电喷雾质谱半定量方法研究制川乌配伍后的大鼠肠内菌生物转化

利用建立的代谢前后生物碱成分的电喷雾质谱半定量分析方法,通过分析各药对共煎液经大鼠肠内菌群代谢后主要生物碱的含量变化,对制川乌与浙贝母、法半夏、白蔹分别配伍后共煎液中双酯型、单酯型及脂型生物碱的肠内菌生物转化进行了深入研究,研究结果表明,配伍后中药浙贝母增加共煎液中双酯型生物碱含量,法半夏降低共煎液中双酯型生物碱含量,而白蔹对共煎液中双酯型生物碱含量影响不大;在代谢过程中,大鼠肠内菌群能够将复方中双酯型生物碱转化为脂型生物碱,从而达到中药配伍的减毒增效目的。本文通过化学方法和肠内菌代谢研究证明了制川乌配伍及代谢的机理。     

川乌与半夏配伍禁忌的化学研究

应用高效液相色谱和电喷雾质谱技术,通过分析生、制川乌单煎及其与生、法半夏共煎液中乌头碱类生物碱的含量和转变途径,发现生半夏在与川乌共煎时对乌头类剧毒生物碱的转化表现出明显的抑制作用,而法半夏却能很好的促进乌头碱类剧毒生物碱的转化,说明川乌与半夏是否可以共用与半夏的炮制与否和炮制方法存在着密切联系.     

"甘草附子汤"和"术附汤"肠内生物转化的电喷雾质谱研究

以电喷雾质谱法作为研究方法,以内标化合物为切入点,对复方中双酯型、单酯型及脂型生物碱的生物转化进行了深入研究,建立了电喷雾质谱对代谢前后生物碱成分的半定量分析方法.分析了中药复方甘草附子汤和术附汤经大鼠肠内菌群代谢主要生物碱的含量变化,研究结果表明,配伍中药甘草和白术可以有效地降低共煎液中双酯型生物碱含量,在代谢过程中...     

生附片化学成分的HPLC/ESI-MSn研究

采用高效液相色谱与电喷雾质谱联用技术, 对生附片的化学成分进行了系统的研究. 并辅以提取离子色谱方法, 发现微量的化学成分. 通过保留时间, 质荷比及多级串联质谱数据, 共鉴定了48个成分, 其中双酯型生物碱8个, 单酯型生物碱7个, 脂型生物碱29个. 其中双酯型生物碱是生附片中的主要成分, 而单酯型和脂型生物碱的含量和种类较少.     

生附片化学成分的HPLC/ESI-MSn研究

采用高效液相色谱与电喷雾质谱联用技术, 对生附片的化学成分进行了系统的研究. 并辅以提取离子色谱方法, 发现微量的化学成分. 通过保留时间, 质荷比及多级串联质谱数据, 共鉴定了48个成分, 其中双酯型生物碱8个, 单酯型生物碱7个, 脂型生物碱29个. 其中双酯型生物碱是生附片中的主要成分, 而单酯型和脂型生物碱的含量和种类较少.     

超高效液相色谱-质谱结合主成分分析方法研究制川乌配伍前后的肠内菌代谢差异

利用超高效液相色谱-质谱联用( UPLC-MS)技术结合主成分分析方法研究制川乌单煎液、制川乌与白芍、制川乌与防己共煎液在大鼠肠内菌中的代谢差异。采用SIMCA-P软件,以肠内菌代谢后乌头类生物碱的相对含量为变量进行主成分( PCA)分析。在主成分得分图中,制川乌单煎液与制川乌-白芍、制川乌-防己共煎液均可以明显区分,说明制川乌单煎液与制川乌-白芍、制川乌-防己共煎液的肠内菌生物转化存在显著差异。通过主成分分析载荷图及独立样本t检验,从制川乌-白芍组得到7种差异显著的标志物,从制川乌-防己组得到6种标志物,其中制川乌-白芍组有4种标志物经肠内菌代谢后含量高于制川乌组,而制川乌-防己组有1种化合物含量高于制川乌组,两组中其它标志物含量低于制川乌组。这些标志物可能是制川乌配伍前后药效差异的物质基础。     

