本文選題：延胡索 + 生物堿�。� 參考：《重慶三峽學(xué)院》2017年碩士論文

【摘要】：本文選取9個(gè)不同生境(生態(tài)環(huán)境)延胡索(Corydalisyanhusuo)為研究對(duì)象,分別采用原子吸收光譜法(AAS)、氣相色譜(GC)法、高效液相色譜(HPLC)法和生物化學(xué)方法測(cè)定了延胡索種子和栽培土壤中4種重金屬含量以及8種有機(jī)氯農(nóng)藥殘留量,以及6種生物堿的含量和3種核苷含量,不同土壤理化指標(biāo)和土壤酶活性,同時(shí)分析探討了生境中不同生態(tài)因子與延胡索有效成分間的關(guān)系,通過(guò)對(duì)不同生境下延胡索質(zhì)量控制和安全性的研究,以期為延胡索藥材的規(guī)范化栽培、保護(hù)利用提供參考依據(jù)。本研究主要結(jié)論如下:1.重金屬含量分析研究。AAS法測(cè)定延胡索藥材及其根際土壤的銅、鉛、鋅、鉻的含量。結(jié)果顯示4種重金屬含量均達(dá)到國(guó)家《藥用植物及制劑進(jìn)出口綠色行業(yè)標(biāo)準(zhǔn)》的要求,如延胡索藥材中銅的含量范圍為2.66～5.55 mg·kg-1,根際土壤中銅的含量范圍為42.35～73.69mg·kg-1。2.農(nóng)藥殘留分析研究。GC法同時(shí)測(cè)定了延胡索藥材和根際土壤中4種六六六同分異構(gòu)體(a-BHC、β-BHC、γ-BHC、d-BHC)和4種滴滴涕同分異構(gòu)體(4,4'-DDD 0,4'-DDT4,4'-DDE 4,4'-DDT)等8種有機(jī)氯農(nóng)藥成分。結(jié)果顯示不同產(chǎn)地延胡索藥材和根際土壤中這8種農(nóng)藥成分都有不同程度的檢出,但含量均未超過(guò)綠色中藥標(biāo)準(zhǔn)的限定。如,a-BHC所有產(chǎn)地都有檢出,4,4'-DDD只在重慶開(kāi)州區(qū)郭家鎮(zhèn)被檢出,含量為 0.007473 mg.kg-1,y-BHC、d-BHC 和 4,4'-DDE 沒(méi)有被檢測(cè)出來(lái)。3.不同生境延胡索6種生物堿含量分析。采用HPLC法同時(shí)測(cè)定延胡索藥材中6種主要生物堿,比較了不同生境延胡索塊莖中6種生物堿含量情況。其中延胡索乙素變動(dòng)范圍為1.18～118μg·mL-1,脫氫紫堇堿變動(dòng)范圍為0.25～200μg·mL-1,鹽酸小僻堿變動(dòng)范圍為0.56～112μg·mL-1,鹽酸巴馬汀變動(dòng)范圍為2.175～87μg·mL-1,鹽酸黃連堿變動(dòng)范圍為2.59～103.6μg·mL-1,原阿片堿變動(dòng)范圍為2.69～26μg·mL-1,峰面積積分值與標(biāo)準(zhǔn)濃度的線性關(guān)系良好,平均加樣回收率在96.19%～97.59%之間,R20.9993。結(jié)果顯示,不同生境延胡索藥材中6種生物堿含量平均值呈現(xiàn)出顯著差異性,同一產(chǎn)地的不同種類(lèi)生物堿的含量平均值也有所不同但同種生物堿所占總生物堿的比例比較接近。如生物堿總含量最高的是浙江省東陽(yáng)市千祥鎮(zhèn)的延胡索,比含量最低的陜西省漢中市周家山鎮(zhèn)的總生物堿多出2倍還要多。4.不同生境延胡索3種核苷含量分析。運(yùn)用HPLC法同時(shí)測(cè)定延胡索藥材中鳥(niǎo)苷、尿苷和腺苷等3種核苷,比較了不同生境延胡索塊莖中3種核苷的含量結(jié)構(gòu)比的差異情況。其中尿苷濃度在12.12～242.40μg·mL-1之間,鳥(niǎo)苷濃度在8.04～160.80μg·mL-1之間,腺苷濃度在6.00～120.00μg·mL-1之間,峰面積積分值與標(biāo)準(zhǔn)濃度的線性回歸關(guān)系良好,回歸方程相關(guān)系數(shù)為R20.9998,平均加樣回收率均在99.63%～104.24%之間,RSD在0.7%～2.3%之間。結(jié)果顯示,不同生境延胡索藥材中3種核苷成分及組成結(jié)構(gòu)比差異性顯著(P0.01);同一產(chǎn)地延胡索藥材的3種成分量及組成結(jié)構(gòu)比也存在明顯差異(P0.05)。其中3種核苷總量較高的是來(lái)自安徽省蕪湖縣花橋鎮(zhèn)延胡索藥材。表明不同產(chǎn)地對(duì)延胡索的核苷含量具有一定的影響。5.土壤理化指標(biāo)含量分析研究。9個(gè)不同種植基地延胡索土壤pH值在4.13～5.48之間,屬于酸性土壤,與各地背景值差異不大。全氮含量范圍在730～2080 mg·kg-1,變異系數(shù)為0.39;有效氮的含量范圍在18.67～242.63mg·kg-1,其變異系數(shù)大于全氮的變異系數(shù),為0.74。全磷含量范圍在0.47～1.12 mg·kg-1,變異系數(shù)為0.35;有效磷的含量范圍在0.03～0.31mg·kg-1,變異系數(shù)為0.37。不同種植基地土壤全氮、有效氮、全磷、有效磷均呈現(xiàn)出分布顯著差異性(P0.01),其中陜西漢中速效氮含量最高,重慶開(kāi)州區(qū)大德速效磷含量最高,表明兩個(gè)產(chǎn)地土壤的肥力較強(qiáng)。6.土壤酶活性分析研究。9個(gè)不同產(chǎn)地的延胡索土壤脲酶活性范圍在57.04～137.32mg·g-1,蔗糖酶活性范圍在1.574～11.82mg·g-1,蛋白酶活性范圍在0.819～10.11mg·g-1,酸性磷酸酶活性范圍在2.683～7.2mg·g-1,中性磷酸酶活性范圍在0.04～3.57mg·g-1,堿性磷酸酶活性范圍在0.007～1.39mg·g-1。不同生境延胡索種植區(qū)的6種土壤酶活性均存在一定的差異性,同一產(chǎn)區(qū)的幾種酶活性也存在差異性,說(shuō)明不同生境延胡索種植區(qū)土壤中氮、磷等轉(zhuǎn)化和供應(yīng)強(qiáng)度存在一定的差異。其中陜西漢中土壤脲酶和蛋白酶活性最高,而土壤蔗糖酶活性最高的是來(lái)自江蘇省通州區(qū)的根際土壤,土壤酸性磷酸酶活性最高的是來(lái)自陜西漢中的延胡索根際土壤,說(shuō)明這些地方的N、C、P轉(zhuǎn)化和供應(yīng)強(qiáng)度大。7.生態(tài)因子與延胡索有效成分的分析。利用雙變量分析法分析了氣候因子、土壤因子、土壤理化指標(biāo)和土壤酶活性與延胡索6種生物堿和3種核苷之間的相關(guān)性,結(jié)果顯示,某些生態(tài)因子與延胡索有效成分間存在一定的顯著相關(guān)性。例如:年均溫與鳥(niǎo)苷、土壤中砂礫比例與鹽酸黃連堿、全磷與原阿片堿相關(guān)系數(shù)|r|均大于0.8,(P0.01)呈極顯著正相關(guān)性。隨著年均溫、土壤中砂礫比例和全磷含量的增大,鳥(niǎo)苷、鹽酸黃連堿和原阿片堿的含量也不斷增大。表明環(huán)境中生態(tài)因子影響著延胡索藥效成分和結(jié)構(gòu)比,可為今后更好的生產(chǎn)和研究延胡索藥材,提供寶貴的資料。
