石油亚砜馏分的色谱法分离

用经典硅胶柱色谱法将石油亚砜ＰＳＯ分离为４个馏分,对其中的第３个馏分进行了高效制备液相色谱的分离制备。通过大量的试验,建立了较好的色谱条件：以甲醇、水混合液为流动相,以Ｃ_（１８）反相柱为固定相,把ＰＳＯ３分离成７个不同的馏分。对其中的两个馏分ＰＳＯ３_ｃ和ＰＳＯ３_ｅ作进一步的色谱分离和纯化后,获得较纯的亚砜馏分,用作结构分析和性能研究。     

高效离子交换液相色谱法分离绿脓杆菌外膜蛋白

利用高效ＤＥＡＥ离子交换液相色谱法分离纯化了绿脓杆菌标准菌株ＰＡＯ１外膜蛋白。流动相为ｐＨ８．０Ｔｒｉｓ－ＨＣｌ缓冲液,色谱柱为ＴＳＫｇｅｌ－ＤＥＡＥ－５ＰＷ（０．７５ｃｍ×７．５ｃｍｉ．ｄ．）。通过纯化外膜蛋白,可以为研究绿脓杆菌外膜通透性与抗生素耐药性之间的关系及缩短研究周期提供有效的方法。     

γ－环糊精－SDS胶束电动色谱法分离十八甲基炔诺酮光学异构体

 研究了ＣＤ－ＳＤＳ胶束电动色谱（ＣＤ－ＭＥＫＣ）对电中性及疏水性的手性化合物的分离，并讨论了不同类型ＣＤ的手性识别作用。使用γ－ＣＤ可以有效地拆分十八甲基块诺酮（ＮＯＧ）旋光异构体。选择了最佳分离条件，通过调节ＣＤ和ＳＤＳ浓度以及其它操作条件可以把ＮＯＧ异构体很好的分离。     

γ－环糊精－SDS胶束电动色谱法分离十八甲基炔诺酮光学异构体

研究了ＣＤ－ＳＤＳ胶束电动色谱（ＣＤ－ＭＥＫＣ）对电中性及疏水性的手性化合物的分离，并讨论了不同类型ＣＤ的手性识别作用。使用γ－ＣＤ可以有效地拆分十八甲基块诺酮（ＮＯＧ）旋光异构体。选择了最佳分离条件，通过调节ＣＤ和ＳＤＳ浓度以及其它操作条件可以把ＮＯＧ异构体很好的分离。     

高效离子交换液相色谱法分离绿脓杆菌外膜蛋白

 利用高效ＤＥＡＥ离子交换液相色谱法分离纯化了绿脓杆菌标准菌株ＰＡＯ１外膜蛋白。流动相为ｐＨ８．０Ｔｒｉｓ－ＨＣｌ缓冲液,色谱柱为ＴＳＫｇｅｌ－ＤＥＡＥ－５ＰＷ（０．７５ｃｍ×７．５ｃｍｉ．ｄ．）。通过纯化外膜蛋白,可以为研究绿脓杆菌外膜通透性与抗生素耐药性之间的关系及缩短研究周期提供有效的方法。     

毛细管电色谱中缓冲液离子强度对ODS柱分离行为的影响

介绍了毛细管电色谱中流动相的选择以及缓冲液的不同离子强度对ＯＤＳ柱分离行为的影响。     

毛细管电色谱中缓冲液离子强度对ODS柱分离行为的影响

 介绍了毛细管电色谱中流动相的选择以及缓冲液的不同离子强度对ＯＤＳ柱分离行为的影响。     

高效液相色谱法测定地榆中金丝桃甙的含量

采用高效液相色谱法测定了地榆中金丝桃甙的含量。色谱柱为Ｓｈｉｍ－ｐａｃｋＣＬＣ－ＯＤＳ柱,以甲醇－０．０２５ｍｏｌ／Ｌ磷酸（５０５０）为流动相,检测波长３７０ｎｍ,测定组分与其它组分的色谱峰达到基线分离。加样回收率平均值为９８．７％,ＲＳＤ为１．９％。为控制中药地榆的内在质量提供了新的测定指标和可靠的方法。     

反相高效液相色谱法同时测定决明子中芦蔡大黄素和大黄素

用反相高效液相色谱法分离并测定了决明子中芦蔡大黄素和大黄素，建立了该中药中芦蔡大黄素，大黄素分离，测定的色谱方法。色谱条件：ＯＤＳ柱，甲醇－水为流动相，检测波长２２３ｎｍ。本研究为决明子的质量评价提供了科学依据。     

反相高效液相色谱法同时测定决明子中芦荟大黄素和大黄素

用反相高效液相色谱法分离并测定了决明子中芦荟大黄素和大黄素，建立了该中药中芦荟大黄素、大黄素分离、测定的色谱方法。色谱条件：ＯＤＳ柱，甲醇－水（８０∶２０Ｖ／Ｖ）为流动相，检测波长２２３ｎｍ。本研究为决明子的质量评价提供了科学依据。     

