Preservative for blood gas detection reagent, preparation method and blood gas detection reagent

Abstract

The invention discloses a preservative for a blood gas detection reagent, a preparation method and the blood gas detection reagent, the preservative comprises N-(1, 3-dimethylol-2, 5-diketone-4-imidazolidinyl)-N, N '-dimethylolurea, 5-chloro-2-methyl-4-isothiazolin-3-one and alpha-mercaptopyridine-N-sodium oxide, the N-(1, 3-dimethylol-2, 5-diketone-4-imidazolidinyl)-N, N'-dimethylolurea, the 5-chloro-2-methyl-4-isothiazolin-3-one, the alpha-mercaptopyridine-N-sodium oxide and the N-(1, 3-dimethylol-2, 5-diketone-4-imidazolidinyl)-N, N '-dimethylolurea are used as the molar ratio of the alpha-mercaptopyridine-N-oxide sodium to the 5-chloro-2-methyl-4-isothiazolin-3-ketone to the alpha-mercaptopyridine-N-oxide sodium is 1: (0.2-20): (0.5-5), and the molar ratio of the alpha-mercaptopyridine-N-oxide sodium to the 5-chloro-2-methyl-4-isothiazolin-3-ketone to the alpha-mercaptopyridine-N-oxide sodium to the 5-chloro-2-methyl-4- The preservative is almost non-toxic, the cell membrane function of microorganisms can be lost through active ingredients in the preservative, the DNA replication and energy metabolism process of the microorganisms is interfered, and finally the purposes of sterilization and corrosion prevention are achieved. Meanwhile, the preservative also plays a role in improving the stability of the detection reagent, the performance index of the detection reagent is ensured, and the blood gas detection reagent containing the preservative can be widely applied to the field of clinical blood gas detection.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of in vitro diagnostic reagents, and relates to a preservative in an in vitro diagnostic reagent and a preparation method thereof, in particular to a preservative for a blood gas detection reagent, a preparation method and a blood gas detection reagent. BACKGROUND

[0002] Blood gas analysis is a key detection means in clinical diagnosis and intensive care, which can quickly evaluate the respiratory function, acid-base balance and electrolyte state of patients, and provide real-time data support for emergency, surgery and chronic disease management. Blood gas analysis mainly determines the H + Concentration and dissolved gas (mainly CO2, O2) in blood of human body through a blood gas analyzer to understand the respiratory function and acid-base balance state of human body, which can directly reflect the lung gas exchange function and acid-base balance state, and the specimen used is usually arterial blood. The blood gas analyzer is provided with a blood gas detection reagent box, which contains a detection reagent with multiple biological active components and a high-sensitivity electrochemical sensor. The specific composition of the detection reagent varies with product types (calibration liquid, quality control liquid, electrode sensor, etc.) and detection items (pH, electrolyte, blood gas, etc.). Generally speaking, the detection reagent mainly includes: potassium, sodium, chlorine, calcium, magnesium, etc. electrolyte solution, organic or inorganic buffer system, oxygen and carbon dioxide gas partial pressure adjusting component, glucose and lactic acid metabolite and biological molecules, etc. The above-mentioned substances are important components in the blood gas detection reagent, and play a crucial role in the accuracy of blood gas analysis.

[0003] However, at the same time, the blood gas detection reagent is easily contaminated by microorganisms due to its rich ion, oxygen and glucose nutrients during long-term exposure to the environment or multiple opening and use processes, which leads to problems such as deterioration of the detection reagent, electrode performance drift or calibration failure, and ultimately seriously affects the performance indicators of the blood gas detection reagent, causing serious deviation of the detection results, and even causing the risk of misdiagnosis. Therefore, an efficient and stable preservative is an important substance to ensure the reliability and service life of the blood gas detection reagent.

