Rapid quantitative detection method for purity of glucaric acid

Abstract

The invention discloses a method for rapidly and quantitatively detecting the purity of glucaric acid, which comprises the following steps: in the presence of a basic catalyst and a static magnetic field, mixing impurity-containing glucaric acid and m-xylene diamine in water and a non-polar organic solvent, and then standing at room temperature for 3-5 minutes for reaction to form a reaction system to be detected; and quantitatively detecting the purity of the glucaric acid according to the fluorescence intensity of the reaction system to be detected. According to the method, the purity of the glucaric acid can be rapidly detected, the sensitivity is high, and the detection limit is low.

Description

Technical Field

[0001] The invention specifically relates to a rapid quantitative detection method for the purity of glucaric acid. Background Art

[0002] Glucaric acid (GA) is a non-toxic glucose derivative with wide application value in medicine, chemical industry, etc. Currently, GA is mainly produced by chemical oxidation methods such as nitric acid oxidation and TEMPO oxidation, but impurities such as gluconic acid and aldonic acid are commonly found in the produced GA.

[0003] The existing GA purity detection methods mainly include high performance liquid chromatography (HPLC), ion chromatography, carbazole colorimetry and gluconate dehydrogenase reaction method. Among them, HPLC has the characteristics of simple sample handling and accurate quantitative analysis, but HPLC cannot completely separate gluconic acid and GA, and thus cannot accurately quantify and analyze. Ion chromatography can separate gluconic acid and glucaric acid and accurately quantify them, but it requires complex pre-treatment of the sample during detection, which is time-consuming and costly. Carbazole colorimetry and glucaric acid dehydrogenase reaction methods cannot accurately perform quantitative analysis on glucaric acid. Summary of the invention

[0004] The main purpose of the present invention is to provide a rapid quantitative detection method for the purity of glucaric acid to overcome the shortcomings of the prior art.

[0005] In order to achieve the above-mentioned invention object, the technical solution adopted by the present invention includes: The embodiment of the present invention provides a rapid quantitative detection method for the purity of glucaric acid, which comprises: S1. In the presence of an alkaline catalyst and a static magnetic field, the impure glucaric acid and m-xylene diamine are mixed in water and a non-polar organic solvent, and then allowed to stand at room temperature for 3 to 5 minutes to react to form a reaction system to be detected; S2. According to the fluorescence intensity of the reaction system to be detected, quantitative detection of the purity of glucaric acid is achieved; Wherein, the magnetic field strength of the static magnetic field is 0.2~1.0 T.

[0006] In one embodiment, the molar ratio of the impure glucaric acid to m-xylene diamine is 1:0.5-2, preferably 1:1.

[0007] In one embodiment, the alkaline catalyst includes sodium bicarbonate, sodium hydroxide, triethylamine or pyridine, etc., preferably sodium bicarbonate.

[0008] In one embodiment, the magnetic field strength of the static magnetic field is 0.3-0.5T.

[0009] In one embodiment, the density of the non-polar organic solvent is greater than that of water. For example, the non-polar organic solvent includes chloroform or dichloromethane.

[0010] In one embodiment, the volume ratio of water to the non-polar organic solvent in step S1 is 1:1-1.5.

[0011] In one embodiment, the parameter conditions used when detecting the fluorescence intensity of the reaction system to be detected include: an excitation wavelength of 212-385 nm, and a detection wavelength of 400-650 nm.

[0012] In one embodiment, the purity of the impure glucaric acid is 90% to 99.999%.

[0013] Furthermore, the method comprises: calculating the impurity content Y in the impurity-containing glucaric acid according to the fluorescence intensity X and Y=(1-2.714 / (XC))×100% of the reaction system to be detected, wherein C=115.2311~120.4763.

[0014] The main detection principle of the present invention is: under static magnetic field conditions, glucaric acid can react and combine rapidly with m-xylenediamine in a short time to generate a strong fluorescent substance because it has more active groups such as carboxyl groups, while gluconic acid, aldonic acid, etc. are difficult to react and combine rapidly with m-xylenediamine because they have fewer active groups and the steric hindrance of m-xylenediamine itself is larger. Therefore, the content of glucaric acid can be detected by detecting the fluorescence intensity of the reaction system.

[0015] Compared with the prior art, the present invention establishes a fluorescence detection method for the purity of glucaric acid, which promotes the selective and preferential combination of m-xylene diamine with glucaric acid through the non-thermal effect of the static magnetic field, etc., to form a fluorescent substance with a stronger signal, and utilizes the change in the fluorescence intensity of the reaction system to achieve quantitative detection of the purity of glucaric acid. The method has high detection sensitivity, good anti-interference ability, and low detection limit, and does not require expensive equipment and reagents. It is simple to operate, has good repeatability, high stability, is safe and environmentally friendly, and provides a new detection path for quantitative detection of the purity of glucaric acid. DETAILED DESCRIPTION

[0016] The technical solution of the present invention is further described in detail below in conjunction with several preferred embodiments. This embodiment is implemented on the premise of the technical solution of the invention, and a detailed implementation method and specific operation process are given, but the protection scope of the present invention is not limited to the following embodiments.

