阅读说明：本技术 谷胱甘肽还原酶测定试剂质控品及制备方法 (Glutathione reductase determination reagent quality control product and preparation method thereof ) 是由 万蒙 邱家明 周惠良 于 2019-09-29 设计创作，主要内容包括：本发明公开了一种谷胱甘肽还原酶测定试剂质控品,其特征在于,包括以下组分的冻干品：磷酸盐缓冲液50～80份,甘氨酸5～10份,黄原胶1～3份,牛血清5～10份,纯品谷胱甘肽还原酶0.1～0.5份,蔗糖2～5份,防腐剂1～2份。质控品为冻干品,能大大提升质控品在使用前的稳定性,易于保存。本发明还公开了上述质控品的制备方法,包括以下步骤：取所述纯品谷胱甘肽还原酶,加入磷酸盐缓冲液,得稀释液；向所述稀释液中加入甘氨酸,保温搅拌,将所得混合溶液等分为第一混合液和第二混合液；向所述第一混合液中加入牛血清和蔗糖,得第一冻干液；向所述第二混合液中加入牛血清、黄原胶和蔗糖,得第二冻干液；将所述第一冻干液和所述第二冻干液混合,冷冻干燥,得质控品。(The invention discloses a glutathione reductase determination reagent quality control product which is characterized by comprising a freeze-dried product of the following components: 50-80 parts of phosphate buffer solution, 5-10 parts of glycine, 1-3 parts of xanthan gum, 5-10 parts of bovine serum, 0.1-0.5 part of pure glutathione reductase, 2-5 parts of sucrose and 1-2 parts of preservative. The quality control product is freeze-dried, can greatly improve the stability of the quality control product before use, and is easy to store. The invention also discloses a preparation method of the quality control product, which comprises the following steps: adding phosphate buffer solution into the pure glutathione reductase to obtain diluent; adding glycine into the diluent, stirring at a constant temperature, and equally dividing the obtained mixed solution into a first mixed solution and a second mixed solution; adding bovine serum and sucrose into the first mixed solution to obtain a first freeze-dried solution; adding bovine serum, xanthan gum and sucrose into the second mixed solution to obtain a second freeze-dried solution; and mixing the first freeze-drying liquid and the second freeze-drying liquid, and freeze-drying to obtain a quality control product.)

1. The quality control product of the glutathione reductase determination reagent is characterized by comprising a freeze-dried product of the following components:

50-80 parts of phosphate buffer solution,

5-10 parts of glycine,

1-3 parts of xanthan gum,

5-10 parts of bovine serum,

0.1-0.5 part of pure glutathione reductase,

2-5 parts of cane sugar,

1-2 parts of a preservative.

2. The glutathione reductase assay reagent quality control product of claim 1, wherein the phosphate buffer solution comprises disodium hydrogen phosphate and sodium dihydrogen phosphate in a mass ratio of 2: 3-6.

3. The glutathione reductase assay reagent quality control product according to claim 1, wherein the preservative comprises sodium azide and sodium glutamate in a mass ratio of 3: 1-2.

4. The method for preparing the glutathione reductase assay reagent quality control product according to any one of claims 1 to 3, characterized by comprising the following steps:

taking the pure glutathione reductase, and adding a phosphate buffer solution for dilution to obtain a diluent;

adding glycine into the diluent, stirring at a constant temperature, and equally dividing the obtained mixed solution into a first mixed solution and a second mixed solution;

adding bovine serum and sucrose into the first mixed solution, uniformly mixing, and standing at a low temperature for 1 hour to obtain a first freeze-dried solution;

adding bovine serum, xanthan gum and sucrose into the second mixed solution, uniformly mixing, and standing for 1 hour under the condition of heat preservation to obtain a second freeze-dried solution;

and mixing the first freeze-drying liquid and the second freeze-drying liquid to obtain a freeze-drying material, and performing freeze drying on the freeze-drying material to obtain the glutathione reductase determination reagent quality control product.

5. The method according to claim 4, wherein the concentration of the phosphate buffer is 50 to 400mmol/L, the concentration of glycine is 50 to 100mmol/L, the concentration of xanthan gum is 100 to 200mmol/L, and the concentration of bovine serum is 30 to 95%.

6. The method according to claim 4, wherein the concentration of glutathione reductase in the diluted solution is 10 to 100U/L.

7. The method according to claim 4, wherein the glycine is added to the diluted solution, and the stirring is performed while maintaining the temperature of the solution at 40 ℃ for 30 minutes.

8. The method according to claim 4, wherein the temperature of the low-temperature standing is 4 ℃ and the temperature of the heat-retaining standing is 20 ℃.

9. The method of claim 4, wherein the step of freeze-drying comprises:

pre-freezing, namely keeping the freeze-dried material at the temperature of-30 to-70 ℃ for 12 to 24 hours;

and (3) performing vacuum freeze drying, namely placing the freeze-dried material subjected to pre-freezing in a negative pressure environment at the temperature of between 20 ℃ below zero and 60 ℃ below zero for 24 to 48 hours.

