Preparation of samples for vitamin b.sub.1.sub.2 and/or...

G - Physics – 01 – N

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G01N 33/82 (2006.01) G01N 33/53 (2006.01)

Patent

CA 1197185

ABSTRACT Vitamin B12 (cobalamin) and/or folate (folic acid) assay tech- niques can be simplified, by eliminating the step of heating or boil- ing the to-be-tested sample prior to its assay. This is accomplished by utilizing compositions and procedures: (1) which substantially completely liberate all vitamin B12 and/or folate from endogenous binding protein without heating or boiling; (2) which substantially completely destroy all endogenous bind- ing protein which is present in the to-be-assayed sample, and which may bind natural and radioactive vitamin B12 or folate both due to its liberation from vitamin Bl2 or folate in the original sample or due to its unbound presence in the original sample; (3) which substantially completely inhibit or block any unde- stroyed endogenous binding protein; and (4) which substantially completely inhibit or destroy any in- trinsic factor-blocking antibodies which may be present in the to-be-assayed sample. In preferred embodiments the protein destroying compositions utilized include both a strong base and a sulfhydral compound which is a strong disulfide protein destroying material while the material of choice to bind with undestroyed endogenous binding protein is a vitamin B12 analogue. The sample is then assayed using known, for example, radioisotope dilution assay techniques.

400429

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