目的:基于气相色谱-质谱(GC-MS)技术结合多元统计分析法对不同栽培地域太子参中挥发性成分组分差异进行研究.方法:样品经气相色谱GC-MS分析,对所得数据进行峰对齐、切割、滤噪等预处理,用主成分分析(PCA)和偏最小二乘法-判别分析(PLS-DA)进行数据处理.结果:太子参江苏句容栽培品位于PC1轴负方向,而福建柘荣栽培品位于PC1轴正方向,说明两个栽培地域太子参样品中的挥发性成分组成差异明显;初步鉴定出两个栽培地域太子参问32种差异显著的化学成分,其中17种共有的差异化学成分呈现不同的变化规律.结论:本实验为揭示生态环境对太子参中挥发性成分合成积累的影响规律提供基础资料.

目的:分析不同产地太子参根际土壤微生物组成差异及不同种质太子参对微生物组成的影响.方法:利用磷脂脂肪酸(PLFA)生物标记法定性、定量分析太子参根际土壤中微生物群落多样性.结果:供试土壤中共检测出92种PLFA,其中i15∶0、i16∶0、16∶0、16∶0 10-Me及18∶1 ω9c含量较高,磷脂脂肪酸总量在江苏句容、福建柘荣两产地土壤中含量较高,4个产地间磷脂脂肪酸种类和比例存在较大差异.结论:太子参根际土壤微生物以细菌为主,还包含放线菌及真菌,不同产地和种质太子参的根际土壤微生物的含量及群落结构均存在较大差异.

目的 分析硫磺熏蒸前后太子参中挥发性成分的变化.方法 用水蒸气蒸馏法提取硫磺熏蒸前后太子参样品中挥发性成分,用GC-MS法对其挥发性成分进行分离和鉴定;数据经预处理后导入SIMCA-P 11.5软件进行主成分分析(PCA)和偏最小二乘法-判别分析(PLS-DA).结果 硫磺熏蒸前后太子参样品能较为明显的分开;初步筛选并鉴定出硫磺熏蒸前后太子参挥发性成分中20种差异显著的化学成分,其中11种共有的差异成分呈现不同的变化规律.结论 为揭示硫磺熏蒸对太子参药材中挥发性成分的影响规律提供基础资料.

OBJECTIVE To investigate the differences in the accumulation and biosynthesis of metabolites in different lactic acid of Pseudostellariae Radix.METHODS 1H-NMR based metabolomic approach combined with multivariate statistical analysis was used to investigate the chemical compositions and find the different metabolites in cultivated Pseudostellariae Radix and its wild-type.RESULTS Thirty-five metabolites were identified in the 1H-NMR spectra,and 15 of which showed remarkable differences.The multivariate statistical analysis showed that the cultivated Pseudostellariae Radix and its wild-type could be distinguished obviously.The contents of isoleucine,linolenic acid,lactic acid,alanine,lysine,glutamic acid,glutamine,acetoacetic acid,succinic acid,γ-aminobutyric acid,phenylalanine,and sucrose in cultivated Pseudostellaria heterophylla were lower than its wild-type,while the cultivated Pseudostellariae Radix contained more xylose,raffinose,and fumaric acid.CONCLUSION This study will provide basic information for exploring the quality formation mechanism and revealing the accumulation and biosynthesis of metabolites in different ecotypes of Pseudostellariae Radix.

目的 考察不同加工方法对何首乌中多元功效物质的影响及其变化规律,以优化及建立何首乌适宜的产地加工方法及其条件.方法 采用胶束电动毛细管色谱二极管阵列检测法(MEKC-DAD)测定12种不同加工方法的何首乌药材中二苯乙烯苷、游离型葸醌、结合型蒽醌及儿茶素;将多元功效物质分析结果标准化处理,进行主成分分析(PCA),对不同加工方法进行综合评价.结果 不同加工方法的何首乌中多元功效物质的量变化呈现一定规律性,二苯乙烯苷的量及结合蒽醌的总量在厚片晒干中最高;游离葸醌的总量在厚片70℃烘干中最高;儿茶素的量在薄片40℃烘干中较高;何首乌药材厚片晒干综合评价指数明显高于其他加工样品.结论 加工方法对何首乌化学成分具有一定的影响,可根据目标成分优选何首乌适宜产地加工方法.