附子配伍过程中二萜类生物碱在Caco-2小肠吸收细胞模型中吸收转运的UPLC/MS研究

利用Caco-2细胞单层模型和超高效液相色谱-三重四极杆电喷雾质谱联用方法, 考察附子中乌头类生物碱提取物的跨膜转运. 在相同浓度的附子乌头类生物碱溶液中, 加入干姜、甘草、半夏及浙贝母的水提物(折合药材质量比例为1∶1), 系统考察了加入前后的表观渗透系数(P_app). 结果表明, 加入配伍药材后附子中11种二萜类生物碱的P_app发生了明显的变化. 干姜、甘草和半夏水提物的加入能够明显地降低二萜类生物碱在小肠中的生物利用度, 尤其抑制其中毒性较大的双酯型生物碱的生物利用度|而浙贝母的水提物却在促进吸收和抑制外排两个方面共同作用, 使二萜类生物碱的生物利用度明显增加.     

草乌中二萜类生物碱的电喷雾串联质谱研究

利用电喷雾质谱技术对传统中药草乌中的二萜类生物碱进行直接分析鉴定.通过实验数据并对比文献发现,草乌中含有单酯型、双酯型、三酯型和脂类等共4种类型生物碱,其中三酯型和脂类生物碱在草乌中为首次发现.由于这些生物碱的结构相似,在电喷雾串联质谱中碎裂方式相同,因此根据电喷雾串联质谱结果确定了这些生物碱的结构.     

基于UPLC/Q-TOF-MS分析附子半夏配伍相反的物质基础

利用超高效液相色谱-飞行时间质谱联用技术(UPLC/Q-TOF-MS)分析附子半夏药对配伍相反的物质基础,从化学成分层次阐释其配伍相反机制.基于UPLC/Q-TOF-MS建立附子半夏药对配伍后生物碱类成分的化学指纹图谱,通过主成分分析法和正交偏最小二乘判别法分析药对配伍在合煎过程中的生物碱类成分的含量变化,找出差异变化显著的化学成分.结果表明正离子模式时附子半夏药对合煎液中次乌头碱,中乌头碱,乌头碱,去氧乌头碱,10-OH-中乌头碱,10-OH-乌头碱等的含量明显增高,而中乌头原碱,去乙酸中乌头原碱,去乙酸次乌头原碱,苯甲酰乌头原碱,苯甲酰次乌头原碱,10-OH-苯甲酰中乌头原碱等含量降低.附子半夏药对配伍应用时双酯型二萜生物碱的含量明显增高,而单酯型二萜生物碱的含量明显降低,这可能是附子半夏药对配伍相反作用的物质基础.     

人参与藜芦配伍化学成分变化的HPLC-ESI-MS与ESI-MS研究

采用电喷雾质谱(ESI-MS)和高效液相色谱与电喷雾质谱联用技术(HPLC-ESI-MS), 对人参与藜芦配伍过程中人参皂苷和藜芦生物碱的变化规律进行了系统研究. 在人参与藜芦配伍的共煎液中鉴定出八种人参皂苷, 其中有六种人参皂苷含量有所降低, Rf和Rb₂的含量基本不变; 此外还鉴定出八种藜芦生物碱, 其中有六种生物碱的含量随人参加入而明显增高. 人参与藜芦配伍, 煎煮液中人参皂苷的含量下降, 藜芦总碱的含量上升. 人参的加入有利于藜芦生物碱的溶出. 因藜芦总碱的毒性较强, 所以人参“反”藜芦具有一定道理.     

电喷雾串联质谱分析附子炮制中的化学成分变化

利用电喷雾质谱方法(ESI-MS)分析了附子加辅料(甘草)炮制前后水煎液中二萜类生物碱在种类和含量方面的变化,通过加入内标化合物,建立了电喷雾质谱的半定量分析方法。此方法具有快速、准确、灵敏的特点,能够更加全面地反映中药配伍炮制过程中多种化学成分的含量变化,并能根据电喷雾串联质谱的分析结果鉴定配伍后产生的新的化学成分,在共煎液中的次乌头碱、中乌头碱和乌头碱的相对含量分别是单煎液中的5.67%、4.05%和4.88%。通过研究附子与甘草的单煎液、共煎液以及药渣中化学成分的变化,揭示了甘草作为辅料,在炮制过程中对附子减毒作用机理。     

