CN108169468B

CN108169468B - Diluent suitable for various blood analyzers and preparation method thereof

Info

Publication number: CN108169468B
Application number: CN201711312375.1A
Authority: CN
Inventors: 韩作湘; 梁净净; 曹丹; 王国锋
Original assignee: Stac Medical Science & Technology Co ltd
Current assignee: Shanghai Lanqiao Biotechnology Co., Ltd
Priority date: 2017-12-12
Filing date: 2017-12-12
Publication date: 2020-04-07
Anticipated expiration: 2037-12-12
Also published as: CN108169468A

Abstract

The invention relates to the technical field of in-vitro diagnostic reagents, in particular to a diluent suitable for various blood analyzers and a preparation method thereof. The diluent comprises a diluent basic formula, 1.0-15.0 g/L of osmotic pressure compensation agent and 0.6-1.5 g/L of buffering agent; the basic formula of the diluent comprises 0.2-1.0 g/L of anticoagulant, 0.3-0.5 g/L of preservative and 0.2-1.5 g/L of stabilizer. The diluent for the blood analyzer improves the stability and the measurement reliability of a blood sample substance by compounding the stabilizer isopropanol and the 1, 2-propylene glycol in the formula, and can be suitable for the blood analyzers of different manufacturers by fixing the basic formula of the diluent and only adjusting the contents of the buffer and the osmotic pressure compensator, thereby simplifying the development steps of in-vitro diagnostic reagents and reducing the research and development cost. And breaks monopoly of reagents of original plants, and reduces the use cost of reagents of all levels of medical institutions.

Description

Diluent suitable for various blood analyzers and preparation method thereof

Technical Field

The invention relates to the technical field of in-vitro diagnostic reagents, in particular to a diluent suitable for various blood analyzers and a preparation method thereof.

Background

Blood cell analysis is the most common item in clinical examination, and blood cells in a blood sample are analyzed and detected by a blood analyzer by extracting the blood sample from a human body, and the detected parameters comprise White Blood Cells (WBC), Red Blood Cells (RBC), Hemoglobin (HGB), Platelets (PLT), Hematocrit (HCT), Mean Corpuscular Volume (MCV) and the like.

The blood analyzer uses a plurality of reagents to assist the detection, wherein a diluent is an indispensable reagent. The diluent for the blood analyzer is mainly used for diluting blood cells and preventing the blood cells from aggregating and adhering; and a certain PH value and osmotic pressure are maintained, so that a proper environment is provided for blood cell detection.

At present, reagents of blood analyzers in the market are various in varieties and are special for special machines, reagents of original factories are often expensive, particularly diluents, the daily work requirement is large, the application cost is very high, and the blood analyzers bring troubles on the cost to vast medical institutions. Therefore, the development of a diluent which has a simple formula and low cost, is suitable for various machine types, can replace the diluent used on the machine of the original factory reagent, and reduces the use cost of the diluent of the hematology analyzer with various levels of medical structures is necessary.

Disclosure of Invention

In order to solve the technical problems, the invention provides a diluent suitable for various blood analyzers and a preparation method thereof.

The technical scheme of the invention is as follows:

the invention provides a diluent suitable for various blood analyzers, which comprises a diluent basic formula, an osmotic pressure compensation agent 1.0-15.0 g/L and a buffering agent 0.6-1.5 g/L;

the basic formula of the diluent comprises 0.2-1.0 g/L of anticoagulant, 0.3-0.5 g/L of preservative and 0.2-1.5 g/L of stabilizer.

Further, the anticoagulant is ethylenediamine tetraacetate, and the concentration of the anticoagulant is 0.2-1.0 g/L.

Further, the anticoagulant is EDTA-K₂。

Further, the preservative is one or more of sodium benzoate, potassium sorbate and potassium hydrogen phthalate, and the concentration of the preservative is 0.3-0.5 g/L; a preferred preservative is potassium hydrogen phthalate.

Further, the stabilizer is one or more of ethanol, ethylene glycol, isopropanol and 1, 2-propylene glycol; the preferable stabilizer is isopropanol and 1, 2-propylene glycol, wherein the concentration of the isopropanol is 0.1-2.0 g/L, and the preferable concentration is 0.5-1.5 g/L; the concentration of 1, 2-propanediol is 0.1 to 1.5g/L, preferably 0.2 to 1.2 g/L.

Further, the osmotic pressure compensator is one or more of sodium chloride, potassium chloride, sodium sulfate and potassium sulfate, and preferably sodium chloride; the concentration is 5.0-10.0 g/L.

