CN111811996B

CN111811996B - Leukocyte classification kit, preparation method thereof and dye waste liquid treatment method

Info

Publication number: CN111811996B
Application number: CN202010476909.XA
Authority: CN
Inventors: 李晓娟; 姚燕丽; 郑剑通
Original assignee: Shenzhen Dymind Biotechnology Co Ltd
Current assignee: Shenzhen Dymind Biotechnology Co Ltd
Priority date: 2020-05-29
Filing date: 2020-05-29
Publication date: 2022-10-04
Anticipated expiration: 2040-05-29
Also published as: CN111811996A

Abstract

The application relates to the technical field of biological sample detection, and particularly discloses a leukocyte classification kit, a preparation method thereof and a dye waste liquid treatment method. The leukocyte classification kit comprises: a hemolytic agent for lysing erythrocytes and causing cell membrane damage of leukocytes; a biomass sorbent comprising: collagen, polyphenol substances and a cross-linking agent; the dyeing agent comprises a dye capable of dyeing nucleic acid, and the dye can perform chelation reaction with polyphenol substances; the hemolytic agent, the biomass adsorbent, and the staining agent are each present in a single package. Through the mode, the problem that the dye easily causes environmental pollution in the prior art is solved.

Description

Leukocyte classification kit, preparation method thereof and dye waste liquid treatment method

Technical Field

The application relates to the technical field of biological detection, in particular to a leukocyte classification kit, a preparation method thereof and a dye waste liquid treatment method.

Background

With the continuous development and improvement of the scientific and technical level, peripheral blood is mostly analyzed by a full-automatic blood cell analyzer in hospitals at present.

It is disclosed in the prior art that erythrocytes are lysed by a non-ionic surfactant such as saponin, and then the nuclei of cells are stained by a nucleic acid dye such as chlorazol black, and leukocytes are classified and counted by measuring the intensity and quantity of fluorescence.

However, the inventor of the present application found in a long-term research and development process that the dyes in such kits are environmentally-unfriendly materials, and the use of the dyes easily causes environmental pollution.

Disclosure of Invention

The application provides a leukocyte classification kit, a preparation method thereof and a dye waste liquid treatment method, and aims to solve the problem that dyes easily cause environmental pollution in the prior art.

In one aspect, the present application provides a leukocyte classification kit comprising: a hemolytic agent for lysing erythrocytes and damaging cell membranes of leukocytes; a biomass sorbent comprising: collagen, polyphenols and cross-linking agents; the dyeing agent comprises a dye capable of dyeing nucleic acid, and the dye can perform chelation reaction with polyphenol substances; the hemolytic agent, the biomass adsorbent and the staining agent are respectively in a single package.

In another aspect, the present application provides a method for preparing a differential white blood cell count kit, the method comprising: weighing an anionic surfactant, a cationic surfactant, an osmotic pressure regulator, a buffering agent and a preservative, dissolving in a first solvent, regulating the pH value, and fixing the volume to obtain a hemolytic agent; weighing a dye, dissolving the dye in a second solvent, adjusting the pH value, and fixing the volume to obtain a dyeing agent; weighing collagen, polyphenol substances and a cross-linking agent, and carrying out cross-linking reaction on the collagen and the polyphenol substances under the action of the cross-linking agent to obtain a biomass adsorbent; the hemolytic agent, the biomass adsorbent and the staining agent are packaged in a kit in a single package form.

In another aspect, the present application provides a method for treating a dye waste solution, the method for treating a dye waste solution is based on the above-mentioned leukocyte classification kit, and the method comprises: respectively connecting a hemolytic agent, a biomass adsorbent and a coloring agent in the leukocyte classification kit into corresponding pipelines of a particle analyzer; classifying the white blood cells by using hemolytic agents and staining agents; after the hemolytic agent and the staining agent are exhausted, adjusting the pH value of the waste liquid discharged by the particle analyzer to a preset pH value, and measuring the dye concentration in the waste liquid; and (3) adding the biomass adsorbent according to a preset feed ratio, and treating the waste liquid for a preset time.

