Extraction and quantification of vitamins A & D in fluid samples
First Claim
1. A method of quantifying vitamin A or D3, or vitamins A and D3, in an aqueous dairy sample within a few hours, comprising the steps of:
- (a) extracting the aqueous dairy sample with an alkaline metal salt and an extraction solvent comprising a mixture of a non-polar organic solvent and a polar organic solvent, to isolate vitamin A or D3, or vitamins A and D3 from the aqueous dairy sample into an organic fraction; and
(b) determining the amount of vitamin D3 or vitamin A, or both vitamin D3 and vitamin A, in the organic fraction,wherein the vitamin is quantified with a monoclonal antibody which specifically binds to the vitamin, and wherein the non-polar organic solvent comprises an aliphatic hydrocarbon having from 4 to 10 carbon atoms, or a mixture thereof, and the polar organic solvent comprises a chloroalkane or ethyl acetate.
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Abstract
The invention discloses monoclonal antibodies for vitamins A (retinol palmitate) and D3 (cholecalciferol); a method for using monoclonal antibodies, and the monoclonal antibodies disclosed herein, in particular, to quantitate these vitamins in fluids such as dairy products, and blood, and also raw or processed agri-food and beverage products. The method involves contacting the sample with a mixture of polar and non-polar organic solvents in combination with inorganic salts to remove fat molecules into an organic fraction, and assaying the organic fraction by immunoassay involving the monoclonal antibodies. The presence of a mixture of non-polar and polar organic solvents increases the separation of vitamins from fat molecules and enables the test samples to be quantified by immunoassay without any further treatment.
15 Citations
10 Claims
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1. A method of quantifying vitamin A or D3, or vitamins A and D3, in an aqueous dairy sample within a few hours, comprising the steps of:
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(a) extracting the aqueous dairy sample with an alkaline metal salt and an extraction solvent comprising a mixture of a non-polar organic solvent and a polar organic solvent, to isolate vitamin A or D3, or vitamins A and D3 from the aqueous dairy sample into an organic fraction; and (b) determining the amount of vitamin D3 or vitamin A, or both vitamin D3 and vitamin A, in the organic fraction, wherein the vitamin is quantified with a monoclonal antibody which specifically binds to the vitamin, and wherein the non-polar organic solvent comprises an aliphatic hydrocarbon having from 4 to 10 carbon atoms, or a mixture thereof, and the polar organic solvent comprises a chloroalkane or ethyl acetate. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification