植物草乌毒蛋白的分离及性质

介绍草乌毒蛋白提取方法。草乌根经ＰＨ７．２５ ＰＢＳ溶液（含９ｇ＝／ＬＮａＣｌ）浸提,浸提液经ＣＭ－ＳＦＦ柱和ＳｅｐｈａｃｒｙｌＳ－２００凝胶过滤柱分离,然后在高效凝胶过滤柱上制德ＡＣＯ毒蛋白,利用光电二极管列检测器的光谱性能确认色谱峰的纯度,并根据标准蛋白的相对分子质量校正曲线求得ＡＣＯ的相对分子质量,用柱后衍生荧光法测定了其氨基酸组成。     

凤凰木种子毒蛋白的高效凝胶过滤色谱分析

介绍用ＨＰＧＦＣ（高效凝胶过滤色谱）对凤凰木种子中提取的３种毒蛋白分别进行色谱分析。并对分离的蛋白峰进行紫外光谱扫描来确认蛋白的纯度。根椐标准相对分子质量曲线,分别得到它们的相对分子质量并与ＳＤＳ－ＰＡＧＦ（十二烷基磺酸钠－聚丙烯酰胺凝胶电泳）所得结果进行比较,用柱后衍生法测定了３个蛋白各自的氨基酸组成。     

重组人干扰素—γ的凝胶色谱分离及其变性剂对分离结果的影响

使用十二烷基硫酸钠，盐酸胍和尿素三种不同的变性剂溶解基因重组人干扰素－Ｇａｍｍａ（ｒ－ＩＦＮ－γ）包含体，然后进行凝胶色谱分离。使用活性，电泳和高效疏水作用色谱三种方法，比较了三种变性剂对分离结果的影响，结果为：尿素溶解包含体后ｒ－ＩＦＮ－γ的分子量与杂质相差较大，盐酸胍溶解液经分离后ｒ－ＩＦＮ－γ比活最高，ＳＤＳ对ｒ－ＩＦＮ－γ的构象影响较大。     

高效凝胶过滤色谱法（光电二极管阵列检测器）分离、测定巴豆毒蛋白

巴豆毒蛋白粗提液在高效蛋白柱上分离，用光电二极管阵列检测器给出光谱信号，判断所需蛋白的峰，并利用光谱图和反相高效液相色谱来确认分离峰的纯度；在高效蛋白柱上测定了巴豆毒蛋白的分子量；利用离子交个邻苯二甲醛柱后衍生法测定了巴豆毒蛋白的氨基酸含量。     

高效凝胶过滤色谱法（光电二极管阵列检测器）分离、测定巴豆毒蛋白

 巴豆毒蛋白粗提液在高效蛋白柱上分离，用光电二极管阵列检测器给出光谱信号，判断所需蛋白的峰，并利用光谱图和反相高效液相色谱来确认分离峰的纯度；在高效蛋白柱上测定了巴豆毒蛋白的分子量；利用离子交个邻苯二甲醛柱后衍生法测定了巴豆毒蛋白的氨基酸含量。     

混合烷基开管电色谱柱的制备和评价

利用溶胶－凝胶（Sol-Gel）技术制备了混合烷基开管毛细管电色谱柱（C8－C13OT－CEC），并考察了其电渗流行为和电色谱性能。研究了流动相中甲醇含量对芳香族中性化合物保留的影响。发现C8－C18OT－CEC柱体现反相分配机理。5种芳香族化合物和4种苯同系物在C8－C13OT－CEC柱上分离良好，同时还考察了分离电压和柱内径对柱效的影响，结果表明高的电压和较小的柱内径能提高柱效。     

OPA柱前衍生反相高效液相色谱法测定氨基酸含量

建立了邻苯二甲醛（ＯＰＡ）手动柱前衍生反相高效液相色谱法测定样品中氨基酸含量的方法。以邻苯二甲醛（ＯＰＡ）／３－巯基丙酸（３－ＭＰＡ）为衍生试剂进行衍生,ＯＤＳ柱分离,３４０ｎｍ检测,在４０ｍｉｎ内１８种氨基酸全部得到基线分离。测定牛血清白蛋白（ＢＳＡ）的氨基酸组成和小鼠血清中的游离氨基酸,取得满意的结果。     

高效液相色谱法测定柴油族组成

在银型磺酸键合硅胶柱上以含苯或环己烯的正己烷为流动相,实现了饱和烃、烯烃的基线分离,以改进的迁移丝式氢火焰离子化检测器（ＭＷ－ＦＩＤ）进行定量检测,同时考察了Ａｇ－ＳＣＸ柱的使用性能及改进的ＭＷ－ＦＩＤ的定量准确性。在此基础上建立了高效液相色谱体系分离柴油族组成（饱和烃、烯烃、芳烃及胶质）。     

