反相高效液相色谱法同时测定桑叶提取物中芸香甙和槲皮素的含量

用Ｐ１００型高效液相色谱系统、ＤＬ－８００色谱工作站、ＹＷＧ－Ｃ１８色谱柱,以甲醇－水－磷酸体系（６０∶４０∶０．４,Ｖ／Ｖ／Ｖ）为流动相分离并同时测定了桑叶提取物中芸香甙与槲皮素的含量。芸香甙、槲皮素含量与峰面积呈良好的线性关系。芸香甙的变异系数为１．９８％,回收率为１０４．０％;槲皮素的变异系数为９．８％,回收率为１０３．２％。     

用高效液相色谱法测定热应激时大鼠肺及肝组织中细胞膜磷脂的变化

应用高效液相色谱法（ＨＰＬＣ）测定了正常大鼠肺及肝组织中细胞膜磷脂的含量及热应激时膜磷脂的含量变化。流动相为甲醇∶乙腈∶８５％磷酸（３∶１００∶１，Ｖ／Ｖ／Ｖ），色谱柱为μ－Ｐｏｒａｓｉｌ柱（３．９ｍｍｉ．ｄ．×３００ｍｍ）。通过测定膜磷脂的变化，可以为阐明机体的发病机理提供可靠的数据     

用高效液相色谱法测定热应激时大鼠肺及肝组织中细胞膜磷脂的变化

 应用高效液相色谱法（ＨＰＬＣ）测定了正常大鼠肺及肝组织中细胞膜磷脂的含量及热应激时膜磷脂的含量变化。流动相为甲醇∶乙腈∶８５％磷酸（３∶１００∶１，Ｖ／Ｖ／Ｖ），色谱柱为μ－Ｐｏｒａｓｉｌ柱（３．９ｍｍｉ．ｄ．×３００ｍｍ）。通过测定膜磷脂的变化，可以为阐明机体的发病机理提供可靠的数据。     

高效液相色谱法同时测定牛肉及其制品中敌草隆、绿麦隆的残留量

介绍了用高效液相色谱同时测定牛肉及其制品中脲类除草剂——敌草隆、绿麦隆残留量的方法。色谱柱为ＳｅｌｅｃｔｏｓｉｌＣ１８柱,流动相为甲醇－水（６０∶４０,Ｖ／Ｖ）,ＵＶ－２４５ｎｍ检测。最小检测量：敌草隆为０．４ｎｇ,绿麦隆为０．５ｎｇ。线性范围均为０．０５～１０ｍｇ／Ｌ。回收率：敌草隆为８７．３４％～８７．６４％,绿麦隆为８８．７８％～９１．９４％。     

反相高效液相色谱法测定人血浆中曲尼司特浓度

报道了用反相高效液相色谱法测定人血浆中曲尼司特浓度。以甲醇－磷酸二氢钾缓冲液（６０∶４０，Ｖ／Ｖ；ｐＨ４．２）为流动相，色谱柱采用ＹＷＧ－Ｃ１８不锈钢柱，紫外３３３ｎｍ波长检测，血浆样品经甲醇沉淀蛋白后直接进样测定。血浆浓度在０．６５～４０ｍｇ／Ｌ范围内线性良好，方法的平均回收率为１００．０％±４．１％。血浆最低检测浓度为０．２ｍｇ／Ｌ。对１０名健康志愿者口服２００ｍｇ曲尼司特后不同时间的血药浓度进行了测定。     

反相高效液相色谱法测定人血浆中曲尼司特浓度

 报道了用反相高效液相色谱法测定人血浆中曲尼司特浓度。以甲醇－磷酸二氢钾缓冲液（６０∶４０，Ｖ／Ｖ；ｐＨ４．２）为流动相，色谱柱采用ＹＷＧ－Ｃ１８不锈钢柱，紫外３３３ｎｍ波长检测，血浆样品经甲醇沉淀蛋白后直接进样测定。血浆浓度在０．６５～４０ｍｇ／Ｌ范围内线性良好，方法的平均回收率为１００．０％±４．１％。血浆最低检测浓度为０．２ｍｇ／Ｌ。对１０名健康志愿者口服２００ｍｇ曲尼司特后不同时间的血药浓度进行了测定。     