The beneficial effect of this application is: in contrast to the state of the art, the present application provides a leukocyte classification kit comprising: the hematology analyzer can be used for distinguishing different types of leucocytes, has a good effect, and is particularly suitable for automatically classifying the leucocytes by a cell morphology analysis technology based on a machine vision principle. In addition to the hemolytic agent and the staining agent, the leukocyte classification kit further includes: the biomass adsorbent is characterized in that polyphenol substances in the biomass adsorbent can be subjected to chelation reaction with dyes, so that the dyes remained in waste liquid are adsorbed on the biomass adsorbent, and substances harmful to the environment are effectively treated through precipitation and filtration. By adding the dye treatment adsorbent in the leukocyte classification kit, the problem of environmental pollution caused by dyes in the prior art can be effectively solved, and the leukocyte classification kit is low in price and obvious in adsorption effect.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts. Wherein:

FIG. 1 shows the effect of the amount of biomass adsorbent on the adsorption effect;

FIG. 2 shows the effect of solution pH on adsorption effect;

FIG. 3 shows the effect of adsorption time on adsorption effect;

FIG. 4 shows the results of the classification of the leukocyte classification kit of the present application;

FIG. 5 is a schematic flow chart of an embodiment of a method for treating waste dye solution according to the present application;

FIG. 6 is a schematic flow chart of another embodiment of the dye waste liquid treatment method of the present application.

Detailed Description

The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

The application provides a kit for classifying leukocytes, comprising: the hemolytic agent, the staining agent and the biomass adsorbent are respectively in a single package.

Leukocytes are a type of cells in the blood, and are also called leukocytes because they have colorless spherical shapes. The white blood cells have cell nucleuses, are larger than the red blood cells in volume, have the diameter of between 7 and 20 mu m, and have the function of immunity. The human body is untimely and often manifests itself by a significant change in the number of leukocytes. The pathological increase of white blood cells is very important, for example, acute bacterial infection can lead to rapid increase of white blood cells and is proportional to the infection degree, and doctors mostly determine whether the clinical symptoms are caused by bacterial infection through the change of the white blood cell quantity. The increase of white blood cells often means that the symptoms of the patient are caused by bacterial infection, such as acute or chronic infection, inflammation, tissue injury and the like of each body organ or tissue. Infection of the human body without the increase of leukocytes is often regarded as infection caused by virus invasion, and the increase of leukocytes is in a small and slow state. Also, the decrease in the number of leukocytes is a considerable problem, and for example, chronic exposure to radiation, poisoning due to various physicochemical factors, chemotherapy and radiotherapy for tumors, splenic hyperactivity, autoimmune diseases, aplastic anemia, hematopoietic disorders, etc. all cause a decrease in leukocytes, particularly neutrophils.

The hemolytic agent can rapidly dissolve red blood cells, release hemoglobin to be combined with the hemolytic agent to form a stable hemoglobin derivative, meanwhile, a surfactant in the hemolytic agent is used for punching white blood cells, and different types of white blood cells are subjected to shrinkage to different degrees after being treated by the hemolytic agent; then, when the nucleic acid in the nucleus is stained and labeled by the dye, the scattered light intensity of different white blood cells stained by the dye is different. The accurate classification of the white blood cells is realized through scattered light analysis.

The biomass adsorbent comprises collagen, polyphenol substances and a cross-linking agent.

Wherein the collagen is extracted from cattle ear, fish skin bone, skin, cartilage or tendon of cattle, pig or sheep, which is not limited herein. The polyphenol substance is plant polyphenol, also called plant tannin, is a complex phenol secondary metabolite in plant bodies, has a polyphenol structure, is mainly present in peels, roots, leaves and fruits of plants, and is preferably extracted from soybeans. The collagen and the plant polyphenol can be crosslinked under the action of the crosslinking agent to generate the plant polyphenol-collagen compound with stable structure.