探讨种质和生态环境因素对太子参中氨基酸类成分积累的影响.采用柱前衍生化高效液相色谱(HPLC)测定太子参中20种游离氨基酸含量,并进行多元统计分析.排序分析表明同一种质不同地域、同一地域不同种质太子参间均存在一定差异;主成分分析显示谷氨酸、苏氨酸、蛋氨酸、异亮氨酸、脯氨酸、γ-氨基丁酸为太子参特征氨基酸,综合评价表明就氨基酸而言江苏句容栽培的ZS2太子参品质较好;单因素方差分析表明栽培地域与其特征氨基酸存在显著相关.生态环境因子是影响太子参中氨基酸含量的主要因素.本研究为深入研究太子参药材的品质形成机制提供基础资料.

Objective To analyze the dynamic changes of phenolic acids and anthraquinones from the aerial parts of Xanthium sibiricum (Xanthii Herba) in different collection periods.Methods To establish an HPLC method for simultaneous determination of chlorogenic acid,neochlorogenic acid,protocatechualdehyde,protocatechuic acid,cryptochlorogenic acid,caffeic acid,1,3-dicaffeoylquinic acid,ferulic acid,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid,aloeemodin,emodin,and chrysophanol in the aerial parts ofX.sibiricum.Results The contents of phenolic acids and anthraquinones in different harvest time showed dynamic changes.In mid July,the content of total phenolic acids was higher.The higher content of total anthraquinones was in late July.The contents of five total phenolic acids were higher in mid July,such as chlorogenic acid,protocatechuic aldehyde,ferulic acid,3,5-dicaffeoylquinic acid,and 1,3-dicaffeoylquinic acid.The content of chlorogenic acid was higher in late June,and the rest of five phenolic acids were higher in mid July.Conclusion This can provide the basis for determining the suitable harvest time of X.sibiricum.

Pseudostellariae Radix (PR) is an important traditional Chinese medicine (TCM), which is consumed commonly for its positive health effects. However, the chemical differences of PR from different cultivated fields and germplasms are still unknown. In order to comprehensively compare the chemical compositions of PR from different cultivated fields, in this study, 1H-NMR-based metabolomics coupled with high performance liquid chromatography (HPLC) were used to investigate the different metabolites in PR from five germplasms (jr, zs1, zs2, sb, and xc) cultivated in traditional fields (Jurong, Jiangsu, JSJR) and cultivated fields (Zherong, Fujian, FJZR). A total of 34 metabolites were identified based on 1H-NMR data, and fourteen of them were found to be different in PR from JSJR and FJZR. The relative contents of alanine, lactate, lysine, taurine, sucrose, tyrosine, linolenic acid, γ-aminobutyrate, and hyperoside in PR from JSJR were higher than that in PR from FJZR, while PR from FJZR contained higher levels of glutamine, raffinose, xylose, unsaturated fatty acid, and formic acid. The contents of Heterophyllin A and Heterophyllin B were higher in PR from FJZR. This study will provide the basic information for exploring the influence law of ecological environment and germplasm genetic variation on metabolite biosynthesis of PR and its quality formation mechanism.

目的 研究不同产地太子参的基因差异表达,分离并鉴定相关差异基因.方法 采用反转录双链DNA扩增片段长度多态性技术分析国内4个主产地太子参差异表达基因片段.结果 筛选6个引物组合进行扩增,从不同产地太子参中获得34条差异表达转录衍生片段(trivially distributed file system,TDFs),对差异片段进行克隆、序列测定,得到21个TDFs核苷酸序列.经过BLASTX程序比对,15个TDFs有其对应的显著同源序列,其中7个TDFs为已知功能的蛋白,这些蛋白主要参与植物的生长发育、抵御病虫害、提高植物抗非生物胁迫的能力.结论 利用cDNA-AFLP技术鉴定出不同产地太子参的差异基因,为揭示太子参药材品质形成的分子机制提供基础资料.

研究不同种源和地域太子参的基因差异表达,分离并鉴定相关差异基因.采用cDNA-AFLP技术分析2个不同种源和地域太子参差异表达基因.筛选6个引物组合进行扩增,从同种源不同地域及同地域不同种源太子参中分别获得44、38条差异表达转录衍生片段(Trivially distributed file system,TDFs),对差异片段进行克隆、序列测定,分别得到27、25个TDFs核苷酸序列.经过BLASTX程序比对,分别有19、16个TDFs有其对应的显著同源序列,其中有7、6个TDFs为已知功能的蛋白,这些蛋白主要参与植物的生长发育、抵御病虫害、提高植物抗非生物胁迫的能力.本研究为揭示太子参药材品质形成的分子机制提供基础资料.

