CN116444402A

CN116444402A - DiDNTB alkali metal salt and preparation method and application thereof

Info

Publication number: CN116444402A
Application number: CN202310362522.5A
Authority: CN
Inventors: 修志明; 王淑红; 赵春影; 范修康; 刘国丽; 田永恒; 昌盛; 彭悦; 刘莉
Original assignee: Jilin Baichun Chemical Technology Co ltd
Current assignee: Jilin Baichun Chemical Technology Co ltd
Priority date: 2023-04-06
Filing date: 2023-04-06
Publication date: 2023-07-18

Abstract

The invention provides a DIDNTB alkali metal salt and a preparation method and application thereof, belonging to the technical field of DIDNTB salification. The alkali metal salt of DIDNTB is DIDNTB lithium salt, DIDNTB sodium salt and DIDNTB potassium salt. The preparation method of the invention comprises the following steps: mixing the DIDNTB and the alkali metal salt in a solvent for salifying reaction to obtain a DIDNTB alkali metal salt crude product; and mixing the DIDNTB alkali metal salt crude product with a solvent for recrystallization to obtain the DIDNTB alkali metal salt. The obtained DIDNTB alkali metal salt overcomes the defect of poor water solubility of the prior DIDNTB, has the advantages of simple preparation process, mild conditions, high product purity, high yield and low cost, can be produced in a large scale, and is beneficial to further application in detection reagents.

Description

DiDNTB alkali metal salt and preparation method and application thereof

Technical Field

The invention relates to the technical field of DIDNTB salification, in particular to a DIDNTB alkali metal salt and a preparation method and application thereof.

Background

5', 5' -dinitro-3 ', 3' -diiodo-3,4,5, 6-tetrabromophenol sulfophthalein, english name 5', 5' -dinoro-3 ', 3' -diiodo-3,4,5, 6-tetrobroom phenol sulfophthalate (DIDNTB) has the following structural formula:

DiDNTB is a protein error indicator and is used to detect microalbumin in urine or blood in the diagnosis of early kidney disease due to its high sensitivity. However, since DIDNTB is poorly soluble in water, a large amount of organic solvents are often used to dissolve in clinical diagnostic reagents, wasting reagents, increasing costs, and being inconvenient to use, affecting further use of DIDNTB.

Therefore, how to overcome the defects of the prior art and increase the water solubility of the DIDNTB so as to achieve the purpose of convenient use is a technical problem which needs to be solved at present.

Disclosure of Invention

The invention aims to provide a DIDNTB alkali metal salt, a preparation method and application thereof, and aims to solve the technical problems that the DIDNTB is poor in water solubility and inconvenient to use.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a DIDNTB alkali metal salt, which has the following structural formula:

wherein M is Li, na or K.

The invention provides a preparation method of DIDNTB alkali metal salt, which comprises the following steps:

1) Salt formation: mixing the DIDNTB and the alkali metal salt in a solvent for salifying reaction to obtain a DIDNTB alkali metal salt crude product;

2) And (5) recrystallizing: and mixing the DIDNTB alkali metal salt crude product with a solvent for recrystallization to obtain the DIDNTB alkali metal salt.

Further, in the step 1), the alkali metal salt is one or more of alkali metal carbonate, alkali metal acetate and alkali metal bicarbonate; the molar ratio of the DIDNTB to the alkali metal salt is 1:0.9-1.8.

Further, in the step 1), the solvent is an aprotic solvent, and the aprotic solvent contains one or more of tetrahydrofuran, acetone, acetonitrile, dioxane, dimethyl sulfoxide, ethyl acetate, dichloromethane, chloroform, carbon tetrachloride, diethyl ether, petroleum ether, n-propyl ether, isopropyl ether, n-butyl ether, isobutyl ether, ethylene glycol dimethyl ether, benzene, toluene and xylene.

Further, in the step 1), the mass-volume ratio of the DIDNTB to the solvent is 1g: 1-18 mL.

Further, in the step 1), the temperature of the salifying reaction is 25-80 ℃, and the time of the salifying reaction is 0.5-13 h.

Further, in the step 2), the recrystallization includes a first recrystallization and a second recrystallization, and the solvent of the first recrystallization includes one or more of ethyl acetate, dichloromethane, chloroform and carbon tetrachloride; the solvent for the second recrystallization comprises one or more of diethyl ether, petroleum ether, n-propyl ether, isopropyl ether, n-butyl ether and isobutyl ether.

