collected in Maoshanmountain of Jiangsu Province, in Zhenjiang of Jiangsu Province, in Nanjing of JiangsuProvince, and in Anhui. The second category includes rhizomes cultured in Zhenjiangand tissue cultured rhizomes cultured in Yixing of Jiangsu Province. Rhizomes of A.lancea (Thunb.) DC. cultured in Hubei Province, rhizomes of A. lancea (Thunb.) DC.subsp. Luotianensis S. L. Hu et X. F. Feng cultured in Hubei Province and A. chinesis(DC.) Koidz. cultured in Shanxi Province are included in the third category. Thiscategory significantly differ from the other ones. Another sample of A. chinesis (DC.)Koidz. collected in Shanxi Province and one sample collected in Inner Mongolia shouldbe classified to the fourth category. Another Inner Mongolia sample is classifiedindividually into the fifth category. To be a conclusion, the chemical constituents ofRhizoma Atractylodises collected from distinct habitats differ from each other. Thechemical compositions are significantly effected by gathering place and habitatcircumstances.更多还原