Releasing agent for detecting content of 25-hydroxyvitamin D in human serum sample and detection method thereof

Abstract

The invention discloses a releasing agent for detecting the content of 25-hydroxyvitamin D in a human serum sample. The releasing agent is prepared from, by mass-to-volume ratio, 0.05-0.3% of sodium dodecyl sulfate, 1-20% of dimethyl sulfoxide and 1-10% of ethyl alcohol; during preparation, the sodium dodecyl sulfate is weighed firstly, after the sodium dodecyl sulfate is dissolved in a small quantity of 0.05 M Tris-NaCl buffer solution, the dimethyl sulfoxide and the ethyl alcohol are added, 0.05 M Tris-NaCl buffer solution is used for constant volume, concentrated hydrochloric acid is used for adjusting the pH value to 7.0, and a releasing agent finished product is obtained. The releasing agent has the advantages that the preparation method is simple, and the components used are less harmful to the environment and organisms; when in detection, the 25-hydroxyvitamin D and the metabolite of the same can be effectively released from binding protein, and thus be fully detected; there isno need to pretreat the sample, the sample can be added with other reagents at the same time, and it is convenient to conduct automatic operation.

Description

technical field [0001] The invention relates to in vitro diagnostic technology, in particular to a release agent for quantitatively detecting the content of 25-hydroxyvitamin D in human serum samples, and also relates to a method for using the release agent to detect the content of 25-hydroxyvitamin D in human serum samples. Background technique [0002] Vitamin D is a steroid hormone of two main types: vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol), which are similar in structure but differ in their side chains. Vitamin D in the human body mainly comes from the conversion of 7-dehydrocholesterol in the skin to vitamin D3 by ultraviolet radiation, and also includes vitamin D3 ingested from fish, meat and other foods and vitamin D2 in dietary supplements. Regardless of the form of vitamin D, it is bound to binding proteins in the blood and transported to the liver, where it is converted to 25(OH)VD, and then converted to 1,25-(OH) in the kidney 2 VD, which is t...

AI Technical Summary

Problems solved by technology

PCT International Application WO2008 / 092917A1 published a method of proteolytic digestion to dissociate 25-hydroxyvitamin D from its binding protein, but this method is cumbersome and time-consuming

Others such as WO2003 / 023391 disclose direct measurement of 25-hydroxyvitamin D by substitution of hydroxylated aromatic carboxylic acids; EP2007 / 140962 proposes the use of one or more amphoteric reagents at pH 3.8 to 4.8 and 5- 30% DMSO, liquid organic amides and optionally 0.5-5% short-chain alcohols release 25-hydroxyvitamin D from the vitamin D-binding protein, the amphoteric compound used in this patent is toxic and dangerous

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