Further, the mass-to-volume ratio of the crude DIDNTB alkali metal salt to the solvent for the first recrystallization is 1g: 1-18 mL; the mass to volume ratio of the crude DIDNTB alkali metal salt to the solvent for the second recrystallization is 1g: 1-18 mL.

Further, in the step 2), the temperature of the recrystallization is 30-80 ℃, and the time of the recrystallization is 0.5-13 h.

The invention provides an application of a DIDNTB alkali metal salt in preparing a trace albumin detection reagent.

The invention has the beneficial effects that:

according to the invention, the DIDNTB alkali metal salt is obtained by salifying and recrystallizing the DIDNTB, so that the problem of poor water solubility of the existing DIDNTB is solved, and the purity of the finally prepared DIDNTB alkali metal salt product reaches 99.0% and the total yield reaches 80%; the preparation method is simple, mild in condition, high in product purity, high in yield and low in cost, can be used for mass production, and is beneficial to further application in microalbumin detection reagents.

Detailed Description

wherein M is Li, na or K.

In the present invention, the alkali metal salt of DIDNTB is preferably a lithium salt of DIDNTB, a sodium salt of DIDNTB, and a potassium salt of DIDNTB.

In the present invention, the preparation process of the DIDNTB alkali metal salt is as follows:

in the present invention, in the step 1), the alkali metal salt is one or several of alkali metal carbonate, alkali metal acetate and alkali metal bicarbonate, preferably alkali metal bicarbonate, and more preferably lithium bicarbonate, sodium bicarbonate or potassium bicarbonate; the molar ratio of the DIDNTB to the alkali metal salt is 1:0.9-1.8, preferably 1:1-1.6, and more preferably 1:1.2-1.4.

In the invention, when the alkali selected in the salt forming reaction in the step 1) is strong alkali, the product becomes black due to the over strong alkali; the alkali metal salt is carbonate, and the product turns to dark green due to stronger alkalinity; when the alkali metal salt is acetate, the reaction time is too long and the reaction is not complete because of the weak alkalinity.

In the invention, if the amount of alkali metal salt selected in the salification reaction is too small and is less than 1.0 equivalent, the reaction is incomplete, and the yield and purity are affected; if the amount of the alkali metal salt is more than 1.6 equivalents, alkali remains, which is not easy to remove and affects the purity.

In the present invention, in the step 1), the solvent is an aprotic solvent, and the aprotic solvent contains one or more of tetrahydrofuran, acetone, acetonitrile, dioxane, dimethyl sulfoxide, ethyl acetate, dichloromethane, chloroform, carbon tetrachloride, diethyl ether, petroleum ether, n-propyl ether, isopropyl ether, n-butyl ether, isobutyl ether, ethylene glycol dimethyl ether, benzene, toluene and xylene, preferably tetrahydrofuran, acetone, acetonitrile and dioxane.

In the invention, when the reaction solvent selected in the salification reaction is a proton solvent, the color of the DIDNTB alkali metal salt is deepened, and the DIDNTB alkali metal salt is not suitable to be adopted; the solvent is aprotic solvent, and the color of the DIDNTB alkali metal salt is unchanged and can be adopted.

In the invention, in the step 1), the mass-volume ratio of the DIDNTB to the solvent is 1g:1 to 18m, preferably 1g:2 to 16mL, more preferably 1g: 5-12 mL.

In the invention, when the volume of the reaction solvent selected for the salification reaction is too small compared with that of the DIDNTB, the reaction is insufficient, and the reaction effect is poor; when the reaction solvent volume is excessively large compared to that of DIDNTB, reagents are wasted and the yield is low.

In the present invention, in the step 1), the temperature of the salification reaction is 25 to 80 ℃, preferably 30 to 75 ℃, and more preferably 35 to 70 ℃; the salt-forming reaction time is 0.5 to 13 hours, preferably 1 to 12 hours, and more preferably 3 to 10 hours.

In the invention, when the reaction temperature adopted in the salification reaction is lower than 30 ℃, the reaction rate is too slow, the time is too long, and the reaction efficiency is low; when the reaction temperature is higher than 75 ℃, the color of the product is too dark, and side reactions are easy to initiate. The reaction time of salification reaction is less than 1 hour, the reaction is incomplete, the yield is low, the purity is low, and the DIDNTB is free and high; the reaction time is longer than 12 hours, the reaction time is too long, the side reaction is more, the impurity is high, and the purity is low.

