Dear colleagues,
I am looking for an easy to establish methodology to measure lipid classes (FFA, TAG, SE, WE, ST, PL) from lipid extracted samples on our HPLC with a binary pump (2 eluents) and equipped with a diode array detector (DAD) or a fluorescence detector (FLD).
Maybe someone is running the same setup in their laboratory or can point me to a published methodology section. Any help is very much appreciated!
Dear Julian,
I think these two articles (attached) may actually help you.
Best wishes
- Mohamed Sabri Bensaad: The two references you provided above to assist Julian with HPLC LIPID separation methods using a DAD or FLD are not applicable to his question.The first reference is not for Lipids at all (wrong sample type). The second uses a detector that he does not have (an evaporative detector).
Julian: You will not find any "easy to establish methodology" by HPLC - UV for Lipid classes. No useful chromophores are present. These are best resolved and detected by HPLC with either Mass (LC-MS) or Evaporative detectors such as ELSD or CAD (e.g. DOI: 10.1080/10826079208017168), not with UV/VIS, as you propose. These are the most sensitive and reliable to use. Note: If sensitivity is not needed in your application, then the evaporative detection method in my above example can also be used with 210nm detection too, as the mobile phase used was intentionally chosen to be UV transparent for comparison. Depending on the column used, a small amount of methanol can also be added to the mobile phase to sharpen the peaks (but methanol absorbs in the low UV range, so only add if sensitivity if OK).
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