摘要 | 为建立五脉绿绒蒿(Meconopsis quintuplinervia Regel)中氨基酸的快速检测方法,阐释环境因子对氨基酸成分及含量的影响,本研究以10-乙基吖啶酮-2-磺酰氯为衍生化试剂,采用高效液相色谱-质谱联用(HPLC-MS)技术对青海省16个居群五脉绿绒蒿中氨基酸成分及含量进行了分析,并采用主成分分析和相关性分析研究了环境因子对氨基酸含量的影响。结果显示:氨基酸衍生物线性回归方程的相关系数为0.998 7~0.999 7,检出限为0.06~1.71 nmoL·L-1,定量限为0.21~5.43 nmoL·L-1,峰面积的相对标准偏差为0.11%~1.89%,表明构建的检测方法较优。在16个居群的五脉绿绒蒿中共检测出17种氨基酸,其中,Phe含量最高;各氨基酸含量的变异系数为28.23%~384.75%。基于氨基酸含量,16个五脉绿绒蒿居群可分为4组,其中,海北藏族自治州祁连县油葫芦沟(P8)居群单独为一组。主成分分析结果显示Orn、Cys、His、Ala为五脉绿绒蒿的特征氨基酸成分。土壤理化特征分析结果显示:P8居群的土壤全盐、有机质、有效氮、有效磷和全氮含量均最高,土壤全钾含量则最低;基于土壤理化性质,海北藏族自治州祁连县扁都口(P7)、P8、果洛藏族自治州班玛县X710(莫林段)(P12)居群分别单独为一组,其余居群聚为一组;主成分分析结果显示:第1主成分中土壤全氮、有机质、全盐含量的载荷较高,第2主成分中土壤全磷和有效钾含量及pH值的载荷较高。相关性分析结果显示:大多数氨基酸含量与土壤有机质、有效氮、全氮和全钾含量显著(P<0.05)或极显著(P<0.01)相关,经度、纬度和海拔对氨基酸含量影响较小。综合分析认为,青海省五脉绿绒蒿中必需氨基酸含量较高,具有较高的研究和应用价值;P8居群的土壤营养较丰富,该居群五脉绿绒蒿中氨基酸种类丰富且含量高;在五脉绿绒蒿的开发利用过程中,需要关注其生长地的土壤微环境。
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Abstract | To establish a rapid detection method for amino acids in Meconopsis quintuplinervia Regel and elucidate the effects of environmental factors on the components and contents of amino acids, 10-ethyl acridine-2-sulfonyl chloride was used as a derivatization reagent, the components and contents of amino acids in M. quintuplinervia from 16 populations in Qinghai Province were analyzed by using highperformance liquid chromatography-mass spectrometry (HPLC-MS) technology, and the effects of environmental factors on contents of amino acids were investigated by using principal component analysis and correlation analysis. The results show that the correlation coefficients of linear regression equations of the amino acid derivatives are 0.998 7-0.999 7, the detection limits are 0.06-1.71 nmol·L-1, the quantitation limits are 0.21-5.43 nmoL·L-1, and the relative standard deviations of peak area are 0.11%-1.89%, indicating that the constructed detection method is relatively good. A total of 17 amino acids are detected in M. quintuplinervia from 16 populations, in which, the content of Phe is the highest; and the coefficients of variation for the contents of amino acids are 28.23%-384.75%. Based on the contents of amino acids, 16 populations of M. quintuplinervia can be divided into 4 groups, in which, the population of Youhulugou, Qilian County, Haibei Tibetan Autonomous Prefecture (P8) is in one group individually. The principal component analysis result shows that Orn, Cys, His, and Ala are the characteristic amino acid components of M. quintuplinervia. The soil physicochemical characteristic analysis result shows that the contents of total salt, organic matter, available nitrogen, available phosphorus, and total nitrogen in soil of the population of P8 are all the highest, while its total potassium content in soil is the lowest; based on the soil physicochemical properties, the populations of Biandukou, Qilian County, Haibei Tibetan Autonomous Prefecture (P7), P8, and X710 (Molin section), Baima County, Golog Tibetan Autonomous Prefecture (P12) are separately clustered into three groups, while the other populations are clustered into one group; the principal component analysis result shows that the loads of contents of total nitrogen, organic matter, and total salt in soil are relatively high in the first principal component, while those of contents of total phosphorus and available potassium and pH value of soil are relatively high in the second principal component. The correlation analysis result shows that the contents of most amino acids are significantly (P<0.05) or extremely significantly (P<0.01) correlated with contents of organic matter, available nitrogen, total nitrogen, and total potassium in soil, while longitude, latitude, and altitude have less influence on amino acid contents. The comprehensive analysis reveals that the contents of essential amino acids in M. quintuplinervia in Qinghai Province are relatively high, and M. quintuplinervia has relatively high research and application values; the nutrients in soil of P8 population are rich, the kinds are abundant and the contents are high of the amino acids in M. quintuplinervia of this population; in the process of development and utilization of M. quintuplinervia, attention should be paid to the soil microenvironment of its habitat.
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关键词 | 五脉绿绒蒿; 氨基酸; 环境因子; 相关性分析 |
Key words | Meconopsis quintuplinervia Regel; amino acid; environmental factor; correlation analysis |
作者 | 冯丹1,2, 舍莉萍3, 宋晓铭1,4, 李佳敏1,4, 孙菁1 |
所在单位 | 1. 中国科学院西北高原生物研究所 青海省青藏高原特色生物资源研究重点实验室 中国科学院藏药研究重点实验室, 青海 西宁 810008;2. 中国科学院大学, 北京 100049; 3. 新疆维吾尔自治区药品审评查验中心, 新疆 乌鲁木齐 841100; 4. 青海民族大学, 青海 西宁 810007 |
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基金项目 | 国家自然科学基金面上项目(32270402); 青海省自然科学基金面上项目(2023-ZJ-919M) |