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Method for detecting residual content of ethylenediamine tetraacetic acid disodium salt in hyaluronic acid

A technology of sodium hyaluronate and hyaluronic acid, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as lack of detection

Pending Publication Date: 2021-09-14
SHANDONG TOPSCI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the current lack of detection of EDTA-2Na content in sodium hyaluronate, the present invention provides a detection method for EDTA-2Na content in sodium hyaluronate, which can quickly and accurately detect the content of EDTA-2Na in sodium hyaluronate samples

Method used

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  • Method for detecting residual content of ethylenediamine tetraacetic acid disodium salt in hyaluronic acid
  • Method for detecting residual content of ethylenediamine tetraacetic acid disodium salt in hyaluronic acid
  • Method for detecting residual content of ethylenediamine tetraacetic acid disodium salt in hyaluronic acid

Examples

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Effect test

Embodiment 1

[0029] Example 1 Content detection of EDTA-2Na in sodium hyaluronate samples

[0030] (1) EDTA-2Na stock solution: Accurately weigh C 10 h 14 N 2 Na 2 o 8 2H 2 O 110.8 mg, put in a 100.0mL volumetric flask, dissolve in water and set the volume to the mark to obtain 1.0mg / mL EDTA-2Na standard stock solution;

[0031] (2) Hyaluronidase solution derived from bovine testis: According to the potency marked in the enzyme manual, weigh an appropriate amount of enzyme, add water to dissolve, and prepare a 100IU / mL hyaluronidase solution.

[0032] (2) Ferric chloride solution: Weigh 1.35 g of ferric chloride, add 30.0 mL of glacial acetic acid to dissolve, and dilute to 100 mL with water to obtain a 0.05 mol / L ferric chloride solution;

[0033] (3) Test solution: Accurately weigh 0.25 g of sodium hyaluronate (average molecular weight 1510 KDa, purity 99.2%), put it in a 150 mL iodine bottle, add 50.0 mL of hyaluronidase solution from bovine testis (enzyme effect price 5000IU), p...

Embodiment 2

[0051] Example 2 Content detection of EDTA-2Na in sodium hyaluronate samples

[0052] (1) EDTA-2Na stock solution: Accurately weigh C 10 h 14 N 2 Na 2 o 8 2H 2 O 110.8 mg, put in a 100.0mL volumetric flask, dissolve in water and set the volume to the mark to obtain 1mg / mL EDTA-2Na standard stock solution;

[0053] (2) Recombinant leech hyaluronidase solution: According to the potency marked in the instructions, weigh an appropriate amount of enzyme, add water to dissolve, and prepare a 200IU / mL hyaluronidase solution.

[0054] (3) Ferric chloride solution: Weigh 1.35 g of ferric chloride, add 30 mL of glacial acetic acid to dissolve, and dilute to 100 mL with water to obtain 0.05 mol / L ferric chloride solution;

[0055] (4) Test solution: Accurately weigh 0.25g of sodium hyaluronate (average molecular weight 2005KDa, purity 101.9%) for test, put it in a 150mL iodine bottle, add 50mL of recombinant leech hyaluronidase solution (enzyme titer 10000IU) , adjust the pH of th...

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Abstract

The invention provides a method for detecting the content of EDTA-2Na (Ethylene Diamine Tetraacetic Acid-2Na) in sodium hyaluronate. The method comprises the following steps: firstly, degrading a sodium hyaluronate sample by adopting hyaluronidase, and then determining the content of the EDTA-2Na by adopting a conventional ferric trichloride derivation method through high performance liquid chromatography. According to the method, specific hyaluronidase is screened, so that the viscosity of a high-concentration sample can be effectively reduced, the column pressure is reduced, and test equipment is protected; and meanwhile, the produced product does not influence the detection of a detection target, and the detection sensitivity can be improved. A more accurate method is provided for production and detection of sodium hyaluronate, and the method has important significance in improving the quality of sodium hyaluronate.

Description

technical field [0001] The invention belongs to the technical field of biological macromolecule impurity detection, relates to a detection method for disodium edetate content, and relates to a detection method for disodium edetate content in hyaluronic acid and its salt. Background technique [0002] Disodium ethylenediaminetetraacetic acid (EDTA-2Na) is an organic compound that can form stable complexes with most metal ions, so it is widely used in various fields such as pharmaceutical industry, chemical industry, and food industry. In the production process of sodium hyaluronate, a small amount of EDTA-2Na is added to remove metal heteroions, but this inevitably introduces EDTA-2Na. Excessive EDTA-2Na will affect the quality of sodium hyaluronate, especially high-purity, medical-grade sodium hyaluronate has higher requirements for impurities. Most of the introduced EDTA-2Na has been removed during the production of sodium hyaluronate, so its residue is very low at ppm lev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/027G01N2030/067
Inventor 刘宝珍刘颖洁王朋田朱伟伟刘守垒王增利刘云刚王海泉秦会利吴建伟
Owner SHANDONG TOPSCI BIO TECH
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