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10 good reasons for adding a mass spectrometer to my UV-HPLC?

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  1. I get molecular weight information on my peak. An MS gives you the mass to charge ratio of any ions it sees, which is a hugely useful piece of information.
    RT and area aren’t enough for identification purposes, but once you add weight information to the equation, things start to become more certain.

  2. Better selectivity – a complex chromatogram can be like trying to see through a forest when you use a UV detector – it just sees trees. We can set the MS to look at the forest and only show trees of an exact height (e.g. 30.8ft), making things much clearer and more visible

  3. Better sensitivity – generally mass spectrometry (MS)is inherently more sensitive in terms of analyte response, but sensitivity is a feature of signal to noise.

  4. Are you seeing the full picture? If using a single wavelength UV detector, we rely on everything having the exact chromaphore that absorbs that wavelength. If the chromaphore is different between the compounds in our chromatogram, we may not be seeing compounds that are present.

  5. You can see what’s under you main peak. If you’ve developed a method, there’s always the risk that something small is lurking under your main peak(s).
    With MS you can detect compounds at levels of 0.1% that fully co-elute with the main peak.
  1. Structural information. Molecular weight information is big information, but once we start dismantling compounds and analyzing the mass of the fragments, we can start to build up a picture of an unknown related substance such as metabolites or degradation products.

  2. Analytes with poor chromaphores. MS doesn’t rely on the compound having a UV chromaphore – we can detect a lot of compounds that have very poor or no UV chromaphore.

  3. It speeds up method development. By tracking our peaks by their molecular weight we can see very clearly how they move when we make changes during development. We can even carry out multiple method developments simultaneously. It also allows us very early on to see if we have any peaks co-eluting that may be missed with UV.

  4. Definitive identification. Retention time matching can be enough for basic confirmation of identity in simple mixtures, retention time combined with molecular weight information gives us significantly more confidence, retention time with molecular weight and with fragmentation spectrum (from an MS/MS capable instrument) is almost definitive.

  5. All MS methods work on UV, not all UV methods work on MS. Develop your method as if it was to be run on MS, even if you use it for UV. You just never know when you may need to go onto an MS to get more info.

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