HPLC-MS/MS�о�̵���彺�������������ܵ��ۻ�ѧ�ɷֵı仯

??OBJECTIVE To identify the chemical components of bile acids from bear bile powder, the intermediate of bear bile powder and Tanreqing capsules by HPLC-MS/MS. METHODS All samples were extracted with 70% methanol(V/V), and an Ultimate XB C₁₈ column (4.6 mm??250 mm, 5 ??m) was employed for separation with acetonitrile-0.1% formic acid as mobile phase in gradient elution. The MS spectrum was acquired in both positive and negative ion mode using ESI ion source. The chemical components were identified by the second mass spectrometric pyrolysis fragments, chromatographic peak retention time and fragmentation regularity summarized from the reference standards and the available literature. RESULTS A total of 33 compounds were successfully identified or tentatively predicted, and six chemical compounds including tauroursodeoxycholic acid, taurochenodeoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid, 7??-hydroxy-3-oxo-5??-cholanic acid and one unknown constituent were finally transferred to Tanreqing capsules through the intermediate of bear bile powder. Moreover, 21 new chemical compounds (major ingredients of free bile acids) were generated during the production process of the intermediate, and 19 components were also detected in Tanreqing capsules. CONCLUSION The investigation of the change of constituents in bear bile powder during Tanreqing capsules production provides a basis for the quality control and evaluation of Tanreqing capsules during production process.     

基于UHPLC-Q-TOF-MS/MS和LTQ-MSn联用技术快速鉴定绞股蓝皂苷类化学成分

目的 建立超高液相色谱-飞行时间高分辨质谱(UHPLC-Q-TOF MS/MS)和超高液相线性离子阱质谱(LTQ-MSⁿ)联用技术对绞股蓝中皂苷类成分快速鉴定的方法。方法 采用Thermo Syncronis C₁₈(2.1 mm×100 mm,1.7 μm)色谱柱,以纯水-乙腈为流动相梯度洗脱,质谱采用电喷雾(ESI)离子源,在负离子模式下采集数据对绞股蓝皂苷类进行成分鉴定。结果 初步鉴定了81种绞股蓝皂苷,其中包括58种已知皂苷和23种可能的未知皂苷结构。结论 UHPLC-Q-TOF-MS/MS可以提供化合物的准确相对分子质量及二级碎片信息,LTQ-MSⁿ可以提供化合物多级质谱信息,该联用技术可以简单、快速鉴定绞股蓝皂苷类成分,并为其质量控制、药效物质基础研究及进一步临床应用提供理论依据和科学研究思路。     

��ЧҺ��ɫ���������÷������������B�е�δ֪����

??OBJECTIVE To establish an HPLC-MS/MS method to identify the unknown impurities in polymyxin B sulfate. METHODS The analysis was performed on Agilent 1260-6550 Q/TOF-MS with a Diamonsil Plus C₁₈ column(4.6 mm??250 mm, 5 ??m). Mobile phase A was 0.01 mol??L^-1 trifluoroacetic acid-acetonitrile(95??5), and mobile phase B was acetonitrile containing 0.1% formic acid. Mobile phase A and B were set at the volume ratio of 79??21 at a flow rate of 1 mL??min^-1 under isocratic elution. The detection wavelength was set at 254 nm. ESI source was used. Positive ion scanning was conducted in the range of m/z 50-1 700 for MS and MS/MS. The unknown components were identified by comparing the MS and MS/MS with the known reference standards like polymyxin B₁ and B₂. The photochemical Paterno-B??chi reaction was performed using a low-pressure mercury lamp as the light source at emission wavelength of 254 nm with acetone/water(50/50, V/V) as the reaction solvent. RESULTS The structures of seven unknown related substances in polymyxin B sulfate were identified. The most abundant impurity was identified to be vinyl polymyxin B₁, for which the double bond was at the end of the fatty acyl residue. CONCLUSION Vinyl polymyxin B₁ is reported for the first time. The method provides a good idea for the identification of related substances in drugs.     

