UPLC-ESI-QTOF/MS快速表征鉴定九华黄精的化学成分

目的采用超高效液相色谱电喷雾电离四极杆飞行时间质谱(UPLC-ESI-QTOF/MS)结合UNIFI软件对九华黄精中的主要化学成分进行快速表征和鉴定。方法采用ZORBAX SB-C_(18)(2.1 mm×100 mm,1.8μm)色谱柱进行分离,柱温30℃,以乙腈-水为流动相进行梯度洗脱,流速为0.3 mL·min^(-1);质谱采用正、负离子MS^(E)的全扫描模式分别采集质谱数据。结果正、负离子下共表征鉴定47个化学成分,包括甾体皂苷类15个、黄酮类8个、有机酸类9个,生物碱类4个,氨基酸类1个,其他类10个,其中3个经对照品鉴定,8种化合物为首次从九华黄精中发现。结论该方法实现了九华黄精中主要化学成分的快速质谱表征和鉴定,为九华黄精的质量评价和药效物质基础研究奠定了基础。

桂枝茯苓胶囊HPLC-ESI-QTOF/MS化学成分分析过程中提取溶剂的优化

采用高效液相色谱电喷雾四极杆飞行时间串联质谱(HPLC-ESI-QTOF/MS)对桂枝茯苓胶囊中的化学成分进行分离和鉴别。运用Agilent Eclipse XDB-C18(4.6 mm×250 mm,5μm)色谱柱,以0.1%甲酸水溶液-乙腈为流动相梯度洗脱;使用ESI离子源,正离子与负离子模式下采集数据。根据Q-TOF/MS正、负离子质谱信息、通过元素组成分析、化合物同位素分布验证和对质谱裂解碎片解析并与相关文献对照,鉴别了桂枝茯苓胶囊中18个化合物。本法为桂枝茯苓胶囊中的化学成分分析提供了一种快速、有效的方法。

UPLC-ESI-QTOF/MS^E结合UNIFI快速分析华蟾素注射液化学成分

采用超高效液相色谱-电喷雾离子源-四级杆飞行时间质谱(UPLC-ESI-QTOF/MS)结合UNIFI天然产物信息平台快速分析华蟾素注射液化学成分。利用QTOF/MS非信息依赖扫描的MS^E模式采集复杂组分质谱信息,将采集的质谱数据文件和自建化学成分库导入UNIFI,进行智能匹配,随后使用MassLynx工作站逐个核对。本实验共辨识76个化学成分,其中蟾蜍二烯羟酸内酯类36个、生物碱类12个(包括1种新成分)、小分子肽类13个、有机酸类7个、酰胺类4个、甾醇类2个、其他类2个。本方法能快速分离、分析华蟾素注射液复杂组分,为该制剂的毒-效成分及质量控制研究奠定了基础。

柔肝胶囊化学成分的UPLC-ESI-QTOF/MS分析

目的:采用液质联用技术(LC-MS)对中药柔肝胶囊(丹参、姜黄等)中的化学成分进行定性分析。方法:运用AC-QUITY UPLC HSS T3(2.1 mm×100 mm,1.8μm)柱,以甲醇-0.1%甲酸为流动相梯度洗脱;质谱使用ESI离子源,正离子与负离子模式下采集数据。结果:通过Q-TOF/MS正、负离子质谱信息及元素组成分析并结合对照品与相关文献数据对照,共鉴定出19个化合物,主要结构类型为:水溶性酚酸类、姜黄素类、皂苷类及异黄酮类化合物。结论:本法对柔肝胶囊的成分鉴定提供一种快速准确的分析方法。

Identification and characterization of phenolics and terpenoids from ethanolic extracts of Phyllanthus species by HPLC-ESI-QTOF-MS/MS

