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- Joined: Fri Aug 05, 2016 4:18 pm
I am currently working on developing an analysis method for Vitamin D2 and D3 in food. I am following the extraction procedure in AOAC method 2012.11, where I performed saponification of the sample (12 g sample + 6 mL Methanol and 4 mL 45% KOH and heat in 75C water bath for 30 mins) then extraction with 20:80 ether: petroleum ether mix, evaporate to dryness then reconstitute with 2 mL methanol. But I am not getting a good recovery. Any suggestion or advice?
HPLC condition,
Column: acclaim C18
Mobile phase: water/methanol (gradient, start with 20/80 water/methanol ramp up to 100% methanol)
UV-Vis detection at 265 nm
column temperature: 30 C