高效液相色谱非多孔型阴离子交换柱分析DNA片段

建立了高效液相色谱非多孔型阴离子交换柱分离ＤＮＡ片段的方法,用ＴＳＫｇｅｌＤＥＡＥ－ＮＰＲ柱、Ｔｒｉｓ－ＨＣｌ缓冲液（ｐＨ９．０）、１．０ｍｏｌ／ＬＮａＣｌ多阶式线性梯度洗脱,分析了核酸分子量参照物ｐＢＲ３２２ＤＮＡ－ＨａｅⅢ,ＬａｍｂｄａＤＮＡ－ＨｉｎｄⅢ及乙肝病毒基因ＰＣＲ产物,探讨了梯度、流速对分离的影响,方法简便、快速、分辨率高     

溶胶-凝胶法制备毛细管硅胶整体柱的研究进展

毛细管硅胶整体柱作为一种新型的分离介质,在色谱领域显示出了强大的生命力。本评述介绍了溶胶-凝胶法制备毛细管硅胶整体柱的方法,重点分析了溶胶-凝胶法制备毛细管硅胶整体柱的影响因素,总结了近几年毛细管硅胶整体柱在高效液相色谱和电色谱中的应用。     

高效凝胶过滤色谱法分离测定豆薯种子蛋白

用高效凝胶蛋白往分离豆薯种子蛋白租提液,结合光电二极管阵列检测器对分离的蛋白峰进行紫外光谱扫描来确认蛋白的纯度,测定了3种蛋白的分子量,采用邻苯二甲醛（OPA）柱后衍生法测定了豆薯种子蛋白的氨基酸含量。     

Liquid chromatographic retention and separation of phenols and related aromatic compounds on reversed phase columns

Summary Isocratic column liquid chromatographic systems with UV absorbance detection at 280 nm have been developed for the separation of 29 phenolics and related compounds.The selectivity was investigated on silica-, carbon- and polymer-based separation columns for the separation of phenolic type of components. The effects of various acetonitrile/buffer mixtures, and pH of the mobile phase, and their impact on the retention of the phenols was assessed. Tables of retention times on the four columns for the 29 phenols with two different acetonitrile/buffer mixtures, together with the retention times at three pHs from 6.5 to 2.3 with varying levels of organic modifier on the LiChrospher RP 18 column are presented.As an application, the analysis of real river water samples from the Ebro river is described using a solid phase extraction step prior to injection into the chromatographic system.     

高效液相色谱法手性拆分(2-戊基-3-苯基-2,3环氧丙烷基)二苯基磷酸酯对映体

采用多糖类手性色谱柱,建立了(2-戊基-3-苯基-2,3环氧丙烷基)二苯基磷酸酯对映体的高效液相色谱手性拆分方法。考察了手性柱类型、流动相组成、流速、柱温等对手性拆分的影响,并对分离机制进行了探讨。结果表明,采用Chiralpak AS-H柱(250×4.6mm,i.d.,5μm),以正己烷-异丙醇(85∶15,V/V)为流动相,在柱温25℃,流速1.0mL/min,检测波长210nm的条件下,(2-戊基-3-苯基-2,3环氧丙烷基)二苯基磷酸酯对映体能达到完全分离,且稳定性和重复性好。该方法也适用于(2-戊基-3-苯基-2,3环氧丙烷基)二苯基磷酸酯类似物的手性拆分。     

灵芝发酵液中蛋白酶抑制剂GLPIA2的纯化及其特性

采用乙醇分级沉淀、凝胶色谱纯化、阴离子交换色谱分离等步骤从灵芝深层发酵液中提取得到蛋白酶抑制剂GLPIA1 与GLPIA2。其中GLPIA2仅在215 nm处有紫外吸收,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳鉴定为单一条带,相对分子质 量为15000。由其氨基酸组成分析谱图可看出,其酸性氨基酸含量较高,碱性氨基酸及芳香族氨基酸含量较低。GLPIA2抑 制剂的底物特异性研究表明,它对天冬氨酸族的胃蛋白酶和酵母蛋白酶A有相对较强的抑制作用。     

Enantiospecific high-performance liquid chromatographic assay with fluorescence detection for the monoamine oxidase inhibitor tranylcypromine and its applicability in pharmacokinetic studies.

In order to be able to measure low concentrations of tranylcypromine enantiomers in biological material, chiral fluorescent derivatization and high-performance liquid chromatography (HPLC) were employed. The internal standard S-(+)-amphetamine and borate-sodium hydroxide buffer pH 11 were added to plasma or urine sample aliquots. o-Phthaldialdehyde was used for precolumn derivatization in combination with the chiral mercaptan N-acetylcysteine. HPLC resolution of the diastereoisomeric derivatives was possible on an octadecylsilane column. The mobile phase consisted of sodium phosphate buffer solution pH 6.5, methanol and tetrahydrofuran. The fluorescence of the eluate was monitored at 344/442 nm. The intra-day coefficients of variation were below 10%, the limit of determination was 0.5 ng/ml. The assay was found to be applicable for routine analyses in a preliminary pharmacokinetic study, in which an oral dose of 20 mg racemic tranylcypromine sulfate was administered to three healthy volunteers. The plasma concentrations were generally low, and those of S-(-)-tranylcypromine significantly exceeded those of the R-(+)-enantiomer. Average maximum concentrations were 57.5 and 6.3 ng/ml for S- and R-tranylcypromine, respectively. While S-tranylcypromine was well detectable within the whole study period (8 h), R-tranylcypromine concentrations fell below the detection limit after 4 h in two out of the three studied volunteers.     