In the present invention, the alkali metal salt of DIDNTB after salt formation mainly contains three types of substances: the first class is large polar salified or non-salified small molecules brought in by the raw material, the second class is free DIDNTB, and the third class is small polar small molecules; the salification reaction not only ensures that the DIDNTB and alkali metal complete salification, but also performs crystallization in a salification solvent, and effectively removes large-polarity salified or non-salified small molecules in the DIDNTB.

In the invention, the DIDNTB with the purity of 90 percent is prepared into a DIDNTB alkali metal crude product by salifying reaction and crystallization, the purity is 94 percent, and the reaction yield is 90 percent.

In the present invention, in the step 2), the recrystallization includes a first recrystallization and a second recrystallization, and the solvent of the first recrystallization includes one or more of ethyl acetate, methylene chloride, chloroform and carbon tetrachloride, preferably ethyl acetate and/or methylene chloride; the solvent for the second recrystallization comprises one or more of diethyl ether, petroleum ether, n-propyl ether, isopropyl ether, n-butyl ether and isobutyl ether, preferably diethyl ether and/or petroleum ether.

In the present invention, step 2) the recrystallization reaction is one time each using two different aprotic solvents; the first recrystallization, the alkali metal salt of the DIDNTB is insoluble in the solvent, and the DIDNTB is easily soluble, so that the free DIDNTB remained in the reaction can be removed; the second solvent recrystallization, the small molecular impurities with small polarity contained in the DIDNTB alkali metal salt are easy to dissolve in the solvent, and the DIDNTB alkali metal salt is insoluble, so that the impurities are removed; therefore, the recrystallization uses two different aprotic solvents to recrystallize once each, and the effect is better.

In the invention, the mass-volume ratio of the crude DIDNTB alkali metal salt to the solvent for the first recrystallization is 1g:1 to 18mL, preferably 1g:2 to 16mL, more preferably 1g: 5-10 mL; the mass to volume ratio of the crude DIDNTB alkali metal salt to the solvent for the second recrystallization is 1g:1 to 18mL, preferably 1g:2 to 16mL, more preferably 1g: 5-10 mL.

In the invention, when the mass-volume ratio of the DIDNTB alkali metal salt crude product to the solvent in the recrystallization reaction is too large, the recrystallization is insufficient, and the purification effect is poor; when the mass-to-volume ratio of the crude DIDNTB alkali metal salt to the solvent is too small, reagents are wasted, and the yield is reduced.

In the present invention, in the step 2), the recrystallization temperature is 30 to 80 ℃, preferably 35 to 75 ℃, and more preferably 40 to 70 ℃; the recrystallization time is 0.5 to 13 hours, preferably 1 to 12 hours, more preferably 5 to 10 hours.

In the invention, the recrystallization reaction temperature is lower than 35 ℃ or the time is less than 1 hour, and the dissolution and dispersion are insufficient; at a recrystallization temperature above 75 ℃ for a period of more than 12 hours, the product is too dark.

In the invention, the purity of the DIDNTB alkali metal salt (purity is 94%) reaches 99.0% after two times of recrystallization of aprotic solvents with different attributes, and the total yield of the two times of recrystallization reaches 88%.

The purity of the finally prepared DIDNTB alkali metal salt product reaches 99.0 percent, and the total yield reaches 80 percent.

The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.

The DIDNTB used in the following experimental examples and examples was a commercially available material with a purity of 90.02%.

Experimental example 1

Comparison of the effects of salt formation reactions

(1) Alkali metal salt

4 parts of DIDNTB is weighed, 104.6g (0.1 mol) of each part is added into a reactor, 0.11mol of alkali metal salt of sodium ions is weighed according to 1.1eq, 300mL of acetonitrile is respectively added into the reactor, the reaction is stirred for 6 hours at 50 ℃, the reaction is completed, the temperature is cooled to room temperature, the filtration is carried out, a filter cake is dried, a crude product is prepared, and the crude product is collected, weighed, and relevant experimental data are shown in table 1.

TABLE 1 selection of alkali metal salts

The data in table 1 indicate that: the sodium hydroxide is used as reaction alkali, the alkalinity is too strong, the appearance of the product is black, and the product does not meet the requirements; the sodium carbonate is used as reaction alkali, the alkalinity is strong, the appearance of the crude product is black and green, excessive alkali is not easy to remove, and the requirement is not met; sodium acetate is used as reaction alkali, the alkalinity is too weak, the reaction is slower, the reaction is insufficient, the appearance of the crude product is orange, and the crude product does not meet the requirements; sodium bicarbonate is used as a reactant, and the product is light green, and has higher yield and higher purity.