[Abstract]:In this paper, 9 different habitats (ecological environment) (Corydalisyanhusuo) were selected as the research object. 4 kinds of heavy metal contents and 8 kinds of organochlorine pesticide residues in Rhizoma Corydalis seed and cultivated soil were determined by atomic absorption spectrometry (AAS), gas chromatography (GC), high performance liquid chromatography (HPLC) and biochemical methods. The contents of alkaloids and 3 kinds of nucleosides, different soil physical and chemical indexes and soil enzyme activities, and the relationship between the different ecological factors and the effective components of the Rhizoma Corydalis in the habitat, and the study on the quality control and safety of the Rhizoma Corydalis in different habitats, in order to provide reference for the standardized cultivation of the rhizoma corydalis and the protection and utilization. The main conclusions of this study are as follows: 1. the content of copper, lead, zinc and chromium in rhizosphere soil of Rhizoma Corydalis and rhizosphere of Rhizoma Corydalis is determined by.AAS method. The results show that the content of 4 kinds of heavy metals all meet the requirements of the national standard for the import and export of medicinal plants and preparations, such as the range of copper in Rhizoma Corydalis is 2.66 to 5.5. 5 mg. Kg-1, the content range of copper in rhizosphere soil is 42.35 ~ 73.69mg. Kg-1.2. pesticide residues analysis and Study on 4 666 isomers (a-BHC, beta -BHC, gamma -BHC, d-BHC) and 4 kinds of organochlorine pesticides (4,4'-DDD 0,4'-DDT4,4'-DDE 4,4'-DDT) and other 8 kinds of organochlorine pesticides in rhizosphere and Rhizosphere Soil The results showed that the 8 Pesticides in rhizosphere and rhizosphere of different habitats were detected in different degrees, but the contents were not more than the standard of green Chinese medicine. For example, all the 8 sources of a-BHC were detected, and 4,4'-DDD was detected only in Guo Jia town, kkai Prefecture, Chongqing, with a content of 0.007473 mg.kg-1, y-BHC, d-BHC and 4,4'-DDE. The content analysis of 6 alkaloids in different habitats of.3. was detected. 