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。     

高效液相色谱法分析生漆多糖中的单糖组成

采用μ－ＢｏｎｄａｐａｋＮＨ２ＴＭ色谱柱,以乙腈∶水∶甲醇（７０∶２５∶５,Ｖ／Ｖ）为流动相,利用示差折光检测器的高效液相色谱法对湖北毛坝大木、毛坝小木和建始３种生漆中漆多糖的单糖组成成分进行了直接分离分析,方法简便、快速、重现性好,回收率在９８．８％～１０３．６％之间,相对标准偏差在５％以下。     

高效液相色谱法测定环境水中痕量4,4′-二氨基联苯、4-硝基酚和苯酚

采用ＰｈｅｎｏｍｅｎｅｘＳｐｈｅｒｅｘＣ１８色谱柱,以含５０ｍｍｏｌ／Ｌ乙酸－乙酸钠缓冲溶液（ｐＨ６．０）的Ｖ（乙腈）∶Ｖ（乙醚）∶Ｖ（水）＝１２∶１０∶７８混合溶液为流动相,用分光光度法检测,检测波长为２７５ｎｍ,在１６ｍｉｎ内实现了４,４′－二氨基联苯、４－硝基酚和苯酚的同时分离测定。检测限分别为０．１４,０．１９和０．０８ｎｇ。标准回收率分别为９８．８５％,９８．１５％和９８．１０％。方法灵敏度高,用于环境水样分析时结果令人满意。     

高效液相色谱法测定皮炎宁酊中间苯二酚和水杨酸的含量

采用高效液相色谱法测定了皮炎宁酊中间苯二酚和水杨酸的含量。色谱柱为ＨｙｐｅｒｓｉｌＯＤＳ柱,流动相为Ｖ（甲醇）∶Ｖ（水）∶Ｖ（乙酸）＝５０∶５０∶０．９。检测波长为２８５ｎｍ。在此条件下,间苯二酚和水杨酸可与其降解产物及杂质完全分开。     

一种规模化蛋白质组分离和鉴定新方法研究及其应用

建立了一种规模化的蛋白质组分离和鉴定新方法。通过对在生命发育过程中具有重要研究价值的人胎肝线粒体蛋白质组的分离分析，表明与毛细管液-质联用的不同分离方法的组合可以增大检测动态范围和分辨率。研究共鉴定了2977个肽段，归属于915种蛋白质。去除批次间冗余后，鉴定的蛋白质为477种，其中291种为唯一蛋白质，186种为蛋白质簇，144种蛋白质明确定位于人胎肝线粒体中。所鉴定蛋白质的分子量分布范围为7000Da～330000 Da，pI值分布在4．0～11．89，克服了两维凝胶电泳在分子量和pH方面的歧视性问题。实验中发现的蛋白质簇以及确定一种蛋白质需要最少肽段数的问题还需要进一步研究。     

TRANSFORMATION OF CULTURED HUMAN FETAL GASTRIC FIBROBLASTS WITH PHYSICO-CHEMICAL AGENTS

Treatment of early passage human fetal gastric fibrohlasts with ultraviolet (UV) light and the chemical carcinogen ethyl nitrosourea (ENU) in succession resulted in an immortally growing cell line, named GTS 8502. The cells of this line display typical transformation characteristics, such as irregularly shaped nuclei, heteroploidization of karyotype and frequent appearance of heteromorphic chromosomes, the enhanced volume ratio of nucleus to cytoplasm, multinucleoli, appearance of microvilli on the surface of the cells and agglutination reaction to lectin concanavalin A. The transformants have high growing and mitotic indices and the ability of focus-formation on monolayers and anchorage independent growth in soft agar medium. Moreover, these cells induced turnouts in nude mice or in immunosuppressed new-born rats through heterotransplantation. The results of various methods including electromicroscopy and histochemical analyses indicate that GTS 8502 cells are of fibroblast origin.Our results thus indi     

Characterization of the metabolites of rosmarinic acid in human liver microsomes using liquid chromatography combined with electrospray ionization tandem mass spectrometry

