多种气相色谱联用技术分析陕西刺五加茎挥发油的化学成分

采用气相色谱/四极杆质谱(GC/qMS)、气相色谱/正交加速飞行时间质谱(GC/oaTOFMS)和气相色谱/傅里叶变换红外光谱(GC/FTIR)联用技术,对一种陕西产刺五加Acanthopanax senticosus (Rupr. et Maxim.) Harms茎挥发油的化学成分进行了分析。基于GC/qMS谱库的检索功能,结合GC/FTIR在结构鉴别上的优势和GC/oaTOFMS对质谱碎片离子精确的质量测定功能,成功地实现了对68个色谱组分的定性分析。与使用单一的联用技术(例如GC/qMS)相比,利用多种色谱联用技术在定性分析上的互补性,可以明显提高对组成复杂的挥发油类样品分析的可靠性。     

气相色谱保留参数用于刺五加果挥发油中萜烯及萜烯氧化物的分类

根据刺五加挥发油中24个挥发性成分在5个程序升温条件下的保留时间,采用基于Levenberg-Marquardt法的软件GC_AB计算得到相应成分的气相色谱保留参数(A, B值)。通过对其中的萜烯及萜烯氧化物之间A, B关系的相关性分析,发现相同碳数的单萜烯、倍半萜烯和其氧化物分别具有良好的A-B线性相关性,但单萜烯与倍半萜烯、单萜烯与其氧化物、倍半萜烯与其氧化物的A-B关系不共线。研究表明,在中草药挥发油之类的复杂未知混合物体系的分析中,通过色谱保留参数进行萜烯及萜烯氧化物的分类是一种很有潜力的辅助定性方法。     

Simultaneous determination and pharmacokinetics of four triterpenoids by ultra high performance liquid chromatography with tandem mass spectrometry after the oral administration of Acanthopanax sessiliflorus (Rupr. et Maxim) Seem extract

A specific, simple, and sensitive ultra high performance liquid chromatography with tandem mass spectrometry method utilizing the Triple Quad system has been developed and validated for the simultaneous determination and pharmacokinetic study of four triterpenoid components of Acanthopanax sessiliflorus in rat plasma. The components are 22‐α‐hydroxychiisanogenin, chiisanogenin, (1R,11α)1,4‐epoxy‐11‐hrdroxy‐3,4‐secolupane‐20(30)‐ene‐3,28‐dioic acid, and 22‐α‐hydroxychiisanoside. Sample preparation involved a liquid–liquid extraction of the analytes with ethyl acetate. Chromatographic separation was accomplished using an Agilent SB‐C₁₈ column (1.8 μm, 2.1 mm × 50 mm) with 2.0 min isocratic elution. The compounds were detected with a triple quadrupole tandem mass spectrometer in multiple reaction monitoring mode and an ESI source in negative mode. The method was linear for all analytes over the investigated range, with all determined correlation coefficients exceeding 0.9906. The limit of quantification of each analyte was lower than 1 ng/mL. The intraday and interday precisions were less than 14.9%, and the accuracy ranged from –10.2 to 11.8%. The mean recoveries of the analytes were higher than 80.0%, and the matrix effects were between 100.4 and 107.1%. These results may contribute to determining the mechanism of action and guiding the clinical application of Acanthopanax sessiliflorus.     

High‐throughput ultra high performance liquid chromatography combined with mass spectrometry approach for the rapid analysis and characterization of multiple constituents of the fruit of Acanthopanax senticosus (Rupr. et Maxim.) Harms

Acanthopanax senticosus (Rupr. et Maxim.) Harms, a traditional Chinese medicine, has been widely used to improve the function of skeleton, heart, spleen and kidney. This fruit is rich in nutrients, but the chemical constituents of Acanthopanax senticosus fruit are still unclear. A rapid method based on ultra high performance liquid chromatography with time‐of‐flight mass spectrometry was developed for the compound analysis of Acanthopanax senticosus fruit in vitro and in vivo. In this study, the Acanthopanax senticosus fruit could significantly increase the weight of immune organs, promote the proliferation of lymphatic T cells, regulate the lymphatic B cell function, and decrease the ability of natural killer cells. A total of 104 compounds of Acanthopanax senticosus fruit including lignans, flavones, triterpenoidsaponins, phenolic acids, and other constituents were identified. Among them, seven chemical compounds were reported for the first time in the Acanthopanax senticosus fruit. Compared with the serum sample of blank and dosed samples, 24 prototype compositions were characterized. The results of our experiment could be helpful to understand the complex compounds of Acanthopanax senticosus fruit in vitro and in vivo for further pharmacological activity studies.     