Phyllanthus species plants are a rich source of phenolics and widely used due to their medicinal properties. A liquid chromatography–tandem mass spectrometry(LC–MS/MS) method was developed using high-pressure liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(HPLC-ESI-QTOFMS/MS) for the identification and characterization of quercetin, kaempferol, ellagic acid and their derivatives in ethanolic extracts of Phyllanthus species. The chromatographic separation was carried out on Thermo Betasil C_8 column(250 mm×4.5 mm, 5 μm) using 0.1% formic acid in water and 0.1% formic acid in methanol as the mobile phase. The identification of diagnostic fragment ions and optimization of collision energies were carried out using 21 reference standards. Totally 51 compounds were identified which include 21 compounds identified and characterized unambiguously by comparison with their authentic standards and the remaining 30 were tentatively identified and characterized in ethanolic extracts of P. emblica, P. fraternus, P. amarus and P.niruri.

瓜蒌皮注射液中化学成分的UPLC/ESI-QTOF-MS分析

目的 :UPLC/ESI-QTOF-MS法鉴定瓜蒌皮注射液中的化学成分。方法 :Waters Acquity UPLC HSS T3色谱柱(2.1 mm×100 mm,1.8μm),以水(A)和乙腈(B)为流动相进行梯度洗脱。采用电喷雾离子源(ESI),正、负两种模式结合MS^E技术测定。所得分子量及碎片信息与文献或已知数据库比对,鉴定化合物的结构。结果:在正负离子模式下,共鉴别出瓜蒌皮注射液化学成分19个,包括7个氨基酸、4个核苷酸、5个黄酮、1个多肽、1个唾液酸和1个含氮化合物。结论 :建立了一种基于UPLC/ESI-QTOF-MS的对瓜蒌皮注射液化学成分快速鉴别的分析方法,为阐明瓜蒌皮注射液的化学物质基础提供依据。

UPLC-ESI-QTOF-MS技术分析苤蓝中的化学成分

采用超高效液相色谱(UPLC)技术,使用C18(2.1×100 mm,1.8μm)色谱柱,用不同比例的乙腈-0.1%甲酸水溶液做流动相梯度洗脱,对苤蓝甲醇提取物的正丁醇萃取相进行分离。以电喷雾电离源负离子模式对色谱流出物进行离子化,飞行时间串联质谱(QTOF-MS)对主要色谱峰进行归属。通过综合分析色谱数据、一级和二级全扫描质谱数据,对苤蓝中的化学成分进行分析鉴定。共从中鉴定出了包括苷类、香豆素类、萜类、生物碱类、有机酸及氨基酸类等在内的19个化合物。苤蓝中化学成分的鉴定,对于后续苤蓝中活性物质的分离及其系统研究可提供有用的参考。

超高效液相色谱-飞行时间质谱联用分析清热凉血方中的化学成分

目的鉴定中药复方清热凉血方浸膏中成分,为其治疗银屑病的物质基础提供科学依据。方法采用超高效液相色谱-飞行时间质谱联用(LC-ESI-QTOF MS)仪分析。色谱柱:5 cm×2.1 mm,1.7μm BEH C18柱,流动相为0.1%的甲酸溶液(A)-0.1%乙腈溶液(B),流速为0.3 mL/min。采用负离子模式与方中主要的单味药对比判断色谱峰的来源药材,继而通过与来源药材化学成分进行精确分子量的对比鉴定化合物。结果样本中检测出30多种物质,共鉴定出22个中药成分,分别为柠檬酸/异柠檬酸(1)、没食子酸(2)、Mudanoside B(3)、儿茶素(6)、羟基芍药苷(7)、短叶苏木酚酸(8)、丹皮酚(10)、牡丹酚新苷(12)、白芍苷R1(13)、芍药苷(14)、鞣花酸(15)、没食子酰芍药苷(16)、五没食子酰葡萄糖苷(17)、Oldenlandoside I(18)、苯甲酰氧芍药苷(19)、牡丹皮苷C(20)、苯甲酰芍药苷(24)、三羟基-十八碳二烯酸(25)、Anguivioside B1/2(26)、三羟基-十八碳二烯酸/9-羟基十八烷二酸(29)、2-羟基-3-甲基蒽醌(30)。结论在清热凉血方主要化学成分中,丹皮酚、芍药苷、白芍苷R1与银屑病的发病机理相关,可能是其活性成分。

