PG, NDGA, BHT, and BHA were extracted with acetonitrile from fat or oil, followed by thin layer chromatographic separation and identification. The four antioxidants were distinctly separated from one another by one-dimensional solvent development of the silica gel adsorbent layer. The chromatography required < 1 hour from spotting to visualization with 2,6-dichloroquinone chlorimide. As little as 1 μg of BHT and 0.5 μg of PG, NDGA, and BHA were detected by this method.
In preparing samples for analysis of antioxidants in fats and oils, melted fat and/or oil is mixed with Celite, blended with acetonitrile, and the acetonitrile is extracted with mixed ethers. The mixed ethers are evaporated, and the oily residue is incorporated with alumina acid. The alumina mixture is placed in a chromatographic column and eluted with aqueous methanol, and the methanolic eluate is extracted into mixed ethers. The ether extract is clean enough for TLC as well as GLC determinations. Recoveries from tallow, lard, and corn oil spiked at 16.7 and 66.7 ppm with BHA and BHT ranged from 80 to 95% for BHA and 91 to 115% for BHT.
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