Analysis of Alkaloids in Coptis chinensis Franch by High Performance Capillary Electrophoresis-Electrospray Ionization Time of Flight Mass Spectrometry

建立了高效毛细管电泳-电喷雾飞行时间质谱联用(HPCE-ESI-TOF/MS)快速定性分析黄连中生物碱类化合物的分析方法.使用未涂层石英毛细管,以50mmol/L乙酸铵-0.5%甲醇溶液(用氨水调至pH=7.2)作为运行缓冲液,分离电压为25kV;鞘液组成为50%甲醇-49.5%水-0.5%乙酸,鞘液流速为4μL/min;质谱选用正离子模式,碰撞电压(Fragmentor)为100V.结果表明,通过各色谱峰紫外光谱和质谱测得精确分子量结果,结合文献,对黄连中7种生物碱进行了鉴定.表明本方法简便、快速,是黄连中生物碱类化合物快速分离、鉴别的有效方法.A new method for the analysis of alkaloids in Coptis chinensis Franch was established by high performance capillary electrophoresis-electrospray ionization time of flight mass spectrometry (HPCE-ESI-TOF/MS). The real samples were separated by an uncoated capillary. 50 mmol/L ammonium acetate containing 0.5% methanol (pH=7.2) was used as the running buffer, and separation voltage was 25 kV. A coaxial sheath flow interface was used as the CE-MS interface, and a 50% methanol-49.5% water- 0.5% acetic acid mixture was used as the sheath liquid with a flow rate of 4 μL/min. The lens voltages in a positive ion mode with a collision induced dissociation (CID) voltage of 100 V were used for ESI-TOF/MS analysis. Seven alkaloids in Coptis chinensis Franch methanol extracts were separated and identified by CE-DAD and CE-ESI-TOF/MS. The coupling of HPCE separation with accurate mass measurement capability of ESI-TOF/MS provides an attractive tool for the identification of alkaloid compounds in Coptis chinensis Franch.国家自然科学基金重点项目(No.20235020);; 青岛“2004将才计划”(No.04-3-JJ-11);; 共建生物医药研发测试中心(No.LS-05-KJZX-76)资助项目

Simultaneous determination of 7 nucleosides in Asterias rollestoni using reversed-phase high performance liquid chromatography

建立了罗氏海盘车中7种核苷化合物的反相高效液相色谱分析测定方法。采用超声波辅助提取,选用两根不同的C18色谱柱串联,以甲醇和0.2%(体积分数)乙酸/水溶液为流动相梯度洗脱分离。优化的色谱条件为:柱温为室温,检测波长为260nM,流速为0.8Ml/MIn,进样量为20μl。结果表明,7种核苷化合物在一定的浓度范围内线性关系良好,次黄嘌呤和胸苷的线性范围为0.65~40Mg/l,尿苷、黄嘌呤和肌苷的线性范围为0.80~40Mg/l,胸腺嘧啶的线性范围为1.15~40Mg/l,鸟苷的线性范围为0.50~40Mg/l。样品中7种核苷化合物的加标回收率为90.00%~105.00%,相对标准偏差为0.72%~3.23%。该方法操作简便、灵敏度高、重复性好,回收率高,适用于罗氏海盘车中7种核苷类成分的同时分析,也可用于罗氏海盘车的质量控制和综合评价。A method for the simultaneous determination of 7 nucleosides in Asterias rollestoni was devel-oped using reversed-phase high performance liquid chromatography ( RP-HPLC) .Analytes were extracted by ultrasonic-assisted extraction and separated on two different C18 columns,which were connected in se-ries,under the gradient elution with the mobile phases of methanol and 0.2% ( v/v) acetic acid/water at room temperature.The chromatographic conditions were as follows: flow rate,0.8 mL/min; detection wavelength,260 nm; injection volume,20 μL.Under the optimized conditions,good linear relationships between the values of mass concentrations and the peak areas of hypoxanthine,uridine,xanthine,thy-mine,inosine,guanosine and thymidine were observed in the ranges of 0.65-40,0.80-40,0.80-40,1.15-40,0.80-40,0.50-40,and 0.65-40 mg/L,respectively.The relative standard devia-tions were around 0.72%-3.23% and the recoveries were around 90.00%-105.00%.The results showed that the developed method is sensitive,accurate and reproducible.It is suitable for the analysis of nucleosides in Asterias rollestoni with high recoveries and it is expected to be used for the quality control and evaluation of Asterias rollestoni.国家自然科学基金项目(20905017);海洋公益性行业科研专项(200705011;200805039);海洋局青年基金项目(2010140);海洋一所基本科研业务专项(GY-022008T32;2010G25);中国科学院实验海洋生物学重点实验室开放基金课