6 main alkaloids in Rhizoma Corydalis were determined by HPLC method. The contents of 6 alkaloids in different habitats were compared. The variation range of tetrahydrocorydine was 1.18 ~ 118 mu g. ML-1, and the variation range of dehydrocorydine was 0.25 to 200 mu g. ML-1. The variation range of acid isolated alkali was 0.56 ~ 112 G. ML-1. The variation range of BA Martin hydrochloric acid was 2.175 ~ 87 mu g. ML-1, and the variation range of coptidine hydrochloride was 2.59 ~ 103.6 G. ML-1. The variation range of the original opioid was 2.69 to 26 mu g. The integral value of peak area was well linear with the standard concentration, and the average recovery rate was between 96.19% and 97.59%, R20. The results of.9993. showed that the average value of 6 alkaloids in different habitats showed significant difference. The average value of different alkaloids in the same origin was different, but the proportion of the same alkaloids was close to the total alkaloids. The highest total alkaloid content was Yung Hu in Dongyang, Zhejiang. The total alkaloids of Zhou Jiashan, Hanzhoung, Shaanxi, Shaanxi Province, are more than 2 times more than the total alkaloids of the 3 kinds of nucleosides in different habitats. 3 kinds of nucleosides, such as guanosine, uridine and adenosine in Rhizoma Corydalis, are measured by HPLC method, and the difference of the content structure ratio of 3 nucleosides in different habitats of Rhizoma Corydalis is compared. The concentration of middle uridine was between 12.12 and 242.40 G. ML-1. The concentration of guanosine was between 8.04 and 160.80 mu g. ML-1. The concentration of adenosine was between 6 and 120 g. ML-1. The linear regression relationship between the peak area integral value and the standard concentration was good. The correlation coefficient of the regression equation was R20.9998, the average recovery rate was 99.63% to 104.24% and RSD in 0.7% to 2.3. The results showed that the composition and composition of 3 kinds of nucleosides in different habitats were significantly different (P0.01). The 3 components and composition of Rhizoma Corydalis in the same origin were also significantly different (P0.05). The total amount of 3 kinds of nucleosides was higher than that of the Rhizoma Corydalis in Huaqiao Town, Wuhu County, Anhui province. The content of nucleoside of Corydalis Corydalis has a certain influence on the analysis of soil physical and chemical indexes in.5.. The pH value of.9 soil of Rhizoma Corydalis in different planting bases is between 4.13 and 5.48, which belongs to acid soil, which is not different from the background values. The total nitrogen content range is 730~2080 mg. Kg-1, the coefficient of variation is 0.39; the content range of effective nitrogen is 18.67 ~ 24. 