Abstract
Radioligand assay for biotin in liver tissues A radioligand assay for biotin in liver tissue is described, 3H-biotin is used as tracer and avidin as binder. The biotin-loaded avidin is separated from free biotin on dextrancoated charcoal, which leaves the avidin—biotin complex in the supernatant liquid. Thus, the avidin—biotin complex can easily be utilized for determination of the radioactivity. Calibration with known additions of biotin in the range 0.25–8.0 ng per assay sample yields a linear logit-log plot. The biotin is extracted from liver tissues by enzymatic proteolysis with papain. This treatment is optimized to liberate the bound forms of the vitamin. Microbiological parallel assays with [Lactobacillus plantarum] were in good agreement with the radioligand assay giving a regression coefficient of 0.974 (n = 44). The coefficient of variation was found to be 4.2% in the range 500–1200 ng of biotin per g of liver tissue (n = 46). The method is simple and reliable and allows the simultaneous analysis of a considerable number of samples.
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