三种混合模式色谱填料色谱性能的比较

以γ-缩水甘油醚氧丙基三甲氧基硅烷为中间活化剂,将3-氨基吡啶、2-巯基苯并咪唑和2-巯基-1-甲基咪唑3种杂环配基键合到粒径为5μm、孔径为30 nm的球形硅胶上,制备3种高效疏水电荷诱导色谱填料。利用溶菌酶、牛血清白蛋白和免疫球蛋白G作为模型蛋白,考察其在3种不同配基色谱柱上的保留行为和分离行为。比较了3种不同性质的模型蛋白在不同色谱柱上的色谱性能的异同。发现以2-巯基-1-甲基咪唑为配基的色谱柱具有良好的p H控制色谱行为和良好的分离性能,初步验证了高效疏水电荷诱导色谱的分离模式及潜在应用。     

疏水性电荷诱导色谱分离抗HER2单克隆抗体

采用疏水性电荷诱导色谱（HCIC）新型生物分离方法,从CHO细胞培养液中高效分离抗HER2单克隆抗体（单抗）。选用典型HCIC介质MEP HyperCel,考察了细胞培养液中单抗稳定性,比较了不同pH下MEP HyperCel介质对抗HER2单抗的吸附性能,发现pH6～9范围内吸附容量均较高,酸性条件下吸附容量显著下降。进一步考察了抗HER2单抗的动态吸附载量,优化了上样和洗脱条件,确定了合适的分离条件,实现了细胞培养液中高效分离单抗,纯度达到94.6%,收率约0.1 mg·（ml料液）^-1。结果表明,HCIC从哺乳动物细胞培养液中分离单抗是切实可行的,为单抗分离提供了新思路。     

一种新的生物分离方法——混合模式吸附层析

混合模式吸附层析是一种新型的生物分离方法,其功能配基兼有两种或两种以上的作用模式,主要为静电和疏水相互作用。在低盐和高盐条件下,都可以实现对目标物的有效吸附,具有明显的耐盐吸附的特性,可避免对料液的额外预处理,从而提高分离效率,减少分离成本,已获得一定的应用。本文对混合模式吸附技术的发展历程、类型及应用现状等进行综述,以推动该技术的发展和进一步应用。     

国外高效液相色谱柱填料生产技术的新发展介绍

介绍了最近几年国外在高效液相色谱填料生产方面的新发展，着重介绍３项专利技术－Ｒｘ技术，Ｓｂ技术和末端带帽技术对填料的色谱行为和性能带来的革命性变化，以及１９９２年发展起来的专门分离手性体的填料。     

银杏萜内酯在反相高效液相色谱中保留行为

以水 -甲醇为流动相和InertsilODS3为固定相 ,系统地研究了银杏萜内酯的反相高效液相色谱上保留行为。研究表明 ,银杏萜内酯容量因子的对数和流动相中甲醇含量呈线形关系 ,其相关系数γ大于 0 .965。     

三种卡马西平血药浓度测定方法比较

探讨3种卡马西平血药浓度测定方法的可行性与测定结果的一致性。分别使用普通高效液相色谱法(HPLC)、高效液相色谱在线固相萃取法(HPLC-on line SPE)、高效液相色谱串联质谱法(HPLC-MS/MS)3种分析方法对46例临床患者血清样本进行检测,并使用SPSS 16.0对检测结果进行统计学分析。3种测定方法,卡马西平在0.5～32.0 mg/L范围内线性关系均良好(R²>0.99),平均回收率均在91.6%～102.2%之间,精密度均小于5%,显著性P=0.446。3种方法均满足卡马西平血药浓度的测定,且3种测定方法的检测结果无显著性差异。     

制备型高效液相色谱分离过程的放大研究（Ｉ）填料尺寸

利用制备型高效液相色谱（ＨＰＬＣ）分离装置,对牛血清白蛋白（ＢＡＳ）和牛血清红蛋白（ＨＧ）进行了制备分离,考察了填料尺寸对柱效和分离度的影响,结果表明,在制备分离过程中,对于难分离物系,可以采用较小直径的色谱填料,以提高分离效率,但在分离度可以满足分离要求的前提下,使用较大直径的色谱填料将更为有利。     

疏水性电荷诱导层析——原理、性能及其应用

介绍了疏水性电荷诱导层析(hydrophobic charge induction chromatography,HCIC)的原理及特点:其功能基团结合了疏水作用和静电相互作用,是一种混合型的双模式层析方法。由于HCIC在蛋白质(特别是抗体)的分离纯化中表现出操作简便、选择性较好等优点,受到了学术界和工业界的关注。对HCIC的技术原理、发展历程、层析行为以及应用现状等进行综述,以期对HCIC的深入研究和应用开发提供指导。     

邻、间、对氨基酚的高效液相色谱分离及间氨基酚的定量测定

<正> 邻氨基酚和对氨基酚是染料和制药工业的重要中间体,间氨基酚用作抗氧剂、显影剂及毛皮染色剂,还用于制备抗结核药对氨基水杨酸。日本国家标准采用气相色谱法测定异构体杂质的含量作为化学法纯度测定的补充。采用高效液相色谱方法则有反相色谱、氰基键合相色谱及吸附色谱等多种体系,但     

生 化 分 离 技 术──制备型高效液相色谱

制备型高效液相色谱（ＨＰＬＣ）是在传统的分析型ＨＰＬＣ的基础上发展起来的一种高效分离纯化技术，主要用于制备高纯度生物活性物质。本文介绍了制备型ＨＰＬＣ的装置组成、制备规模及应用实例。     