几种豆制酱油中代谢产物的非靶向成分解析

【目的】明确4种中日知名品牌酱油中存在显著差异的成分物质,为进一步解析酱油中的未知代谢产物及靶向精准测定酱油中的代谢产物提供理论依据和方法参考。【方法】采用超高效液相色谱串联飞行时间质谱(UPLC-QTOF-MS)进行物质信息采集,并用生物信息学软件MS-DIAL和XCMS对实验数据进行多元变量统计分析。【结果】日产酱油中JS1和JS2的精氨酸质谱丰度分别为3.42×10^7、3.32×10^7,显著高于国产酱油CS1(0.98×10^7)和CS2(1.72×10^7);日产酱油JS1和JS2中的生物碱哈尔满质谱丰度(分别为2.02×10^7、1.58×10^7),显著高于国产酱油CS1(1.16×10^7)和CS2(0.84×10^7)。【结论】日产2种酱油中代谢产物精氨酸、生物碱哈尔满的相对含量显著高于国产2种酱油。

响应面分析法优化黑果枸杞总花色苷提取工艺及结构鉴定

以新疆产黑果枸杞为原料,采用溶剂浸提法提取黑果枸杞总花色苷。通过Box-Behnken响应面实验设计和DesignExpert8.0.6分析软件,以浸提温度、浸提时间、乙醇浓度和液料比四个因素为自变量,浸提液吸光度A为响应值,研究各个变量及其交互作用对黑果枸杞总花色苷浸提率的影响。模拟得到回归方程,并确定最佳工艺条件为:浸提温度58℃、浸提时间74min、乙醇浓度65%、液料比55:1(m L/g)。总花色苷纯化分级后,利用超高效液相色谱-电喷雾串联高分辨质谱(UPLC-DAD-ESI-QTOF-MS/MS)技术对其中流分HG-15进行研究。结果表明,黑果枸杞总花色苷HG-15流分中含有矮牵牛素-3-O-(6-O-对香豆酰)芸香糖苷-5-O-葡萄糖苷(Petunidin-3-O-(6-O-p-coumaryl)-rutinoside-5-O-glucoside)、锦葵素-3-反-对香豆酰芸香糖苷-5-O-葡萄糖糖苷(Malvidin-3-trans-pcoumaryl-rutinoside-5-O-glucoside)、飞燕草素-3-O-(6-O-乙酰)葡萄糖苷(Delphinidin-3-O-(6-Oacetyl)-glucoside)等六种主要花色苷。

基于LC/QTOF-MS/MS法鉴定兰考泡桐花化学成分

目的:初步鉴定兰考泡桐花中的化学成分,为该植物的药效物质基础研究提供数据支持。方法:运用液相色谱与四级杆飞行时间质谱联用技术(LC/QTOF-MS/MS),以0.1%甲酸水(A)-乙腈(B)为流动相,梯度洗脱,在正离子模式下分析。结果:本研究从兰考泡桐花中共鉴定出12个化合物,其中通过与标准品的保留时间和化合物的特征碎片的比对,准确鉴定出5个化合物,分别为芦丁、野漆树苷、木犀草素、芹菜素和熊果酸。另外,通过标准品总结出的裂解规律及相关文献报道,初步鉴定出7个黄酮类成分。结论:本研究基于LC-QTOF/MS的方法对兰考泡桐花所含化学成分进行了表征,为泡桐属的同类植物的物质基础研究提供了有益参考。

Structural characterization of monoterpene indole alkaloids in ethanolic extracts of Rauwolfia species by liquid chromatography with quadrupole time-of-flight mass spectrometry