ObjectiveTo analyze the correlation between index components and climatic factors of Pseudostellariae Radix.MethodsCorrelation analysis ( CA), partial least square (PLS) and PCA ordination were applied to analyze the correlation of index components in Pseudostellariae Radix and climatic factors, and to explore the main climatic factors affecting the accumulation of index components in Pseudostellariae Radix.ResultsThe origin and harvest time had influence on index components. The monthly minimum temperature, monthly average humidity, monthly precipitation and the annual precipitation were the dominant factors influencing the content of index components.ConclusionThrough the positive and negative correlation between climatic factors and index components, it could explain the rationality of traditional harvest time and the reason of high content of heterophyllin B in Pseudostellariae Radix from Jiangsu Jurong.

To explore rapidly the potential chemical markers for differentiating Pseudostellariae Radix from different cultivated fields and germplasms, a method is proposed based on ultra-performance liquid chromatography-triple time-of-flight mass/mass spectrometry (UPLC-Triple TOF-MS/MS) coupled with multivariate statistical analysis. Peak matching, peak alignment, and noise filtering were used in analyzing mass spectrometric data. Accurate m/z value analysis of MS data based on software of database search and MS/MS fragment analysis were applied to identify constituents. The obtained data were statistically analyzed with hierarchical cluster analysis (HCA), principal component analysis (PCA), and partial least squared-discriminant analysis (PLS-DA) to compare the differences among these samples. The PLS-DA loading plot obtained from all mass data showed that 21 compounds were identified as the potential chemical markers to characterize the samples. The results provide experimental data to reveal the influence of ecological environments and germplasms on metabolite biosynthesis in Pseudostellariae Radix.

基于气相色谱-质谱(GC-MS)结合多元统计分析法对不同采收期太子参中挥发性成分组分差异进行研究.样品经GC-MS分析,对所得数据进行峰对齐、切割、滤噪等预处理,用主成分分析(PCA)和偏最小二乘法-判别分析(PLS-DA)进行数据处理.结果显示,不同采收期太子参样品能较为明显的分开,且传统采收期7月初样品与其他采收期样品化学组成差异较大;初步筛选并鉴定出不同采收期太子参间35种差异显著的化学成分,其中10种共有的差异化学成分且呈现不同的变化规律.本实验为揭示不同采收期太子参中挥发性成分动态变化规律及确定其药材合理采收期提供基础资料.

目的:基于顶空气相色谱-质谱(HSGC-MS)技术,建立了分析不同产地来源的苍耳草和苍耳子中挥发性成分的方法.方法:采用HSGC-MS技术对苍耳草和苍耳子20批样品进行测定,通过NIST05标准质谱检索库对主要化合物进行鉴定,数据处理用SIMCA-P 11.5软件进行主成分分析.结果:主成分得分图显示,苍耳草及苍耳子样品间的差异得到明显区分,通过载荷图和T检验筛选出差异显著的11种标志物丙醛(Propanal,tR/min1.64)、二甲基硫醚(Dimethyl sulfide,tR/min 1.73)、2-甲基丙醛(Propanal,2-methyl-,tR/min 1.83)、正丁醛(Butanal,tR/min 1.95)、2,3-丁二酮(2,3-Butanedione,tR/min 2.07)、2-甲基丁醛(Butanal,2-methyl-,tR/min 2.48)、丙二酸(Propanedioic acid,tR/min 2.74)、1-氯戊烷(Pentane,1-chloro-,tR/rnin 3.62)、己醛(Hexanal,tR/min 4.62)、庚醛(Heptanal,tR/min 7.97)、a-蒎烯(1R.alpha-pinene,tR/min 9.53).结论:从苍耳草与苍耳子挥发性成分差异的角度,可为揭示二者药性、药效差异的物质基础提供依据.

