Ascorbic acid in lymphocytes: cell preparation and liquid-chromatographic assay.
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.
D001205:Ascorbic Acid / Q000097:blood*; D002469:Cell Separation / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D007962:Leukocytes / Q000032:analysis*; D009000:Monocytes / Q000032:analysis; D009504:Neutrophils / Q000032:analysis; D010659:Phenylhydrazines; D012016:Reference Values
D010659:Phenylhydrazines; C004787:2,4-dinitrophenylhydrazine; D001205:Ascorbic Acid