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Clinical Chemistry 28: 2165-2169, 1982;
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Clinical Chemistry, Vol 28, 2165-2169, Copyright © 1982 by American Association for Clinical Chemistry

Ascorbic acid in lymphocytes: cell preparation and liquid- chromatographic assay

W Lee, P Hamernyik, M Hutchinson, VA Raisys and RF Labbe

Measurements of ascorbic acid concentration in leukocytes by "high- performance" liquid chromatography (HPLC) provides better nutritional assessment, leading to better management, particularly of presymptomatic and critically ill patients. This procedure includes a simple, reproducible cell-separation technique that requires no more than 2 mL of whole blood. Cell populations are separable with greater than 95% purity and greater than 99% viability. Ascorbic acid is assayed by HPLC. The vitamin can be reproducibly quantified in concentrations as low as 0.1 microgram/mL of cell extract. The chromatographic procedure is very rapid, analysis being completed within 15 min after specimen preparation. The assay is suitable also for urine and protein-free filtrates of plasma and of other biological materials. Reference intervals for plasma, mononuclear leukocytes, and polymorphonuclear leukocytes were established. A preliminary clinical evaluation revealed that hospital patients were at a greater risk of ascorbic acid deficiency than expected.


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