中药紫花地丁黄酮碳苷类化学成分的超高效液相色谱-电喷雾离子源-四极杆飞行时间串联质谱研究

采用超高效液相色谱-电喷雾离子源-四极杆飞行时间串联质谱(UPLC-ESI-QTOF-MS/MS)方法,以中药紫花地丁中分离得到的5个黄酮碳苷化合物为对照品,研究了黄酮二糖碳苷类化合物在ESI-QTOF-MS/MS质谱中的裂解规律。研究进一步证实了文献报道的黄酮二糖碳苷C-6位取代糖基较C-8位糖更易优先发生裂解;但同时发现结构中如果含有不稳定构象的糖基取代时,该规律不再适用。通过对照品的质谱裂解规律,并结合文献和高分辨质谱数据,成功表征和鉴定了紫花地丁中21个黄酮二糖碳苷类以及4个黄酮氧苷类成分。该方法能快速、灵敏、准确地对中药中微量黄酮碳苷类成分的结构进行表征和鉴定。     

中药复方小续命汤抗阿尔茨海默病有效成分组中6种有效成分的同时测定

建立了用高效液相色谱分析中药复方小续命汤抗阿尔茨海默病有效成分组中各成分的方法,并将其应用于此有效成分组的质量控制。采用ODS色谱柱,以水-甲酸-乙二胺(流动相A，体积比为100∶0.1∶0.1)和甲醇-甲酸(流动相B，体积比为100∶0.05)为流动相,采用梯度洗脱，流速1 mL/min，检测波长240 nm。在上述色谱条件下,可分离测定有效成分组中的6种成分。6种成分均具有良好的线性；其回收率分别为:芍药苷99.1%,升麻苷99.6%,黄芩苷98.4%,5-O-甲基维斯阿米醇苷99.9%,防己诺林碱99.6%,粉防己碱102.0%;相对标准偏差（RSD）分别为1.3%,1.4%,0.4%,0.8%,0.2%,1.4%。所建立的方法简便、快捷,可用于小续命汤有效成分组的质量控制。     

UPLC-Q-TOF-MS/MS法分析鉴定鸡血藤中药复方提取物的化学成分

应用超高效液相色谱-四极杆-飞行时间串联质谱(UPLC-Q-TOF-MS/MS)技术定性鉴别鸡血藤中药复方提取物的化学成分。采用Waters ACQUITY UPLC BEH C₁₈色谱柱(100 mm×2.1 mm,1.7μm),以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,电喷雾离子源正、负离子模式扫描。通过质谱数据库、化合物裂解规律,并结合相关文献进行鉴别。共鉴别出82种化合物,以黄酮类、苯丙素类、萜类化合物为主,包括黄酮类22种、苯丙素类24种、萜类25种、酚酸类4种、甾体类6种、氨基酸类1种,各药材组成复方后首次发现的新化学成分有10种。UPLC-Q-TOF-MS/MS技术为鉴别鸡血藤中药复方中化学成分提供了简便、快速、准确的方法,鉴定得到的各个化学成分涵盖了组方中各味药材的主要成分,可为该复方的质量标准及药效物质研究提供实验依据和理论基础。     

超高效液相色谱-电喷雾串联质谱法分离鉴定烟草中5种糖苷类香味前体物质

采用超高效液相色谱-电喷雾串联质谱法(UPLC-ESI MS/MS)并结合气相色谱-质谱法分离鉴定了烟草中5种主要的糖苷类香味前体物质。烟样经甲醇提取、XAD-2柱净化,得到初步纯化的糖苷,在pH 5条件下将其酶解,释放出糖苷配基。采用气相色谱-质谱分析并通过标准谱库检索确定了5种挥发性苷元;然后通过电喷雾质谱(负离子模式)确定糖苷母离子并作碎片离子扫描(MS2),确定了5种糖苷类香味前体物质的存在形式;最后采用UPLC-ESI MS/MS,以甲醇和乙酸-乙酸铵水溶液为流动相,通过RP-C18柱分离,在多反应监测(MRM)模式下,鉴定了烟草中5种主要的糖苷类香味前体物质,为应用液相色谱-质谱分析缺乏标准样品的糖苷类香味前体物质奠定了基础。     

"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...     

