阅读说明：本技术 一种总铁结合力检测试剂盒及其制备方法 (total iron binding force detection kit and preparation method thereof ) 是由 袁嘉扬 张凤 于 2019-09-12 设计创作，主要内容包括：本发明涉及生物技术领域,涉及一种总铁结合力检测试剂盒,尤其涉及一种总铁结合力检测试剂盒的制备方法,包括试剂R1的制备以及试剂R2的制备,将配液罐放在磁力搅拌器上,使溶液保持中速转动状态,边搅拌边投入Tris缓冲液、亚铁嗪、抗坏血酸、第二缓冲液,搅拌30分钟至物料完全溶解,溶液清澈透明配液罐底部无沉淀后,然后定容至最终体积。本发明的优点是：本试剂盒是采用分光光度法以亚铁嗪为底物进行总铁结合力的检测,在酸性介质中与转铁蛋白结合的血清铁从转铁蛋白中解离出来,再被还原剂还原为Fe2+,后者与亚铁嗪生成紫红色化合物,通过分光光度计检测562nm处吸光度值检测血清、血浆样品中的总铁含量,线性范围可做到50-400μmol/L。(the invention relates to the technical field of biology, and relates to a total iron binding force detection kit, in particular to a preparation method of the total iron binding force detection kit, which comprises the steps of preparing a reagent R1 and preparing a reagent R2, wherein a liquid preparation tank is placed on a magnetic stirrer, so that a solution keeps a medium-speed rotation state, Tris buffer solution, ferrous oxazine, ascorbic acid and a second buffer solution are added while stirring, the stirring is carried out for 30 minutes until materials are completely dissolved, and after no precipitate is left at the bottom of the clear and transparent liquid preparation tank, the solution is subjected to constant volume until the final volume is reached. The invention has the advantages that: the kit adopts a spectrophotometry method to detect the total iron binding force by taking the ferrochrome as a substrate, serum iron bound with transferrin in an acidic medium is dissociated from the transferrin and then reduced into Fe2+ by a reducing agent, the Fe2+ and the ferrochrome generate a mauve compound, the total iron content in serum and plasma samples is detected by detecting the absorbance value at 562nm by a spectrophotometer, and the linear range can reach 50-400 mu mol/L.)

1. A total iron binding force detection kit is characterized in that: the reagent comprises a reagent R1 and a reagent R2, wherein each component and concentration of the reagent R1 comprises:

the concentrations of the components of the reagent R2 include:

2. the total iron binding force detection kit according to claim 1, characterized in that: the first preservative and the second preservative comprise one of sodium azide, Proclin-950, Proclin-300 and thimerosal.

3. The total iron binding force detection kit according to claim 1, characterized in that: the pH value of the reagent R1 is between 4.00 and 5.00.

4. The total iron binding force detection kit according to claim 1, characterized in that: the pH value of the reagent R2 is between 7.00 and 8.30.

5. The method for preparing a total iron binding force detection kit according to any one of claims 1 to 4, wherein the total iron binding force detection kit comprises: the method comprises the following steps: A) reagent R1 preparation:

adding 70% of purified water into a liquid preparation tank, placing the liquid preparation tank on a magnetic stirrer, turning on a power switch of the magnetic stirrer, adjusting a stirrer to a medium-speed rotation speed to keep the solution in a medium-speed rotation state, adding Tris buffer solution, thiourea, hydroxylamine hydrochloride, ammonium ferrous sulfate, TritonX-100 and a first preservative while stirring, stirring for 10-30 minutes until all materials are completely dissolved, adjusting the pH value to 4.00-5.00 after no precipitate is left at the bottom of the clear and transparent liquid preparation tank of the solution, and then fixing the volume to the final volume;

filtering the dissolved preparation solution through a microporous filter membrane according to the operation rules of a microporous filter, storing the filtered solution in a finished product tank, and marking;

B) reagent R2 preparation:

adding 70% of purified water into a liquid preparation tank, placing the liquid preparation tank on a magnetic stirrer, turning on a power switch of the magnetic stirrer, adjusting a stirrer to a medium-speed rotation speed to keep the solution in a medium-speed rotation state, adding Tris buffer solution, ferrous oxazine, ascorbic acid and a second buffer solution while stirring, stirring for 10-30 minutes until the materials are completely dissolved, adjusting the pH value to 7.00-8.30 after no precipitate is left at the bottom of the clear and transparent liquid preparation tank, and then fixing the volume to the final volume;

And filtering the dissolved preparation solution through a microporous filter membrane according to the operation rules of the microporous filter, and storing the filtered solution in a finished product tank for marking.