[0004] The common preservatives in the current blood gas detection reagent mainly include sodium azide, thiomersal, benzoic acid / sodium benzoate, etc., which inhibit microbial growth to ensure the stability of the detection reagent and the accuracy of the detection result. Among them, sodium azide can release azide ions in aqueous solution, and azide ions have strong oxidizing property and reactivity, which can react with biological macromolecules such as proteins and nucleic acids in microorganisms, thereby inhibiting or killing microorganisms. It has the advantages of broad-spectrum antibacterial and low price, but its toxicity is strong, which can interfere with some enzymes (such as horseradish peroxidase HRP) or metal ion binding reagents, affecting the performance of the reagent. In addition, sodium azide is flammable and explosive, which poses a safety hazard in transportation and storage. The preservative mechanism of thiomersal is related to the release of mercury ions. Mercury ions can react with sulfhydryl (-SH) in microorganisms to form irreversible sulfur-mercury bonds, thereby destroying the enzyme system and protein structure of microorganisms, interfering with their metabolic activity, and leading to death or loss of reproductive ability. It has good antibacterial effect, but the potential toxicity of mercury easily leads to environmental problems. Sodium benzoate can interfere with the enzyme system of microorganisms, especially hinder the oxidative phosphorylation process in bacterial intracellular, thereby inhibiting the growth and reproduction of microorganisms. It can also change the membrane permeability of microorganisms, block the entry of nutrients and the excretion of metabolic products, thereby reducing the growth rate of microorganisms. However, it is only suitable for acid liquid reagents (such as some buffers). It can be seen that the existing preservatives have the technical problems of high toxicity, limited application range, and inhibition of metal ions in the detection reagent, which damages the detection performance of the reagent and affects the actual detection accuracy.

[0005] Therefore, it is necessary to further improve the preservatives in the blood gas detection reagent in the prior art. SUMMARY

[0006] To this end, the technical problem to be solved by the present application is that the preservatives in the existing blood gas detection reagent have strong toxicity, inhibit metal ions in the diagnostic reagent, damage the reagent and affect the actual detection accuracy, and have a small application range. Therefore, a preservative for blood gas detection reagent with low toxicity, which can improve the stability of the detection reagent and has a wide application range, a preparation method and a blood gas detection reagent are proposed.

[0007] To solve the above technical problems, the technical scheme of the present application is as follows:

[0008] The present application provides a preservative for blood gas detection reagent, comprising: N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dimethylol urea, 5-chloro-2-methyl-4-isothiazolin-3-one, α-mercapto pyridine-N-oxide sodium, wherein the molar ratio of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dimethylol urea, 5-chloro-2-methyl-4-isothiazolin-3-one, α-mercapto pyridine-N-oxide sodium is 1:(0.2-20):(0.5-5).

[0009] As preferred, the molar ratio of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dimethylol urea, 5-chloro-2-methyl-4-isothiazolin-3-one, α-mercapto pyridine-N-oxide sodium is 1:(0.2-5):(0.5-2).

[0010] As preferred, a polar organic solvent is further included.

[0011] As preferred, the polar organic solvent is one of dimethyl sulfoxide, ethanol, ethylene glycol, ethyl acetate, and methanol.

[0012] The present application provides a method for preparing the preservative for blood gas detection reagent as claimed in any one of claims 1-4, comprising the following steps:

[0013] Preparation of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dimethylol urea solution, weighing N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dimethylol urea and dissolving it in a polar organic solvent to obtain N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dimethylol urea solution;

[0014] Preparation of 5-chloro-2-methyl-4-isothiazolin-3-one solution, weighing 5-chloro-2-methyl-4-isothiazolin-3-one and dissolving it in a polar organic solvent to obtain 5-chloro-2-methyl-4-isothiazolin-3-one solution;

[0015] Mixing the N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dimethylol urea solution, 5-chloro-2-methyl-4-isothiazolin-3-one solution and the α-mercapto pyridine-N-oxide sodium uniformly to obtain the preservative.

[0016] As preferred, the concentration of the N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dimethylol urea solution is 0.5-5 mM.

[0017] Preferably, the concentration of the 5-chloro-2-methyl-4-isothiazolin-3-one solution is 0.5-5 mM.

[0018] The third aspect of the present application provides a blood gas detection reagent, which comprises the preservative.

[0019] The above technical solution of the present application has the following advantages compared with the prior art:

[0020] The preservative for blood gas detection reagent provided by the present application comprises N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one, and α-thiopyridine-N-oxide sodium, wherein the molar ratio of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one, and α-thiopyridine-N-oxide sodium is 1:(0.2-20):(0.5-5). The preservative has low toxicity and almost no toxicity, can cause the loss of cell membrane function of microorganisms through the effective components in the above components, and then interfere with the DNA replication and energy metabolism process of microorganisms, and finally achieve the purpose of sterilization and preservation. At the same time, the preservative also plays a role in improving the stability of the detection reagent, ensuring the performance indicators of the detection reagent, and the blood gas detection reagent with the preservative can be widely used in the field of clinical blood gas detection. DETAILED DESCRIPTION

[0021] In order to make the content of the present application more easily understood, the present application will be further clearly and completely explained according to specific embodiments of the present application. The following embodiments are only a part of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor belong to the scope of protection of the present application.

