Quantitative Determination of Vitamin B12 in Blood by Radioassay

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Since the last year we studied on quantitative determination of vitamin B12 in blood by competitive binding analysis using a sephadex intrinsic factor complex as a binder and obtained some findings as follows: A standard curve obtained by diluting a standard vitamin B12dilution (1600 pg/ml) was slightly sigmoid in the range from 25 to 1600 pg/ml. When the same sample was analysed in duplicate to check reproducibility, the variation coefficient was low, and the method was thought applicable to the clinical determination. Changes of vitamin B12 in Blood by dilution of serum and recovery rates of vitamin B12 were studied. No fixed correlation between the observed values and the dilution rates was observed and mean recovery rate was 99.92%. The normal range of vitamin B12in serum obtained with normal 50 subjects is 564.6 ±261.3 (210-1200) pg/ml. After injecting 100 mg of tetrahydrofolate (FAH4) intramuscularly to normal subjects and patients with megaloblastic anemia, aplastic anemia, nephrosis and Banti's syndrome, changes of vitamin B12in blood were studied. In patients with aplastic anemia, nephrosis and Banti's syndrome, significantly low values were obtained 2 to 4 hours after injection. Though this method has to be studied further in some aspects, it is simple and many samples can be analyzed in relatively short time. For this reason it is considered that the method would be widely applicable.
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