Importance of HPLC detector

Basic questions from students; resources for projects and reports.

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Hey all,
How important is LC detector for method development is I have LC-MS? I got mixed suggestion on it. One of the suggestions was detector is must for LC development and he highly recommended to buy. The other expert said LC-MS itself has detector so I do not need to by LC detector. I do not know which is the correct suggestion.
I personally think it is reasonable not to buy LC detector as LC-MS has mass detector. But at some point I think it is better to use LC detector for method development before shooting everything into mass spec.

What will be the advantages of having LC-detector?

Hoping to get good suggestions from the experts here.

Thank you.

The mass spec *is* a detector for HPLC (the mass spectrometrists prefer to think of the HPLC as an intake system for the mass spec, but that's another discussion :lol: )

Presumably you are referring to a UV-vis absorbance detector (either a diode-array type or a variable wavelength detector). I would definitely want to have one because it is in many ways complementary to the MS. It's a passive detector (MS is an active detector) so you can run it in series with the MS and it gives you a look at compounds that may not ionize well (or at all) under your MS conditions.
-- Tom Jupille
LC Resources / Separation Science Associates
+ 1 (925) 297-5374
I like having a PDA in addition to a mass spec for various reasons:
(1) A PDA spectrum can still be quite diagnostic for a class of chemical. If you are asked, for example, to find all the flavonoids present at significant level in a plant sample, then PDA is very valuable because every flavonoid peak will have a very similar spectrum, and it's much quicker to look at each significant UV peak to check its spectrum looks sensible, than to examine every mass peak to see whether it has a mass and fragmentation pattern appropriate for a flavonoid.
(2) UV absorbance is very stable, while ionisation efficiency changes over the course of running samples. If you cannot obtain isotopically-labelled internal standards (a very common problem!), then external standard quantification based on PDA may be more reliable than external standard quantification based on MS. You can use the MS to confirm identity, and PDA to quantify.
(3) There are many situations where a whole class of compound will have the same response per unit concentration when measured by UV absorbance, but not by MS. This is approximately true of things like anthocyanins, where the visible absorbance changes only slightly when extra groups (glycosylation etc.) are added to the anthocyanin. In these cases, if you can only obtain commercial standards for one or two members of the class of compound, you can still estimate the absolute concentrations of other members by assuming the same response in PDA. You can't do this in mass spec, where the response is likely to be very different.
(4) UV absorbance is a really convenient way to check the performance of your pump, using acetone-spiked solvent versus solvent!

(A little note on dual detector measurements, PDA + MS; I tend to check that the data approximately correlate. If they don't, I assume that the PDA has detected the wrong thing, because PDA isn't as specific as MS. If they do, I assume that PDA has measured the correct thing, and probably more reliably than MS).

Despite what I've written, I do have one instrument without a PDA: they're much more limited when used on high-sensitivity triple quad systems, because the amount of analyte necessary for detection in PDA is so many orders of magnitude above the optimum concentration for MS. It's hard to find a range of concentrations where both detectors are doing a good job.
Thank you all.
4 posts Page 1 of 1

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