(2) Alkali metal salt charging equivalent

5 parts of DIDNTB is weighed, 104.6g (0.1 mol) of each part is added into a reactor, then 0.9eq, 1.0eq, 1.2eq, 1.6eq and 1.8eq of sodium bicarbonate are respectively measured, respectively and sequentially added into the reactor, 300mL of dioxane is respectively added, the mixture is stirred at 55 ℃ for 2 hours, after the reaction is finished, the mixture is cooled to room temperature and filtered, a filter cake is dried, a crude product is prepared, and the crude product is collected, weighed and relevant experimental data are shown in Table 2.

TABLE 2 selection of alkali metal salt dosing equivalents

The data in table 2 shows that: the feeding ratio of sodium bicarbonate to DIDNTB is 0.9eq, and the reaction is incomplete; the feed ratio is 1.8eq, the alkali is excessive in the system, the color of the crude product is deepened, and the excessive alkali is not easy to remove; the feed ratio is 1.0 to 1.6eq, the yield is higher, the purity is higher, and the effect is better.

(3) Salifying solvent

Weighing 5 parts of DIDNTB, adding 104.6g (0.1 mol) of each part into a reactor in sequence, weighing 1.1eq of sodium bicarbonate 5 parts, respectively adding tetrahydrofuran, acetone, acetonitrile, dioxane and 450mL of methanol into the reactor in sequence, respectively reacting at 40 ℃ for 10 hours, cooling to room temperature after the reaction is finished, filtering, drying a filter cake to obtain a crude product, collecting, weighing, and obtaining relevant experimental data shown in Table 3.

TABLE 3 selection of salt-forming solvents

The data in table 3 indicate that: when the salifying reaction solvent is tetrahydrofuran, acetone, acetonitrile and dioxane, the yield can reach more than 90 percent, and the purity is more than 94 percent; the reaction solvent is methanol (proton solvent), and compared with the reaction solvent, the reaction solvent has lower yield and lower purity.

(4) The salt-forming solvent is used in an amount

Weighing 5 parts of DIDNTB, adding 104.6g (0.1 mol) of each part into a reactor, respectively weighing 1.2eq of sodium bicarbonate 5 parts, respectively adding into the reactor, respectively adding acetonitrile according to the mass-volume ratio of 1:1, 1:2, 1:6, 1:16 and 1:18 of the DIDNTB and the solvent, stirring and reacting for 6 hours at 50 ℃, cooling to room temperature after the reaction is finished, filtering, drying a filter cake, preparing a crude product, collecting and weighing. The relevant experimental data are shown in table 4.

TABLE 4 selection of the amount of salifying solvent used

The data in table 4 indicate that: the mass-volume ratio of the DIDNTB to the acetonitrile is 1g to 1mL, the reaction is incomplete, and the purity of the product is low; when the mass-volume ratio is 1g to 18mL, the yield is too low; when the mass volume ratio is 1 g:2-16 mL, the product yield is high, the purity is high, and the effect is good.

(5) Salt formation temperature

5 parts of DIDNTB is weighed, 104.6g (0.1 mol) of each part is added into a reactor, 1.1eq of sodium bicarbonate is respectively weighed, the mixture is respectively added into the reactor, 500mL of acetonitrile is respectively added, the mixture is stirred and reacted for 6 hours at 25 ℃, 30 ℃,50 ℃, 75 ℃, 80 ℃ respectively, filtration is carried out after the reaction is finished, and filter cakes are dried and weighed to obtain crude products. The relevant experimental data are shown in table 5.

TABLE 5 selection of salt formation temperatures

The data in table 5 shows that: the reaction temperature is 25 ℃, the obtained crude product is orange, the reaction is incomplete, the yield is low, and the purity is low; the reaction temperature is 80 ℃, the obtained crude product is dark green, the side reaction is increased, the impurity is high, the yield is low, and the purity is low; the reaction temperature is 30-75 ℃, the product is light green, the yield is high, the purity is high, and the effect is good.

(6) Salt formation reaction time

5 parts of DIDNTB is weighed, 104.6g (0.1 mol) of each part is respectively added into a reactor, 1.1eq of sodium bicarbonate is respectively weighed, the mixture is added into the reactor, 500mL of dioxane is respectively added, the mixture is stirred and reacted for 0.5 hour, 1 hour, 6 hours, 12 hours and 13 hours at 50 ℃, after the reaction is finished, the mixture is cooled to room temperature, filtered, and filter cakes are dried to prepare crude products, collected and weighed. The relevant experimental data are shown in table 6.