�߷���ȿ���Һ��ɫ����������ͬʱ�ⶨ��ͬ��λ�Ͳ�ͬ�ӹ���������ˏ�а�����ͺ�����ɷֺ���

??OBJECTIVE To establish an UFLC-QTRAP-MS/MS method for simultaneous determination of 15 kinds of amino acids and 12 kinds of nucleosides and nucleobases in different parts of Metaplexis japonica(Thunb.) Makino processed by different methods. METHODS The analysis was carried out on a Waters XBridge Amide column(2.1 mm??100 mm,3.5 ??m) with elution by mobile phase of 0.2% formic acid in acetonitrile-0.2% formic acid in water at a flow rate of 0.6 mL??min^-1. The column temperature was maintained at 30 ??. The target compounds were analyzed by positive ion multiple reaction monitoring(MRM) mode. RESULTS Twenty-seven multiple constituents showed good linearity(r??0.997 3) in the ranges of the tested concentrations. The average recoveries of the 27 components were 98.17%-100.75% with relative standard deviations of 0.90%-2.49%. CONCLUSION The established method is accurate and precise, which provides a reliable solution for selecting the parts and the optimal processing method of Metaplexis japonica(Thunb.)Makino.     

���ڳ���ЧҺ��ɫ��-�߷ֱ����׼�����������̵����ע��Һ�ͽ��һ�ѧ�ɷֵĶ��Բ���

??OBJECTIVE To identify rapidly the chemical constituents in Tanreqing injection and Tanreqing capsules by ultra-performance liquid chromatography with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS^E).METHODS The separation was performed on a Waters Acquity UPLC BEH C₁₈ column (1.0 mm??100 mm, 1.7 ??m) with acetonitrile-0.1% formic acid as mobile phase in gradient elution. ESI ion source was employed in negative ion mode. The differences of chemical compositions between the two preparations and the sources of these compounds were illustrated based on their retention time, accurate mass measurements and the mass fragments by comparison with those in the literature or database and the reference standards.RESULTS A total of 111 compounds including 13 unknown components were identified or tentatively characterized. Among these compounds, 14 were derived from Scutellariae Radix (SR) intermediate, 36 were from Bear Bile Powder(BBP) intermediate, 7 were from Caprae Hircus Cornu(CHC) intermediate, 34 were from Lonicerae japonicae Flos(LJF) intermediate and 22 were from Forsythiae Fructus(FF) intermediate. Moreover, quinic acid and rutin were simultaneously detected in LJF and FF intermediates, 28 constituents were unambiguously confirmed by their reference standards. However, 71 compounds were observed both in injection and capsules, while 24 compounds were only found in Tanreqing injection and 16 compounds only in the capsules.CONCLUSION The differences of chemical constituents between Tanreqing injection and capsules are effectively characterized by UPLC/Q-TOF-MS^E method, which will facilitate the quality control of the two preparations.     

��˿����Ƽ��ز�λ�ڴ���Ѫ��ͷ���еĴ�л�ɷַ���

??OBJECTIVE To identify the metabolites of Cuscuta chinensis in the serum and feces of rats with estrogenic estrogen, and to provide the basis for elucidating the pharmacological basis of its efficacy. METHODS Using the HPLC-ESI-MS/MS technique, the molecular weight and molecular formula of the compounds were preliminarily estimated by the first-order mass spectrometry, and the chemical constituents of serum and feces were characterized by the retention time of the standard, the fragment ion information, the fragmentation law of mass spectrometry and the reference data. RESULTS Five prototypic components and 4 metabolites were identified in the serum. A total of 11 chemical constituents were identified in the feces after administration, including 5 prototype components and 6 metabolites. CONCLUSION Through the comparative analysis between serum and feces, the main metabolic pathways of Cuscuta chinensis compound are deduced, which provides reference for the basic research on the estrogenic substance of Cuscuta chinensis.     

����UFLC-Triple TOF-MS/MS�����������ӻ�ѧ�ɷ�

??OBJECTIVE To qualitatively analyze the major chemical constituents of Magnoliae Officinalis Cortex by UFLC-Triple TOF-MS/MS. METHODS The analysis was carried out on an Agilent ZORBAX SB-C₁₈ column(4.6 mm ??250 mm,5 ??m)with gradient elution of 0.1% formic acid-acetonitrile under both positive and negative ionization modes of ESI, respectively. The chemical components of Magnoliae Officinalis Cortex were identified by analyzing the accurate mass of molecular and product ions provided by UFLC-Triple TOF-MS/MS, comparison with reference standards and the literature data. RESULTS A total of 24 compounds were identified or tentative presumed in Magnolia Officinalis Cortex, including 12 lignans, 9 alkaloids, and 3 phenylpropanoid glycosides. CONCLUSION This study provides the foundation and support for study on material foundation of efficacy and the overall assessment of quality of Magnoliae Officinalis Cortex.     