基于UHPLC-QTOFMS的黄芩汤化学成分和大鼠入血成分分析北大核心CSCD

建立了一种超高效液相色谱-四极杆飞行时间质谱(UHPLC-QTOF MS)联用鉴定黄芩汤化学成分及大鼠灌胃给药后入血成分的方法。采用ACQUITY UPLC XBridge BEH C_(18)色谱柱(2.1 mm×100 mm,2.5µm),以0.1%甲酸水和0.1%甲酸乙腈为流动相梯度洗脱;使用电喷雾离子源正负离子模式采集碎片信息。依据质谱裂解规律,结合对照品、文库比对分析,初步鉴定了黄芩汤中68种化学成分和35种入血成分,并讨论了黄芩素、黄芩苷、芍药苷、甘草苷等的治疗潜力。该方法高效、稳定、灵敏,可为黄芩汤的药效物质基础研究提供参考,也可用于其他复方的成分分析,有助于提升传统中药中小分子药物发现的效率。     

高效液相-四极杆飞行时间串联质谱分析黄蜀葵花中黄酮醇类化合物

以中药材黄蜀葵花为分析对象,采用高效液相-电喷雾/四极杆-飞行时间串联质谱,识别中药中黄酮醇类化合物.通过解析紫外光谱和二级质谱,识别了17个黄酮醇糖苷和2个黄酮醇苷元,用对照品对照及分析酸水解液的方法进行了验证;并探讨了黄酮醇类化合物的电喷雾/串联质谱(ESI/MS/MS)的裂解方式;在此分析基础上,研究了HPLC指纹图谱和多指标含量测定方法,方法学考察结果表明符合测定要求;采用金丝桃苷、杨梅素和槲皮素作参照,以折算分子量的方法计算相对含量,扩展中药质量控制多指标化的范畴.     

高效液相色谱/电喷雾-离子阱-飞行时间质谱分析鉴定中药虎杖中的主要化学成分

建立了快速、准确鉴别中药虎杖中化学成分的液相色谱-质谱法。采用高效液相色谱/电喷雾-离子阱-飞行时间质谱(HPLC/ESI-IT-TOF MS)对蒽醌类以及羟基二苯乙烯类对照品,包括大黄素、大黄酚、大黄素甲醚、大黄酸、芦荟大黄素和虎杖苷进行了分析,总结其多级裂解规律。建立了虎杖甲醇提取物的液相色谱分离条件及质谱检测条件,根据负离子模式下获得的各组分多级质谱数据,对比对照品碎裂特征并参考文献,对主要色谱峰进行指认,共鉴别了10个化合物,包括白藜芦醇-4′-O-葡萄糖苷、虎杖苷、大黄素-8-O-葡萄糖苷、白藜芦醇、决明松-8-O-葡萄糖苷、大黄素-1-O-葡萄糖苷、决明松-8-O-(6′-乙酰基)葡萄糖苷、大黄素甲醚-8-O-葡萄糖苷、大黄素甲醚-8-O-(6′-乙酰基)葡萄糖苷和大黄素,其中决明松-8-O-(6′-乙酰基)葡萄糖苷和大黄素甲醚-8-O-(6′-乙酰基)葡萄糖苷为虎杖中新发现的成分。研究结果表明,在中药化学成分研究工作中,采用电喷雾-离子阱-飞行时间质谱可提高中药化学成分的分析效率并有利于新化合物的发现和鉴别。     

基于UPLC-Q-TOF MS鉴定复方香薷水化学成分

采用超高效液相色谱-四极杆串联飞行时间质谱（UPLC-Q-TOF MS）技术对中药复方香薷水的化学成分进行快速分析鉴定。通过色谱峰响应程度和分离效果优化复方香薷水的前处理方法和仪器条件;采用Agilent Phe Hex(150 mm×2.1 mm,1.9μm)色谱柱进行分离,以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,柱温35℃,流速0.3 mL/min;通过正、负两种离子模式采集复方香薷水的质谱数据,得到精确质量数和二级碎片,结合对照品、Pubchem、安捷伦中药化合物质谱数据库和文献报道等信息对其化合物进行分析鉴定。鉴定出复方香薷水的112个成分,其中黄酮类22个,苯丙素类20个、有机酸18个、萜类14个、生物碱11个、氨基酸5个以及其他类型化合物22个。该研究全面阐述了复方香薷水的化学物质基础,为其质量控制和药效物质研究提供了理论依据。     

超高效液相色谱-四极杆飞行时间质谱法快速鉴定甘舒诺丹胶囊化学成分

采用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF/MS）法对甘舒诺丹胶囊的化学成分进行快速鉴定。根据质谱数据结合自建数据库,利用相对分子质量、碎片离子信息以及裂解规律等方式鉴定甘舒诺丹胶囊中的化学成分,应用UNIFI 2.0软件的二元比较功能实现了各化合物的归属。从甘舒诺丹胶囊中鉴定出76种化学成分,包括21种醌类成分,18种黄酮类成分,11种有机酸类成分,7种萜类成分,3种香豆素类成分,1种木脂素类成分,15种其它成分,全面反映了甘舒诺丹胶囊中具有药理活性的化学物质和各个药材的特征性成分,为中药复方胶囊化学成分的快速鉴定、药效物质基础的发现以及临床的合理应用提供了数据参考。     