TABLE 6 selection of salt formation reaction times

The data in table 6 shows that: the reaction time is 0.5 hour, the obtained crude product is orange red, the reaction is incomplete, the yield is low, and the purity is low; the reaction time is 13 hours, the obtained crude product is dark green, the side reaction is increased, the impurity is high, the yield is low, and the purity is low; the reaction time is 1-12 hours, the obtained crude product is light green, and the yield and purity are high.

Experimental example 2

Comparison of the recrystallization Effect

Salt forming reaction: weighing 4183g (4.0 mol) of DIDNTB total, adding 369.6 g (4.4 mol) of sodium bicarbonate with the weight-volume ratio of 1.1eq into a reactor, adding 12000mL of acetonitrile total according to the weight-volume ratio of 1:3 of the DIDNTB and the solvent, stirring for reaction for 6 hours at 50 ℃, cooling to room temperature after the reaction is finished, filtering, drying a filter cake, collecting 3905 g of a DIDNTB sodium salt crude product, and obtaining 91.6% yield and 94.36% purity; the next step was used.

(1) Selection of the first recrystallization solvent

Taking 500 g of the crude DIDNTB sodium salt prepared in the previous step, dividing the crude DIDNTB sodium salt into 5 parts, adding 100 g of each part into a reactor, and adding 400mL of ethyl acetate, dichloromethane, chloroform, carbon tetrachloride and methanol respectively. Wherein, stirring at 45 ℃ for 5 hours respectively, stopping stirring, cooling to room temperature, filtering, drying a filter cake to obtain a product, collecting and weighing. The relevant experimental data are shown in table 7.

TABLE 7 selection of first recrystallization solvent

The data in table 7 indicate that: the first recrystallization solvent is ethyl acetate, dichloromethane, chloroform or carbon tetrachloride, the purity is above 97%, and the yield is above 95%; the solvent is proton solvent methanol, the yield is lower, and the purity is not improved.

Selection of the second recrystallization solvent

First recrystallization: taking 900 g of the DIDNTB sodium salt crude product prepared in the previous step, adding the crude product into a reactor, adding 3600mL of ethyl acetate, stirring for 5 hours at 45 ℃, stopping stirring, cooling to room temperature, filtering, drying a filter cake to prepare a product, collecting 860 g, and obtaining 95.6% of yield and 97.18% of purity for the next step of experiment;

and (3) secondary recrystallization: 800 g of DIDNTB sodium salt prepared in the last step is divided into 8 parts, 100 g of each part is added into a reactor respectively, diethyl ether, petroleum ether, n-propyl ether, isopropyl ether, n-butyl ether, isobutyl ether, ethylene glycol dimethyl ether and ethanol (proton solvent) are added respectively, the volume is 600mL, stirring is carried out at 35 ℃ for 6 hours, after stopping stirring, cooling to room temperature, filtering, and drying a filter cake to obtain a pure product. And (5) collecting and weighing. The relevant experimental data are shown in table 8.

TABLE 8 selection of the second recrystallization solvent

The data in table 8 indicate that: the second recrystallization solvent is diethyl ether, petroleum ether, n-propyl ether, isopropyl ether, n-butyl ether, isobutyl ether and ethylene glycol dimethyl ether, the yield of the obtained pure product is more than 94%, and the purity is more than 99.0%; the yield of the ethylene glycol dimethyl ether is lower, and the purity is not obviously improved; other protic solvents, such as ethanol, give product yields of less than 90.0% and reduced purity after recrystallization.

(3) The amount of recrystallization solvent used

Taking 500 g of the coarse DIDNTB sodium salt prepared in the previous step, dividing the coarse DIDNTB sodium salt into 5 parts, adding 100 g of each part into a reactor respectively, then adding ethyl acetate into the mixture according to the mass-volume ratio of the coarse DIDNTB sodium salt to the recrystallization solvent of 1:1, 1:2, 1:6, 1:16 and 1:18 respectively, stirring for 5 hours at 50 ℃, stopping stirring, filtering, drying a filter cake, weighing, and obtaining the product, wherein relevant experimental data are shown in Table 9.

