I am trying to analyse fortified extruded rice by HPLC. My column is C18. Is there any established method to do that?
Rice is typically not fortified with vitamin A (or pro-vitamin A; a fat soluble vitamin).
If you fortified it, did you fortify it with retinol? retinol acetate? retinol palmitate?
You might want to think about why you want to do this.
These are all hexane soluble and should extract out the vitamin A. Blow a portion of it down with a stream of nitrogen and bring back to volume with a mixture of methanol:chloroform (9:1). Inject a portion that is suitable with the diameter of your column. For mobile phases, try high concentrations of methanol: water (85;15 or 90:10). On a C18 column, you should have your peak within 5 minutes. For detection, either UV at 313 or 328 nm. For better and specific detection use a fluorescence detector at Ex=450nm and Em=324nm. AOAC has method for infant formula, 2011.07. If you need to digest your material, then this method will apply. You need to do recovery studies to find out if the method is working. Because your material is so polar (starch and water), a straight extraction might work. Good Luck.
Read this manual, you will able to take an idea, This manual provide more methods and details about vitamin analysis
http://www.inmu.mahidol.ac.th/aseanfoods/doc/ASEAN%20Manual%20of%20Food%20Analysis.pdf
Rice is typically not fortified with vitamin A (or pro-vitamin A; a fat soluble vitamin).
If you fortified it, did you fortify it with retinol? retinol acetate? retinol palmitate?
You might want to think about why you want to do this.
These are all hexane soluble and should extract out the vitamin A. Blow a portion of it down with a stream of nitrogen and bring back to volume with a mixture of methanol:chloroform (9:1). Inject a portion that is suitable with the diameter of your column. For mobile phases, try high concentrations of methanol: water (85;15 or 90:10). On a C18 column, you should have your peak within 5 minutes. For detection, either UV at 313 or 328 nm. For better and specific detection use a fluorescence detector at Ex=450nm and Em=324nm. AOAC has method for infant formula, 2011.07. If you need to digest your material, then this method will apply. You need to do recovery studies to find out if the method is working. Because your material is so polar (starch and water), a straight extraction might work. Good Luck.
Yes Jack Sir is right
Vitamin A has unsaturation. it can be detected by UV-VIS spectroscopy using methanol as a sovent
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