Chromatography Forum

Hi Everyone,

I am using Agilent 1200 with openLAB CDS for the analysis that I am doing. I am plotting a standard calibration curve but I observed that the peak area does not increase as the concentration increases. The peak area provided by the integration output does not follow a defined order. Does anyone know how to solve this problem? I hope to hear from someone soon.

Sounds like you might be loading the same level instead of different levels (e.g. 1-5) to make up a calibration table. Check your individual data files and make sure you are loading the correct signal (e.g. specific single wavelength) for each std.

I am not loadin the same signal. I always check out the integration output and then plot the graph in excel

Please provide the detailed method conditions, column dimensions, detector setting, Rt's, and peak area values for all of the stds. No way to help unless you provide more detailed information.

I have observed that the integration was going from base to valley and valley to valley because the analyte and internal standard was not resolved. I have sorted that out and it should be working properly now. Thank you for the response.

On another note, does anyone know any free software that I can use to analyse my data for HPLC method valideation?

Glad you solved it. Poor quality integration settings are something that we would never have been able to identify (without seeing) or help you with over the web. These forums have limited use in troubleshooting problems that could be easily solved in seconds/minutes on-site. If there is a chromatographer at your location then you will get the quickest and maybe the best responses to these types of basic questions working with them.

Hi Itsme,

Not sure what you exactly are needing with respect to HPLC data and analytical method validation software. What are you interested in analyzing?


You can find some examples of software ResearchGate...not all of these are Free, I'd say.