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Establishment of the model of vascular endothelial cell membrane chromatography and its preliminary application

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Chinese Science Bulletin 2007年第7期52卷 922-928页

作者： LI YiPing he langchong Department of Pharmacy School of Medicine Xi'an Jiaotong University Xi'an 710061 China

A model of vascular endothelial cell membrane chromatography was established by using an ECV304 cell membrane stationary phase (ECV304 CMSP) prepared by immobilizing the ECV304 cell membrane onto the surface of silica carrier. The surface and chromatographic characteristics of ECV304 CMSP were studied. The active component from Caulophyllum robustum was screened by using the model of vascular endothelial cell membrane chromatography. The interaction between the active component and membrane receptor was determined by using a replace experiments. The effect of the active component was tested by using tube formation of ECV304 cell. The results indicated that the model of ECV304 cell membrane chromatograph (ECV304 CMC) can stimulate the interaction between drug and receptor in vitro and the retention characteristics of taspine as active component was similar to that of model molecule in the model of ECV304 CMC. And therefore, taspine acted on VEGFR2 and inhibited the tube formation of ECV304 cell induced by VEGF. This model can be used to screen definite active component as a screening model.

关键词：血管内皮细胞细胞膜色谱法模型血管发生活性成分筛选应用

Establishment of the model of white blood cell membrane chromatography and screening of antagonizing TLR_4 receptor component from Atractylodes macrocephala Koidz

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Science China(Life Sciences) 2006年第2期49卷 182-189页

作者： LI Cuiqin he langchong School of Medicine Xi'an Jiaotong University Xi'an 710061 China School of Medicine Xi'an Jiaotong University Xi'an 710061 China

A model of white blood cell membrane chromatography (WB-CMC) was established to screen active component from Atractylodes macrocephala Koidz. The component can antagonize Toll-like receptor 4 (TLR4); inhibit inflammatory reaction. In the model of WB-CMC, cell membrane stationary phase (CMSP) was prepared by immobilizing the rabbit white blood cell membrane (WBCM) onto the surface of silica carrier; taxinol was used as a model molecule. The active component which can act on WBCM; its receptor (such as TLR4) as an effective target in A. macrocephala was determined by using a replacement experiment. The anti-inflammatory effects of the active component were tested by using pharmacological methods in vivo. The results indicated that the retention characteristics of atractylenolide I as active component was similar to that of taxinol in the model of WB-CMC. And so, atractylenolide I acted on the WBCM; TLR4; its anti-inflammatory activity was related with antagonizing TLR4. Therefore, the interaction between the active component; WBCM; its receptor can be simulated by the model of WB-CMC in vitro. This model can be used to screen active components; to study effective characteristics for acting on definite targets.

关键词： white blood cell membrane chromatography, TLR4 receptor, atractylenolide I, anti-inflammatory effect.

Enzymatic activity and chromatographic characteristics of the cell membrane immobilized on silica surface

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Chinese Science Bulletin 1999年第9期44卷 826-831页

作者： langchong he Guangde Yang Xindu Geng NW Univ Xian Inst Modern Separat Sci Xian 710069 Peoples R China Xian Med Univ Coll Pharm Xian 710061 Peoples R China

A new cell membrane preparation, silica carrier cell membrane (CCM), has been presented, in which cell membrane was immobilized on silica which acted as carrier by the adsorption. The surface characteristics of the silica CCM have been tested by a scanning electron microscope and surface energy spectrometer. Comparison has been made of the changeable rule of enzymatic activity of rabbit red cell membrane in three states: the silica CCM, the suspension and the precipitation cell membranes, with temperature and time. The result showed that the silica CCM still keeps the same enzymatic activity as the other two states of cell membranes. The interactions between calcium antagonists and rabbit red and cardiac muscle cell membranes, and the stereoscopic interactions between dihydropridines and rabbit cerebellar cell membrane have been investigated by the chromatographic system of the silica CCM as a stationary phase. From the foregoing results, the chromatographic parameters obtained from the system would

关键词： cell membrane preparation silica CCM enzymatic activity affinity chromatography enantiomers.