Quantitative evaluation main of the components in Paeoniae Radix Alba–Atractylodis Macrocephalae Rhizoma herbal pair by high‐performance liquid chromatography

The aim of this study was to investigate the influence of compatibility on the contents of main compounds in Paeoniae Radix Alba and Atractylodis Macrocephalae Rhizoma. Ten compounds were separated on an Inertsil ODS‐SP Extend C₁₈ column (250 mm × 4.6 mm, 5 μm) and detected by a diode array detector with the mobile phase consisting of aqueous phosphoric acid (0.1%, v/v; A) and acetonitrile (B) by linear gradient elution. All analytes showed good linearity over a wide concentration range (r² ≥ 0.9989). The limits of detection and quantification were <8.10 and 10.80 μg/mL, respectively. The intra‐ and interday variations were <4.36%. The average recoveries were observed from 94.90 to 103.38%, with relative standard deviation ranging from 1.23 to 3.15% for the analytes. The established method was reliable enough for global quality evaluation of Paeoniae Radix Alba, Atractylodis Macrocephalae Rhizoma, and their co‐decoctions.     

色谱指纹谱用于中药大黄抗肿瘤活性成分的筛选

采用液相色谱-质谱联用方法分析了中药大黄经过SD大鼠肝匀浆体外代谢前后的指纹谱中色谱峰面积、保留值的差异。指出5种游离型蒽醌化合物在SD大鼠肝匀浆体外代谢体系中只有大黄酚发生代谢反应转化为芦荟大黄素。考察了体外代谢条件下,肝匀浆浓度与代谢时间对大黄酚转化及其代谢产物的影响。SD大鼠体外抗肿瘤试验表明,大黄代谢物对于人宫颈癌(HeLa)细胞的抑制活性略高于其提取物。通过比较芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的活性,并结合大黄酚的体外代谢反应的考察,解释了大黄代谢物对肿瘤细胞活性的抑制率的提高是由大黄酚的代谢产物芦荟大黄素浓度的增加引起的。     

液质联用技术研究人参与干姜或赤芍配伍前后人参皂苷及其抗氧化活性的变化

采用高效液相色谱与电喷雾质谱联用技术(HPLC-ESI-MS),对不同质量比的人参与干姜或赤芍配伍过程中人参皂苷的变化进行了研究,发现随加入的干姜量增加,共煎液中的人参皂苷含量依次降低;少量的赤芍可以使各皂苷的溶出量增加;同时测定了人参单煎液、人参与干姜、赤芍共煎液中正丁醇提取物和水提物的抗氧化活性。 以抗坏血酸(500 μmol/L)作对照,人参与干姜、赤芍配伍溶液的抗氧化活性比人参单煎液要好,同时人参与干姜、赤芍共煎液中正丁醇提取物的FRAP(铁离子还原/抗氧化能力测定)值分别为1562.29和2969.78 μmol/L,高于人参与2种药单煎液（1260.27和2502.07 μmol/L）之和。     

Structural elucidation and identification of alkaloids in Rhizoma Coptidis by electrospray ionization tandem mass spectrometry

Simple, convenient, sensitive and accurate analytical methods are needed for the structural characterization and identification of alkaloid components in Rhizoma Coptidis in traditional Chinese herbal medicine, which has important bioactivity. In this work, the identification of alkaloid compounds in Rhizoma Coptidis was investigated by obtaining molecular mass information using electrospray ionization mass spectrometry (ESI-MS). Multi-stage tandem mass spectrometric (ESI-MS(n)) data for the alkaloid compounds were used for detailed structural characterization, then structure information was obtained by comparison of the fragmentation mechanisms of both alkaloids in Rhizoma Coptidis and standard samples of berberine, palmatine, coptisine and jatrorrhizine by MS. Based on the results obtained, the structure of a novel compound was elucidated. The results of the experiments demonstrate that ESI-MS(n) is a sensitive, selective and effective tool for the rapid determination of alkaloids in Rhizoma Coptidis.     

