Methods for isolation, use and analysis of ferritin
This invention provides methods of isolating ferritin from plant and animal material. The isolated ferritin can be administered to humans or animals in need of iron, and can be used to treat or supplement iron deficiency. The isolated ferritin can be used in industrial applications, such as increasing the iron content in heat-processed food or beverages. The methods of the invention also include quantitation of iron derived from plant or animal ferritin.
1. A method for determining an amount of ferritin iron from a legume in a sample comprising the steps of:
(a) first, providing a sample comprising a legume;
(b) second, determining the total amount of iron in the sample;
(c) third, preparing an aqueous soluble extract of the sample to make a soluble sample extract;
(d) fourth, determining the total amount of iron in the soluble sample extract;
(e) fifth, determining the amount of legume ferritin protein in the soluble sample extract using an automated quantitative immunological analysis, wherein the automated quantitative immunological analysis is capillary electrophoresis, and wherein the automated quantitative immunological analysis is conducted utilizing an antibody specific to the legume ferritin protein; and
(f) sixth, determining the amount of ferritin iron in the sample using the amount of total iron in the soluble sample extract divided by the amount of the legume ferritin protein in the soluble sample extract, multiplied by 0.75, multiplied by the total amount of iron in the sample, thereby determining the amount of ferritin iron from the legume in the sample.
2. The method of claim 1 wherein the sample is selected from the group consisting of a nutraceutical, a food, a dietary supplement and combinations thereof.
3. The method of claim 1 wherein the sample is a processed material.
4. The method of claim 1 wherein the sample is a natural material.
5. The method of claim 1 wherein the sample is from a waste stream from the processing of legumes.
6. The method of claim 1 wherein the soluble extract is prepared from the sample by mechanical separation.
7. A method for determining an amount of ferritin iron from a legume in a nutraceutical sample comprising the steps of:
(a) first, providing a nutraceutical sample comprising a legume;
(b) second, determining the total amount of iron in the nutraceutical sample;
(c) third, preparing an aqueous soluble extract of the nutraceutical sample to make a soluble sample extract;
(d) fourth, determining the total amount of iron in the soluble sample extract;
(e) fifth, determining the amount of legume ferritin protein in the soluble sample extract using an automated quantitative immunological analysis, wherein the automated quantitative immunological analysis is capillary electrophoresis, and wherein the automated quantitative immunological analysis is conducted utilizing an antibody specific to the legume ferritin protein; and
(f) sixth, determining the amount of ferritin iron in the nutraceutical sample using the amount of total iron in the soluble sample extract divided by the amount of legume ferritin protein in the soluble sample extract, multiplied by 0.75, multiplied by the total amount of iron in the sample, thereby determining the amount of ferritin iron from the legume in the nutraceutical sample.