[0017] Unless otherwise specified, the experimental materials used in the following examples can be purchased from conventional biochemical reagent companies.

[0018] Example 1 A series of glucaric acid samples with known purity (99.999wt%, 99.99wt%, 99.0wt%, 95wt%, 90wt%, 85wt%, 80wt%, 75wt%) were taken as standard samples. The tests were performed according to the following methods: S1. Mix about 2.5 mmoL of standard sample, about 2.6 mmoL of m-xylenediamine and about 0.5 g of sodium bicarbonate in a mixture of water and chloroform (about 20 mL) in a volume ratio of about 1:1, and then let it stand in a static magnetic field with an intensity of about 0.3 T at room temperature for about 5 minutes to react to form a reaction system to be detected.

[0019] S2. Detecting the fluorescence intensity of the reaction system to be detected, wherein the detection excitation wavelength adopted is 212-385 nm, and the detection wavelength is 400-650 nm.

[0020] Based on the test results, a standard relationship curve between the fluorescence intensity X of the standard sample and the glucaric acid content Y therein was established, and the corresponding standard calculation formula was Y=(1-2.714 / (X-115.2311))×100%.

[0021] Example 2 A series of glucaric acid samples with known purity (99.999wt%, 99.99wt%, 99.0wt%, 95wt%, 90wt%, 85wt%, 80wt%, 75wt%) were taken as standard samples. The tests were performed according to the following methods: S1. Mix about 2.5 mmoL of standard sample, about 2.6 mmoL of m-xylenediamine and about 0.5 g of sodium bicarbonate in a mixture of water and chloroform (about 20 mL) with a volume ratio of about 1:1, and then let it stand for about 3 minutes in a static magnetic field with an intensity of about 0.5 T at room temperature to react to form a reaction system to be detected.

[0022] S2. Detecting the fluorescence intensity of the reaction system to be detected, wherein the detection excitation wavelength adopted is 212-385 nm, and the detection wavelength is 400-650 nm.

[0023] Based on the test results, a standard relationship curve between the fluorescence intensity X of the standard sample and the glucaric acid content Y therein was established, and the corresponding calculation formula was Y = (1-2.714 / (X-120.4763)) × 100%.

[0024] Application examples: Five commercially available glucaric acid samples were tested according to the methods of Examples 1 and 2. The average test results of each sample using the method of Example 1 were 92.317%, 90.674%, 95.127%, 91.284%, and 92.367%, respectively. The average test results of each sample using the method of Example 2 were 92.315%, 90.681%, 95.121%, 91.282%, and 92.370%, respectively. The test results were basically consistent.

[0025] In addition, referring to the literature (Chin J Anal Lab, 2013, 32(5): 20-24), the above five glucaric acid samples were tested by reverse phase ion pair liquid chromatography, and the test results were basically consistent with the above test results, proving the effectiveness of the method of the present application.

[0026] It should be understood that the technical solution of the present invention is not limited to the above-mentioned specific implementation cases. Any technical deformation made according to the technical solution of the present invention without departing from the scope of protection of the purpose of the present invention and the claims shall fall within the protection scope of the present invention.

Claims

1. A rapid quantitative detection method for the purity of glucaric acid, characterized in that: include: S1. In the presence of an alkaline catalyst and a static magnetic field, the impure glucaric acid and m-xylene diamine are mixed in water and a non-polar organic solvent, and then allowed to stand at room temperature for 3 to 5 minutes to react to form a reaction system to be detected; S2. According to the fluorescence intensity of the reaction system to be detected, quantitative detection of the purity of glucaric acid is achieved; Wherein, the magnetic field strength of the static magnetic field is 0.2~1.0 T.

2. The method according to claim 1, characterized in that: The molar ratio of the impure glucaric acid to m-xylene diamine is 1:0.5-2.

3. The method according to claim 1, characterized in that: The alkaline catalyst includes sodium bicarbonate, sodium hydroxide, triethylamine or pyridine.

4. The method according to claim 1, characterized in that: The magnetic field strength of the static magnetic field is 0.3~0.5T.

5. The method according to claim 1, characterized in that: The non-polar organic solvent has a density greater than that of water.

6. The method according to claim 5, characterized in that: The non-polar organic solvent includes chloroform or dichloromethane.

7. The method according to claim 1, characterized in that: In step S1, the volume ratio of water to the non-polar organic solvent is 1:1-1.

5.

8. The method according to claim 1, characterized in that: The parameters used to detect the fluorescence intensity of the reaction system to be detected include: an excitation wavelength of 212-385 nm, and a detection wavelength of 400-650 nm.

9. The method according to claim 1, characterized in that: The purity of the impure glucaric acid is 90% to 99.999%.

10. The method according to claim 1, characterized in that include: The impurity content Y in the impurity-containing glucaric acid is calculated according to the fluorescence intensity X and Y=(1-2.714 / (XC))×100% of the reaction system to be detected, wherein C=115.2311~120.4763.