HPLC troubleshooting and method optimization


Everybody who ever worked in analytics knows that the work doesn’t always run smoothly. Problems often appear when you need them the least. Even excellently maintained devices can suddenly show changes. For analytical systems like a HPLC, that means: Always schedule more time than necessary.

A second guiding principle could be: You never stop learning in analytics! The plethora of “troubleshooting guides”, tips and tricks and comments on the internet or in books might indicate that there is a solution for every problem. In reality, this might often be a different case: There can be different causes for an occurring problem, for example a changed peak form, fluctuations in pressure, bad isolation or altered retention times – or an inconvenient combination of different causes. Inversely, it can happen that the same cause can lead to different, simultaneously occurring problems. In both cases you need to find the cause(s) first. Only then you can go on to solve the problem.

Experience and the resulting insider knowledge are often missing during troubleshooting and subsequent problem resolution. The so-called “troubleshooting guides” often point in the right direction. But in most cases, these guides stop right at the point where it gets important. The reason for this is, on the hand, that every HPLC system has to be viewed individually, while a general “troubleshooting guide” can only look at the HPLC device in a general way. On the other hand, details in the implementation or the hardware are seen as unimportant or basics are seen as self-evident and aren’t included in troubleshooting. However, a systematic, device-specific approach is necessary for HPLC troubleshooting to find the cause(s) and solve the problem.

Does the HPLC device run smoothly, but you still feel like you don’t have enough time? Maybe you have an especially long analysis time – this might be necessary in some cases, for example if you have to comply with a standardized method description or if you have to isolate hundred different substances. Maybe you just have adopted an old method from your colleagues that had been established at a 20 year old HPLC device. In such cases, you should think about method optimization. Maybe you have newer hardware and a better column – then the analysis time can be shortened with just a few steps. This saves you time and money for solvents.