[0022] Embodiment 1

[0023] The present embodiment provides a preservative for blood gas detection reagent, which comprises N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one, and α-thiopyridine-N-oxide sodium, and the molar ratio of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one, and α-thiopyridine-N-oxide sodium is 1:2:1.

[0024] The preservative for blood gas detection reagent provided in the embodiment uses N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazoline-3-ketone, and α-thiopyridine-N-sodium oxide as raw material components. The above raw materials are stable in nature, inexpensive, easy to obtain, and environmentally friendly, and have a preservative effect. N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea can slowly release formaldehyde in an aqueous environment. Formaldehyde is an effective antibacterial agent that can penetrate the cell wall of microorganisms, cross-link biological macromolecules such as proteins and nucleic acids, thereby destroying the metabolic function of microorganisms, leading to the death or inability of microorganisms to reproduce, and achieving the effect of killing microorganisms, thereby achieving the effect of preservation. After 5-chloro-2-methyl-4-isothiazoline-3-ketone contacts microorganisms, it will rapidly react with the thiol groups in the cells of the microorganisms, interfere with the normal folding and function of proteins, cause the loss of cell membrane function of the microorganisms, and then cause the death of the microorganism cells. α-Thiopyridine-N-sodium oxide prevents cell division and proliferation by interfering with the DNA replication and energy metabolism of microorganisms, thereby preventing them from continuing to carry out life activities. The preservative provided in the embodiment can also break the chemical bonds of proteins in bacterial, fungal, and algal cells, destroy their cell structure, and then destroy the cell membrane and cell wall functions of microorganisms, causing the contents of the microorganisms to leak, leading to cell death. In addition, the effective components in the preservative can also penetrate the cell membrane, interfere with the enzyme system in the cell, especially coenzyme A related to energy metabolism, thereby inhibiting the metabolic activity of microorganisms. The components in the above preservative work synergistically to have good bactericidal effect. The preservative only inhibits the metabolic link of bacteria and fungi, has strong bacteriostatic ability, provides a preservative effect, and thus improves the stability of the detection reagent. However, the preservative has no effect on most ionic substances and other components in the detection reagent, does not interfere with the performance of the detection reagent, ensures the detection accuracy of the detection reagent, and can be applied to blood gas detection reagents to improve the preservative performance of blood gas detection reagents.

[0025] To form an aqueous environment, the preservative for blood gas detection reagent provided in the embodiment further includes an organic solvent. N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea and 5-chloro-2-methyl-4-isothiazoline-3-ketone are dissolved in the organic solvent. In the embodiment, the organic solvent is preferably ethanol.

[0026] The embodiment also provides a method for preparing the above-mentioned preservative for blood gas detection reagent, which comprises the following steps:

[0027] Preparation of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dihydroxymethyl urea solution, an appropriate amount of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dihydroxymethyl urea is weighed and dissolved in polar organic solvent ethanol to obtain N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dihydroxymethyl urea solution, and in this embodiment, the concentration of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dihydroxymethyl urea is 2.5 mM.

[0028] Preparation of 5-chloro-2-methyl-4-isothiazolin-3-one solution, an appropriate amount of 5-chloro-2-methyl-4-isothiazolin-3-one is weighed and dissolved in polar organic solvent ethanol to obtain 5-chloro-2-methyl-4-isothiazolin-3-one solution, and in this embodiment, the concentration of 5-chloro-2-methyl-4-isothiazolin-3-one is 3 mM.

[0029] Mix the N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidyl)-N,N'-dihydroxymethyl urea solution and 5-chloro-2-methyl-4-isothiazolin-3-one solution obtained in the above steps with α-sodium mercaptopyridine-N-oxide to obtain the preservative.

[0030] This embodiment also provides a blood gas detection reagent, which comprises the preservative prepared by the above steps. Specifically, the blood gas detection reagent provided by this embodiment is a two-component reagent, which comprises reagent R1 and reagent R2. The reagent R1 and the reagent R2 are stored in the same environment and in separate bags. When used, the reagent R1 and the reagent R2 are respectively calibrated to a blood gas detection instrument to obtain a standard curve of the blood gas detection reagent, so as to test a clinical sample. The specific components and proportions of the blood gas detection reagent are shown in Table 1 below. The reagent R1 and the reagent R2 are prepared by mixing the components with process water to obtain the corresponding concentrations, and adjusting the pH value.