10. The method of claim 9, wherein the vacuum freeze-drying step comprises:

placing the freeze-dried material in an environment with the atmospheric pressure of 5Pa, and keeping the temperature of the environment at-40 to-60 ℃ for 24 to 36 hours;

and (3) placing the freeze-dried material in an environment with the atmospheric pressure of 15Pa, keeping the environment at the temperature of-20 to-30 ℃ for 8 to 12 hours.

Technical Field

The invention relates to the technical field of biochemical detection, in particular to a glutathione reductase determination reagent quality control product and a preparation method thereof.

Background

Glutathione Reductase (GR) is contained in blood, and its content is measured by a glutathione reductase measuring reagent.

In the process of producing the glutathione reductase determination reagent, a sample needs to be calibrated by adopting a calibrator, a calibration system can be generated by utilizing the calibrator, and then a quality control product is adopted to detect whether the calibration system is qualified or not, so that the quality control product is required to have higher precision and stability. At present, most of quality control products and calibration products are the same components, and the components are liquid pure glutathione reductase, buffer solution and stabilizer.

Because the precision and the stability of the quality control product have higher requirements, the existing liquid quality control product is not easy to store, and can quickly deteriorate after being unsealed, thereby seriously affecting the detection result.

Disclosure of Invention

The invention aims to provide a stable and easily-preserved glutathione reductase determination reagent quality control product.

A glutathione reductase assay reagent quality control product comprises a freeze-dried product of the following components:

50-80 parts of phosphate buffer solution,

5-10 parts of glycine,

1-3 parts of xanthan gum,

5-10 parts of bovine serum,

0.1-0.5 part of pure glutathione reductase,

2-5 parts of cane sugar,

1-2 parts of a preservative.

The invention has the beneficial effects that: the quality control product is a freeze-dried product, can greatly improve the stability of the quality control product before use, and is easy to store; the phosphate buffer solution can facilitate the preparation and molding of the quality control product, the preservative can prolong the quality of the quality control product, the sucrose can prevent the inactivation of the pure glutathione reductase in the preparation process, and the glycine and the xanthan gum can keep the overall activity of the quality control product.

In addition, the quality control product of the reagent for measuring the glutathione reductase provided by the invention can also have the following additional technical characteristics:

further, the phosphate buffer solution comprises disodium hydrogen phosphate and sodium dihydrogen phosphate, and the mass ratio of the disodium hydrogen phosphate to the sodium dihydrogen phosphate is 2: 3-6.

Further, the preservative comprises sodium azide and sodium glutamate in a mass ratio of 3: 1-2.

The invention also aims to provide a preparation method of the glutathione reductase assay reagent quality control product, which comprises the following steps:

(1) taking the pure glutathione reductase, and adding a phosphate buffer solution for dilution to obtain a diluent;

(2) adding glycine into the diluent, stirring at a constant temperature, and equally dividing the obtained mixed solution into a first mixed solution and a second mixed solution;

(3) adding bovine serum and sucrose into the first mixed solution, uniformly mixing, and standing at a low temperature for 1 hour to obtain a first freeze-dried solution;

(4) adding bovine serum, xanthan gum and sucrose into the second mixed solution, uniformly mixing, and standing for 1 hour under the condition of heat preservation to obtain a second freeze-dried solution;

(5) and mixing the first freeze-drying liquid and the second freeze-drying liquid to obtain a freeze-drying material, and performing freeze drying on the freeze-drying material to obtain the glutathione reductase determination reagent quality control product.

Further, the concentration of the phosphate buffer solution is 50-400 mmol/L, the concentration of glycine is 50-100 mmol/L, the concentration of xanthan gum is 100-200 mmol/L, and the concentration of bovine serum is 30-95%.

Further, the concentration of the glutathione reductase in the diluent is 10-100U/L.

Further, glycine is added into the diluent, and in the step of stirring at a constant temperature, the stirring at the constant temperature is performed for 30 minutes while keeping the temperature of the solution at 40 ℃.

Further, the temperature of the low-temperature standing is 4 ℃, and the temperature of the heat preservation standing is 20 ℃.

Further, the freeze-drying step comprises:

pre-freezing, namely keeping the freeze-dried material at the temperature of-30 to-70 ℃ for 12 to 24 hours;

and (3) performing vacuum freeze drying, namely placing the freeze-dried material subjected to pre-freezing in a negative pressure environment at the temperature of between 20 ℃ below zero and 60 ℃ below zero for 24 to 48 hours.

Further, the vacuum freeze drying step is as follows:

placing the freeze-dried material in an environment with the atmospheric pressure of 5Pa, and keeping the temperature of the environment at-40 to-60 ℃ for 24 to 36 hours;

and (3) placing the freeze-dried material in an environment with the atmospheric pressure of 15Pa, keeping the environment at the temperature of-20 to-30 ℃ for 8 to 12 hours.

Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.

Detailed Description

In order to make the objects, features and advantages of the present invention comprehensible, specific embodiments accompanied with examples are described in detail below. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.