Molecular mass spectrometric identification of superoxide dismutase in the liver of mice <Emphasis Type="Italic">Mus musculus</Emphasis> and <Emphasis Type="Italic">Mus spretus</Emphasis> using a metallomics analytical approach

This paper reports the identification and quantification of superoxide dismutase in the liver of Mus musculus and Mus spretus mice using a metallomics analytical approach. The approach consisted of using orthogonal chromatographic systems coupled to ICP–MS and UV detectors. Size-exclusion fractionation of the cytosolic extracts was followed by anion-exchange chromatographic separation of Cu- and Zn-containing species. After purification then tryptic digestion, Cu- and Zn-containing superoxide dismutase was identified by nESI-QqTOF. The MS–MS spectra of doubly charged peptides, with the Mascot searching engine, were used to obtain the sequence of the protein.     

Purification of pro- and eukaryotic superoxide dismutases by charge-controlled hydrophobic chromatography

The process of purifying superoxide dismutases was simplified using charge-controlled hydrophobic chromatography on 10-carboxydecyl Sepharose. In only one chromatographic step following ammonium sulphate precipitation, Fe-containing superoxide dismutase from Pseudomonas putida and Cu,Zn-containing superoxide dismutase from bovine erythrocytes were purified with an overall yield of about 70% to electrophoretic homogeneity. The specific activities of the crystalline enzyme preparations were expressed in McCord and Fridovich units and were 3000 and 3200 U/mg, respectively.     

Quantitative determination of SC-68328 in dog plasma using flow injection and tandem mass spectrometry

A flow injection/tandem mass spectrometric assay was developed to quantitate SC-68328 in dog plasma using its stable isotopic analog [13C4]SC-68328 as an internal standard (IS). Since SC-68328, a manganese-based superoxide dismutase mimetic, is very unstable, very polar and adheres to silica-based high-performance liquid chromatographic columns, the analyte and IS were derivatized to their bis-isothiocyanate forms followed by a liquid-liquid extraction with methylene chloride and analyzed using positive ion electrospray mass spectrometric detection. SC-68328 was quantitated using the peak-height ratio of SC-68328 to its IS using MS/MS in the multiple reaction monitoring mode. The lower limit of quantitation of the assay was 0.25 microg ml(-1) SC-68328 in dog plasma with an inter-day precision of 11.8% and an accuracy of 113% (n = 12). Acceptable precision and accuracy were also obtained for concentrations in the calibration curve range (0.25-10 microg ml(-1) SC-68328 in dog plasma).     

Absolute quantification of superoxide dismutase in cytosol and mitochondria of mice hepatic cells exposed to mercury by a novel metallomic approach

In the last years, the development of new methods for analyzing accurate and precise individual metalloproteins is of increasing importance, since numerous metalloproteins are excellent biomarkers of oxidative stress and diseases. In that way, methods based on the use of post column isotopic dilution analysis (IDA) or enriched protein standards are required to obtain a sufficient degree of accuracy, precision and high limits of detection. This paper reports the identification and absolute quantification of Cu,Zn-superoxide dismutase (Cu,Zn-SOD) in cytosol and mitochondria from mice hepatic cells using a innovative column switching analytical approach. The method consisted of orthogonal chromatographic systems coupled to inductively coupling plasma-mass spectrometry equipped with a octopole reaction systems (ICP-ORS-MS) and UV detectors: size exclusion fractionation (SEC) of the cytosolic and mitochondrial extracts followed by online anion exchange chromatographic (AEC) separation of Cu/Zn containing species. After purification, Cu,Zn-SOD was identified after tryptic digestion by molecular mass spectrometry (MS). The MS/MS spectrum of a doubly charged peptide was used to obtain the sequence of the protein using the MASCOT searching engine. This optimized methodology reduces the time of analysis and avoids the use of sample preconcentration and clean-up procedures, such as cut-off centrifuged filters, solid phase extraction (SPE), precipitation procedures, off-line fractions insolates, etc. In this sense, the method is robust, reliable and fast with typical chromatographic run time less than 20 min. Precision in terms of relative standard deviation (n = 5) is of 3–5% and detection limits is 0.21 ng Cu g⁻¹.     