Rosmarinic acid (RA) is a phenolic acid originally isolated from the herb medicine Rosmarinus officinalis. The purpose of this study was to identify the metabolites of RA. RA was incubated with human liver microsomes in the presence of β-nicotinamide adenine dinucleotide phosphate tetrasodium salt and/or uridine diphosphate glucuronic acid using glutathione (GSH) as a trapping agent. After 60-min incubation, the samples were analyzed using high-resolution liquid chromatography tandem mass spectrometry. Under the current conditions, 14 metabolites were detected and identified. Our data revealed that RA was metabolized through the following pathways: the first pathway is the oxidation of catechol to form ortho-quinone intermediates, which react with GSH to form mono-GSH adducts (M1, M2, and M3) and bis-GSH adducts (M4 and M5); the second pathway is conjugation with glucuronide to yield acylglucuronide (M7), which further reacts with GSH to form RA-S-acyl-GSH adduct (M9); the third pathway is hydroxylation to form M10, M11, and M12, which further react with GSH to form mono-GSH adducts (M13 and M14); the fourth pathway is conjugation with GSH through Michael addition (M6); the fifth pathway is conjugation with glucuronidation, forming M8, which is the major metabolic pathway of RA.     

Copper (II)-Surfactant Complex and Its Nano Analog as Potential Antitumor Agents

The in vitro anticancer activity of copper cetyl trimethyl ammonium bromide (Cu-CTAB)-loaded cyclodextrin nanoparticles on Ehrlich ascites carcinoma (EAC), colon cancer cells (HCT 116), liver cancer cells (HepG-2), breast cancer cells (MCF-7), and cervix cancer cells (Hela) was investigated using MTT due assay. Cyclodextrin nanoparticles loaded with Cu-CTAB exerted in vitro anticancer activity against the previous human cancer cell lines comparable to the activity of free non-entrapped in nanoparticles macro-particle Cu-CTAB. The nano analog was synthesized by physical loading using grinding with ball mill. The ratio between Cu-CTAB and cyclodextrin oligosaccharide was 1 Cu-CTAB: 3 cyclodextrin. The particle size of the nano derivative was determined using the transmitted electron microscope (TEM).     

Steep pulsed electric fields modulate cell apoptosis through the change of intracellular calcium concentration

A steep electric pulsed field with low intensity (150–250 V/cm) and relative long time (10 min) was applied to adherent liver cancer cell line SMMC-7721 and the liver cell line HL-7702. Results showed that the electric field with intensity of 200 and 250 V/cm could trigger cell apoptosis, whereas the SMMC-7721 cell was more sensitive to the electric stimulation than the HL-7702 cell. Laser Scanning Confocal Microscope (LSCM) was used to measuring the real-time change of cytosolic free Ca²⁺ concentration. When cells were exposed electric pulses with 100 V/cm intensity for 10 min, there was no significant change of intracellular calcium concentration. With the intensity increased to 200 and 250 V/cm, intracellular calcium concentration decreased significantly. Results demonstrated the relationship between the apoptosis and change of intracellular calcium concentration. And the steep electric pulsed field can be used to the cancer therapy.     

System to screen and purify active ingredients from herbal medicines using hydrogel-modified human umbilical vein endothelial cell membrane chromatography coupled with semi-preparative high-performance liquid chromatography-offline-high-performance liquid chromatography-mass spectrometry

Analytical screening and validation systems based on a combination of cell membrane chromatography and two-dimensional chromatography-tandem mass spectrometry are incapable of providing prepared samples containing the active ingredients found in traditional Chinese medicine; therefore, these samples cannot be directly used in subsequent studies. In this study, a semi-preparative cell membrane chromatography column was developed using a hydrogel-modified carrier and human umbilical vein endothelial cells to optimize prepared conditions, such as hydrogel polymerization, cell fragmentation, and cell membrane volume. This increased the binding ratio of membrane protein and carrier to 15.79 mg/g. The column was systematically evaluated using multitarget tyrosine kinase inhibitors that displayed good specificity and reproducibility. Subsequently, using the column coupled with a semi-preparative high-performance liquid chromatography-offline-high-performance liquid chromatography-mass spectrometry system, 15 active ingredients were screened and purified from Indigo naturalis, and five main components were identified: l -lysine, oxyresveratrol, tryptanthrin, isorhamnetin, and indirubin. Furthermore, the pharmacological effects of the ingredients were confirmed using cell proliferation and apoptosis assays. Results revealed potent proliferation-inhibiting and apoptosis-promoting abilities on human chronic myelogenous leukemic cells and human promyelocytic leukemic cells (p < 0.001). Overall, the system presented screening and purification functions that could be used to prepare I. naturalis samples acting on the epidermal growth factor receptor and vascular endothelial cell growth factor.     