2.63mg kg-1, its variation coefficient is greater than the variation coefficient of total nitrogen, the range of total phosphorus content of 0.74. is 0.47 ~ 1.12 mg. Kg-1, the coefficient of variation is 0.35; the content range of effective phosphorus is 0.03 ~ 0.31mg kg-1, the coefficient of variation is total nitrogen of soil of different planting base of 0.37., available nitrogen, total phosphorus and available phosphorus all show significant difference in distribution (P0.01). The content of quick acting nitrogen in Shaanxi Hanzhoung was the highest, and the content of quick acting phosphorus in dad in Chongqing district was the highest, which showed that the soil enzyme activity of strong.6. soil in two habitats was analyzed. The urease activity of Rhizoma Corydalis in different habitats was 57.04 to 137.32mg. G-1, the activity range of invertase was 1.574 to 11.82mg. G-1, and the activity range of protease was 0.819. 10.11mg / g-1, acid phosphatase activity range from 2.683 to 7.2mg. G-1, neutral phosphatase activity range from 0.04 to 3.57mg. G-1, alkaline phosphatase activity range from 0.007 to 1.39mg. G-1. in different habitats of the cultivated area of the 6 soil enzyme activity is different, the same production area of the enzyme activity is also different. There are certain differences in the transformation and supply intensity of nitrogen and phosphorus in the soil of different habitats. Among them, the activity of soil urease and protease in Shaanxi Hanzhoung is the highest, but the highest activity of soil invertase is from the rhizosphere soil of Tongzhou District in Jiangsu Province, and the highest soil acid phosphatase activity is from the rhizosphere of the Rhizoma Corydalis in Hanzhoung, Shaanxi. Soil, explain the analysis of N, C, P transformation and strong.7. ecological factors and the effective components of Rhizoma Corydalis in these places. The correlation between climate factors, soil factors, soil physicochemical indexes and soil enzyme activity with 6 alkaloids and 3 kinds of nucleosides of the Rhizoma Corydalis was analyzed by bivariate analysis. There is a significant correlation between the effective components of the cable. For example, the annual temperature and guanosine, the proportion of gravel in the soil and the corioline hydrochloride, the total phosphorus and the primary opiate correlation coefficient |r| are more than 0.8, and (P0.01) has a very significant positive correlation. It shows that the ecological factors in the environment affect the composition and structure ratio of Rhizoma Corydalis, and can provide valuable information for the better production and study of Rhizoma Corydalis in the future.

【學(xué)位授予單位】：重慶三峽學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S567.239

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