Screening epidermal growth factor receptor antagonists from Radix et Rhizoma Asari by two‐dimensional liquid chromatography

Radix et Rhizoma Asari is a traditional Chinese medicine, and has many pharmacological effects, such as calming, analgesia, anti‐inflammation, antiarrhythmic, antihypertensive, antivirus, etc. But few studies have screened the active compounds from extracts of Radix et Rhizoma Asari for tumor therapy. In this study, a two‐dimensional liquid chromatography system was built to screen active compounds acting on epidermal growth factor receptor (EGFR) from Radix et Rhizoma Asari. The screening result showed that asarinin from Radix et Rhizoma Asari was the targeted component that could act on EGFR specificity. The competitive binding assay and molecular docking assay results showed asarinin binding with EGFR in similar manner as with gefitinib, which was used as a positive control drug. Then the antitumor effect of asarinin was studied through cell growth assay in vitro. The results showed that gefitinib and asarinin could inhibit highly expressed EGFR cell growth in a dose‐dependent manner in the range of dose from 0.10 to 102.4 μM. This two‐dimensional liquid chromatography system will be a useful method in drug discovery from natural medicinal herbs for searching potential antitumor candidates.     

Determination and pharmacokinetics of 6,7-dimethoxycoumarin in rat plasma after intragastric administration of different decoctions of yinchenhao tang

A simple and sensitive reversed-phase high-performance liquid chromatography method with UV detection is developed and validated for the determination of 6,7-dimethoxycoumarin in rat plasma and comparative analysis of its pharmacokinetics after intragastric administration of 6,7-dimethoxycoumarin and three different decoctions of yinchenhao tang. The plasma samples are deproteinated with acetonitrile. The components are separated on a Kromasil C18 column (250 x 4.6 mm, 5 microm,) with methanol-1% acetic acid solution-tetrahydrofuran (30:63:7, v/v/v) as the mobile phase, and the UV detector is set at 340 nm. Coumarin is used as an internal standard. The linear calibration curve is obtained in the concentration range of 25-2500 ng/mL. The lower limit of quantitation of the method is 25 ng/mL. The intra- and inter-day precision are less than 12%, and the accuracy determined with relative error ranges from -2.9% to 1.7%. The data obtained from rat plasma are analyzed with Topfit 2.0 Pharmacokinetic Software. With pharmacokinetic analysis, the main parameters after intragastric administration of 6,7-dimethoxycoumarin, Herba Artemisiae Scopariae decoction, Artemisiae Scopariae decoction plus Radix et Rhizoma Rhei and Fructus Gardeniae decoction, yinchenhao tang are as follows: T(1/2) is 0.29, 1.30, 1.07, and 1.75 h, AUC(-->t) is 919.1, 1215.0, 2035.3, and 2537.9 ng-h/mL, AU(0-->) is 928.5, 1325.9, 2094.4, and 2612.6 ng x h/ mL, respectively.     

苦黄注射液中大黄蒽醌含量测定方法改进

按苦黄注射液原质量标准（草案）中测定方法,大黄蒽醌用氯仿提取,阴性试验结果为苦黄注射液大黄蒽酯标示量的１６．５０％（ｎ＝５）,结果偏高;改用石油醚提取,降为６．１５％,重复性试验的ＲＳＤ为１．４４％（ｎ＝５）,加标回收率为１０１．８％（ｎ＝５）,显色在１－４ｈ内稳定,表明该法是可行的。     

Chemical composition differentiation of Shen‐Shuai‐Ning granule between combined decoction and separated decoction using HPLC‐DAD‐ESI‐QTOF‐MS

Shen‐Shuai‐Ning (SSN) granule, a traditional Chinese medicine formula, is widely used in clinical practice for treating chronic renal failure. However, its detailed chemical profile is unknown. Here, HPLC‐ESI‐QTOF‐MS was employed for the systematic chemical analysis of SSN. A total of 52 compounds were identified and the characteristic ions of the compounds were described. Furthermore, chemical consistency between the combined decoction and the separated decoction of SSN was evaluated using HPLC‐DAD. A chemical comparison between two preparations of SSN granule (combined decoction and separated decoction of Coptides Rhizoma) indicated a significant difference in the content of many compounds, including salvianolic acid A, salvianolic acid B, berberine, palmatine and epiberberine. As a result, separated decoction of Coptides Rhizoma would lead to a significantly decrease in depsides in Salviae Miltiorrhizae Radix et Rhizoma and an increase in alkaloids in Coptidis Rhizoma.