《色谱柱分类与选择-内部培训资料》.ppt

* * Now that we have seen how one can measure the selectivity difference between different chromatogaphic conditions, we will use this technique to measure the selectivity difference between different solvents. Then we will look into the same parameter for column chemistry and pH. * * * * Now that we have seen how one can measure the selectivity difference between different chromatogaphic conditions, we will use this technique to measure the selectivity difference between different solvents. Then we will look into the same parameter for column chemistry and pH. * * A change of the mobile phase pH affects only analytes with ionizable functional groups, such as amines, carboxylic acids or phenols. The good news is that most pharmaceuticals as well as their metabolites contain ionic functions. This means that the retention pattern can be influenced by pH changes. pH changes have a strong effect on retention. The retention factor may change by a factor of 10, sometimes up to 30 between the ionized form and the non-ionized form of an analyte. More importantly, strong selectivity effects are caused by changes in the pH. In the next few slides, we will examine this in more detail. * * To summarize the changes caused by changes in pH: Both the retention and the selectivity of a separation are influenced by pH. One can get a drastic change in the retention factor going from the non-ionic form to the ionic form of the analyte. Typical values are around 10-fold, but a 30 fold change in retention is not unusual. This change is equivalent to a change in solvent composition of around 20%. Without question, a pH change results in the largest changes in the selectivity of a separation. The conclusion from all of this is that pH is the most powerful tool in methods development. * * As stated on the slide….. * * 5min内，5%ACN-95%ACN，1，4ML/MIN 10MM 甲酸胺 PH=3.0,醋酸胺7.0，碳酸氢胺10.0 With the tools that I have outlined, how do we complete this work? We recommend the steps outlined in this flo