不同生境下延胡索品质与安全性研究

发布时间：2018-05-12 18:31

本文选题：延胡索 + 生物碱　；参考：《重庆三峡学院》2017年硕士论文

【摘要】：本文选取9个不同生境(生态环境)延胡索(Corydalisyanhusuo)为研究对象,分别采用原子吸收光谱法(AAS)、气相色谱(GC)法、高效液相色谱(HPLC)法和生物化学方法测定了延胡索种子和栽培土壤中4种重金属含量以及8种有机氯农药残留量,以及6种生物碱的含量和3种核苷含量,不同土壤理化指标和土壤酶活性,同时分析探讨了生境中不同生态因子与延胡索有效成分间的关系,通过对不同生境下延胡索质量控制和安全性的研究,以期为延胡索药材的规范化栽培、保护利用提供参考依据。本研究主要结论如下:1.重金属含量分析研究。AAS法测定延胡索药材及其根际土壤的铜、铅、锌、铬的含量。结果显示4种重金属含量均达到国家《药用植物及制剂进出口绿色行业标准》的要求,如延胡索药材中铜的含量范围为2.66～5.55 mg·kg-1,根际土壤中铜的含量范围为42.35～73.69mg·kg-1。2.农药残留分析研究。GC法同时测定了延胡索药材和根际土壤中4种六六六同分异构体(a-BHC、β-BHC、γ-BHC、d-BHC)和4种滴滴涕同分异构体(4,4'-DDD 0,4'-DDT4,4'-DDE 4,4'-DDT)等8种有机氯农药成分。结果显示不同产地延胡索药材和根际土壤中这8种农药成分都有不同程度的检出,但含量均未超过绿色中药标准的限定。如,a-BHC所有产地都有检出,4,4'-DDD只在重庆开州区郭家镇被检出,含量为 0.007473 mg.kg-1,y-BHC、d-BHC 和 4,4'-DDE 没有被检测出来。3.不同生境延胡索6种生物碱含量分析。采用HPLC法同时测定延胡索药材中6种主要生物碱,比较了不同生境延胡索块茎中6种生物碱含量情况。其中延胡索乙素变动范围为1.18～118μg·mL-1,脱氢紫堇碱变动范围为0.25～200μg·mL-1,盐酸小僻碱变动范围为0.56～112μg·mL-1,盐酸巴马汀变动范围为2.175～87μg·mL-1,盐酸黄连碱变动范围为2.59～103.6μg·mL-1,原阿片碱变动范围为2.69～26μg·mL-1,峰面积积分值与标准浓度的线性关系良好,平均加样回收率在96.19%～97.59%之间,R20.9993。结果显示,不同生境延胡索药材中6种生物碱含量平均值呈现出显著差异性,同一产地的不同种类生物碱的含量平均值也有所不同但同种生物碱所占总生物碱的比例比较接近。如生物碱总含量最高的是浙江省东阳市千祥镇的延胡索,比含量最低的陕西省汉中市周家山镇的总生物碱多出2倍还要多。4.不同生境延胡索3种核苷含量分析。运用HPLC法同时测定延胡索药材中鸟苷、尿苷和腺苷等3种核苷,比较了不同生境延胡索块茎中3种核苷的含量结构比的差异情况。其中尿苷浓度在12.12～242.40μg·mL-1之间,鸟苷浓度在8.04～160.80μg·mL-1之间,腺苷浓度在6.00～120.00μg·mL-1之间,峰面积积分值与标准浓度的线性回归关系良好,回归方程相关系数为R20.9998,平均加样回收率均在99.63%～104.24%之间,RSD在0.7%～2.3%之间。结果显示,不同生境延胡索药材中3种核苷成分及组成结构比差异性显著(P0.01);同一产地延胡索药材的3种成分量及组成结构比也存在明显差异(P0.05)。其中3种核苷总量较高的是来自安徽省芜湖县花桥镇延胡索药材。表明不同产地对延胡索的核苷含量具有一定的影响。5.土壤理化指标含量分析研究。9个不同种植基地延胡索土壤pH值在4.13～5.48之间,属于酸性土壤,与各地背景值差异不大。全氮含量范围在730～2080 mg·kg-1,变异系数为0.39;有效氮的含量范围在18.67～242.63mg·kg-1,其变异系数大于全氮的变异系数,为0.74。全磷含量范围在0.47～1.12 mg·kg-1,变异系数为0.35;有效磷的含量范围在0.03～0.31mg·kg-1,变异系数为0.37。不同种植基地土壤全氮、有效氮、全磷、有效磷均呈现出分布显著差异性(P0.01),其中陕西汉中速效氮含量最高,重庆开州区大德速效磷含量最高,表明两个产地土壤的肥力较强。6.土壤酶活性分析研究。9个不同产地的延胡索土壤脲酶活性范围在57.04～137.32mg·g-1,蔗糖酶活性范围在1.574～11.82mg·g-1,蛋白酶活性范围在0.819～10.11mg·g-1,酸性磷酸酶活性范围在2.683～7.2mg·g-1,中性磷酸酶活性范围在0.04～3.57mg·g-1,碱性磷酸酶活性范围在0.007～1.39mg·g-1。不同生境延胡索种植区的6种土壤酶活性均存在一定的差异性,同一产区的几种酶活性也存在差异性,说明不同生境延胡索种植区土壤中氮、磷等转化和供应强度存在一定的差异。其中陕西汉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来自江苏省通州区的根际土壤,土壤酸性磷酸酶活性最高的是来自陕西汉中的延胡索根际土壤,说明这些地方的N、C、P转化和供应强度大。7.生态因子与延胡索有效成分的分析。利用双变量分析法分析了气候因子、土壤因子、土壤理化指标和土壤酶活性与延胡索6种生物碱和3种核苷之间的相关性,结果显示,某些生态因子与延胡索有效成分间存在一定的显著相关性。例如:年均温与鸟苷、土壤中砂砾比例与盐酸黄连碱、全磷与原阿片碱相关系数|r|均大于0.8,(P0.01)呈极显著正相关性。随着年均温、土壤中砂砾比例和全磷含量的增大,鸟苷、盐酸黄连碱和原阿片碱的含量也不断增大。表明环境中生态因子影响着延胡索药效成分和结构比,可为今后更好的生产和研究延胡索药材,提供宝贵的资料。
[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.

【学位授予单位】：重庆三峡学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S567.239

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