hplc色谱柱选择与维护保养.pptVIP

The 样品溶剂 must be considered whenever you see poor 色谱峰峰形, especially if you have compounds that elute early in the run. It is possible to see 峰形前伸ing from a strong 样品溶剂. The 峰形前伸ing can look just like a fronting peak or it could possibly be a partially split peak like the one we see here, or I have seen it split more into two unresolved peaks. In this example the 样品溶剂 is 30 uL of 乙腈 in the A chromatogram. The 流动相 is only 18% 乙腈, so this is a lot stronger. The result is a split peak on the first peak. Notice the second peak as well. That peak is much broader 和tails a little. This would probably just be noticed as less 柱效 than would be expected. ? If the 样品溶剂 is the 流动相 the result is two very sharp peaks with excellent 色谱峰峰形. The 进样体积 here is 30 uL, but be aware that this can occur with even lower 进样体积s depending on elution times 和流动相 composition. ? These two things, 样品 solvent strength 和corresponding 进样体积 as well as 样品 load on the 色谱柱 are the two primary factors of 色谱峰峰形 problems. 样品 solubility, though not specifically shown here affects both of these 和therefore 色谱峰峰形. Start by dissolving your 样品 in whatever solvent it is most soluble 和then dilute to match your 流动相. Do this to obtain good solubility 和limit 色谱峰峰形 problems. 1.????? Why Use 含缓冲液的流动相? This sl内径e shows分离 of a mixture of ac内径ic和neutral 待测组份 carried out at pH 7.0. These are non-内径eal conditions f或analyzing these ac内径s, but were chosen f或demonstration purposes. The分离 on left shows the分离 using pH-adjusted水 in 流动相. This was prepared with a minimum of phosphoric ac内径和钾hydrox内径e solution. pH of deionized和organic-free水 can 改变a 批号 on a daily basis, as a function of source 或br和of purifier, 时间 of year, amount of dissolved carbon diox内径e, 等, 等 All ac内径s elute very early as they are charged和the硅胶surface is negatively charged at this pH. last two 组份 色谱峰 tail more than is desirable at this pH. This 色谱峰峰形 can be improved by 增加 缓冲液浓度 to 35-50 mM 或by adding 10 mM triethylamine to 流动相. Not only i