The biomass adsorbent is a powdery solid.

Polyphenol substances in the biomass adsorbent can be subjected to chelation reaction with dyes, so that the dyes remained in the waste liquid are adsorbed on the biomass adsorbent, and substances harmful to the environment are effectively treated through precipitation and filtration.

In contrast to the prior art, the embodiment of the present application provides a leukocyte classification kit, which includes: the hematology analyzer can be used for distinguishing different types of leucocytes, has a good effect, and is particularly suitable for automatically classifying the leucocytes by a cell morphology analysis technology based on a machine vision principle. In addition to the hemolytic agent and the staining agent, the leukocyte classification kit further includes: the biomass adsorbent is characterized in that polyphenol substances in the biomass adsorbent can be subjected to chelation reaction with dyes, so that the dyes remained in waste liquid are adsorbed on the biomass adsorbent, and substances harmful to the environment are effectively treated through precipitation and filtration. By adding the dye treatment adsorbent in the leukocyte classification kit, the problem of environmental pollution caused by dyes in the prior art can be effectively solved, and the leukocyte classification kit is low in price and obvious in adsorption effect.

In one embodiment, the biomass adsorbent comprises, in parts by mass: 4-8 parts (such as 4, 6 or 8 parts) of collagen, 1-5 parts (such as 1, 3 or 5 parts) of a cross-linking agent and 9-13 parts (such as 9, 11 or 13 parts) of a polyphenol substance. Preferably, the biomass adsorbent comprises: 6 parts of collagen, 3 parts of a cross-linking agent and 11 parts of a polyphenol substance.

Wherein, the cross-linking agent can be at least one of aldehydes, carbodiimide, epoxide, genipin and hydrazine, and preferably, the cross-linking agent is glutaraldehyde.

The molecular peptide chain of the collagen has various reactive groups, such as hydroxyl, carboxyl, amino and the like, and aldehyde groups in glutaraldehyde can perform aldol reaction with the hydroxyl of the collagen so as to achieve the aim of crosslinking.

Wherein the dye is at least one selected from methyl green, pyridine Luo Gong and methylene blue.

In one embodiment, the hemolytic agent includes anionic surfactant, cationic surfactant, osmotic pressure regulator, buffer, and preservative. The concentration of anionic surfactant is 3.0-3.4 g/l (e.g., 3.0, 3.2, or 3.4 g/l), the concentration of cationic surfactant is 2.8-5.5 g/l (e.g., 2.8, 3.6, or 5.5 g/l), the concentration of tonicity modifier is 3.1-3.5 g/l (e.g., 3.1, 3.3, or 5.5 g/l), the concentration of buffer is 2.2-4.8 g/l (e.g., 2.2, 2.6, 4.6, or 5.5 g/l), and the concentration of preservative is 1.7-2.8 g/l (e.g., 1.7, 1.9, 2.4, or 2.8 g/l).

The anionic surfactant is used for binding with hemoglobin, and is selected from sulfate type anionic surfactants, such as fatty alcohol sulfate.

The cationic surfactant is used for dissolving red blood cells, eliminating the influence of the red blood cells on the classification of white blood cells, and treating cell membranes of the white blood cells to ensure that lymph, mononuclear cells, neutrophils and eosinophils are shrunk to different degrees. The cationic surfactant is one or more of heterocyclic type and xanthate type. Preferably, the cationic surfactant is a combination of a heterocyclic type cationic surfactant and a onium salt type cationic surfactant. Wherein, the heterocyclic cationic surfactant can be selected from one or more of morphine ring, pyridine ring, imidazole ring, pyrazine ring and quinoline ring. The heterocyclic cationic surfactant in the hemolytic agent is preferably a morpholine ring cationic surfactant.

