The use of octadecyl-bonded microparticulate silica in the separation of free and bound fractions during saturation analysis of vitamin D metabolites

The use of octadecyl-bonded microparticulate silica to separate free and bound fractions during the saturation analysis of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D has been investigated. A slurry of octadecyl-bonded silica in an appropriate incubation buffer was prepared and used in parallel...

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Bibliographic Details
Published in:Journal of pharmaceutical and biomedical analysis 1992-09, Vol.10 (9), p.631-637
Main Authors: Coldwell, R.D., Nolan, J., Trafford, D.J.H., Mawer, E.B., Makin, H.L.J.
Format: Article
Language:English
Subjects:
Animals
Binding, Competitive
Biological and medical sciences
Calcifediol - isolation & purification
Calcifediol - pharmacokinetics
Calcitriol - isolation & purification
Calcitriol - pharmacokinetics
Cattle
Charcoal
Chemistry
Chemistry, Analytical
Chick Embryo
Cytosol - metabolism
dextran-coated charcoal
Dextrans
General pharmacology
Humans
Life Sciences & Biomedicine
Medical sciences
Octadecyl-bonded microparticulate silica
Pharmaceutical technology. Pharmaceutical industry
Pharmacology & Pharmacy
Pharmacology. Drug treatments
Physical Sciences
Protein Binding
Radioligand Assay
saturation analysis
Science & Technology
Silicon Dioxide
Temperature
Thymus Gland - metabolism
Vitamin D - isolation & purification
Vitamin D - pharmacokinetics
vitamin D metabolites
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Summary:The use of octadecyl-bonded microparticulate silica to separate free and bound fractions during the saturation analysis of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D has been investigated. A slurry of octadecyl-bonded silica in an appropriate incubation buffer was prepared and used in parallel with a conventional dextran-coated charcoal suspension in several assay procedures. Standard curves, non-specific binding and plasma values were compared. A competitive protein binding assay for 25-hydroxyvitamin D and two radioreceptor assays and one radioimmunoassay for 1,25-dihydroxyvitamin D were investigated. In most cases the octadecyl-bonded silica preparation gave the more favourable results; its action was rapid, time- and temperature-independent, and it produced low non-specific binding and higher B 0 values in all the assays examined. It was in our hands easier to use than dextran-coated charcoal. The use of octadecyl-bonded silica is recommended as an efficient agent for the separation of free and bound fractions in the saturation analysis of vitamin D metabolites.
ISSN:0731-7085
1873-264X
DOI:10.1016/0731-7085(92)80090-A