[0031] Table 1

[0032]

[0033] Example 2

[0034] The embodiment provides a preservative for blood gas detection reagent, the preservative comprising N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium, and the molar ratio of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium is 1:0.2:0.5.

[0035] The preservative for blood gas detection reagent provided by the embodiment further comprises an organic solvent, and N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea and 5-chloro-2-methyl-4-isothiazolin-3-one are dissolved in the organic solvent, and in the embodiment, the organic solvent is preferably dimethyl sulfoxide.

[0036] The embodiment further provides a method for preparing the preservative for blood gas detection reagent, and the method comprises the following steps.

[0037] A solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is prepared, an appropriate amount of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is weighed and dissolved in a polar organic solvent dimethyl sulfoxide to obtain a solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, and in the embodiment, the concentration of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is 0.5 mM.

[0038] A solution of 5-chloro-2-methyl-4-isothiazolin-3-one is prepared, an appropriate amount of 5-chloro-2-methyl-4-isothiazolin-3-one is weighed and dissolved in a polar organic solvent dimethyl sulfoxide to obtain a solution of 5-chloro-2-methyl-4-isothiazolin-3-one, and in the embodiment, the concentration of 5-chloro-2-methyl-4-isothiazolin-3-one is 0.5 mM.

[0039] The solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, the solution of 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium obtained in the above steps are uniformly mixed to obtain the preservative.

[0040] The embodiment further provides a blood gas detection reagent, which comprises the preservative prepared in the above steps.

[0041] Embodiment 3

[0042] The embodiment provides a preservative for blood gas detection reagent, the preservative comprising N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium, and the molar ratio of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium is 1:20:5.

[0043] The preservative for blood gas detection reagent provided by the embodiment further comprises an organic solvent, and N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea and 5-chloro-2-methyl-4-isothiazolin-3-one are dissolved in the organic solvent, and in the embodiment, the organic solvent is preferably ethylene glycol.

[0044] The embodiment further provides a method for preparing the preservative for blood gas detection reagent, and the method comprises the following steps.

[0045] A solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is prepared, an appropriate amount of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is weighed and dissolved in a polar organic solvent ethylene glycol to obtain the solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, and in the embodiment, the concentration of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is 5 mM.

[0046] A solution of 5-chloro-2-methyl-4-isothiazolin-3-one is prepared, an appropriate amount of 5-chloro-2-methyl-4-isothiazolin-3-one is weighed and dissolved in a polar organic solvent ethylene glycol to obtain the solution of 5-chloro-2-methyl-4-isothiazolin-3-one, and in the embodiment, the concentration of 5-chloro-2-methyl-4-isothiazolin-3-one is 5 mM.

[0047] The solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, the solution of 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium obtained in the above steps are uniformly mixed to obtain the preservative.

[0048] The embodiment further provides a blood gas detection reagent, which comprises the preservative prepared in the above steps.

[0049] Embodiment 4

[0050] The embodiment provides a preservative for blood gas detection reagent, the preservative comprising N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium, and the molar ratio of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium is 1:5:2.

[0051] The preservative for blood gas detection reagent provided by the embodiment further comprises an organic solvent, and N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea and 5-chloro-2-methyl-4-isothiazolin-3-one are dissolved in the organic solvent, and in the embodiment, the organic solvent is preferably ethyl acetate.

[0052] The embodiment further provides a method for preparing the preservative for blood gas detection reagent, and the method comprises the following steps.

[0053] A solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is prepared, an appropriate amount of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is weighed and dissolved in a polar organic solvent ethyl acetate to obtain a solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, and in the embodiment, the concentration of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is 2 mM.

[0054] A solution of 5-chloro-2-methyl-4-isothiazolin-3-one is prepared, an appropriate amount of 5-chloro-2-methyl-4-isothiazolin-3-one is weighed and dissolved in a polar organic solvent ethyl acetate to obtain a solution of 5-chloro-2-methyl-4-isothiazolin-3-one, and in the embodiment, the concentration of 5-chloro-2-methyl-4-isothiazolin-3-one is 2.5 mM.

[0055] The solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, the solution of 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium obtained in the above steps are uniformly mixed to obtain the preservative.

