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Title: SUPERCRITICAL FLUID EXTRACTION AND REVERSED-PHASE LIQUID CHROMATOGRAPHY METHOD FOR FAT-SOLUBLE VITAMINS AND CAROTENOIDS

Author
item Burri, Betty
item Neidlinger, Terry
item LO, ANNIE
item Wong, Monica

Submitted to: International Symposium on High Performance Liquid Phase Separations
Publication Type: Abstract Only
Publication Acceptance Date: 8/1/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: We developed a supercritical fluid procedure for extracting retinol, retinyl esters, tocopherols, and the carotenoids from vitamin supplements and liver tissue. The supercritical fluid extracts could be injected onto the HPLC column without further pretreatment. Samples were extracted using a Prepmaster coupled to an Accutrap (Suprex, Pittsburgh, PA). Sample size was one supplement capsule diluted 1:1000 with hexane, or 0.1 gm minced liver mixed with 1 gm Hydromatrix (Varian, Harbor City, CA); packed into a 5 mL sample container. Samples were eluted with SFC grade CO2 at 400 ATM, 40oC, 1 mL/min flow rate. Extracts were eluted into 1.5 mL hexane (restrictor 65 deg C). Thirty uL aliquots were injected onto a reversed-phase HPLC column (Resolve C18 column and guard column; Waters Chromatography, Milford, MA). Chromatography was run on a System Gold gradient HPLC (Beckman, Fullerton, CA) with diode array detector; using a modification of a previously described method (Peng & Peng, Cancer Epidemiol. Biomark. Prevent. 1:375-382, 1992). Extraction efficiency was >85% for each analyte, CV (for extraction and chromatography, n=5) ranged from 4% for alpha-carotene to 9% for lutein zeaxanthin. This method is less time-consuming and labor intensive then traditional saponification and extraction procedures for fat soluble vitamins.