EGFR/cell membrane chromatography-online-high performance liquid chromatography/mass spectrometry method for screening EGFR antagonists from Radix Angelicae Pubescentis

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Science China Chemistry 2010年第11期53卷 2357-2362页

作者： SiCen Wang Meng Sun YanMin Zhang Jie Zhang langchong he Xi An Jiao Tong Univ Sch Med Minist Educ Key Lab Environm & Genes Related Dis Xian 710061 Peoples R China

The intracellular kinase domains of the epidermal growth factor receptor(EGFR) in some tumor cells are significant targets for drug *** have developed a new EGFR cell membrane chromatography(EGFR/CMC)-online-high performance liquid chromatography/mass spectrometry(HPLC/MS) method for screening anti-EGFR antagonists from medicinal herbs such as Radix Angelicae *** this study,the heK293 EGFR cells with high expression of EGFR were used to prepare cell membrane stationary phase(CMSP) in the EGFR/CMC *** retention fractions on the EGFR/CMC model were directly analyzed by combining a 10 port columns switcher with a HPLC/MS system *** a result,osthole from Radix Angelicae Pubescentis was found to be the active component acting on EGFR like dasatinib as the control *** was a good relationship between their inhibiting effects on EGFR secretion and heK293 EGFR cell growth in *** new EGFR/CMC-online-HPLC/MS method can be applied for screening anti-EGFR antagonists from TCMs,for instance,Radix Angelicae *** will be a useful method for drug discovery with natural medicinal herbs as a leading compound resource.

关键词： EGFR cell membrane chromatography high performance liquid chromatography/mass spectrometry anti-EGFR antagonists Radix Angelicae Pubescentis

Development of the model of pancreatic β-cell membrane chromatography and its chromatographic characteristics

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Chinese Science Bulletin 2005年第21期50卷 2427-2431页

作者： YANG Guangde he langchong BIAN Xiaoli ZHAO Liang Department of Pharmacy School of Medicine Xi'an Jiaotong University Xi'an 710061 China

A new model of pancreatic β-cell membrane chromatography has been established by using a β-cell membrane stationary phase (β-CMSP) prepared by immobi-lizing the β-cell membrane onto the surface of silica carrier. The protein level and K+, Na+-ATP enzymatic activity of the β-CMSP were detected respectively. The surface characteris-tics of the β-CMSP were tested by using the scanning elec-tron microscope and surface energy spectrometer. In this model, the column (10 mm × 2 mm, I.D.) packed with β-CMSP, 25 mmo1/L ammonium sulfate buffer solution (pH 7.4) as mobile phase with the flow rate of 0.2 mL/min at 37 ± 0.5℃ were used in the following studies. The retention char-acteristics of the sulfonylureas (gliquidone, glibenclamide, gliclazide and glipizide) were investigated under the chro-matographic conditions above. The affinities of the sulfony-lureas on β-cell membrane and receptor will be expressed by using the logk′ values (the logarithm of capacity factor of a solute) in the model. The correlation of the affinity with the pharmacological effect of the sulfonylureas was analyzed also.

关键词：胰腺细胞细胞隔膜色析法低血糖症蛋白质水平

卵类糖蛋白手性柱拆分钙离子拮抗剂尼卡地平对映体

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分析化学 2005年第12期33卷 1779-1782页

作者：傅强贺浪冲西安交通大学药学系西安交通大学药学系西安710061

研究了在卵类糖蛋白(OGCH I)手性柱上影响钙离子拮抗剂尼卡地平对映体拆分的主要因素,建立了尼卡地平对映体的拆分方法。氨丙基硅胶在0.2 mol/L磷酸钠缓冲液(pH 6.6)中与OGCH I反应,制备的固定相装填于10 cm不锈钢柱中,用高效液相色谱... 详细信息

研究了在卵类糖蛋白(OGCH I)手性柱上影响钙离子拮抗剂尼卡地平对映体拆分的主要因素,建立了尼卡地平对映体的拆分方法。氨丙基硅胶在0.2 mol/L磷酸钠缓冲液(pH 6.6)中与OGCH I反应,制备的固定相装填于10 cm不锈钢柱中,用高效液相色谱法拆分尼卡地平对映体。优化的色谱条件为:0.05 mol/L磷酸钠缓冲液(pH 5.5)-甲醇(65∶35),流速0.2 mL/m in,紫外检测波长236 nm,柱温:25℃。在此条件下,尼卡地平对映体可得到良好分离。通过改变有机改性剂的种类和比例、流动相的pH值、缓冲溶液的浓度以及流速等色谱条件,确定了影响尼卡地平对映体拆分的主要因素。

