Dear Collegues,
I'm starting a new project and I need to know what is the maximum concentration for a sample that have to be processed by analytical HPLC.
Thank you so much for the answers!
Luca
Jack has provided an excellent response. Maximum sample concentration is not determined by "HPLC". HPLC is the name of an analytical technique. The specific method(s) used and system configuration will determine any maximums, as Jack noted. *Besides, samples can be diluted, and this is why a scientific HPLC sample "Loading Study" is run on the selected method (the results tell us what the maximum load is AND if the signal response is in range. It will be different for different samples).
Please consult with someone local, who is an experienced analytical HPLC chromatographer about your project and once a method has been found or chosen, any sample requirements or limits can be established.
The sample loading, and the concentration, will be limited by the range on your detector, and the chromatography itself.
Detectors have a minimum level of detection, and also a limit of quantitation which determined the minimum sample that can be detected or quantitated. Likewise, they also have a maximum level beyond which either more compound can't be detected, or the detector can't provide meaningful data to determine the amount of sample detected.
The chromatography is limited by large peaks encroaching onto potential impurities, causing the detector to treat them as a single compound.
There is much more to the topic, more than can be answered in a fast reply on social media. There are courses that cover this very well. Start with the manufacturer of your HPLC equipment, and look for local experts in your university. It seems like a simple question, but it really is very complex.
Jack has provided an excellent response. Maximum sample concentration is not determined by "HPLC". HPLC is the name of an analytical technique. The specific method(s) used and system configuration will determine any maximums, as Jack noted. *Besides, samples can be diluted, and this is why a scientific HPLC sample "Loading Study" is run on the selected method (the results tell us what the maximum load is AND if the signal response is in range. It will be different for different samples).
Please consult with someone local, who is an experienced analytical HPLC chromatographer about your project and once a method has been found or chosen, any sample requirements or limits can be established.
Depends on your injector, detector, column.... For an analytical HPLC (depending on your molecule) it is probably ~1 mg. For a process HPLC which has a much thicker column it is ~100 mg.
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