Rauwolfia species(Apocynaceae) are medicinal plants well known worldwide due to its potent bioactive monoterpene indole alkaloids(MIAs) such as reserpine,ajmalicine,ajmaline,serpentine and yohimbine.Reserpine,ajmalicine and ajmaline are powerful antihypertensive,tranquilizing agents used in hypertension.Yohimbine is an aphrodisiac used in dietary supplements.As there is no report on the comparative and comprehensive phytochemical investigation of the roots of Rauwolfia species,we have developed an efficient and reliable liquid chromatography-tandem mass spectrometry(LC–MS/MS) method for ethanolic root extract of Rauwolfia species to elucidate the fragmentation pathways for dereplication of bioactive MIAs using highperformance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry(HPLC–ESI–QTOF–MS/MS) in positive ion mode.We identified and established diagnostic fragment ions and fragmentation pathways using reserpine,ajmalicine,ajmaline,serpentine and yohimbine.The MS/MS spectra of reserpine,ajmalicine,and ajmaline showed C-ring-cleavage whereas E-ring cleavage was observed in serpentine via Retro Diels Alder(RDA).A total of 47 bioactive MIAs were identified and characterized on the basis of their molecular formula,exact mass measurements and MS/MS analysis.Reserpine,ajmalicine,ajmaline,serpentine and yohimbine were unambiguously identified by comparison with their authentic standards and other 42 MIAs were tentatively identified and characterized from the roots of Rauwolfia hookeri,Rauwolfia micrantha,Rauwolfia serpentina,Rauwolfia verticillata,Rauwolfia tetraphylla and Rauwolfia vomitoria.Application of LC–MS followed by principal component analysis(PCA) has been successfully used to discriminate among six Rauwolfia species.

Antioxidant activity and identification of bioactive compounds from leaves of Anthocephalus cadamba by ultra-performance liquid chromatography/electrospray ionization quadrupole time of flight mass spectrometry

Objective:To evaluate the antioxidant potential of different extract/fractions of Anthocephalus cadamba（A.cadamba）（Roxb.） Miq.（Rubiaceae） and study the tentative identification of their active constituents.Methods:The extract/fractions were screened for antioxidant activity using various in vitro assays viz.DPPH assay,ABTS assay,superoxide anion radical scavenging assay, reducing power assay and plasmid DNA nicking assay.Total phenolic content of extract/fractions was determined by colorimetric method.An ultra-performance LC-electrospray-quadrupoletime of flight mass spectrometry method was used to analyse the active constituents of extract/ fractions of A.cadamba.Results:The ethyl acetate fraction was found to be most active fraction in all the assays as compared to other extract/fractions.The IC50 value of ethyl acetate fraction （ETAC fraction） was 21.24μg/mL,1.12μg/mL,9.68μg/mL and S7.81μg/mL in DPPH assay, ABTS assay,reducing power assay and superoxide scavenging assay respectively.All the extract/ fractions also showed the potential to protect the plasmid DNA（pBR322） against the attack of hydroxyl radicals generated by Fenton’s reagent.The bioactive compounds were identified by UPLC-ESI-QTOF-MS,by comparing the mass andλmax with literature values.Conclusions:The potential of the extract/fractions to scavenge different free radicals in different systems indicated that they may be useful therapeutic agents for treating radical-related pathologic damage.

Chemical composition, antiparasitic and cytotoxic activities of aqueous extracts of Ziziphus joazeiro Mart

Objective: To compare the in vitro antiparasitic activity of aqueous extracts from Ziziphus joazeiro leaves and stem bark against Trypanosoma cruzi, Leishmania braziliensis, and Leishmania infantum, as well as to evaluate its cytotoxicity in mammalian cells, in addition to identifying the chemical composition of the extracts. Methods: Ziziphus joazeiro leaf and stem bark aqueous extracts were prepared by cold extraction maceration and subjected to ultra-efficient liquid chromatography coupled to a quadrupole/time of flight system. The susceptibility assays used Trypanosoma cruzi CLB5 strains and promastigote forms of Leishmania braziliensis and Leishmania infantum for antiparasitic activity of the extracts. Moreover, mammalian fibroblasts NCTC clone 929 were used for cytotoxicity analysis. Results: Terpenoid compounds, flavonoids and phenolic acid were identified in extracts. The stem bark aqueous extracts presented more significant results in terms of antiparasitic activity compared with the leaf aqueous extracts, especially against Leishmania braziliensis and Leishmania infantum promastigote forms with an IC50 < 500 μg/mL. The cytotoxicity evaluation showed moderate toxicity of the stem bark aqueous extracts, which is relevant information for the rational use of this plant part since it is widely used by the population. Conclusions: These preliminary results may contribute to the formulation of new therapeutic agents against this group of neglected diseases, so further investigations are required to delineate the mechanisms of action mainly of the aqueous extract of stem bark of Ziziphus joazeiro.