关键词：尼卡地平手性分离卵类糖蛋白高效液相色谱

一些酸性药物在正相硅胶/反相洗脱色谱中保留机理研究

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药学学报 1998年第1期33卷 42-47页

作者：贺浪冲耿信笃西安医科大学药学院西北大学现代分离研究所

用液相色谱法中“溶质计量置换保留模型”（ＳＤＭＲ）的两个线性公式，分析了正相硅胶／反相洗脱色谱体系中酸性药物分子与固定相和流动相分子间的相互作用机理，研究了流动相ｐＨ值、离子强度、温度和强溶剂浓度改变对溶质保留的影... 详细信息

用液相色谱法中“溶质计量置换保留模型”（ＳＤＭＲ）的两个线性公式，分析了正相硅胶／反相洗脱色谱体系中酸性药物分子与固定相和流动相分子间的相互作用机理，研究了流动相ｐＨ值、离子强度、温度和强溶剂浓度改变对溶质保留的影响。从而阐明了酸性药物的保留机理为一种以反相色谱为主的与正相色谱共存的混合保留机理。此方法可以作为药物制剂和中药材中酸性药物成分的分离测定方法。

关键词：酸性药物液相色谱反相洗脱分离药物成分

Determination of ribavirin by flow injection chemiluminescence

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Journal of Nanjing Medical University 2006年第5期20卷 266-270页

作者： Juqing Jin Yuhai Tang langchong he Hong Yao Yuanyuan Sun School of Science . Xi＇an Jiaotong University Xi＇an 710061 China School of Medicine Xi＇an Jiaotong University Xi＇an 710061 China Pharmacy Faculty of Fujian Medical University Fuzhou 350004 China

Objective： To establish a rapid and precise continuous flow-injection chemiluminescence （CL） method to determine ribavirin. Methods： The ribavirin could restrain strongly CL reaction of luminol in sodium hydroxide （NaOH） solution with potassium chlorate （KClO3）. The different experimental parameters affecting the CL intensity were studied carefully. Results： Under optimum conditions （The concentrations of luminol, KClO3, and NaOH were at 0.1 mmol/L, 0.5μmol/L, and 0.2 mol/L, respectively）, the linear range of the working curves was 0.01-7.00μg/ml with a detection limit of 0.004μg/ml. Conclusion： The method is simple, rapid and sensitive, and successfully applied to the determination of ribavirin in pharmaceutical preparations and biological fluids.

关键词： flow injection analysis chemiluminescence ribavirin

葛根中有效部位及有效成分的高效液相色谱分析

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西安交通大学学报（医学版） 2005年第3期26卷 216-219页

作者：朱丽华贺浪冲西安交通大学医学院天然药物研究与工程中心陕西西安710061

目的建立全面控制葛根及其制剂的定性和定量分析方法。方法用反相高效液相色谱法(RPHPLC),以甲醇水(42∶58)为流动相在250nm波长下检测,对葛根有效部位(醋酸乙酯提取部位)进行指纹图谱定性分析,对葛根有效部位中的有效成分葛根素和大豆... 详细信息

目的建立全面控制葛根及其制剂的定性和定量分析方法。方法用反相高效液相色谱法(RPHPLC),以甲醇水(42∶58)为流动相在250nm波长下检测,对葛根有效部位(醋酸乙酯提取部位)进行指纹图谱定性分析,对葛根有效部位中的有效成分葛根素和大豆苷元进行定量测定,并对11种不同来源的葛根药材进行分析测定。结果在定性分析条件下,葛根有效部位的HPLC指纹图谱中有5个共有峰,可以作为其定性鉴别的指标峰;在以葛根素和大豆苷元为定量指标分析中,其方法的回收率分别为98.0%和95.4%,相对标准偏差(RSD)分别为1.30%和1.61%,葛根药材中葛根素和大豆苷元的含量分别在0.213%～3.626%和0.012%～0.049%范围内。结论以葛根有效部位指纹图谱定性分析和有效成分定量分析方法,可以准确有效地控制其质量。

关键词：葛根有效部位有效成分 RP-HPLC 指纹图谱