[0056] The embodiment further provides a blood gas detection reagent, which comprises the preservative prepared in the above steps.

[0057] Embodiment 5

[0058] The embodiment provides a preservative for blood gas detection reagent, the preservative comprises N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium, and the molar ratio of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium is 1:15:3.5.

[0059] The preservative for blood gas detection reagent provided by the embodiment further comprises an organic solvent, and N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea and 5-chloro-2-methyl-4-isothiazolin-3-one are dissolved in the organic solvent, and in the embodiment, the organic solvent is preferably methanol.

[0060] The embodiment further provides a method for preparing the preservative for blood gas detection reagent, and the method comprises the following steps.

[0061] A solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is prepared, an appropriate amount of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is weighed and dissolved in a polar organic solvent methanol to obtain the solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, and in the embodiment, the concentration of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea is 1 mM.

[0062] A solution of 5-chloro-2-methyl-4-isothiazolin-3-one is prepared, an appropriate amount of 5-chloro-2-methyl-4-isothiazolin-3-one is weighed and dissolved in a polar organic solvent methanol to obtain the solution of 5-chloro-2-methyl-4-isothiazolin-3-one, and in the embodiment, the concentration of 5-chloro-2-methyl-4-isothiazolin-3-one is 0.8 mM.

[0063] The solution of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazole alkyl)-N,N'-dihydroxymethyl urea, the solution of 5-chloro-2-methyl-4-isothiazolin-3-one and α-thiopyridine-N-oxide sodium obtained in the above steps are uniformly mixed to obtain the preservative.

[0064] The embodiment further provides a blood gas detection reagent, which comprises the preservative prepared in the above steps.

[0065] Experimental example

[0066] 1. Test the detection results of the blood gas detection reagent provided in Example 1 on the detection samples when the preservative is added and not added.

[0067] The blood gas detection reagent added with the preservative provided in Example 1 and the blood gas detection reagent without the preservative were determined by BG5000E blood gas biochemical analyzer, and the determination parameters and methods were as follows: when the sample bin temperature was 37℃, the reagent R1 and reagent R2 in the blood gas determination reagent were used to calibrate the instrument, 200μL sample was used, the current change was determined by the electrode on the instrument, and the results of K + , Na + , Cl - , Ca 2+ and pH test items in the sample were calculated. + + - 2+ The K + , Na + , Cl - , Ca 2+ and pH values of 15 clinical samples were recorded, and the test results are shown in Table 2.

[0068] Table 2

[0069]

[0070]

[0071] The test results show that the blood gas detection reagent added with the preservative provided in the application has no significant difference in the determination of the K + , Na + , Cl - , Ca 2+ content and pH value of the sample compared with the blood gas detection reagent without the preservative, which shows that the preservative provided in the application does not affect the detection performance of the blood gas detection reagent.

[0072] 2. Preservative effect test

[0073] The blood gas detection reagent without the preservative provided in the application (reagent A), the blood gas detection reagent added with the conventional Proclin preservative (reagent B) and the blood gas detection reagent added with the preservative provided in Example 1 of the application (reagent C) were coated in the culture medium and placed in a 37℃ incubator for 1-8 weeks, and visual confirmation was made to see if there was bacterial growth, and the test results are shown in Table 3.

[0074] Table 3

[0075]

[0076] From the above test results, it can be seen that the blood gas detection reagent without the addition of the preservative has bacterial growth within one week, the blood gas detection reagent with the addition of the conventional Proclin preservative has bacterial growth starting from the sixth week, and the blood gas detection reagent with the addition of the preservative provided in the application has no bacterial proliferation within 8 weeks.

[0077] 3. Determination of the preservative effect of the preservative with different proportions

[0078] The preservative is prepared according to the preparation method provided in the application under different proportions of raw materials, and the preservative includes N-(1, 3-dihydroxymethyl-2, 5-diketone-4-imidazole alkyl)-N, N'-dihydroxymethyl urea (hereinafter referred to as A), 5-chloro-2-methyl-4-isothiazoline-3-ketone (hereinafter referred to as B) and α-mercapto pyridine-N-oxide sodium (hereinafter referred to as C). 36 groups of test culture media are prepared, and the components of the test culture media are shown in Table 4 as follows:

[0079] Table 4

[0080]

[0081] The test culture media are prepared by mixing the above components with purified water, stirring uniformly and adjusting the pH value. The 36 groups of test culture media with the addition of the preservative with different proportions are stored in a culture box at 37°C for 4 weeks, and then the bacterial confirmation experiment is performed on the test culture media after storage, and the test results are shown in Table 5 as follows:

[0082] Table 5

[0083]

[0084]

[0085] The above 36 groups of test culture media are observed at the first week, the third week and the fourth week, and no bacterial growth is found in the culture media. It can be seen that the preservative provided in the application has good bacteriostatic effect within the above molar ratio range.