TABLE 9 selection of the amount of recrystallization solvent used

The data in table 9 indicate that: the mass-volume ratio of the DIDNTB sodium salt crude product to the ethyl acetate is 1g to 1mL, stirring and dispersing are insufficient, the purity of the product is low, and the purity is not obviously improved; when the mass-volume ratio is 1g to 18mL, the yield of the product is too low; when the mass volume ratio is 1 g:2-16 mL, the yield is higher, and the purity is higher.

(4) Recrystallization temperature

Taking 500 g of the DIDNTB sodium salt crude product prepared in the previous step, dividing the crude product into 5 parts, adding 100 g of each part into a reactor respectively, adding 400mL of ethyl acetate respectively, stirring for 9 hours at 30 ℃, 35 ℃,50 ℃, 75 ℃ and 80 ℃, stopping stirring, cooling to room temperature, filtering, drying a filter cake, and obtaining a product, collecting and weighing. The relevant experimental data are shown in table 10.

Table 10 selection of recrystallization temperature

The data in table 10 shows that: the recrystallization temperature is 30 ℃, the recrystallization is insufficient, and the purity is low; the reaction temperature is 80 ℃, the color of the obtained product is deepened, the purity is reduced, and the yield is reduced; the reaction temperature is 35-75 ℃, the color of the product meets the requirements, and the yield and the purity are high.

(5) Recrystallization time

Taking 500 g of the DIDNTB sodium salt crude product prepared in the previous step, dividing the crude product into 5 parts, adding 100 g of each part into a reactor respectively, adding 400mL of ethyl acetate respectively, stirring for 0.5 hour, 1 hour, 7 hours, 12 hours and 13 hours respectively, cooling to room temperature after stopping stirring, filtering, drying a filter cake, obtaining a product, collecting and weighing. The relevant experimental data are shown in table 11.

TABLE 11 selection of recrystallization times

The data in table 11 shows that: the recrystallization time is 0.5 hour, the recrystallization is insufficient, and the purity is low; the recrystallization time is 13 hours, the color of the obtained product is deepened, the purity of the product is low, and the yield is reduced; the recrystallization time is 1-12 hours, the color of the product meets the requirements, and the yield and purity are high.

Example 1

Preparation of DiDNTB lithium salt

Salt forming reaction: 104.6g (0.1 mol) of DIDNTB is weighed and added into a reactor, 8.2 g (0.12 mol) of 1.2eq lithium bicarbonate is added, 300mL of acetone is added, the mixture is stirred and reacted for 8 hours at 40 ℃, after the reaction is finished, the mixture is cooled to room temperature, filtered, a filter cake is dried and collected, 96.8 g of DIDNTB lithium salt crude product is obtained, the yield is 92.3%, and the purity is 94.13%.

And (5) recrystallizing:

first recrystallization: 96.8 g of crude product obtained in the previous step is added into a reactor, 200mL of dichloromethane is added, stirring is carried out at 40 ℃ for 4 hours, stirring is stopped, cooling to room temperature is carried out, filtering is carried out, a filter cake is dried, 91.5 g of the product is collected, the yield is 94.5%, and the purity is 97.32%.

And (3) secondary recrystallization: 91.5 g of the product obtained in the last step is added into a reactor, 400mL of isopropyl ether is added, stirring is stopped for 1 hour at 55 ℃, stirring is stopped, cooling is carried out to room temperature, filtering is carried out, a filter cake is dried and collected, and the obtained DIDNTB lithium salt pure product is obtained, 86.5 g is weighed, the yield is 94.5%, the total yield is 82.4%, and the purity is 99.13%.

Example 2

Preparation of DiDNTB sodium salt

1) Salt forming reaction: 104.6g (0.1 mol) of DIDNTB is weighed and added into a reactor, 9.2 g (0.11 mol) of 1.1eq sodium bicarbonate is added, 300mL of acetonitrile is added, the mixture is stirred at 40 ℃ for reaction for 8 hours, after the reaction is finished, the mixture is cooled to room temperature, filtered, and a filter cake is dried and collected, 99.3 g of DIDNTB sodium crude product is obtained, the yield is 93.2%, and the purity is 95.15%.

2) And (5) recrystallizing:

first recrystallization: 99.3 g of crude product obtained in the previous step is added into a reactor, 200mL of dichloromethane is added, stirring is carried out at 40 ℃ for 4 hours, stirring is stopped, cooling to room temperature is carried out, filtering is carried out, a filter cake is dried, 93.8 g is collected, the yield is 94.5%, and the purity is 97.32%.