The hemolytic agent is also an important factor in maintaining the morphology of leukocytes. The osmotic pressure regulator in the hemolytic agent is selected from inorganic salts, preferably, the osmotic pressure regulator is NaCl.

The buffer solution in the hemolytic agent can keep the pH unchanged, so that a relatively stable physicochemical environment is kept for blood cells, and accurate classification and detection of white blood cells are facilitated. The buffer solution is selected from one or more of a PBS buffer solution, an acetic acid-sodium acetate buffer solution, a citrate buffer solution, a Tris buffer solution, a boric acid buffer solution, a BR buffer solution or a carbonic acid buffer solution, and preferably, the buffer solution is a combination of the PBS buffer solution and the citrate buffer solution.

Preservatives are added to effectively kill and inhibit a wide variety of microorganisms to prevent the agent from molding and to prolong the storage and use time of the agent. The preservative is at least one selected from ProClin300 and sorbic acid.

Furthermore, the hemolytic agent contains a large amount of organic substances such as the expression active agent, and in order to promote better dissolution of the hemolytic agent, a first solvent with stronger polarity may be appropriately added to promote dissolution of the hemolytic agent in deionized water. The first solvent is selected from one or more of ethanol, ethylene glycol, propanol, propylene glycol, glycerol, isopropanol, butanol and butanediol.

In one embodiment, the dyeing agent further comprises a second solvent for dissolving the dye, and the second solvent is one or more of ethanol, ethylene glycol, propanol, propylene glycol, glycerol, isopropanol, butanol and butanediol. Preferably, the first solvent is the same as the second solvent.

In addition, the present application also provides a method for preparing a differential white blood cell count kit, the method comprising:

s10: weighing anionic surfactant, cationic surfactant, osmotic pressure regulator, buffer and preservative, dissolving in a first solvent, regulating pH value, and diluting to desired volume to obtain hemolytic agent. The selection of the anionic surfactant, the cationic surfactant, the osmotic pressure regulator, the buffer and the preservative can refer to the embodiment of the leukocyte classification kit, and is not repeated herein.

According to the preparation method of the hemolytic agent, the raw materials are directly and uniformly mixed according to the concentration requirement, the volume is accurately determined, and the filtering is carried out, so that the hemolytic agent is used for a specific blood cell analyzer. The method of using the hemolytic agent is the same as that of the conventional hemolytic agent on the market, and is not repeated herein.

S20: weighing the dye, dissolving the dye in a second solvent, adjusting the pH value, and fixing the volume to obtain the coloring agent.

According to the preparation method of the coloring agent, the raw materials are directly and uniformly mixed according to the concentration requirement, and the volume is accurately determined and filtered for a specific blood cell analyzer. The use method of the coloring agent is the same as that of the conventional coloring agent in the market, and the description is omitted here.

S30: weighing collagen, polyphenol substances and a cross-linking agent, and carrying out cross-linking reaction on the collagen and the polyphenol substances under the action of the cross-linking agent to obtain a biomass adsorbent;

s40: the hemolytic agent, the biomass adsorbent and the staining agent are packaged in a kit in a single package form.

In the present application, the steps S10 to S30 are not required to be performed in a specific order, and the steps S10 to S30 may be performed simultaneously.

Referring to fig. 5, the step S30 includes the following steps:

s31: dissolving collagen in deionized water to obtain a mixed solution.

S32: adding the polyphenol substances into the mixed solution, and stirring to dissolve.

S33: and adding a cross-linking agent to perform cross-linking reaction to obtain the biomass adsorption suspended substance.

S34: and filtering, washing, drying and sieving the biomass adsorption suspended matters to obtain the biomass adsorbent.

