I want to measure glucosylceramide or lactosylceramide quantity by spectrometer or HPLC.
Dear Kartik,
There are several methods you can choose from depending on your goal.
For instance, you can screen glyco(sphingo)lipids in your samples by cheap and simple spectrophotometric methods, such as the orcinol and DNS methods, designed to determine the sugar content in your sample and that will allow you to choose samples of interest (in case you have many to quantitate).
For a more accurate estimate on the amount of glyco(sphingo)lipid classes you can decide for a normal-phase (or HILIC) HPLC method coupled with UV detector. This will require a prior derivatization step of the sugar moiety, and then you´ll be able to detect the HexCer and LacCer (and even ganglioside) classes by looking at the retention times (as these classes will exhibit different chromatographic behaviour).
In case you're interested in quantifying individual molecular species of HexCer and LacCer then the high resolution LC-MS is the best option, and also the most expensive.
Hope this helps,
Ana
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