Further, the buffer is one or more of phosphate buffer, Tris-HCl buffer and Good's buffer; preferably Tris-HCl buffer; the concentration is 0.6-1.5 g/L.

The pH value of the diluent prepared by the invention is 7.0-9.0.

In a preferred embodiment of the present invention, the diluent is prepared by determining the contents of anticoagulant, stabilizer and preservative, adjusting the buffer to maintain the pH of the reagent consistent with the factory reagent, and adjusting the osmotic pressure compensating agent to maintain the average red blood cell volume (MCV) of the on-machine test consistent with the factory reagent based on the on-machine results of the blood analyzer. The preparation process includes determining the basic recipe of diluent, and regulating osmotic pressure compensator and buffering agent to make the diluent match with corresponding blood analyzer.

The anticoagulant can chelate with some positive ions (especially calcium and iron ions) in the blood sample after being added to the blood sample to form stable chelate, thereby preventing blood coagulation. The preservative ensures that the substances are not humus or rotten, and further ensures the stability of the diluent. The stabilizing agent is used for maintaining the osmotic pressure and potential balance of the blood sample, so that the stability of each cell and component in the blood sample is improved, and meanwhile, the solution after the blood sample is diluted presents a stable hydroelectric layer, and the uniformity and stability of the whole solution are ensured. The role of the osmolality compensator in the diluent is to further ensure the concentration of the whole blood sample diluent, thereby maintaining the osmolality of the diluent. The buffer is used to adjust the pH of the diluent.

The invention has the beneficial effects that: the diluent for the blood analyzer improves the stability and the measurement reliability of a blood sample substance by compounding the stabilizer isopropanol and the 1, 2-propylene glycol in the formula, and can be suitable for the blood analyzers of different manufacturers by fixing the basic formula of the diluent and only adjusting the contents of the buffer and the osmotic pressure compensator, thereby simplifying the development steps of in-vitro diagnostic reagents and reducing the research and development cost. And breaks monopoly of reagents of original plants, and reduces the use cost of reagents of all levels of medical institutions.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below. It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments.

The purity of the reagents used in the examples of the present invention was analytical grade.

The diluent comprises the following components: EDTA-K₂0.2-1.0 g/L of isopropanol, 0.5-1.5 g/L of isopropanol, 0.2-1.2 g/L of 1, 2-propylene glycol, 5.0-10.0 g/L, Tris-HCl, 0.6-1.5 g/L of sodium chloride and 0.3-0.5 g/L of potassium hydrogen phthalate.

The invention first determines EDTA-K₂Isopropanol, 1, 2-propanediol and potassium hydrogen phthalate, and secondly adjusting the Tris-HCl content to enable the dilution to be analysed with different bloodThe pH value of the instrument is matched, and the amount of sodium chloride serving as an osmotic pressure compensator is adjusted according to the on-machine result of the adopted blood analyzer, so that the average red blood cell volume (MCV) detected on the on-machine instrument is consistent with that of the original reagent. Finally, the diluent is used for analyzing and detecting blood cells on a blood cell analyzer, and the applicability of the diluent to different models is evaluated according to the correlation between the diluent and the detection result of the original factory reagent. The test is carried out by taking ABXMICROS 60, ABX Pentra 80 and MEK-6318 models as examples, but is not limited to the three models. Meanwhile, in order to verify that the reliability and the stability of the detection data of the diluent are better than those of the diluent added with only one stabilizer by compounding the stabilizer isopropanol and the 1, 2-propylene glycol, the invention carries out the following embodiments.

The components were accurately weighed according to the following basic formulation in table 1, the PH of the solution was controlled by adding different amounts of Tris-HCl buffer, and the osmolality and conductivity of the solution were adjusted by adding sodium chloride to base solutions of different PH. After dissolving in sequence, using purified water to fix the volume, and filtering by using a 0.2 mu m microporous filter membrane to obtain the diluent for the blood analyzer. According to different PH and sodium chloride, the diluent formula suitable for different blood analyzers is determined.

TABLE 1

Example 1

A base solution having PH =7.1 was prepared according to the above table, and then 6.0g of sodium chloride was added to the base solution. The osmotic pressure of example 1 was 325 mOsm/kg; the conductivity was 18.9mS/cm.

Fresh blood of 5 healthy people is taken, the diluent of the invention in the example is used for testing on an ABX MICROROS 60 blood analyzer by using the original factory diluent and the invention diluent respectively, and blood cell analysis and detection results are compared. The results of the experiment are shown in table 2.

