精选现代色谱分析-hplc1.pptVIP

Snyder溶剂分组三角形 * 正相色谱流动相的选择 由于固定相是极性的，P值增大，洗脱能力增加，样品k值降低。 分离时，首先选择合适P值的溶剂，使样品k值在1－10。常用溶剂是在烷烃中加入一种极性较大的溶剂为极性调节剂，如在正己烷中加入氯仿。 确定极性参数P后，若分离不好，则改用其它组别的强溶剂。 * 反相色谱流动相的选择 由于固定相是非极性的，P值增大，洗脱能力降低，样品k值增大。 反相色谱一般以水作为基础溶剂，再加入一定量与水互溶的有机调节剂，常用的有甲醇、乙腈、四氢呋喃等。 为了适应不同溶质的需求，有时需加酸碱或缓冲溶液。常用的酸为乙酸、磷酸，碱用胺类，缓冲溶液常用磷酸盐或醋酸盐缓冲液。 * * * * * * * * Now lets cover some good column practices that will help you to increase your column lifetime. Filter solvents before you use them. The column is the HPLC systems best filter, so make sure the column isnt performing this task. Pretreat samples that contain strongly retained compounds of no interest. This means to remove any sample matrix, or contaminants, or incompatible analytes to the method/column. Solid phase extraction is good for this clean up. Avoid extreme column temperatures, greater than 60 degrees, or check the vendor limits. Some columns are now made for higher temperatures. Flush/clean the column often with strong solvents. Also use gradient cleaning. Keep the mobile phase pH between 3 and 7 unless otherwise indicated by the column vendor (polymeric phases). If it is necessary to do this, use a precolumn and expect a shorter lifetime. Always use fresh buffer solutions and aqueous mobile phases or treat them with sodium azide, this can be dangerous. Water solutions grow bugs or bacteria, these will contaminate and permanently modify the stationary phase. In line with the last point, prepare columns for storage by rinsing all of the buffer out of the column, then store in the appropriate solvent, at least 50% organic. This will keep bacteria from growing on the column. Make sure to rinse the buffer off first and with water, as many buffer salts precipitate in 100% organic. Then cap the column tightly to keep air out. Avoid physically mishandling the columns. This includes banging, dropping, overtightening fittings, and hitting your lab partner. * Here is a schematic illustrating the difference between a pre-column and a guard colum