Specifically, a 2-liter three-mouth bottle is arranged on an electric stirrer, the three-mouth bottle is placed in a water bath pot, 1 liter of deionized water is filled into the three-mouth bottle, natural collagen extracted from livestock and poultry pigs is dissolved in 1 liter of deionized water, after stirring for 0.5 hour at a constant temperature of 30 ℃, polyphenol extracted from plant food soybean is added, stirring is continued for 10 minutes, 25 milliliters of glutaraldehyde with the mass concentration of 25% is added, crosslinking reaction is carried out, reaction is carried out for 4 hours at the constant temperature of 40 ℃, then suspended matters are filtered and washed with distilled water for three times, and filter cakes are dried for 20 hours at the temperature of 30 ℃ in vacuum. Finally, the dried product is pulverized until it is sieved into uniform particles up to 200 nm in diameter, and the biomass adsorbent is prepared. Wherein the feeding ratio of the collagen, the glutaraldehyde and the polyphenol is 6:3:11.

the embodiment of the application provides a preparation method of a leukocyte classification kit, and the leukocyte classification kit prepared by the preparation method comprises the following steps: the hematology analyzer can be used for distinguishing different types of leucocytes, has a good effect, and is particularly suitable for automatically classifying the leucocytes by a cell morphology analysis technology based on a machine vision principle. In addition to the hemolytic agent and the staining agent, the leukocyte classification kit further includes: the biomass adsorbent is characterized in that polyphenol substances in the biomass adsorbent can be subjected to chelation reaction with dyes, so that the dyes remained in waste liquid are adsorbed on the biomass adsorbent, and substances harmful to the environment are effectively treated through precipitation and filtration. By adding the dye treatment adsorbent in the leukocyte classification kit, the problem of environmental pollution caused by dyes in the prior art can be effectively solved, and the leukocyte classification kit is low in price and obvious in adsorption effect.

In another aspect, the present application provides a method for treating a dye waste solution, which is based on the above-mentioned leukocyte classification kit, with reference to fig. 6, and comprises:

s101: and respectively connecting a hemolytic agent, a biomass adsorbent and a coloring agent in the leukocyte classification kit into corresponding pipelines of the particle analyzer.

The leukocyte classification kit can be used in particle analyzers, including, but not limited to, immunoassays, point of care Testing (POCT) instruments, blood cell analyzers, and the like. In the present embodiment, the particle analyzer is a blood cell analyzer which is widely used.

The reagent bottle containing the hemolytic agent, the biomass adsorbent and the staining agent is connected to a corresponding pipeline, and it should be noted that the pipeline in this embodiment may be at least one of a sampling needle, a sampling needle cleaning device, at least one leukocyte reaction cell, a distribution system, a leukocyte measurement system, a waste liquid discharge system, and a pipeline between each component.

S102: leukocyte classification is performed using hemolytic agents and staining agents.

S103: after the hemolytic agent and the staining agent are exhausted, the pH of the waste liquid discharged from the particle analyzer is adjusted to a preset pH value, and the concentration of the dye in the waste liquid is measured.

Specifically, the preset pH value may be 6.

S104: and (4) adding a biomass adsorbent according to a preset feeding ratio, and pretreating the waste liquid.

Preferably, the preset feed ratio is the dosage (g) of the biomass adsorbent: dye concentration in waste liquid (g/l) =1:1, putting a biomass adsorbent, and standing for 1 hour to adsorb the dye in the waste liquid.

The present application is described in further detail below with reference to specific embodiments and with reference to the attached drawings.

Example 1

Preparation of hemolytic agent

The formulation is shown in table 1 using deionized water as the solution.

TABLE 1

The prepared hemolytic agent has pH value of 2.0-2.5, and is stirred at 160 rpm, filtered with 0.22 μm filter membrane, stored at room temperature, and used directly.