Structural characterization and identification of major constituents in jitai tablets by high-performance liquid chromatography/diode-array detection coupled with electrospray ionization tandem mass spectrometry

In the present study a universally applicable HPLC-DAD/ESI-MS/MS method was developed for carrying out the comprehensive characterization of Jitai tablets (JTT). Based on the ESI-MSⁿ fragmentation patterns of the reference standards, a total of 101 components were identified or tentatively characterized by comparing their retention times, UV and MS spectra with those of reference standards or through the matching of empirical information with those of published components in the in-house library. The characteristic fragmentation pattern of alkaloids, phenolic acids, tanshinones, flavonoid glycosides, cyanogenic glycosides, ginsenosides, 2-(2-phenylethyl) chromones, phthalides and gingerol-related compounds were tentatively elucidated using structurally-relevant product ions. It was observed that neutral losses of C₉H₁₀O₃ and C₉H₈O₂ were the characteristic product ions of scopola alkaloids. Neutral fragment mandelonitrile was the characteristic ion of cyanogenic glycosides. To our knowledge, tropylium ion and C₄H₂O unit were the characteristic ions of 2-(2-phenylethyl) chromone, which resulted from the Retro-Diels-Alder (RDA) cleavage of the C ring. The results indicated that the developed analysis method could be employed as a rapid, effective technique for structural characterization of chemical constituents in TCM. This work is expected to provide comprehensive information for the quality evaluation and pharmacokinetic studies of JTT.     

On-line identification of sugarcane (Saccharum officinarum L.) methoxyflavones by liquid chromatography-UV detection using post-column derivatization and liquid chromatography-mass spectrometry

Sugarcane (Saccharum officinarum L., Gramineae) bagasse and leaves were investigated for their flavonoid content and transgenic sugarcane ("Bowman-Birk" and "Kunitz") was compared with non-modified ("control") plants. Analyses were carried out by high-performance liquid chromatography coupled to diode array UV detection (LC/UV), also using post-column addition of shift reagents, and tandem MS (atmospheric pressure chemical ionization-MS/MS and collision-induced dissociation-MS). On-line UV and MS data demonstrated the presence of methoxyflavone glycosides and aglycones in a total of seven compounds. Three naturally occurring flavones glycosides and two unusual erythro- and threo-diastereoisomeric flavolignan 7-O-glucosides were identified together with their aglycones.     

七叶一枝花中薯蓣皂苷的分离及结构鉴定研究

本文利用高效液相色谱-串联质谱联用方法研究了七叶一枝花中的薯蓣皂苷。实验采用高效液相色谱分离了七叶一枝花中的3种薯蓣皂苷;通过与电喷雾质谱联用获得了这几种化合物的分子量信息;再用MS/MS获得了这几种化合物进一步的结构信息。采用此方法可快速分析鉴定从七叶一枝花中分离得到的薯蓣皂苷。     

Identification and determination of glycosides in tobacco leaves by liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry

HPLC-atmospheric pressure chemical ionization MS (HPLC-APCI-MS) was used to screen and identify glycosides in tobacco leaf. MS/MS and MS3 and photodiode array (PDA) detection were also used in the characterization. A total of 12 glycosides were found and four of them were identified based on their abundant [M + H]+ ions, UV spectra, and MS/MS analysis and they are scopolin, rutin, quercetin-3-glycoside, and kaempferol-3-rutinoside. Analytical characteristics of the method were investigated. The contents of these glycosides were obtained and compared based on the relative peak area to the internal standard in seven kinds of tobacco leaf.     

Neutral loss scan in complement with high-resolution MS/MS: Combination of detection methods for flavonoid and limonoid glycosides analysis

In this study, neutral loss scan and high-resolution MS/MS were used in combination to detect and tentatively identify various flavonoid and limonoid glycosides in navel orange albedo, juice, peel and pulp. These compound classes are of research interest due to their flavour and bioactive properties, and although flavonoid glycosides have been previously studied in other food matrices, to the best of our knowledge, neutral loss scans have not been used for the elucidation of limonoid glycosides. Neutral loss masses of 120, 162 and 308 Da were selected for the detection of hexose, rutinose and neohesperidose-substituted flavonoids, whereas 197 Da was explored for limonoid glycosides due to their tendency to form ammonium adducts. Fragmentation patterns obtained from targeted MS/MS were then used to differentiate rutinose and neohesperidose substituents as well as flavonoid subclasses of flavones, flavanones and flavonols. Additionally, high-resolution MS/MS was also used for the identification of aglycones by accurate mass (to four decimal places), allowing for the differentiation of aglycones with similar unit masses but different chemical formulas. In total, 19 flavonoid glycosides and six limonoid glycosides were detected. This workflow allows for a rapid screening of flavonoid and limonoid glycosides in citrus, which can be further extended to other food products such as tea.     

