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研究了在反相高效液相色谱中的烷基醇-甲酸-水、烷基醇-三氟醋酸-水和烷基醇-磷酸盐-水三种流动相,9种生物大分子的分离体系中Z的表征。发现当蛋白质完全变性时,Z值与其分子量呈正比,而与置换剂分子大小呈反比。Z值会因流动相中离子对试剂的存在而减小,且浓度愈大,减小程度愈甚。对于在异丙醇-水溶液中的不同种类离子对试剂而言,对蛋白构象变性影响的减小顺序为44％(V/V)甲酸>0.05mol/L磷酸盐缓冲液(pH<2.5)>0.1％(V/V)三氟醋酸。
The separation system of three kinds of biomacromolecules, alkyl alcohol - formic acid - water, alkyl alcohol - trifluoroacetic acid - water and alkyl alcohol - phosphate - water three mobile phases in reversed phase high performance liquid chromatography Characterization of Z. It was found that when the protein is completely denatured, the Z value is proportional to its molecular weight and inversely proportional to the size of the displacer molecule. The Z value decreases due to the presence of ion-pair reagents in the mobile phase, and the greater the concentration, the greater the reduction. For the different kinds of ion-pairing reagents in isopropanol-water solution, the reduction in protein conformational degeneration was in the order of 44% (V / V) Formic acid> 0.05 mol / L Phosphate buffer 0.1% (V / V) trifluoroacetic acid.