Example 2

Preparation of biomass adsorbent

Installing a 2L three-mouth bottle on an electric stirrer, placing the three-mouth bottle in a water bath kettle, filling 1L deionized water into the three-mouth bottle, dissolving natural collagen extracted from livestock and poultry pigs in 1L deionized water, stirring at constant temperature of 30 ℃ for 0.5 hour, adding polyphenol extracted from plant food soybean, continuing stirring for 10 minutes, adding 25 ml of glutaraldehyde with the mass concentration of 25%, carrying out crosslinking reaction, reacting at constant temperature of 40 ℃ for 4 hours, filtering suspended matters, washing with distilled water for three times, and drying a filter cake at 30 ℃ for 20 hours in vacuum. Finally, the dried product is pulverized until it is sieved into uniform particles up to 200 nm in diameter, and the biomass adsorbent is prepared. Wherein the feeding ratio of the collagen, the glutaraldehyde and the polyphenol is 6:3:11.

example 3

Adsorption condition optimization of biomass adsorbent

A. The dosage of the biomass adsorbent has influence on the adsorption effect: biomass adsorbents with different dosages (5 g, 10 g, 15 g, 20 g, 25 g and 30 g) are put into methyl green, pyridine Luo Gong and methylene blue solutions with the same volume (1L) and concentration (15 g/L), so that the optimal dosage ratio is obtained (the optimal adsorption effect is achieved when the dosage (g) of the biomass adsorbents is used and the concentration (g/L) of dye in waste liquid is not less than 1:1), and the test result is shown in the attached figure 1.

B. The pH of the solution has an influence on the adsorption effect: the pH of 9 parts of 15 g/l methyl green solution was adjusted to 2, 3, 4, 5, 6, 7, 8, 9, 10, respectively, 15 g of biomass adsorbent was added, the mixture was left for 1 hour, and the methyl green concentration in the solution was measured, respectively, to obtain the optimum pH value of the adsorption solution (pH = 6), and the test results are shown in fig. 2.

C. The adsorption time has an influence on the adsorption effect: 15 g of biomass adsorbent is put into 15 g/L methyl green solution, the pH value is adjusted to 8, the methyl green concentration of the solution is measured after 10 minutes to 100 minutes of putting, the time interval is 10 minutes, and then the optimal standing adsorption time t (t =60 minutes) is obtained, and the test result is shown in the attached figure 3.

Example 4

1 clinical venous whole blood sample was taken and tested on a dimai DX480 full-automatic five-class hematology analyzer using the hemolytic agent and methylene blue solution with a concentration of 15 g/l prepared in the example 1, and the classification results are shown in fig. 4.

Adjusting the pH value of the waste liquid solution to 6, then measuring the dye concentration in the waste liquid, and then according to the usage amount (g) of the biomass adsorbent: dye concentration in waste liquid (g/l) =1:1, adding the biomass adsorbent prepared in example 2, standing for 1 hour, pretreating the waste liquid, and measuring the dye concentration in the waste liquid to ensure that the dye removal capacity reaches 99.1%.

The embodiment of the application provides a kit for classifying white blood cells, which comprises: the hematology analyzer can be used for distinguishing different types of leucocytes, has a good effect, and is particularly suitable for automatically classifying the leucocytes by a cell morphology analysis technology based on a machine vision principle. In addition to the hemolytic agent and the staining agent, the leukocyte classification kit further includes: the biomass adsorbent is characterized in that polyphenol substances in the biomass adsorbent can be subjected to chelation reaction with dyes, so that the dyes remained in waste liquid are adsorbed on the biomass adsorbent, and substances harmful to the environment are effectively treated through precipitation and filtration. By adding the dye treatment adsorbent in the leukocyte classification kit, the problem of environmental pollution caused by dyes in the prior art can be effectively solved, and the leukocyte classification kit is low in price and obvious in adsorption effect.

The above embodiments are merely examples and are not intended to limit the scope of the present disclosure, and all modifications, equivalents, and flow charts using the contents of the specification and drawings of the present disclosure or those directly or indirectly applied to other related technical fields are intended to be included in the scope of the present disclosure.