Simultaneous Determination of Phenolics and Polymethoxylated Flavones in Citrus Fruits by Ultra-High Performance Liquid Chromatography Coupled with Triple-Quadrupole Mass Spectrometry (UHPLC-QqQ-MS)

Phenolic and polymethoxylated flavones are important bioactive components in citrus fruit. Here, a rapid and sensitive method based on ultra-high performance liquid chromatography coupled with triple-quadrupole mass spectrometry (UHPLC-QqQ-MS) was developed for the simultaneous determination of phenolic and polymethoxylated flavones in the peels and pulp of mandarins, tangelos, and oranges. Three phenolic acids and eight flavonoids, including polymethoxylated flavones, were separated and determined using positive and negative ion modes in a single chromatographic run of only 11?min using the multiple reaction monitoring detection mode. The method was validated with high recoveries from 96.1% to 103.5%, good precision with interday relative standard deviations less than or equal to 7.3%, intraday relative standard deviations ≤2.64%, low limits of detection from 1.0 to 18?µg L^–1, and low limits of quantitation in the range from 3.0 to 61?µg L^–1. The application of this UHPLC-QqQ-MS/MS method to the citrus extracts of three cultivars showed that mandarin fruits contained the highest total amounts of the 11 analytes, followed by tangelos and oranges. This study provides a reliable and quantitative method that can be used for the development of functional products and quality evaluation of citrus fruits.     

应用高效液相色谱和电喷雾质谱联用技术分析鉴定车前草中的苯乙醇苷化合物

本文采用高效液相色谱与电喷雾质谱联用技术在线分析鉴定了车前草提取物中的三种苯乙醇苷化合物。实验采用反相C18色谱柱,0.2%的醋酸水溶液和乙腈梯度洗脱,车前草中的苯乙醇苷化合物得到很好的分离。在电喷雾质谱负离子条件下,获得了三种苯乙醇苷化合物的分子离子峰,分子量信息,进一步通过质谱的源内CID技术得到相应化合物的结构信息。通过得到的这些信息与文献中的已知化合物或标准品对照从而推断出化合物的结构。     

Development of a Liquid Chromatography Tandem Mass Spectrometry Method for Identification of Flavonoids in Ginkgo biloba

A rapid, selective and sensitive method for analysis of trace flavonoids and its glycoside derivatives in ginkgo has proposed. Ultrasonic‐assisted extraction of sample preparation was adopted to extract trace flavonoids in ginkgo leaf and its processed product. The compounds were identified using liquid chromatography negative electrospray ionization triple quadrupole tandem mass spectrometry (MS/MS). The neutral loss scan mode of MS/MS was used to screen flavonoid compounds and those compounds with acid group, or having rhamnosyl, glucosyl, or coumaroyl moiety in the samples. The successive data‐dependent product ion scan mode of MS/MS was used to identify the structure of the components. The analytical results represented three aglycone flavonoids and seven flavonoid glycosides in ginkgo. The method detection limits were evaluated for the analytes analyzed in the range of 0.88 to 2.67 μg/mL.     

Identification and quantification of eight flavones in root and shoot tissues of the medicinal plant huang-qin (Scutellaria baicalensis Georgi) using high-performance liquid chromatography with diode array and mass spectrometric detection

A method of analysis of eight flavones using high-performance liquid chromatography (HPLC)-diode array detection (DAD)-mass spectrometry (MS) in root and aerial tissues of the medicinal plant Scutellaria baicalensis was developed. The identity of the analytes was confirmed using retention time, UV-vis and mass spectral comparisons to commercial standards. Both UV-vis and mass spectral patterns were characterized for glycosylated flavones. Two additional flavone glycosides were tentatively identified as chrysin-7-glucuronide and wogonoside, but not quantified. Greenhouse and in vitro-grown tissues were analyzed with flavone concentrations ranges of 0.14-150 and 0.030-1.7 microg/mg for greenhouse root and shoot tissue, respectively, and 0.0068-6.4 and 0.082-1.5 microg/mg for in vitro-grown roots and shoots, respectively.