TABLE 2 comparison of the results of blood cell analysis and measurement of the reagent of the present invention on ABX MICROROS 60 blood analyzer with the results of the reagent of the original factory

Example 2

A base solution having PH =8.0 was prepared according to the above table, and 7.2g of sodium chloride was added to the base solution. The osmotic pressure of example 2 was 340 mOsm/kg; the conductivity was 18.2mS/cm.

Fresh blood of 5 healthy people is taken, the diluent of the invention of the second embodiment is used for testing on an ABX Pentra 80 blood analyzer by using the original factory diluent and the invention diluent respectively, and the analysis and detection results of blood cells are compared. The results of the experiment are shown in table 3.

TABLE 3 comparison of the result of blood cell analysis and detection of the reagent of the present invention in ABX Pentra 80 blood analyzer with the experimental result of the reagent of the original factory

Example 3

A base solution having PH =7.3 was prepared according to the above table, and then 7.0g of sodium chloride was added to the base solution. The osmotic pressure of example three was 330 mOsm/kg; the conductivity was 18.15mS/cm.

Fresh blood of 5 healthy people is taken, the diluent of the invention of the third embodiment is used for testing on an MEK-6318 blood analyzer by using the original factory diluent and the invention diluent respectively, and the differential counting result of the leucocytes is compared. The results of the experiment are shown in table 4.

TABLE 4 comparison of the results of blood cell analysis and detection of the reagent of the present invention in MEK-6318 blood analyzer with the results of the reagent experiment in the factory

Example 4

The diluent formulations are shown in table 5.

TABLE 5

The diluent A, B, C suitable for the three blood analyzers was prepared according to the above table, and the results of analyzing and detecting blood cells were compared with the reagents of the present invention of the first, second, and third examples. The comparative results are shown in Table 6.

Table 6 example 4 comparison of the results of the assay of the inventive reagents with the blood cells of example 1, example 2 and example 3 on a hematology analyzer.

As can be seen from the table, the pH of the solution is controlled within the range of 7.0-9.0, and the diluent prepared according to the invention can completely replace the original reagent to carry out blood cell analysis and detection on blood analyzers such as ABX MICROROS 60, ABX Pentra 80, MEK-6318 and the like. And by compounding the stabilizer isopropanol and the 1, 2-propylene glycol, the reliability and the stability of the detection data of the diluent are superior to those of the stabilizer only added with the isopropanol. In conclusion, the diluent for different blood analyzers provided by the invention has the advantages of simple operation, easy realization and lower cost of the prepared reagent, and can meet the requirements of most blood analyzers on analysis and detection of blood cells in the market.

Claims

1. The diluent suitable for various blood analyzers is characterized by comprising a diluent basic formula, an osmotic pressure compensation agent 1.0-15.0 g/L and a buffering agent 0.6-1.5 g/L;

the basic formula of the diluent comprises 0.2-1.0 g/L anticoagulant, 0.3-0.5 g/L preservative and 0.2-1.5 g/L stabilizer;

the stabilizer is isopropanol and 1, 2-propylene glycol, and the concentration of the isopropanol is 0.1-2.0 g/L; the concentration of the 1, 2-propylene glycol is 0.1-1.5 g/L;

the anticoagulant is ethylenediamine tetraacetate, and the concentration of the anticoagulant is 0.2-1.0 g/L;

the osmotic pressure compensator is one or more of sodium chloride, potassium chloride, sodium sulfate and potassium sulfate.

2. The diluent of claim 1, wherein the anticoagulant is EDTA-K₂。

3. The diluent of claim 1, wherein the preservative is one or more of sodium benzoate, potassium sorbate and potassium hydrogen phthalate, and the concentration is 0.3-0.5 g/L.

4. The diluent of claim 3, wherein the preservative is potassium hydrogen phthalate.

5. The diluent according to claim 1, wherein the concentration of the isopropanol is 0.5-1.5 g/L; the concentration of the 1, 2-propylene glycol is 0.2-1.2 g/L.

6. The diluent of claim 1, wherein the osmotic pressure compensator is sodium chloride; the concentration is 5.0-10.0 g/L.

7. The diluent of claim 1, wherein the buffer is one or more of a phosphate buffer, a Tris-HCl buffer, and a Good's buffer.

8. The diluent according to any one of claims 1 to 7, wherein the pH of the diluent is 7.0 to 9.0.