Identification and quantification of metallothionein isoforms and superoxide dismutase in spiked liver extracts using HPLC-ESI-MS offline coupling and HPLC-ICP-MS online coupling

A two-dimensional chromatographic method for the characterization of metallothionein isoforms (MT) and superoxide dismutase (SOD) in spiked liver extracts was developed for the optimization of extraction procedures from liver samples. Element-specific detection (ICP-MS) and molecule-specific detection (ESI-MS) were applied for maximum species information. A special focus was laid on the quantitative data evaluation (species stoichiometry, calibration with and without matrix, recovery), which is neglected in most MT/SOD publications with hyphenated techniques. Linearity, precision (residual standard deviation of calibration curves <10%), and detection limits (<0.6 mg L(-1) for MT isoforms and 13 mg L(-1) for SOD) prove the suitability of the method for quantification. An alternative quantification is proposed for the extension towards other lesser or even unknown trace element species, especially the native porcine MT and SOD.     

色谱法中的谱带和色谱峰

对色谱法中的谱带作了具体的定义。详细论述了谱带与色谱峰之间的关系,指出分析谱带是研究色谱现象的基础。色谱法中的色谱峰并不局限于色谱图中的色谱峰,只要是组分的分析谱带,都可以形成一个色谱峰与之相对应。     

半制备型柱与饼性能的比较

 通过比较具有相同柱几何体积的半制备柱和饼的性能 ,发现在同样的条件下 ,用液相方法分离生物大分子时 ,在分离效率和蛋白活性回收方面 ,饼和柱的差异不大 ;由于饼中的填料较柱中的装填更为紧密和均匀 ,当流动相流速增加时 ,饼不仅反向压力上升慢 ,而且质量和体积负载也比柱高。该结果表明使用饼的分离不仅具有可与高效液相 (HPLC)媲美的分离效率 ,而且还具有中、低压分离的高柱负载 ,这有利于蛋白保持高的生物活性 ,为饼的工业化应用奠定了基础。     

Correction of retention time shifts for chromatographic fingerprints of herbal medicines

In this study, the combination of chemometric resolution and cubic spline data interpolation was investigated as a method to correct the retention time shifts for chromatographic fingerprints of herbal medicines obtained by high-performance liquid chromatography-diode array detection (HPLC-DAD). With the help of the resolution approaches in chemometrics, it was easy to identify the purity of chromatographic peak clusters and then resolve the two-dimensional response matrix into chromatograms and spectra of pure chemical components so as to select multiple mark compounds involved in chromatographic fingerprints. With these mark components determined, the retention time shifts of chromatographic fingerprints might be then corrected effectively. After this correction, the cubic spline interpolation technique was then used to reconstruct new chromatographic fingerprints. The results in this work showed that, the purity identification of the chromatographic peak clusters together with the resolution of overlapping peaks into pure chromatograms and spectra by means of chemometric approaches could provide the sufficient chromatographic and spectral information for selecting multiple mark compounds to correct the retention time shifts. The cubic spline data interpolation technique was user-friendly to the reconstruction of new chromatographic fingerprints with correction. The successful application to the simulated and real chromatographic fingerprints of two Cortex cinnamomi, fifty Rhizoma chuanxiong, ten Radix angelicae and seventeen Herba menthae samples from different sources demonstrated the reliability and applicability of the approach investigated in this work. Pattern recognition based on principal component analysis for identifying inhomogenity in chromatographic fingerprints from real herbal medicines could further interpret it.     

Information theory applied to chromatographic fingerprint of herbal medicine for quality control

At present, the construction of chromatographic fingerprints plays an important role in the quality control of complex herbal medicines. In this work, information theory was applied to obtain chromatographic fingerprints with good performance. Moreover, according to the characteristics of the chromatographic fingerprints obtained, some modifications of the calculation of the information content were conducted. In comparison with the information content from several chromatographic fingerprints obtained, reliable chromatographic fingerprints with a high separation degree and uniform concentration distribution of chemical components could be determined. The successful application of information theory with modification to simulated chromatographic fingerprints together with real herbal medicines such as Rhizoma chuanxiong and Ginkgo biloba from different sources demonstrated clearly that the proposed method to determine chromatographic fingerprints was reasonable and reliable and it was user-friendly. Chromatographic fingerprints determined with high separation degrees and uniform concentration distribution of chemical ingredients might also chemically represent characteristic components of herbal medicines for quality control.     

一种新型核壳型色谱固定相在高效液相色谱中的评价及应用

采用扫描电镜、透射电镜和氮气吸附法对制备的新型核壳型色谱固定相进行了表征,结果表明该固定相单分散性好、表面放射状孔道壳层结构均一、孔径分布窄。对该核壳材料的表面进行C18键合修饰,考察其基本色谱性能,色谱柱的理论塔板数超过150000块/m,色谱峰峰形对称,甲苯与乙苯的保留因子之比为1.45,亚甲基选择性优异。将该核壳材料应用于汽车尾气中醛酮类化合物的检测,在优化的色谱条件下,2,4-二硝基苯肼（DNPH）衍生的醛酮类化合物在15 min内获得了较好的分离效果。该核壳型C18色谱固定相具有分离速度快、选择性好、柱效高等特点,适于复杂样品的高效、快速分离分析。