亲和层析介质非特异性吸附的定量表征研究

针对亲和层析介质的非特异性吸附,基于脉冲响应法,建立了定量表征方法。以牛血清白蛋白、酵母发酵液和CHO细胞培养液作为典型杂质,考察了四种蛋白A亲和层析介质和两种基质微球,在不同pH和盐浓度条件下非特异性吸附。发现介质和基质对杂质均有不同程度的吸附,在酸性条件下非特异性吸附相对较强。对于不同料液,存在不同的杂质吸附机制,可通过静电、疏水作用或两者共同作用结合。两种介质和基质的比较结果表明,琼脂糖基介质的非特异性吸附主要依赖其基质微球与杂质间相互作用,而聚甲基丙烯酸酯基介质对杂质的吸附作用强于其基质微球。结果表明,本文建立的非特异性吸附定量表征方法切实可行,可用于量化表征介质非特异性吸附,探究相关作用机制,为介质研发提供新的分析方法和依据。     

Molecularly imprinted monolithic material for the extraction of three organic acids from Salicornia herbacea L

A molecularly imprinted monolithic material was designed and prepared by in situ thermally initiated copolymerization for the extraction of protocatechuic acid, caffeic acid, and ferulic acid from Salicornia herbacea L. Field emission scanning electron microscopy and offline solid‐phase extraction (SPE) were investigated for the characterization of this material. The extract samples were loaded onto and passed through the monolithic material; the target compounds were selectively retained on the material, whereas other interfering substances were quickly washed out. Chromatographic analysis was conducted on a C₁₈ column with ultraviolet detection at 270 nm, and an eluting solution consisting of acetonitrile, water, and acetic acid (14/86/0.5 v/v/v, pH 5.0) was used as the mobile phase at a flow rate of 0.8 mL/min. The linearity was confirmed in the concentration ranges of 0.10–200.00, 0.20–400.00, and 0.30–600.00 μg/mL for protocatechuic acid, caffeic acid, and ferulic acid, respectively, with r² greater than 0.9997. The SPE recoveries of the three organic acids ranged from 71.08 to 81.02%, and the coefficient of variation (precision) was 3.08–5.70%. This method is simple, economical, and specific and has been used successfully in the extraction of three organic acids from S. herbacea L. This cartridge can be used as a potential tool for the extraction of drugs from natural plants. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

氯吡脲的高效液相色谱分析

建立了采用高效液相色谱法,经C18不锈钢柱,以甲醇∶水=60∶40为流动相,检测波长为265nm,对氯吡脲进行分析,该方法的线性相关系数为0.9998%,RSD=1.0953%,回收率范围94.59～101.63%,平均值98.11%,精密度1.23%,该方法操作简单,分析速度快,毒性小,可操作性强,适合于水果中氯吡脲的残留分析。     

Characterization of ethylene‐propylene copolymers with high‐temperature gradient adsorption liquid chromatography and CRYSTAF

Blends of linear polyethylene (PE) and isotactic polypropylene (iPP) with different average molar masses and a series of ethylene‐propylene (EP) copolymers with different chemical composition as well as blends of PE, Ipp, and EP copolymers were separated using a carbon‐column packing (Hypercarb®) and gradients of 1‐decanol or 2‐ethyl‐1‐hexanol → 1,2,4‐trichlorobenzene (TCB). The separation is based on full adsorption of linear PE on the carbon sorbent at temperature 160°C. However, iPP is not adsorbed and elutes in size exclusion mode. The random EP copolymers have been adsorbed in the column packing and separated according to their average chemical composition after application of the gradient starting with alcohol and ending with pure TCB. The elution volumes of the copolymers depended linearly on the average concentration of ethylene in the copolymers. The HPLC elution profiles were correlated with the CRYSTAF elution profiles. In contrast to CRYSTAF, fully amorphous polyolefin samples were separated with the high‐temperature adsorption liquid chromatography. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Separation of polyolefins based on comonomer content using high‐temperature gradient adsorption liquid chromatography with a graphitic carbon column

This report describes the application of a recently developed polyolefin characterization tool based upon gradient adsorption high‐temperature liquid chromatography (HT‐LC) using a graphitic carbon stationary phase to polyolefin homopolymer and previously unreported copolymer systems. Polyolefin‐based materials find utility in a broad range of applications and are differentiated by parameters such as molecular weight and comonomer content. Polymer comonomer distribution is commonly determined by crystallinity‐based separations (ATREF, CRYSTAF). These techniques, however, are time consuming. In addition, some semicrystalline polymers undergo cocrystallization, impacting the techniques' universal utility. Adsorption‐based HT‐LC can ideally overcome the limitations of crystallinity‐based separations, shedding new light on the composition of randomly‐polymerized polyolefins. In this report the basic separation capability of the adsorption HT‐LC technique, using a graphitic carbon column, is demonstrated for poly (ethylene‐co‐octene) and poly(ethylene‐co‐propylene) systems and compared with select precipitation/redissolution HT‐LC and ATREF results. Select results in this paper are also compared and contrasted to other recent publications on similar separations of polyolefins. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

高效液相色谱法测定化妆品中的肌肽含量

建立了高效液相色谱—二极管阵列检测器(HPLC-DAD)测定化妆品中肌肽含量的方法。样品在破乳剂存在的条件下,用纯水经超声波提取30 m i n,在3 000 r/m i n条件下离心1 0 m in,采用Prevail~(TM)Carbohydrate ES色谱柱(250 mm×4.6 mm,5μm),以甲醇与乙酸水溶液(pH 3.5)作为流动相,以1.0 mL/min进行等度洗脱,检测波长为210 nm,柱温25℃。结果显示,肌肽在0.2~20.0 mg/L呈良好的线性关系,相关系数为0.999 9,保留时间和峰面积的相对标准偏差均小于0.5%,加标回收率为93.33%~108.64%,检出限为0.02mg/L,定量限为0.07mg/L。