And (3) secondary recrystallization: 93.8 g of the product obtained in the last step is added into a reactor, 400mL of isopropyl ether is added, stirring is stopped for 1 hour at 55 ℃, stirring is stopped, cooling to room temperature is carried out, filtering is carried out, a filter cake is dried and collected, and the obtained DIDNTB sodium salt pure product is obtained, 89.2 g is weighed, the yield is 95.1%, the total yield is 83.7%, and the purity is 99.34%.

Example 3

Preparation of DiDNTB Potassium salt

1) Salt forming reaction: 104.6g (0.1 mol) of DIDNTB is weighed and added into a reactor, 10.0 g (0.1 mol) of 1.0eq potassium bicarbonate is added, 300mL of acetone is added, the mixture is stirred and reacted for 8 hours at 40 ℃, after the reaction is finished, the mixture is cooled to room temperature, filtered, a filter cake is dried and collected, and 99.3 g of a crude product of the DIDNTB potassium salt is prepared, the yield is 91.8%, and the purity is 95.21%.

2) And (5) recrystallizing:

first recrystallization: 99.3 g of crude product obtained in the previous step is added into a reactor, 200mL of dichloromethane is added, stirring is carried out at 40 ℃ for 4 hours, stirring is stopped, cooling to room temperature is carried out, filtering is carried out, a filter cake is dried, 92.5 g of the product is collected, the yield is 93.2%, and the purity is 97.63%.

And (3) secondary recrystallization: 92.5 g of the product obtained in the last step is added into a reactor, 400mL of isopropyl ether is added, stirring is stopped for 1 hour at 55 ℃, stirring is stopped, cooling is carried out to room temperature, filtering is carried out, a filter cake is dried and collected, and the obtained DIDNTB potassium salt pure product is obtained, 88.0 g is weighed, the yield is 95.1%, the total yield is 81.3%, and the purity is 99.13%.

Test example:

the lithium salt of DIDNTB, the sodium salt of DIDNTB, the potassium salt of DIDNTB and DIDNTB obtained in examples 1, 2 and 3 were tested for solubility, and the comparative results of the related data are shown in table 12:

TABLE 12 solubility of alkali metal salt of DIDNTB obtained in examples 1 to 3

Definition of name terms for solubility representation:

is very soluble: means that 1g (mL) of solute can be dissolved in less than 1mL of solvent;

easily soluble: means that 1g (mL) of solute can be dissolved in 1-less than 10mL of solvent;

dissolving: means that 1g (mL) of solute can be dissolved in 10-less than 30mL of solvent;

slightly soluble: means that 1g (mL) of solute can be dissolved in 30-less than 100mL of solvent;

slightly dissolving: means that 1g (mL) of solute can be dissolved in 100-less than 1000mL of solvent;

very slightly dissolving: dissolving means that 1g (mL) of solute can be dissolved in 1000-less than 10000mL of solvent;

hardly soluble or insoluble: dissolution means that 1g (mL) of solute is not completely dissolved in 10000mL of solvent;

it can be seen in table 12 that the lithium, sodium and potassium salts of DIDNTB significantly increased in water solubility.

Example 4

Application of DIDNTB lithium salt in detection reagent

1) Preparation of the solution: preparing a color development liquid: 30mg of DIDNTB lithium salt and 0.3g of surfactant are respectively weighed, added into 100mL of distilled water and stirred uniformly for later use; preparing albumin standard solution: weighing human serum albumin, dissolving with distilled water, and preparing into concentration gradients of 10, 25, 50, 100 and 200mg/L respectively for later use; preparing a buffer solution: preparing 100mL of citric acid-sodium citrate buffer solution with pH of 3.0, and uniformly stirring for later use;

2) Determination of albumin content: and respectively adding 4.8mL of buffer solution, 100 mu L of chromogenic solution and 100 mu L of 5 concentrations of human serum albumin standard solution into 6 colorimetric tubes, respectively adding 100 mu L of protein standard solution to be detected, establishing a standard curve, and measuring the content of micro albumin in a sample to be detected.