HPLC-DAD/ELSD-ESI-TOF/MS��������Ƭ�л�ѧ�ɷּ����ָ�궨��ָ��ͼ���о�

??OBJECTIVE To analyze and identify the compositions using HPLC-DAD/ELSD-ESI-TOF/MS for establishing the multi index quantatitive HPLC fingerprint of the Jinming Tablet, in order to providing scientific basis for its quality control.METHODS The Agilent SB C₁₈ column (4.6 mm??250 mm, 5 ??m) was used with a mobile phase of acetonitrile-0.2% acetic acid in gradient elution, the flow rate was 0.8 mL??min^-1, the sample voume was 30 ??L, the column temperature was maintaine at 25 ??, the detection wavelength was set at 280 nm. ESI-TOF/MS positive and negative ion mode scanning was used to qualitatively analyze the components in methanol water extract from Jinming Tablet.RESULTS Twenty compounds in Jinming Tablet extract could be primarily identified by HPLC-DAD on-line detection, in which five compounds were determined; based on the analysis of multiple batches of samples, the multi index chromatographic fingerprint was established, meanwhile combined with similarity analysis to achieve the quality evaluation of Jinming Tablet.CONCLUSION The HPLC-DAD/ELSD fingerprint film analysis method for Jinming Tablet is established, which could provide reference for the quality control of the material basis of other compound.     

基于UPLC-ESI-Q-TOF-MS/MS技术快速鉴定彝族药双参的化学成分

目的 应用超高效液相色谱-电喷雾/四极杆飞行时间串联质谱(UPLC-ESI-Q-TOF-MS/MS)技术快速鉴定双参药材的化学成分。方法 采用Acquity UPLC BEH C₁₈色谱柱(2.1 mm×100 mm, 1.7μm)分离,以体积分数0.1%甲酸水溶液(A相)-乙腈(B相)为流动相进行梯度洗脱,流速0.4 mL·min^-1,采用电喷雾离子源(ESI),在正、负离子模式下采集数据,扫描范围m/z 100～2 000,通过与对照品的保留时间及质谱数据信息对比,并结合精确相对分子质量、质谱裂解规律、质谱数据库(ChemicalBook, ChemSpider, Pubmed等)及相关文献,对双参的化学成分进行快速鉴定。结果 从彝族药双参中共鉴定出59个化合物,其中包括11个环烯醚萜类、23个有机酸类、14个三萜类、4个糖苷、其他类(1个香豆素类、1个生物碱类、2个酯类、1个多酚类、2个核苷类),其中通过与对照品对比鉴定了8个化合物,分别为绿原酸、隐绿原酸、异绿原酸A、异绿原酸B、异绿原酸C、马钱苷、马钱苷酸和樟芽菜苷。结论 该研究首次...     

���鲹��ѧ�ɷּ����PC12ϸ���ı��������о�

??OBJECTIVE To study the chemical constituents from the rhizome of Drynaria fortunei and the protective effects of them on PC12 cells induced by A??_25-35. METHODS The compounds were isolated and purified by silica gel, Sephadex LH-20, ODS column chromatography, and their structures were identified on basis of spectroscopic methods, such as MS and NMR. PC12 cells were treated with A??_25-35 to establish the Alzheimer' s disease models. The compounds of different concentrations were added into culture medium to detect the protection. MTT assay was used to detect cell vitality and to observe the protective effects of compounds on PC12 cells induced by A??_25-35. RESULTS Nine compounds were isolated and identified as naringin(1), neoeriocitrin(2), 5,7-dihydroxychromone-7-neohesperidoside(3), (E)-4-O-??-D-glucopyranosyl caffeic acid(4), kaempferol(5), luteolin(6), protocatechoic acid(7), psoralen(8), and ??-sitosterol(9) . The cell experiments were performed on the compounds 1-8 and the results showed they can promote the proliferation of PC12 cells. The cell vitality increase with concentration rising, and the difference is statistically significant (P<0.05). CONCLUSION Compounds 1-8 play an important role in protecting A??_25-35-induced injury in PC12 cells and they are the main active components of Drynaria fortunei in the protection of central nervous function.