建立HPLC同时测定苍耳子、苍耳草药材中绿原酸、新绿原酸、原儿茶醛、原儿茶酸、隐绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、阿魏酸、3,4-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸、芦荟大黄素、大黄素及大黄酚含量的方法.采用Agilent ZORBAX SB-C18色谱柱(250 ×4.6 mm,5μm),以甲醇(A)-0.1％甲酸水溶液(B)为流动相,梯度洗脱,流速1.0 mL/min,柱温35℃,检测波长254 nm,进样量10 μL.14种成分的浓度与峰面积的线性关系良好(r＞0.9931);加样回收率为97.18％～101.09％.苍耳类药材中酚酸和蒽醌类14种成分的含量存在差异,其中绿元素、原儿茶酸含量较高,芦荟大黄素、大黄素、大黄酚含量较低.该方法简单、准确,重现性较好,可用于苍耳类药材内在质量的评价和控制.

分析何首乌中10种核苷类成分,探讨10种核苷类成分含量对总核苷含量的影响程度.采用超高效液相色谱-串联四极杆-线性离子阱质谱技术测定何首乌中10种核苷类成分的含量,应用逐步回归分析方法得到10种核苷类成分含量对总核苷含量影响程度的最优方程.结果显示何首乌核苷类成分中以尿苷、腺嘌呤、鸟苷、胞苷含量较高;逐步回归分析显示尿苷、胞苷和胸苷含量对总核苷含量的影响最为显著.尿苷、胞苷和胸苷含量可标示总核苷作为何首乌药材质量评价的一项指标,为何首乌药材质量的多指标评价体系构建提供科学依据.

This work was designed to investigate proteins differentially expressed in cultivated Pseudostellaria heterophylla and its wild type using i TRAQ proteomics approach. The extracted proteins were digested using FASP method and identified by i TRAQ coupled with LC-MS/MS technology and then analyzed by Protein Pilot 5.0 search engine. Proteins differentially expressed were searched through comparison of relatively quantified proteins. The analysis was conducted using GO(gene ontology), KEGG and STRING. A total of 3 775 proteins were detected, among them, 3 676 proteins can be quantified, of which 127 proteins were up-regulated and 205 were down-regulated in cultivated Pseudostellaria heterophylla. We found 71 significantly differentially expressed proteins for further analysis. These proteins were classified into nine categories: heat shock proteins, transferases, oxidoreductases, lyases, isomerases, ligases, hydrolases, tubulin and translocases. The results indicated that the carbohydrate and cellular amino acids metabolism of cultivated Pseudostellaria heterophylla were weaker than its wild type and its ability of responding to stress was much stronger. GWD1, PHS1, GBE1, PGM, and BAM1 are the important proteins to regulate sucrose; met E and CYS are the key proteins that regulate amino acids in cultivated and wild Pseudostellaria heterophylla. This will provide the basic information for exploring the cause of secondary metabolites differences in different ecotype of Pseudostellaria heterophylla and the protein mechanism of its quality formation.

建立苍耳草药材中无机元素的电感耦合等离子体质谱(ICP-MS)分析方法,测定苍耳草不同产地及商品药材12个样品中24种无机元素的含量,用SPSS16.0对数据进行主成分分析及相关性分析.检测结果显示苍耳草药材中24种元素之间有一定的相关性,Ca、K、Mg、P、Fe的含量较高,重金属及有害元素的含量应引起重视;主成分分析选出5个主因子,得出Ca、K、Sr、Fe、Al、V、Mg、Ba、Mn是苍耳草的特征无机元素.本实验为苍耳草药材的质量控制及安全性评价提供依据.

建立衍生化气相色谱-质谱联用(GC-MS)同时测定苍耳子不同产地及商品药材中14种脂肪酸含量的方法.采用HP-5MS(30.0 m×250 μm×0.25 μm)毛细管色谱柱,程序升温,初温:80℃,以10℃/min升至195℃,保持2 min,再以3℃/min升至230℃,保持2 min.进样口温度250℃,分流比10∶1,进样量1.0 μL,载气为氮气,流速为10 mL/min,对苍耳子不同产地及商品药材脂肪油的甲酯化样品采用SIM模式进行分析.14种脂肪酸的响应峰面积与其相应浓度的线性关系良好(r ＞0.9948),加样回收率为97.02％～100.75％.本方法准确、专属,重复性好,能有效测定苍耳子药材中含有的脂肪酸,为苍耳子药材内在质量的综合评价提供依据.