CN110568206A - total iron binding force detection kit and preparation method thereof - Google Patents
total iron binding force detection kit and preparation method thereof Download PDFInfo
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- CN110568206A CN110568206A CN201910868124.4A CN201910868124A CN110568206A CN 110568206 A CN110568206 A CN 110568206A CN 201910868124 A CN201910868124 A CN 201910868124A CN 110568206 A CN110568206 A CN 110568206A
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/90—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving iron binding capacity of blood
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Abstract
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.
Description
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:
The first embodiment is as follows:
the total iron binding force detection kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises the following components in percentage by weight: tris buffer solution with the concentration of 80mmol/L, thiourea with the concentration of 30mmol/L, hydroxylamine hydrochloride with the concentration of 150mmol/L, ammonium ferrous sulfate with the concentration of 5mmol/L, TritonX-100 with the concentration of 0.2 percent, Proclin-950 with the concentration of 0.8 ml/L;
The concentrations of the components of the reagent R2 include: tris buffer with the concentration of 30mmol/L, ferrioxazine with the concentration of 20mmol/L, ascorbic acid with the concentration of 10mmol/L, Proclin-950 with the concentration of 0.8 ml/L.
the pH value of the reagent R1 is 4.00.
the pH value of the reagent R2 is 7.00.
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 minutes until all materials are completely dissolved, adjusting the pH value to 4.00 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 minutes until the materials are completely dissolved, adjusting the pH value to 7.00 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.
example two:
the total iron binding force detection kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises the following components in percentage by weight: tris buffer solution with the concentration of 120mmol/L, thiourea with the concentration of 45mmol/L, hydroxylamine hydrochloride with the concentration of 220mmol/L, ammonium ferrous sulfate with the concentration of 20mmol/L, TritonX-100 with the concentration of 1.0 percent, Proclin-300 with the concentration of 1.2 ml/L;
the concentrations of the components of the reagent R2 include: tris buffer with the concentration of 55mmol/L, ferrioxazine with the concentration of 35mmol/L, ascorbic acid with the concentration of 30mmol/L, Proclin-300 with the concentration of 1.2 ml/L.
The pH of the reagent R1 was 4.5.
The pH of the reagent R2 was 7.6.
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 20 minutes until all materials are completely dissolved, adjusting the pH value to 4.5 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 20 minutes until the materials are completely dissolved, adjusting the pH value to 7.6 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.
Example three:
the total iron binding force detection kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises the following components in percentage by weight: tris buffer solution with the concentration of 160mmol/L, thiourea with the concentration of 60mmol/L, hydroxylamine hydrochloride with the concentration of 300mmol/L, ammonium ferrous sulfate with the concentration of 30mmol/L, TritonX-100 with the concentration of 2.0 percent, and thimerosal with the concentration of 2.0 ml/L;
the concentrations of the components of the reagent R2 include: tris buffer with the concentration of 80mmol/L, ferrochromine with the concentration of 50mmol/L, ascorbic acid with the concentration of 50mmol/L, and thimerosal with the concentration of 2.0 ml/L.
The pH value of the reagent R1 is 5.00.
The pH of the reagent R2 was 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 30 minutes until all materials are completely dissolved, adjusting the pH value to 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 30 minutes until the materials are completely dissolved, adjusting the pH value to 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.
example four:
The product was calibrated using the standards, and the results are shown in table 1, and the calibration curve is shown in fig. 1.
Table 1: scaling the result
The linear range can be 50-400 mu mol/L.
The precision CV is shown in Table 2:
precision CV < 5%.
In summary, 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.
the above embodiments are merely illustrative of the technical ideas and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and not to limit the protection scope of the present invention. All equivalent changes and modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention.
Claims (5)
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.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111122882A (en) * | 2020-01-02 | 2020-05-08 | 四川纳海川生物科技有限公司 | Total iron detection kit and preparation method thereof |
| CN111141913A (en) * | 2020-01-02 | 2020-05-12 | 四川纳海川生物科技有限公司 | Ceruloplasmin detection kit and preparation method thereof |
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| US4308027A (en) * | 1978-11-08 | 1981-12-29 | R.C.C. Societa' Ricerche Di Chimica Clinica S.R.L. | Method and composition for direct determination of iron in blood serum |
| WO1995013544A1 (en) * | 1992-09-30 | 1995-05-18 | Daiichi Pure Chemicals Co., Ltd. | Reagent for determining unsaturated iron-binding capacity |
| WO2001081930A2 (en) * | 2000-04-26 | 2001-11-01 | Dade Behring Inc. | Reagent and method for "serum iron" assay in plasma |
| CN101226152A (en) * | 2007-01-16 | 2008-07-23 | 温州医学院 | Automatic analysis method and liquid stabilising agent for blood serum total ferro combining ability |
| CN109613280A (en) * | 2018-12-29 | 2019-04-12 | 中拓生物有限公司 | A kind of serum iron determination kit and its preparation method and application |
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2019
- 2019-09-12 CN CN201910868124.4A patent/CN110568206A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4308027A (en) * | 1978-11-08 | 1981-12-29 | R.C.C. Societa' Ricerche Di Chimica Clinica S.R.L. | Method and composition for direct determination of iron in blood serum |
| WO1995013544A1 (en) * | 1992-09-30 | 1995-05-18 | Daiichi Pure Chemicals Co., Ltd. | Reagent for determining unsaturated iron-binding capacity |
| WO2001081930A2 (en) * | 2000-04-26 | 2001-11-01 | Dade Behring Inc. | Reagent and method for "serum iron" assay in plasma |
| CN101226152A (en) * | 2007-01-16 | 2008-07-23 | 温州医学院 | Automatic analysis method and liquid stabilising agent for blood serum total ferro combining ability |
| CN109613280A (en) * | 2018-12-29 | 2019-04-12 | 中拓生物有限公司 | A kind of serum iron determination kit and its preparation method and application |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111122882A (en) * | 2020-01-02 | 2020-05-08 | 四川纳海川生物科技有限公司 | Total iron detection kit and preparation method thereof |
| CN111141913A (en) * | 2020-01-02 | 2020-05-12 | 四川纳海川生物科技有限公司 | Ceruloplasmin detection kit and preparation method thereof |
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