The Determination of Folic Acid (Pteroylmonoglutamic Acid) in Fortified Products by Reversed Phase High Pressure Liquid Chromatography

Abstract

A reversed phase HPLC method was developed for the separation and determination of pteroylglutamic acid (PGA) in fortified foods. Extraction was carried out by heating with phosphate-citrate buffer, pH 8.0 containing ascorbate, and incubation with papain at 40°C for 4 hrs. The extracts were purified and concentrated on a short DEAE column which was rinsed with phosphate buffer, pH 7.0, of increasing molarity. PGA was eluted with 0.1M phosphate buffer, pH 7.0, containing 0.5M NaCl. The eluants were chromatographed on a Spherisorb ODS 10 μm column (250 × 4.6 mm) using a 30 min linear gradient of 2% to 30% acetonitrile in 0.1M acetate buffer, pH 4.0, at 1 ml/min and an absorbance detector at 280 nm. The coefficients of variation on analysis of 8 replicate samples of a milk and soy protein based infant formulas were 5.9% (at 4.6 ng/50 μl inject) and 6.8% (at 1.8 ng/50 μl inject) respectively.     

反相高效液相法同时检测3种探针药物

 用反相高效液相同时测定血清中咖啡因、氨苯砜、氯唑沙宗探针药物的质量浓度,以乙腈 磷酸盐缓冲体系(含0 02mol/L的磷酸二氢钾和0 02mol/L的三乙胺,pH6 5)(体积比为25∶75)为流动相,以安替比林为内标,经C18柱(250mm×4 6mmi d ,5 0μm)分离,紫外检测器检测,使3种探针药物得到较好的分离,并且在有效血药浓度范围内线性良好。该法简便、快速,能够为临床安全有效的用药提供科学的依据。     

Use of the simplex method to optimize the mobile phase for the micellar chromatographic separation of inorganic anions

The sequential simplex strategy has been used to optimize the mobile phase used for separation of inorganic anions by micellar chromatography on a C(18)- micro Bondapak column, with absorption detection at 230 nm. The amount of acetonitrile and the concentration of phosphate buffer (pH 6.0) were chosen for optimization. The optimum mobile phase was found to be 38% acetonitrile in 18.2 mmol L(-1) phosphate buffer (pH 6.0) containing 10 mmol L(-1) cetyltrimethylammonium bromide (CTAB); this optimum was achieved within seven experiments. The separation of the five anions (nitrite, nitrate, iodide, thiocyanate, and thiosulfate) was accomplished in 18 min.     

Simultaneous determination of caffeine, theophylline and theobromine in human plasma by on-line solid-phase extraction coupled to reversed-phase chromatography

A reversed-phase liquid chromatographic column switching system was described for the determination of caffeine (CF), theophylline (TH) and theobromine (TB) in human plasma with a direct injection procedure. A short protein-coated mu Bondapak CN silica pre-column (20 x 3 mm, i.d.) was used for enrichment of the drugs and clean up from weakly retained plasma components using phosphate buffer saline pH 7.4. After washing step, the retained drugs were flushed into a reversed-phase column (5 microm TSK gel ODS-80 TM, 150 x 4.6 mm i.d.) with a mobile phase of methanol-0.01 M phosphate buffer, pH 3.5 (30:70, v/v) for the final separation. The eluent was monitored with a UV detector at 275 nm. The resulting chromatograms showed no interference from endogenous plasma components. A linear relationship between the concentration of drug and peak height was confirmed in the range of 0.5-20 microg/mL for all drugs. High extraction recoveries from plasma ranging from 96.12 to 100.32% were achieved. Validation of the method was examined performing intra- and inter-day accuracy and precision and was found to be satisfactory. The coefficients of variation of the three drugs were less than 3% for intra-day and less than 4% for inter-day run assays.     

阴离子交换整体柱对蛋白质的分离与纯化

分别用乙二胺、二乙胺、三乙胺将自制的以甲基丙烯酸缩水甘油酯(GMA)为单体、乙二醇二甲基丙烯酸酯(EDMA)为交联剂的整体柱修饰为弱、强阴离子交换整体柱。考察了该整体柱的性能,选择出分离蛋白质(牛血清白蛋白、溶菌酶和谷胱甘肽)的最佳实验条件,并在最佳分离条件下考察了这些蛋白质在整体柱上的色谱行为和该整体柱对纤维素降解酶的分离纯化情况。实验结果表明,该整体柱性能良好,可以实现对纤维素降解酶的快速分离与纯化。同时,实验也证明采用梯度洗脱可以实现对某些蛋白质的分离纯化。