亲和色谱_硼酸亲和色谱预柱_多维液相色谱_直接进样_

硅胶基硝化苯硼酸亲和色谱材料的合成及应用

http://www.100md.com 《分析化学》 2006年第10期

亲和色谱,硼酸亲和色谱预柱,多维液相色谱,直接进样,,亲和色谱，硼酸亲和色谱预柱，多维液相色谱，直接进样,1引言,2实验部分,3结果及讨论,Refere

     摘要合成了硅胶基硝化苯硼酸亲和色谱材料，首先对间氨基苯硼酸进行硝基化，制得3氨基4硝基苯硼酸功能基团，通过 γ缩水甘油醚氧丙基三甲氧基硅烷把功能基团键合到硅胶基体上，在20.7 MPa压力下装成亲和色谱预柱(35 mm×4.6 mm i.d.)。用该预柱通过六通阀后接ODS分析柱(250 mm×4.6 mm i.d.)，构成中心切割二维HPLC。该系统能对含有顺二羟基结构的化合物进行在线富集，实现生物复杂样品直接进样分离分析。使用该系统对尿中多种修饰核苷进行了分离分析，以pH值7.95的0.25 mol/L NH4Ac 碱性缓冲液把实际尿样(100 μL)中核苷保留在预柱上，生物大分子无保留通过，再切换六通阀，以pH 4.50的25 mmol/L KH2PO4酸性洗脱液把保留在预柱上的核苷洗脱，进样到下一级ODS分析柱柱头上聚焦，然后用梯度洗脱法(pH 4.50的25 mmol/L KH2PO4 缓冲液与体积比60∶40的甲醇水梯度混合构成洗脱液)完成核苷在ODS分析柱上的分离(紫外检测260 nm)， 11种目标核苷的分离分析取得了良好的定性定量结果。

    关键词亲和色谱，硼酸亲和色谱预柱，多维液相色谱，直接进样

    Synthesis and Application of a Nitroboronic Acidsubstituated Silica for Affinity Chromatography

    Li Fanglou, Zhao Xinjie, Xu Guowang

    (National Chromatographic Resarch & Analysis Center, Dalian Institute of Chemical Physics,Chinese Academy of Sciences, Dalian 116023)

    Abstract A boronic acidsubstituted silica was synthesized by using a γglycidoxypropyltrimethoxysilane as the spacer arm and using a 3amino4nitrobenzeneboronic acid as the ligand. The 3amino4nitrobenzeneboronic acid was modified from maminobenzeneboronic acid. Then, boronic acidsubstituted silica was packed in a stainlesssteel column (35 mm×4.6 mm i.d.) under the pressure of 2.07 MPa as an affinity precolumn. A coupledcolumn liquid chromatographic method for the direct analysis of 11 urinary nucleosides is described. Efficient online cleanup and concentration of 11 nucleosides from urine samples were obtained by the affinity precolumn as the first column (extraction column) and an ODS column (250 mm×4.6 mm i.d.) as the second column (analytical column). The mobile phases applied consisted of 0.25 mol/L ammonium acetate (pH 7.95) on extraction column, and of 25 mmol/L potassium dihydrogen phosphate (pH 4.5) on analytical column, respectively. Determination of urinary nucleosides was performed on analytical column by using a linear gradient elution comprising 25 mmol/L potassium dihydrogen phosphate (pH 4.5) and methanolwater (60∶40, V/V) with UV detection at 260 nm. Calibration plots of 11 standard nucleosides show good RSD, good recovery and excellent linearity (r > 0.995). ......

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