HPLC-ESI-QTOF-MS法测定减肥保健食品中16种食欲抑制剂

建立高效液相色谱-电喷雾离子化-四级杆串联飞行时间质谱法(HPLC-ESI-QTOF-MS)高通量快速测定减肥保健食品中16种食欲抑制剂的含量。样品经甲醇超声萃取、Oasis MCX固相萃取柱净化,以甲醇和有乙酸铵溶液为流动相,梯度洗脱,采用QTOF-MS质谱仪进行分析。结果表明:16种食欲抑制剂在1.0~100.0 ng/mL范围内具有良好的线性关系,相关系数≥0.9995;方法检出限在0.03~0.30μg/kg,方法定量限在0.10~1.00μg/kg;加标回收率在91.7%~99.2%,相对标准偏差在1.1%~3.9%。

复方紫灵胶囊化学成分的UPLC-ESI-QTOF-MS-MS分析

目的:采用超高效液相色谱-电喷雾质谱法对复方紫灵胶囊中化学成分进行分析。方法:运用ACQUITY BEH C18超高效液相色谱柱(2.1 mm×100 mm,1.7μm),以乙腈-0.1%甲酸水溶液为流动相梯度洗脱;质谱使用ESI离子源,正离子与负离子模式下采集数据;运行时间20 min。结果:通过QTOF-MS-MS正、负离子质谱信息及元素组成分析并结合对照品与相关文献数据对照,共鉴定出30个化合物。结论:该方法对复方紫灵胶囊的成分鉴定提供了一种快速准确的分析方法。

UPLC/ESI-QTOF-MS-based metabolomics survey on the toxicity of triptolide and detoxication of licorice

Triptolide(TP) from Tripterygium wilfordii has been demonstrated to possess anti-inflammatory, immunosuppressive, and anticancer activities. TP is specially used for the treatment of awkward rheumatoid arthritis, but its clinical application is confined by intense side effects. It is reported that licorice can obviously reduce the toxicity of TP, but the detailed mechanisms involved have not been comprehensively investigated. The current study aimed to explore metabolomics characteristics of the toxic reaction induced by TP and the intervention effect of licorice water extraction(LWE) against such toxicity. Obtained urine samples from control, TP and TP + LWE treated rats were analyzed by UPLC/ESI-QTOF-MS. The metabolic profiles of the control and the TP group were well differentiated by the principal component analysis and orthogonal partial least squares-discriminant analysis. The toxicity of TP was demonstrated to be evolving along with the exposure time of TP. Eight potential biomarkers related to TP toxicity were successfully identified in urine samples. Furthermore, LWE treatment could attenuate the change in six of the eight identified biomarkers. Functional pathway analysis revealed that the alterations in these metabolites were associated with tryptophan, pantothenic acid, and porphyrin metabolism. Therefore, it was concluded that LWE demonstrated interventional effects on TP toxicity through regulation of tryptophan, pantothenic acid, and porphyrin metabolism pathways, which provided novel insights into the possible mechanisms of TP toxicity as well as the potential therapeutic effects of LWE against such toxicity.