[0086] The preservative formed by the components of N-(1, 3-dihydroxymethyl-2, 5-diketone-4-imidazole alkyl)-N, N'-dihydroxymethyl urea, 5-chloro-2-methyl-4-isothiazoline-3-ketone and α-mercapto pyridine-N-oxide sodium has not only the strong preservative effect as shown in the above test results, but also avoids the problems of the traditional sodium azide and mercuric sulfide preservative which brings harm to the environment and does not meet the goal of green chemistry, and overcomes the technical problem of the limited use of benzoic acid / sodium benzoate (only applicable to acidic solution). The preservative provided in the application is applicable to alkaline solution and has a wider application range, so that the blood gas detection reagent with the preservative can be widely applied to clinical blood gas detection.

[0087] Obviously, the above embodiments are merely example for clearly illustrating but not limitation to the embodiments. Based on the above description, other different forms of changes or variations can be made by those skilled in the art. Here, all the embodiments need not and can not be enumerated. The obvious changes or variations derived from the above description are still within the protection scope of the present application.

Claims

1. A preservative for blood gas testing reagents, characterized in that, include: N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidinyl)-N,N'-dihydroxymethylurea, 5-chloro-2-methyl-4-isothiazolin-3-one, and α-mercaptopyridine-N-oxide sodium, wherein the molar ratio of N-(1,3-dihydroxymethyl-2,5-diketone-4-imidazolidinyl)-N,N'-dihydroxymethylurea, 5-chloro-2-methyl-4-isothiazolin-3-one, and α-mercaptopyridine-N-oxide sodium is 1:(0.2-20):(0.5-5).

2. The preservative for blood gas testing reagents according to claim 1, characterized in that, The molar ratio of N-(1,3-dihydroxymethyl-2,5-dione-4-imidazolidinyl)-N,N'-dihydroxymethylurea, 5-chloro-2-methyl-4-isothiazolin-3-one, and α-mercaptopyridine-N-oxide sodium is 1:(0.2-5):(0.5-2).

3. The preservative for blood gas testing reagents according to claim 1 or 2, characterized in that, It also includes polar organic solvents.

4. The preservative for blood gas testing reagents according to claim 3, characterized in that, The polar organic solvent is one of dimethyl sulfoxide, ethanol, ethylene glycol, ethyl acetate, and methanol.

5. A method for preparing a preservative for a blood gas testing reagent as described in any one of claims 1-4, characterized in that, Includes the following steps: To prepare N-(1,3-dihydroxymethyl-2,5-dione-4-imidazolidinyl)-N,N'-dihydroxymethylurea solution, weigh N-(1,3-dihydroxymethyl-2,5-dione-4-imidazolidinyl)-N,N'-dihydroxymethylurea and dissolve it in a polar organic solvent to obtain N-(1,3-dihydroxymethyl-2,5-dione-4-imidazolidinyl)-N,N'-dihydroxymethylurea solution; To prepare a 5-chloro-2-methyl-4-isothiazolin-3-one solution, weigh 5-chloro-2-methyl-4-isothiazolin-3-one and dissolve it in a polar organic solvent to obtain the 5-chloro-2-methyl-4-isothiazolin-3-one solution. The N-(1,3-dihydroxymethyl-2,5-dione-4-imidazolidinyl)-N,N'-dihydroxymethylurea solution, the 5-chloro-2-methyl-4-isothiazolin-3-one solution, and the α-mercaptopyridine-N-oxide sodium were mixed evenly to obtain the preservative.

6. The preparation method according to claim 5, characterized in that, The concentration of the N-(1,3-dihydroxymethyl-2,5-dione-4-imidazolidinyl)-N,N'-dihydroxymethylurea solution is 0.5-5 mM.

7. The preparation method according to claim 5, characterized in that, The concentration of the 5-chloro-2-methyl-4-isothiazolin-3-one solution is 0.5-5 mM.

8. A blood gas testing reagent, characterized in that, Includes the preservatives for blood gas testing reagents as described in any one of claims 1-4.

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