Technical Field

The invention relates to the technical field of biology, relates to a total iron binding force detection kit, and particularly relates to a preparation method of the total iron binding force detection kit.

background

the determination of the total iron binding force is a routine inspection item, and a plurality of clinical data show that the total iron binding force is reduced due to diseases such as iron-deficiency anemia and acute hepatitis, and the total iron binding force is increased due to diseases such as liver cirrhosis, nephropathy, uremia and hemochromatosis.

At present, the clinical determination of total iron binding force mainly adopts an atomic absorption method through light magnesium carbonate adsorption and various complexation photometry. The atomic absorption method not only needs a special expensive instrument, but also needs precipitation and centrifugation, has high detection cost and complicated operation steps, and is difficult to popularize clinically.

Disclosure of Invention

The purpose of the invention is: aiming at the defects, the total iron binding force detection kit and the preparation method thereof are provided.

In order to achieve the purpose, the invention adopts the technical scheme that:

a total iron binding force detection kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises the following components in concentration:

the concentrations of the components of the reagent R2 include:

The first preservative and the second preservative comprise one of sodium azide, Proclin-950, Proclin-300 and thimerosal.

The pH value of the reagent R1 is between 4.00 and 5.00.

the pH value of the reagent R2 is between 7.00 and 8.30.

A preparation method of a total iron binding force detection kit comprises the following steps: A) reagent R1 preparation:

adding 70% of purified water into a liquid preparation tank, placing the liquid preparation tank on a magnetic stirrer, turning on a power switch of the magnetic stirrer, adjusting a stirrer to a medium-speed rotation speed to keep the solution in a medium-speed rotation state, adding Tris buffer solution, thiourea, hydroxylamine hydrochloride, ammonium ferrous sulfate, TritonX-100 and a first preservative while stirring, stirring for 10-30 minutes until all materials are completely dissolved, adjusting the pH value to 4.00-5.00 after no precipitate is left at the bottom of the clear and transparent liquid preparation tank of the solution, and then fixing the volume to the final volume;

Filtering the dissolved preparation solution through a microporous filter membrane according to the operation rules of a microporous filter, storing the filtered solution in a finished product tank, and marking;

B) reagent R2 preparation:

adding 70% of purified water into a liquid preparation tank, placing the liquid preparation tank on a magnetic stirrer, turning on a power switch of the magnetic stirrer, adjusting a stirrer to a medium-speed rotation speed to keep the solution in a medium-speed rotation state, adding Tris buffer solution, ferrous oxazine, ascorbic acid and a second buffer solution while stirring, stirring for 10-30 minutes until the materials are completely dissolved, adjusting the pH value to 7.00-8.30 after no precipitate is left at the bottom of the clear and transparent liquid preparation tank, and then fixing the volume to the final volume;

And filtering the dissolved preparation solution through a microporous filter membrane according to the operation rules of the microporous filter, and storing the filtered solution in a finished product tank for marking.

Compared with the prior art, the invention achieves the technical effects that: the kit adopts a spectrophotometry method to detect the total iron binding force by taking the ferrochrome as a substrate, serum iron bound with transferrin in an acidic medium is dissociated from the transferrin and then reduced into Fe2+ by a reducing agent, the Fe2+ and the ferrochrome generate a mauve compound, the total iron content in serum and plasma samples is detected by detecting the absorbance value at 562nm by a spectrophotometer, and the linear range can reach 50-400 mu mol/L.

Drawings

FIG. 1 is a graph of the calibration using the standard in example 4 of the present invention. Where the X-axis represents standard concentration and the Y-axis represents dOD.

Detailed Description

the invention is further described with reference to the following figures and examples: