HPLC-DAD-ESI-MS^n法鉴定茯苓中的三萜成分

目的:应用HPLC-DAD-ESI-MSn方法鉴定茯苓(Poria cocos(Schw.)Wolf)中的三萜成分。方法:采用Hedera ODS-2(250 mm×4.6 mm ID,5μm)柱,以乙腈-水(0.1%甲酸)为流动相,流速1.0 mL.min-1,柱温30℃,二极管阵列(DAD)和电喷雾质谱(ESI-MSn)联合检测。结果:从茯苓中鉴定了13个,推测了8个三萜类化学成分。结论:应用HPLC-DAD-ESI-MSn方法能简单、快速地鉴定茯苓中的三萜成分。

Rapid characterization of 96 chemical constituents in Citri Reticulatae Folium(leaves of ‘Fuju') using HPLC-DAD-ESI-MS^n

In order to systematically investigate the chemical constituents of Citri Reticulatae Folium （leaves of ＇Fuju＇）, an analytical method that included high-performance liquid chromatography, diode array detection, electrospray ionization, and ion-trap time-of-flight mass spectrometry （HPLC-DAD-ESI-MSn） was used to separate and identify the individual chemical components of Citri Reticulatae Folium. As a result, 96 compounds were tentatively identified in this study： including 31 phenolic acids, 4 flavonoid aglycones, 6 flavonoid mono-O-glycosides, 10 flavonoid-O-diglycosides, 5 flavonoid mono-C-glycosides, 5 flavonoid di-C-glycosides, 6 flavonoid O,C-glycosides, 5 （3-hydroxy-3-methylglutaryl） glycosyl flavonoids, 1 flavan-3-ol, and 2 alkaloids. In addition, 21 polymethoxy flavonoids （PMFs） were identified in this paper. Among these compounds, 52 compounds, which were previously found in other Citrus plants, have been identified for the first time in Citri Reticulatae Folium. 15 compounds have not been previously found in the Citrus genus were identified. Moreover, 9 potentially new compounds have also been detected in this paper. This is the first report of the full characterization of chemical components of Citri Reticulatae Folium （leaves of＇Fuju＇） by HPLC-DAD-ESI-MSn.

中药库拉索芦荟化学成分的液质定性分析

为阐明中药库拉索芦荟(Aloe barbadensis)叶的汁液浓缩干燥物的化学成分,该研究采用HPLC-DAD-ESI-IT-TOF-MS n技术,结合对照品对比和文献检索,对其进行系统的定性分析。以水(A)-乙腈(B)为流动相进行梯度洗脱,流速为1.0 mL·min^(-1),质谱使用ESI离子源,采用负离子模式分析液质数据。结果表明:(1)首次阐明中药库拉索芦荟中蒽醌类(芦荟大黄素、大黄素甲醚、大黄素-8-O-β-D-吡喃葡萄糖苷)、蒽酮类(芦荟素A、芦荟糖苷A)、色酮类(芦荟新苷D、7-O-甲基芦荟新苷A、altechromone A、芦荟苦素、芦荟新苷G、芦荟新苷C)、α-吡喃酮类(芦荟宁A、芦荟宁B)四类成分的主要化合物的裂解途径。蒽醌类化合物的裂解途径以失去CO_(2)和CO为主,蒽酮类化合物的裂解途径以己糖苷的裂解和失去CO为主,色酮类化合物的裂解途径以己糖苷的裂解和酯基的水解为主,α-吡喃酮类的裂解途径主要包括己糖苷的裂解、CO_(2)和H_(2)O的丢失等。(2)共检测到168种化学成分,参考相关文献、数据库和对照品数据共鉴定/指认了其中的78种化学成分,包括3种蒽醌类成分、29种蒽酮类成分、35种色酮类成分、7种α-吡喃酮类成分、4种其他类成分;78种化学成分中有23种为新发现的库拉索芦荟叶的化学成分,其中aloinoside D、isoeleutherin、ethylidene-aloenin等14种成分具有抗菌、抗炎或清除自由基等药理活性。该研究结果进一步丰富了中药库拉索芦荟的化学成分信息,为芦荟的药效物质基础研究及质量控制奠定了基础。

Stability and degradation of hydroxysafflor yellow A and anhydrosafflor yellow B in the Safflower injection studied by HPLC-DAD-ESI-MS^n

Safflower is a popular Chinese medicinal plant and Safflower injection is extensively used for the clinical treatment of cerebrovascular and cardiovascular diseases. In this study, HPLC-DAD-ESI-MSn was utilized to study the stability and degradation of the two major but chemically unstable bioactive compounds hydroxysaffior yellow A and anhydrosaffior yellow B, in Safflower injection. The impact of light irradiation, temperature, and pH on the stability of these two compounds were studied. The results showed that hydroxysafflor yellow A and anhydrosafflor yellow B could degrade at high temperature （〉60 ℃） or extreme pHs （pH ≤ 3.0 or 〉7.0）, but not under light irradiation. The common degradation product was p-coumaric acid. Chemical structures of the other degradation products were characterized by LC-MS. Hypothetical degradation pathways were proposed. In addition, ADP-induced platelet aggregation tests showed that the degradation of anhydrosaffior yellow B could reduce the anticoagulation activities of Safflower injection. Our results suggest that temperature and pH are critically important for the preparation and storage of Safflower injection.

野地瓜茎正丁醇萃取物中主要抗氧化活性成分的筛选

探清野地瓜茎不同极性萃取物的抗氧化活性,筛选并鉴定抗氧化活性较高萃取物的主要化学成分。通过系统溶剂法对野地瓜茎提取物进行萃取,获得石油醚、二氯甲烷、乙酸乙酯、正丁醇相提取物及剩余上层被萃取水相提取物;采用1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除能力、2,2’-联氮基-双-(3-乙基苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除能力和总还原力(TRPA)试验综合评价其不同极性萃取物的抗氧化活性;结合在线高效液相色谱—质谱—二苯基三硝基苯肼(HPLC-DAD-ESI/MS n-DPPH)快速筛选并鉴定活性较高萃取物中的抗氧化活性成分。结果表明:正丁醇萃取物具有较强的抗氧化活性(P<0.05),随着质量浓度的增大呈明显的剂量依赖效应,正丁醇萃取物对DPPH自由基、ABTS自由基清除能力的半数抑制浓度(IC 50)分别为(23.28±0.21),(76.30±1.13)μg/mL。经液相色谱、质谱、文献报道和对照品的综合分析,从野地瓜茎正丁醇萃取物中筛选的3种抗氧化活性化合物为5-O-咖啡酰奎宁酸、3-O-咖啡酰奎宁酸和4-O-咖啡酰奎宁酸。

基于高效液相色谱-二极管阵列光谱检测-电喷雾离子化质谱联用鉴定铁皮石斛叶中8种黄酮碳苷化合物及裂解规律研究

目的采用高效液相色谱-二极管阵列光谱检测-电喷雾离子化质谱联用技术对铁皮石斛叶中黄酮碳苷类成分进行分析和鉴别。方法用XB C18色谱柱(4.6 mm×250 mm,5μm);0.4%甲酸水溶液-甲醇为流动相,梯度洗脱,流速为1.0 mL.min-1;柱温30℃;DAD扫描范围190～400 nm;检测波长335 nm。Agilent电喷雾离子阱多级质谱仪;负离子检测模式;毛细管电压4 500 V;干燥气温度350℃,干燥气流量12.0 L.min-1,雾化气压力241 kPa;扫描范围m/z 50～600。结果首次从铁皮石斛叶中快速发现和鉴定了苷元均为芹菜素的8种黄酮二碳糖苷类化合物,单糖均连接在C-6位和C-8位。并探讨了黄酮碳苷化合物电喷雾质谱(ESI-MSn)特征碎裂规律。结论该方法灵敏度高、分离度好,适用于铁皮石斛叶化学成分的快速鉴定。黄酮二碳糖苷ESI-MSn特征碎裂规律为快速发现和鉴定黄酮碳苷类化合物有一定借鉴作用。

羌活属植物在不同海拔引种的生态适宜性研究

采用人工引种栽培方式研究不同海拔梯度下羌活与宽叶羌活的生长状况、地下生物量和有效成分含量的变化规律,筛选羌活属植物的最佳种植区域。结果表明:(1)从栽培植株成活率和生物量等指标来看,羌活的生态适应范围是2 600~4 100 m;宽叶羌活生态适应海拔高度是1 700~3 600 m。(2)从药用部位有效成分含量与总量来看,羌活有效成分含量与总量较高的海拔范围是2 600~3 600 m,而宽叶羌活有效成分含量与总量较高的海拔则是1 700~3 600 m。综上所述,羌活生态和品质亚最适宜范围为2 600~3 600 m;宽叶羌活对应区域较宽,为1 700~3 600 m。该结果将为羌活属植株的野生驯化与人工栽培提供科学依据。

Characterization of chemical constituents and in vivo metabolites of Kai-Xin-San prescription by HPLC/DAD/ESI-MS^n

This work aims to elucidate the chemical constituents of Kai-Xin-San（KXS） and its metabolites in rat plasma.KXS extracts were separated on an Agilent HPLC SB-C 18 column,analyzed by ion-trap tandem mass spectrometry and high-accuracy qTOF mass spectrometry in negative ion mode.A total of 39 compounds,including 11 ginsenosides,14 Polygala saponins,5 sucrose esters,8 oligosaccharide esters and 1 xanthone were characterized from KXS.Fifteen of them were confirmed by reference standards.No constituents were detected from Poria or Acori Tatarinowii Rhizoma.After oral administration of KXS（7 g/kg）,10 ginsenosides and 18 Polygala compounds were detected in rat plasma.This study indicates that ginseng saponins,Polygala saponins and saccharide esters could be the major effective components of KXS prescription.

基于HPLC-DAD-ESI-IT-TOF-MS^n技术的小叶莲化学成分分析

应用HPLC-DAD-ESI-IT-TOF-MSn技术对小叶莲甲醇提取物中的主要化学成分进行定性分析。采用Agilent Zorbax SB-C18（4.6 mm×250 mm,5μm）色谱柱,以0.1%甲酸水溶液（A）-乙腈（B）为流动相进行梯度洗脱,流速为1.0 mL·min^-1。质谱使用ESI离子源,正、负离子分析模式采集数据,通过高分辨质谱解析,并结合相关文献和对照品数据共指认了54个化合物,包括木脂素类成分18个和黄酮类成分36个,鉴定了其中50个化合物,其中27个化合物为小叶莲中首次报道。HPLCDAD-ESI-IT-TOF-MSn技术能快速准确地鉴定小叶莲中所含的主要成分,研究结果为控制小叶莲药材质量、阐释其药效物质和作用机制提供了科学依据,同时对进一步开发和利用小叶莲资源具有指导意义。