What solvent should I use for a carotenoid injection into an HPLC system (after drying sample).
Is HPLC solvent for animal carotenoids different from other organisms, for example vegetable or phytoplankton?
There are more than 500 carotenoids.
HPLC solvent system depends on nature of compound to be analyzed.
Prior to HPLC analysis sample preparation is very crucial step for carotenoid analysis. Generally, reverse phase HPLC on C18 column is preferred. The most important part for selecting mobile phase is polarity, viscosity and volatility. Many solvent systems have been suggested for carotenoids analysis. Primary solvents are methanol and acetonitrile. Sometimes, small amount of modifiers such as THF can be added to improve the resolution.
Consider non-aqueous reverse phase (Snyder, L.R.; Kirkland, J.J.; Glajch, J.L.; Practical HPLC Method Development, 2nd Ed,
John Wiley & Sons, 1997, p.264 ).
About HPLC system I need information about:
Best mobile phase used for separation on C18 columns? Bonded phase?
Solvent for use sample ? flow? and more detaile about analysis of sea anemone carotenoids?
You'd probably want an end-capped C18 bonded to silica. Avoid polymeric reverse phase for this application since the polymer may swell in the solvents. Flow is whatever the manufacturer specifies for the column.
For my prep run, I found methanol/ dichloromethane solvent system worked well: http://www.isco.com/WebProductFiles/Applications/101/Poster_and_Paper_Reprints/Facile_Isolation_of_Carotenoid_Antioxidants.pdf
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