姜黄素固体脂质纳米粒和微胶囊的制备、表征及体内药动学的比较研究

目的探究固体脂质纳米粒和微胶囊2种制剂方法对姜黄素理化表征的影响,并对其在SD大鼠体内的药动学行为进行定性定量研究。方法分别采用薄膜-超声分散法和均质乳化-喷雾干燥法制备姜黄素固体脂质纳米粒(curcumin solid lipid nanoparticles,Cur-SLN)和姜黄素微胶囊(curcumin microcapsules,Cur-MC),并对其外观形态、粒度分布、ζ电位、包封率、载药量和体外释放度进行表征;通过超高效液相色谱串联飞行时间质谱(UPLC-QTOF-ESI-MS/MS)法鉴定ig后大鼠血浆中的代谢产物,采用超高效液相色谱串联三重四级杆质谱(UPLC-ESI-MS/MS)法测定血药浓度,比较药动学参数和生物利用度。结果2种制剂方法均可显著降低姜黄素的粒径,使其均匀分布,Cur-SLN和Cur-MC的平均粒径分别为(184.3±7.9)、(415.3±10.3)nm,ζ电位分别为(-48.1±0.9)、(-16.4±0.4)m V,Cur-SLN和Cur-MC可将姜黄素的体外释药率从原料药的51.12%分别提高至87.79%和83.02%。ig后大鼠血浆代谢物主要为姜黄素及其葡萄糖醛酸化代谢物(curcumin glucuronide,Cur-glu),与游离姜黄素(free curcumin,Cur-F)组相比,2种水溶姜黄素制剂的药时曲线下面积(AUC_(0~t))、达峰浓度(C_(max))等药动学参数显著提高,Cur-SLN组姜黄素和Cur-glu的相对生物利用度分别增加4.31和4.63倍,Cur-MC组分别增加3.19和2.73倍。结论2种制剂均可显著提高姜黄素的口服吸收生物利用度,且固体脂质纳米粒优于微胶囊。

半夏白术天麻汤在大鼠血浆中的药物代谢研究

目的:研究半夏白术天麻汤在大鼠血浆中的药物代谢。方法:采用大鼠灌胃给药方式,建立液相色谱-电喷雾-四极杆-飞行时间/质谱(UHPLC-ESI-Q-TOF/MS)分析方法,研究半夏白术天麻汤在大鼠血浆代谢产物及代谢途径。结果:大鼠灌胃给予半夏白术天麻汤后,鉴定的33种代谢产物在大鼠血浆中依次发生Ⅰ相和Ⅱ相代谢反应,主要的代谢方式为氧化、还原、水解、亚硫酸化、葡萄糖醛酸化、甲基化和乙酰化。结论:本研究采用UHPLC-ESI-Q-TOF/MS法研究半夏白术天麻汤方药物主要成分在大鼠血浆中的代谢产物及代谢途径,为其多成分协同增效发挥作用的药效物质基础和临床应用提供科学依据。

不同产地丹参茎叶UPLC指纹图谱与化学模式识别研究

目的研究并建立不同产地丹参茎叶UPLC指纹图谱,采用电喷雾离子源-四极杆-飞行时间质谱(ESI-QTOF/MS)对其特征峰进行定性研究,为丹参地上茎叶资源的综合开发利用提供科学依据。方法采用UPLC法构建丹参茎叶的指纹图谱,色谱条件为ACQUITY UPLC BEH C18(100 mm×2.1 mm,1.7μm)色谱柱,以乙腈-0.1%甲酸水为流动相梯度洗脱,体积流量0.4 m L/min,检测波长280 nm,柱温35℃。质谱检测采用负离子模式;电压3.0 k V;离子源温度120℃;雾化温度400℃;雾化气800 L/h。同时应用相似度评价、聚类分析和主成分分析(PCA)对实验数据进行处理,以分析5个不同产地12批丹参茎叶样品间的相似性及差异性,并通过UPLC/Q-TOF/MS对丹参茎叶共有峰进行了定性分析。结果获得丹参茎叶UPLC特征图谱,共有峰13个。12批丹参茎叶的相似度为0.823~0.997,聚类分析、PCA结果与相似度分析结果一致。结论丹参茎叶UPLC指纹图谱的构建和化学模式的识别为其质量控制和丹参非药用部位利用价值的挖掘提供科学依据。