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

Clin Chem. 1982 Oct;28(10):2165-9.

Abstract

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.

MeSH terms

  • Ascorbic Acid / blood*
  • Cell Separation / methods
  • Chromatography, High Pressure Liquid / methods
  • Humans
  • Leukocytes / analysis*
  • Monocytes / analysis
  • Neutrophils / analysis
  • Phenylhydrazines
  • Reference Values

Substances

  • Phenylhydrazines
  • 2,4-dinitrophenylhydrazine
  • Ascorbic Acid