核桃楸皮挥发油化学成分分析

采用水蒸汽蒸馏法提取核桃楸皮中的挥发油,用气相色谱／质谱联用法,鉴定了39种化合物,其中分离出烃类(26种,71．80％)、酮类(3种,10．83％)、醇类(6种,7．96％)、呋喃类(1种,5．79％)、酚类(1种,1.99％)、肟类(1种,0.95％),酯类(1种,0．71％)7大类化合物,有3类(7种,33．93％)为已知药用成分,为进一步评价其质量和开发新药提供了基础数据。     

气相-质谱分析北细辛根和根茎中的挥发性成分

 采用 质谱联用技术对北细辛根和根茎的挥发性成分进行分离分析 ,鉴定出了 36种化学成分 ,主要成分为十五烷、甲基丁香酚、黄樟醚、3 ,5 二甲氧基甲苯等。对照北细辛根和根茎的分析结果可以发现 ,两者的挥发性成分基本相同 ,但含量有较大差异 ,这为北细辛的临床合理用药及质量控制标准的确定提供了实验依据。     

全二维气相色谱/飞行时间质谱分析不同产地的川芎挥发油

建立了川芎挥发油的全二维气相色谱/飞行时间质谱(GC×GC/TOF MS)指纹图谱,并结合聚类分析评价了川芎药材的质量。4种萜类和苯酞类物质在DB-Petro×DB-17柱系统上实现了明显的族组分分离。从新都市的1个样品中分离出375种组分,其中相似度、反相似度皆大于800的化合物215个;根据质谱库检索和保留指数验证,或参照标准化合物及文献报道,从中定性了43个化合物。结合偏最小二乘法-判别分析,可以很好地将4个产区的挥发油样品加以区分,并找出20种差异最大的化合物,其中包括4种苯酞类物质。在此基础上,进一步应用正交偏最小二乘法,关联化学成分和抗氧化活性,结果发现,苯酞类物质(如藁本内酯、川芎内酯A和新蛇床内酯)对川芎挥发油样品地区差异的影响最大,其中彭州产川芎样品中苯酞类物质的含量最高。本文通过高分辨色谱技术研究了相邻产地的药物化学组成差异,并将化学指纹及生物学活性相关联,建立了系统的中药质量评价及活性化合物筛选研究模式。     

Variation of the Lignan Content of Schisandra chinensis (Turcz.) Baill. and Schisandra sphenanthera Rehd. et Wils.

A rapid and convenient extraction technique followed by HPLC analysis has been developed for determination of lignans in Schisandra chinensis (Turcz.) Baill. and Schisandra sphenanthera Rehd. et Wils. Under the optimized conditions the lignans schisandrin, schisantherin, deoxyschisandrin, and γ-schisandrin were extracted from ten samples collected from different regions in China and then quantified by HPLC. The lignan content of the ten samples is different. S. sphenanthera Rehd. et Wils. is richer in deoxyschisandrin but there is less schisandrin and γ-schisandrin is not found. S. chinensis (Turcz.) Baill. is usually richer in γ-schisandrin, the maximum amount being 4.263 mg g⁻¹, although one sample contained none. This method is simple, rapid, sensitive, and repeatable, and is suitable for identification and differentiation of samples from different regions of China.     