Identification of Bohai Sea crude oil based on characteristic fingerprinting by MSPD-RP-HPLC

建立渤海原油的高效液相色谱(HPlC)特征指纹图谱,为渤海原油的鉴别提供一种新方法。以氧化铝作为基质固相分散剂进行样品处理,采用HPlC法进行分析,双反相C18色谱柱串联,以乙腈-水为流动相,二元线性梯度洗脱,流速为1.0 Ml/MIn,柱温为20℃,双波长检测226 nM、254 nM,进样量10μl;采用所发展的方法对10批渤海原油样品进行分析,建立渤海原油的HPlC特征指纹图谱,结合相似度分析用于渤海原油的鉴别。该方法具有很好的精密度、重复性和稳定性,10批渤海原油特征指纹图谱有13个共有峰,结合相似度分析可以用于渤海原油样品的鉴别,说明HPlC特征指纹图谱是渤海原油鉴别的有力工具。Based on the high performance liquid chromatography(HPLC) characteristic fingerprinting,a new method for crude oil identification of Bohai Sea was developed.The sample pretreatment was done using matrix solid-phase dispersion,and Al2O3 was used as the dispersant.The chromatographic fingerprints were determined by injecting 10 L sample solution each time on two reverse phase C18 columns in series with the gradient elution solvent system composed of water and acetonitrile.The flow rate was 1.0 mL/min,the column temperature was maintained at 20℃ and the signals were acquired at 226 nm and 254 nm.The characteristic fingerprints of crude oil were established by analysis of ten representative samples derived from Bohai Sea,and identification was carried out by the similarity analysis of the characteristic fingerprints.The method was ideally suited for the fingerprinting of Bohai Sea crude oil with good precision,repeatability and stability.13 peaks were selected as the common peaks of the characteristic fingerprint and used for the similarity analysis,which could be applied to distinguish crude oil of Bohai Sea from that of other sources.The characteristic HPLC fingerprinting technique thus was a powerful tool for the identification of crude oil originated from Bohai Sea.海洋公益性行业科研专项(200705011;201005034-3);中国海监技术支撑体系项目;2008年海洋环境保护及节能减排专

Analysis of alkaloids in Coptis chinensis Franch by high performance capillary electrophoresis-electrospray ionization time of flight mass Spectrometry

A new method for the analysis of alkaloids in Coptis chinensis Franch was established by high performance capillary electrophoresis-electrospray ionization time of flight mass spectrometry (HPCE-ESI-TOF/MS). The real samples were separated by an uncoated capillary. 50 mmol/L ammonium acetate containing 0.5% methanol (pH=7.2) was used as the running buffer, and separation voltage was 25 kV. A coaxial sheath flow interface was used as the CE-MS interface, and a 50% methanol-49.5% water- 0.5% acetic acid mixture was used as the sheath liquid with a flow rate of 4 mu L/min. The lens voltages in a positive ion mode with a collision induced dissociation (CID) voltage of 100 V were used for ESI-TOF/MS analysis. Seven alkaloids in Coptis chinensis Franch methanol extracts were separated and identified by CE-DAD and CE-ESI-TOF/MS. The coupling of HPCE separation with accurate mass measurement capability of ESI-TOF/MS provides an attractive tool for the identification of alkaloid compounds in Coptis chinensis Franch

长江江峡库区鱼类资源现状的研究

1997-2000年在长江三峡库区的巴南和万州江段共监测1292船次，统计渔获物4096.38kg，生物学测定2464尾，共采集鱼类30种，分属8科24属。目前库区渔获物组成和结构与20世纪70年代相比发生了较大变化，白甲鱼、中华倒刺（鱼+巴）、岩原鲤等在渔获物中比例已经很少，铜鱼资源也明显下降，（鱼+管）、（鱼+感）、鳊等在渔获物中已基本消失，而南方鲇、鲤、黄颡鱼等比例相对上升，渔获物中高龄鱼个体减少，低龄鱼及幼鱼个体比重增加。与70年代资料相比，三峡区鱼类资源已呈衰退趋势