反相高效液相法测定大鼠肝组织中DNA的碱基含量

 利用反相高效液相法 ,采用SupelcosilLC 18柱 (2 5 0mm× 4 6mmi d ,5 μm) ,以甲醇 0 0 5mol/LKH2 PO4缓冲液 (体积比为 2 0∶80 )为流动相 ,流速 0 8mL/min ,在 2 5 4nm波长处检测 ,对生活在高原 (海拔 2 3km)的习服大鼠肝组织中脱氧核糖核酸 (DNA)的碱基含量进行了检测 ,发现各碱基在DNA中所占的比例是相对稳定的 :腺嘌呤 (A) 2 8 8%、鸟嘌呤 (G) 2 3 3%、胞嘧啶 (C) 17 4 %、胸腺嘧啶 (T) 2 5 3% ,并利用内标法对DNA甲基化水平进行了测定。     

Electroanalytical Method for Quantification of Hepatocellular Carcinoma Cells as Charge Transport Barriers in Culture Media

Here we report a strategy in which electroanalytical method was used to quantify human hepatic carcinoma cell (HepG2) density in culture media and their interaction with biomolecules. The cyclic voltammogram response of vitamin and amino acids redox mediators containing culture media had shown distinct oxidation peak at ?0.63 V along with a low intensity peak at +0.67 V. Both oxidation and reduction peak current of culture media were gradually decreased with an increase in cell number indicating their role as charge transport barrier at electrode surface. The difference between cathodic and anodic peak potential was also decreased with the addition of cells. The oxidation peak disappeared in CV response, with the addition of optimum cell number in culture media, indicating the adsorption of redox mediators at cell surface. CV response of fetal bovine serum (FBS) containing cell suspension showed presence of reduction and oxidation peaks of culture media in CV. This indicates stronger possibility of binding of serum proteins with cells and release of redox mediators in culture media. The chemical interactions of cells with FBS was further confirmed by the FTIR and UV‐Vis spectroscopy. The viability, adhesion and proliferation responses of the cells were found to be normal. The reported electroanalytical method may be applied in the future for rapid quantification of cell density and confirmation of interactions among cells and biomolecules.     

高效液相色谱法测定人血清中鬼臼乙叉甙浓度

建立了测定血清中鬼臼乙叉甙浓度的反相高效液相色谱方法。血清样品以乙酸乙酯提取,加表鬼臼毒噻吩糖甙作内标,以甲醇∶水（５５∶４５）为流动相,紫外２５４ｎｍ检测,最低检测浓度０．１ｍｇ／Ｌ,在药物浓度为０．５～５０ｍｇ／Ｌ范围内有良好的线性关系,Ｙ＝０．０１６０＋０．０３４０Ｘ,ｒ＝０．９９９１,回收率达９５．０％。日内、日间精密度分别为４．１６％和４．７６％（ｎ＝５）。用所建立的方法测定了患者的血清浓度,结果表明,方法简便、灵敏。     

高效液相色谱法测定人血浆中的芦氟沙星

 建立了测定人血浆中芦氟沙星质量浓度的高效液相色谱法 ,血浆用二氯甲烷提取 3次 ,以 UltrasphereODS(4.6mm i.d.× 2 5 0 mm)为色谱柱 ,流动相为甲醇 -1 0 mmol/L 溴化四丁铵 -三乙胺 (体积比为 3 2∶ 68∶0 .5 ) ,用磷酸调 p H2 .8,检测波长 2 95 nm,流速为 1 .2 m L/min,以培氟沙星为内标。血浆中芦氟沙星的线性范围为 0 .1～ 1 0 mg/L ,最低检测质量浓度为 0 .0 5 mg/L ,回收率为 99.7% ,日内、日间 RSD分别为 2 .3 3 %和3 .83 %。方法简便、快速、准确 ,适用于人血浆中芦氟沙星质量浓度的测定。