Browsing by Author "Hefferan, Theresa E."
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- Separation and quantitation of B-6 vitamers in rat plasma by high performance liquid chromatographyHefferan, Theresa E. (Virginia Polytechnic Institute and State University, 1983)A high performance liquid chromatographic method was developed for the separation and quantitation of the B-6 vitamers in rat plasma. The separation time was under 20 minutes. The analyses were accomplished using a reverse phase C₁₈ column; a binary system of methanol and heptane sulfonic acid, the ion pair; and a fluorescence detector using 300 nm excitation and 375 nm emission filters. The minimum detectable quantity was determined to be 5 ng for PL and PN and 4 ng for PM. Samples were extracted using potato acid phosphatase; phosphorylated forms of the B-6 vitamers were converted to nonphosphorylated. The B-6 vitamers were measured in plasma from rats whjch had intakes of vitamin B-6 ranging from 7% - 551% of the basal requirement for the vitamin. PL was found to be the predominate B-6 vitamer in plasma from animals consuming <50% B-6 basal requirement and PM for the lowest intake group. PL and total B-6 values were significantly correlated (r = 0.99, p < 0.0001). Significant differences (p < 0.01) in plasma PL and total B-6 concentrations were observed for rats with inadequate and adequate B-6 intakes. The method seemed to be a sensitive indicator of vitamin B-6 status in rats thus indicating potential for use with humans.