Example 5

Application of DIDNTB sodium salt in detection reagent

1) Preparing a solution: preparing a color development liquid: 40mg of DIDNTB sodium salt and 0.3g of surfactant are respectively weighed and dissolved in 100mL of distilled water, and the mixture is stirred uniformly for later use; preparing albumin standard solution: weighing human serum albumin, dissolving with distilled water, and preparing into concentration gradients of 10, 25, 50, 100 and 200mg/L respectively for later use; preparing a buffer solution: preparing 100mL of citric acid-sodium citrate buffer solution with pH of 3.0, and uniformly stirring for later use;

2) Determination of albumin content: and respectively adding 4.8mL of buffer solution into 6 colorimetric tubes, respectively adding 100 mu L of chromogenic solution, respectively adding 100 mu L of 5 concentrations of human serum albumin standard solution, respectively adding 100 mu L of protein standard solution to be detected, establishing a standard curve, and measuring the trace albumin content in a sample to be detected.

Example 6

Application of DIDNTB potassium salt in detection reagent

1) Preparing a solution: preparing a color development liquid: 50mg of DIDNTB potassium salt and 0.3g of surfactant are respectively weighed and dissolved in 100mL of distilled water, and the mixture is stirred uniformly for later use; preparing albumin standard solution: weighing human serum albumin, dissolving with distilled water, and preparing into concentration gradients of 10, 25, 50, 100 and 200mg/L respectively for later use; preparing a buffer solution: preparing 100mL of citric acid-sodium citrate buffer solution with pH of 3.0, and uniformly stirring for later use;

2) Determination of albumin content: and respectively adding 4.8mL of buffer solution into 6 colorimetric tubes, respectively adding 100 mu L of chromogenic solution, respectively adding 100 mu L of 5 concentration human serum albumin standard solutions, respectively adding 100 mu L of protein standard solution to be detected, establishing a standard curve, and measuring the content of trace albumin in a sample to be detected.

From the above examples, the present invention provides a DIDNTB alkali metal salt, and a preparation method and application thereof. According to the invention, the DIDNTB alkali metal salt is obtained by salifying and recrystallizing the DIDNTB, so that the problem of poor water solubility of the existing DIDNTB is solved, and the purity of the finally prepared DIDNTB alkali metal salt product reaches 99.0% and the total yield reaches 80%; the preparation method is simple, mild in condition, high in product purity, high in yield and low in cost, can be used for mass production, and is beneficial to further application in micro-albumin detection reagents.

The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims

1. A DIDNTB alkali metal salt, characterized in that the DIDNTB alkali metal salt has the structural formula:

wherein M is Li, na or K.

2. The process for preparing a DIDNTB alkali metal salt according to claim 1, comprising the steps of:

3. The method according to claim 2, wherein in the step 1), the alkali metal salt is one or more of alkali metal carbonate, alkali metal acetate and alkali metal hydrogencarbonate; the molar ratio of the DIDNTB to the alkali metal salt is 1:0.9-1.8.

4. The method according to claim 2 or 3, wherein in the step 1), the solvent is an aprotic solvent, and the aprotic solvent contains one or more of tetrahydrofuran, acetone, acetonitrile, dioxane, dimethyl sulfoxide, ethyl acetate, dichloromethane, chloroform, carbon tetrachloride, diethyl ether, petroleum ether, n-propyl ether, isopropyl ether, n-butyl ether, isobutyl ether, ethylene glycol dimethyl ether, benzene, toluene, and xylene.

5. The method according to claim 4, wherein in the step 1), the mass-to-volume ratio of the di-dntb to the solvent is 1g: 1-18 mL.

6. The method according to claim 2, 3 or 5, wherein in the step 1), the salt forming reaction is carried out at a temperature of 25 to 80 ℃ for a time of 0.5 to 13 hours.

7. The method according to claim 6, wherein in the step 2), the recrystallization comprises a first recrystallization and a second recrystallization, and the solvent of the first recrystallization comprises one or more of ethyl acetate, methylene chloride, chloroform and carbon tetrachloride; the solvent for the second recrystallization comprises one or more of diethyl ether, petroleum ether, n-propyl ether, isopropyl ether, n-butyl ether and isobutyl ether.

8. The preparation method according to claim 7, wherein the mass-to-volume ratio of the crude DIDNTB alkali metal salt to the solvent for the first recrystallization is 1g: 1-18 mL; the mass to volume ratio of the crude DIDNTB alkali metal salt to the solvent for the second recrystallization is 1g: 1-18 mL.

9. The method according to claim 7 or 8, wherein in the step 2), the recrystallization temperature is 30 to 80 ℃ and the recrystallization time is 0.5 to 13 hours.

10. Use of the alkali metal salt of DIDNTB according to claim 1 for the preparation of a microalbumin detection reagent.