Rapid quantification of four major bioactive alkaloids in Corydalis decumbens (Thunb.) Pers. by pressurised liquid extraction combined with liquid chromatography-triple quadrupole linear ion trap mass spectrometry

A new method based on pressurised liquid extraction (PLE) followed by liquid chromatography-triple quadrupole linear ion trap mass spectrometry (LC-QTrap-MS) analysis has been developed for the identification and quantification of four major alkaloids in extracts of Corydalis decumbens (Thunb.) Pers. PLE extractions were performed using 90% ethanol; temperature was set at 100 °C and pressure at 1500 psi. HPLC analysis was performed on a Waters XBridge™ C₁₈ column (150 mm × 2.1 mm i.d., 3.5 μm) eluted by a mobile phase of acetonitrile and 0.2% acetic acid. Data acquisition was carried out in multiple reaction monitoring transitions (MRMs) mode, monitoring two MRM transitions to ensure an accurate identification of target compounds in the samples. Additional identification and confirmation of target compounds were performed using the enhanced product ion modus (EPI) of the linear ion trap. The novel LC-QTrap-MS platform offers the best sensitivity and specificity for characterization and quantitative determination of the four alkaloids in C. decumbens (Thunb.) Pers. and fulfils the quality criteria for routine laboratory application.     

Analysis of cysteine-containing proteins using precolumn derivatization with <Emphasis Type="Italic">N</Emphasis>-(2-ferroceneethyl)maleimide and liquid chromatography/electrochemistry/mass spectrometry

N-(2-Ferroceneethyl)maleimide (FEM) is introduced as an electroactive derivatizing agent for thiol functionalities in proteins. Using appropriate reaction conditions, the derivatization is completed within five minutes and no unspecific labeling of free amino functions is observed. Liquid chromatography/electrochemistry/mass spectrometry was used to detect the reaction products. The reagent is a useful tool for determining the number of free thiol groups or the total number of free and disulfide-bound thiol groups in proteins. The electrochemical cell provides additional information, because the increase in mass spectrometric response upon electrochemical oxidation of the neutral ferrocene to the charged ferrocinium groups is monitored. The method was successfully applied to the analysis of native proteins and their tryptic digests. Dedicated to Prof. Werner Engewald on the occasion of his 70th birthday.     

纳升级反相液相色谱-串联质谱法分析锦灯笼提取物中的蛋白质

通过水提、酸沉法得到锦灯笼果实提取物,其中蛋白质含量为188 mg/g(以提取物干重计),共含有18种氨基酸,其中8种人体必需氨基酸占氨基酸总量的31%。基于鸟枪法蛋白质组学的分析方法,用纳升级反相液相色谱-串联质谱(nano-RPLC-MS/MS)系统分析锦灯笼果实提取物中蛋白质的酶解产物,结合数据库检索,共鉴定得到60种蛋白质;通过生物信息学分析,得到锦灯笼提取物中的蛋白质具有催化活性、抗氧化活性、酶调节活性、养分贮液囊活性、运输活性、结合活性六大生物活性,其中鉴定到与抗氧化相关的蛋白质有3种,为锦灯笼中蛋白质的功能性质的进一步研究奠定了基础。     

Lipid analysis by thin-layer chromatography--a review of the current state

High-performance thin-layer chromatography (HPTLC) is a widely used, fast and relatively inexpensive method of separating complex mixtures. It is particularly useful for smaller, apolar compounds and offers some advantages over HPLC. This review gives an overview about the special features as well as the problems that have to be considered upon the HPTLC analysis of lipids. The term "lipids" is used here in a broad sense and comprises fatty acids and their derivatives as well as substances related biosynthetically or functionally to these compounds. After a short introduction regarding the stationary phases and the methods how lipids can be visualized on an HPTLC plate, the individual lipid classes will be discussed and the most suitable solvent systems for their separation indicated. The focus will be on lipids that are most abundant in biological systems, i.e. cholesterol and its derivates, glycerides, sphingo- and glycolipids as well as phospholipids. Finally, a nowadays very important topic, the combination between HPTLC and mass spectrometric (MS) detection methods will be discussed. It will be shown that this is a very powerful method to investigate the identities of the HPTLC spots in more detail than by the use of common staining methods. Future aspects of HPTLC in the lipid field will be also discussed.