Some problems associated with assay of 25-hydroxycalciferol in human serum. 1977

R S Mason, and S Posen

Methods available for the assay of 25-hydroxycalciferol in human serum are evaluated and compared to one another. Ethanol was chosen for use in the initial extraction procedure and rat-kidney cytosol as the binding protein, although good alternative methods are also available. We used silicic acid for chromatography and found this an essential step. Reproducibility was increased when, after "bound" and "free" material were separated, an aliquot of the supernate was pipetted into the counting vial instead of the entire supernatant fluid being decanted. Beta-lipo-protein added to the assay system was of no advantage; added bovine serum albumin interfered with the assay by giving rise to high blank values. With ethanol extraction, silicic acid chromatography, rat kidney cytosol and separation on dextran-coated charcoal, sera from normal subjects showed a mean 25-hydroxycalciferol concentration of 28.5 microng/liter (range, 13.1 to 43.9) during the fall season. Coefficients of variation for a control serum were 4.9% (intra-assay) and 10.9% (interassay).

UI MeSH Term Description Entries
D007668 Kidney Body organ that filters blood for the secretion of URINE and that regulates ion concentrations. Kidneys
D008722 Methods A series of steps taken in order to conduct research. Techniques,Methodological Studies,Methodological Study,Procedures,Studies, Methodological,Study, Methodological,Method,Procedure,Technique
D010018 Osteomalacia Disorder caused by an interruption of the mineralization of organic bone matrix leading to bone softening, bone pain, and weakness. It is the adult form of rickets resulting from disruption of VITAMIN D; PHOSPHORUS; or CALCIUM homeostasis. Adult Rickets,Rickets, Adult
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D012016 Reference Values The range or frequency distribution of a measurement in a population (of organisms, organs or things) that has not been selected for the presence of disease or abnormality. Normal Range,Normal Values,Reference Ranges,Normal Ranges,Normal Value,Range, Normal,Range, Reference,Ranges, Normal,Ranges, Reference,Reference Range,Reference Value,Value, Normal,Value, Reference,Values, Normal,Values, Reference
D001798 Blood Proteins Proteins that are present in blood serum, including SERUM ALBUMIN; BLOOD COAGULATION FACTORS; and many other types of proteins. Blood Protein,Plasma Protein,Plasma Proteins,Serum Protein,Serum Proteins,Protein, Blood,Protein, Plasma,Protein, Serum,Proteins, Blood,Proteins, Plasma,Proteins, Serum
D002021 Buffers A chemical system that functions to control the levels of specific ions in solution. When the level of hydrogen ion in solution is controlled the system is called a pH buffer. Buffer
D002606 Charcoal An amorphous form of carbon prepared from the incomplete combustion of animal or vegetable matter, e.g., wood. The activated form of charcoal is used in the treatment of poisoning. (Grant & Hackh's Chemical Dictionary, 5th ed) Activated Charcoal,Actidose,Actidose-Aqua,Adsorba,Carbomix,Charbon,CharcoAid,CharcoCaps,Charcodote,Formocarbine,Insta-Char,Kohle-Compretten,Kohle-Hevert,Kohle-Pulvis,Kohle-Tabletten Boxo-Pharm,Liqui-Char,Norit,Ultracarbon,Charcoal, Activated
D002850 Chromatography, Gel Chromatography on non-ionic gels without regard to the mechanism of solute discrimination. Chromatography, Exclusion,Chromatography, Gel Permeation,Chromatography, Molecular Sieve,Gel Filtration,Gel Filtration Chromatography,Chromatography, Size Exclusion,Exclusion Chromatography,Gel Chromatography,Gel Permeation Chromatography,Molecular Sieve Chromatography,Chromatography, Gel Filtration,Exclusion Chromatography, Size,Filtration Chromatography, Gel,Filtration, Gel,Sieve Chromatography, Molecular,Size Exclusion Chromatography
D003600 Cytosol Intracellular fluid from the cytoplasm after removal of ORGANELLES and other insoluble cytoplasmic components. Cytosols

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