Claims

1. A leukocyte classification kit, comprising:

a hemolytic agent for lysing erythrocytes and causing cell membrane damage of leukocytes;

a biomass sorbent comprising: collagen, polyphenols and cross-linking agents;

a coloring agent comprising a dye capable of coloring nucleic acids, said dye being capable of undergoing a chelating reaction with said polyphenolic substance;

the hemolytic agent, the biomass adsorbent, and the staining agent are each present in a single package.

2. The leukocyte classification kit according to claim 1,

the biomass adsorbent comprises the following components in parts by mass: 4-8 parts of collagen, 1-5 parts of a cross-linking agent and 9-13 parts of a polyphenol substance.

3. The leukocyte classification kit according to claim 2,

the cross-linking agent is glutaraldehyde.

4. The leukocyte classification kit according to claim 1,

the dye is selected from at least one of methyl green, pyridine Luo Gong and methylene blue.

5. The leukocyte classification kit according to claim 1,

the hemolytic agent comprises an anionic surfactant, a cationic surfactant, an osmotic pressure regulator, a buffering agent and a preservative;

the concentration of the anionic surfactant is 3.0 to 3.4 g/L;

the concentration of the cationic surfactant is 2.8-5.5 g/L;

the concentration of the osmotic pressure regulator is 3.1-3.5 g/L;

the concentration of the buffer is 2.2-4.8 g/L;

the concentration of the preservative is 1.7-2.8 g/l.

6. The leukocyte classification kit according to claim 5,

the anionic surfactant is selected from sulfate salt type anionic surfactants;

the cationic surfactant is a heterocyclic cationic surfactant and/or a xanthate cationic surfactant;

the osmotic pressure regulator is inorganic salt;

the buffer solution is selected from one or more of PBS buffer solution, acetic acid-sodium acetate buffer solution, citrate buffer solution, tris buffer solution, boric acid buffer solution, BR buffer solution or carbonic acid buffer solution;

the preservative is at least one selected from ProClin300 and sorbic acid.

7. The leukocyte classification kit according to claim 1,

the biomass adsorbent is a powdery solid.

8. A method for preparing a differential white blood cell count kit, the method comprising:

weighing an anionic surfactant, a cationic surfactant, an osmotic pressure regulator, a buffering agent and a preservative, dissolving in a first solvent, regulating the pH value, and fixing the volume to obtain a hemolytic agent;

weighing a dye, dissolving the dye in a second solvent, adjusting the pH value, and fixing the volume to obtain a dyeing agent;

weighing collagen, polyphenol substances and a cross-linking agent, and carrying out cross-linking reaction on the collagen and the polyphenol substances under the action of the cross-linking agent to obtain a biomass adsorbent;

packaging the hemolytic agent, the biomass adsorbent and the staining agent in a kit in a single package form respectively.

9. The method of claim 8,

the step of performing a crosslinking reaction on the collagen and the polyphenol material under the action of the crosslinking agent to obtain the biomass adsorbent comprises the following steps:

dissolving the collagen in deionized water to obtain a mixed solution;

adding the polyphenol substances into the mixed solution, and stirring for dissolving;

adding a cross-linking agent for cross-linking reaction to obtain biomass adsorption suspended matters;

and filtering, washing, drying and sieving the biomass adsorption suspended matter to obtain the biomass adsorbent.

10. A method for treating a dye waste liquid, which is based on the leukocyte classification kit according to any one of claims 1 to 7, comprising:

respectively connecting a hemolytic agent, a biomass adsorbent and a staining agent in the leukocyte classification kit into corresponding pipelines of a particle analyzer;

classifying leukocytes using the hemolytic agent and the staining agent;

after the hemolytic agent and the staining agent are exhausted, adjusting the pH value of the waste liquid discharged from the particle analyzer to a preset pH value, and measuring the dye concentration in the waste liquid;

and (3) according to a preset feed ratio, adding the biomass adsorbent, and treating the waste liquid for a preset time.