长江三峡库区鱼类资源现状的研究

1997— 2 0 0 0年在长江三峡库区的巴南和万州江段共监测 1 2 92船次 ,统计渔获物40 96.38kg,生物学测定 2 4 64尾 ,共采集鱼类 30种 ,分属 8科 2 4属。目前库区渔获物组成和结构与 2 0世纪 70年代相比发生了较大变化 ,白甲鱼、中华倒刺、岩原鲤等在渔获物中比例已经很少 ,铜鱼资源也明显下降 ,、鳡、等在渔获物中已基本消失 ,而南方鲇、鲤、黄颡鱼等比例相对上升 ,渔获物中高龄鱼个体减少 ,低龄鱼及幼鱼个体比重增加。与 70年代资料相比 ,三峡库区鱼类资源已呈

长江江峡库区鱼类资源现状的研究

1997-2000年在长江三峡库区的巴南和万州江段共监测1292船次，统计渔获物4096.38kg，生物学测定2464尾，共采集鱼类30种，分属8科24属。目前库区渔获物组成和结构与20世纪70年代相比发生了较大变化，白甲鱼、中华倒刺（鱼+巴）、岩原鲤等在渔获物中比例已经很少，铜鱼资源也明显下降，（鱼+管）、（鱼+感）、鳊等在渔获物中已基本消失，而南方鲇、鲤、黄颡鱼等比例相对上升，渔获物中高龄鱼个体减少，低龄鱼及幼鱼个体比重增加。与70年代资料相比，三峡区鱼类资源已呈衰退趋势

长江江峡库区鱼类资源现状的研究

1997-2000年在长江三峡库区的巴南和万州江段共监测1292船次，统计渔获物4096.38kg，生物学测定2464尾，共采集鱼类30种，分属8科24属。目前库区渔获物组成和结构与20世纪70年代相比发生了较大变化，白甲鱼、中华倒刺（鱼+巴）、岩原鲤等在渔获物中比例已经很少，铜鱼资源也明显下降，（鱼+管）、（鱼+感）、鳊等在渔获物中已基本消失，而南方鲇、鲤、黄颡鱼等比例相对上升，渔获物中高龄鱼个体减少，低龄鱼及幼鱼个体比重增加。与70年代资料相比，三峡区鱼类资源已呈衰退趋势

On-line Screening and Identification of Free Radical Scavenging Compounds in Water Extract of Tea by High Performance Liquid Chromatography-Mass Spectrometry-2,2-Diphenyl-1-Picrylhydrazyl

将基于在线高效液相色谱-二苯基三硝基苯肼(HPlC-dPPH)快速筛选自由基清除剂的方法与电喷雾飞行时间质谱(ESI-TOf/MS)结合,建立了茶叶粗提物中抗氧化活性成分在线筛选与鉴别的方法。本方法是在HPlC色谱柱分离后进行分流,一路进入ESI-TOf/MS用于各化合物的快速鉴别,另一路流出液与稳定的自由基dPPH混合,实现在线筛选自由基清除剂的作用。本方法用于茶水中抗氧化成分的快速筛选与鉴别,筛选出11个具有明显dPPH自由基清除作用的化合物,通过ESI-TOf/MS在线分析获得的质谱信息,结合相关文献和数据库,实现了各化合物的快速鉴别。11个化合物分别为茶氨酸、THEOgAllIn、没食子儿茶素、茶碱、色氨酸、表没食子儿茶素、表没食子儿茶素没食子酸酯、表儿茶酚、没食子酸酯、没食子儿茶素没食子酸酯及儿茶素没食子酸酯。本方法效率高、稳定性好,是复杂天然产物中抗氧化剂快速筛选与鉴别的有力工具。A new method based on the HPLC on-line scavenging diphenyl-1-picrylhydrazyl(DPPH·) radical activity and ESI-Tof/MS for the rapid screening and identification of radical scavengers from water extract of tea was developed.Water extract of tea was separated by HPLC.The outlet of HPLC flow cell was connected to a split valve in order to divert a flow to ESI-Tof/MS for on-line rapid identification of the antioxidants in water extract of tea,and another stream react post-column with the DPPH solution for on-line screening the antioxidants in water extract of tea.The results showed that eleven compounds in tea have scavenging radical activity,and all the eleven compounds including theanine,theogallin,gallocatechin grade,theophyline,tryptophan,epigallocatechin,epigallocatechin-3-gallate,table catecholc,gallate,gallocatechin-3-gallate and catechin gallate were identified by ESI-Tof/MS according to literatures and data base.The method has also been used for several other natural products successfully.This method is reliable,efficient and it is a powerful tool for rapid screening and identification of free radical scavenging compounds in complex natural products.国家自然科学基金项目(No.20675021);福建省科技厅茶叶质量控制项目(No.2006N0043);海洋公益性行业科研专项(No.200705011);海洋一所基本科研业务专项(No.GY-022008T32)资