摘要
系统建立了化学合成小分子肽(O8肽)的反相HPLC分离分析方法,考察了色谱填料种类、流动相组成及梯度条件对样品分离的影响。通过从分析型到半制备型分离线性放大过程中分离条件与分离效果的关系的探讨,建立了等度洗脱条件下较大规模地分离制备该小肽的方法。试验结果表明:硅胶基质的色谱柱比聚合物基质的色谱柱更适合于分离该小肽,而粒径(5μm,10μm)对分离的影响不大;较缓的梯度条件可以明显改善分离,但所需分离时间延长。在制备分离中应综合考虑分离效果与运行时间的关系。
A reversed_phase HPLC method for the purification and determination of a small synthetic pepˉtide(O8peptide)is reported.The effects of the stationary phase,particle size,mobile phase composition and gradient program on sample separation were investigated.In order to scale up to a semi_preparative scale,the performance of this separation method under various experimental conditions has been exˉplored.Under isocratic elution conditions,the scaled_up separation of this O8peptide was achieved.Our experimental results indicated that silica_based column is more suitable than polymer_based column for the separation of this peptide,whereas the particle size of adsorbents has little influence on separation.The flatter the gradient is,the greater the resolution is,but also the longer the separation time should be.Consequently,a compromise should be made between high resolution and reasonable separation time.
出处
《分析测试学报》
CAS
CSCD
北大核心
2004年第4期37-39,共3页
Journal of Instrumental Analysis
