For determining Furosine compound of rice flour I have to do RP-HPLC. Reference article suggest that in sample preparation-
after acid hydrolysis, the filtrates(0.50 ml) were applied to a Millipore SEP-PAK Clartridge (C18, 500 mg, Altech,Belgium), prewetted with water(0.50 ml) and eluted with 3.0 M of 2.0 ml HCl, then go for HPLC.
My problem I am confused that do I have to activate the cartridge before follow this reference? But they don't mention it any article!
and what do they mean by prewetted ? is it at first wetted by water then have to pass 0.50 ml sample, then eluted with 3.0 M,2.0 ?
Activation (or conditioning) step is usually necessary. However, some cartridges do not require this step so I suggest you follow the protocol provided by the manufacturer (Or, you may ask the author for their procedure).
"Pre-wetting" (I'm more familiar with "Equilibration step"): the wetting solution before sample loading is mostly water for RP, but it must have similar solvent strength and pH as your sample so that: (1) residual conditioning solvent can be washed out, and (2) maximize the analyte retention.
For more info: https://www.sigmaaldrich.com/analytical-chromatography/sample-preparation/spe/reversedphase-methodology.html
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