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A method for determining trace sugar impurities in amino acid raw materials

A technology for amino acids and APIs, applied in measurement devices, material separation, analysis of materials, etc., can solve the problems of large temperature influence, complicated operation steps, low detector sensitivity, etc., and achieve high precision, good repeatability, and high sensitivity. Effect

Active Publication Date: 2021-11-16
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sugars have no characteristic ultraviolet absorption characteristics, and can only be detected by ultraviolet detectors after derivatization with chromophoric groups. The operation steps are cumbersome when using ultraviolet detectors.
Although the differential refractive index detector can directly measure carbohydrates, the detector has low sensitivity, is greatly affected by temperature, and is not compatible with gradient elution

Method used

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  • A method for determining trace sugar impurities in amino acid raw materials
  • A method for determining trace sugar impurities in amino acid raw materials
  • A method for determining trace sugar impurities in amino acid raw materials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: A pretreatment method for enriching sugar in amino acid raw materials

[0035] (1) Resin pretreatment: Accurately weigh 10.0g of 732 cation exchange resin, place it in a 250ml Erlenmeyer flask, wash with water until the effluent is colorless, treat with 70mL of 5% (v / v) HCl overnight, wash with water until neutral ; Treat with an equal volume of 5% (m / v) NaOH overnight, wash with water until neutral; finally treat with 70mL of 5% (v / v) HCl overnight, then wash with water until neutral, and set aside.

[0036] (2) Amino acid sample pretreatment: Take 1 part of the pretreated resin in a 250mL conical flask, accurately weigh 0.20g of the amino acid sample and dissolve it in 100mL water, add the solution to the conical flask, and place it on a magnetic stirrer at room temperature Low-speed static adsorption until the amino acid is completely adsorbed. The solution was filtered, and the filtrate was rotary evaporated to about 30 mL, and then lyophilized at -80°C,...

Embodiment 2

[0037] Embodiment 2: the selection of pretreatment condition

[0038] (1) Choice of resin type:

[0039] Referring to the method of Example 1, the mixed solution of arginine and glucose (the concentration of arginine in the mixed solution is 2g / mL, and the concentration of glucose is 1mg / mL) is processed, the difference is that the 732 cationic resin is replaced by macroporous Adsorption resin, D113 weak acid type cation exchange resin and D001 type cation exchange resin, other conditions are the same as embodiment 1, utilize the ninhydrin method to investigate the adsorption situation of amino acid, the results are shown in figure 1 .

[0040] The adsorption of amino acid and sugar impurities by macroporous adsorption resin is similar, and the recovery rate of sugar is too low. Depend on figure 1 It can be seen that the adsorption of amino acids by D113 resin is completed within 4.5 hours, and the adsorption capacity is poor. D001 type resin can complete the adsorption of...

Embodiment 3

[0044] Example 3: A method for detecting sugar in amino acid raw materials

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Abstract

The invention discloses a method for measuring trace carbohydrate impurities in amino acid raw materials, belonging to the technical field of drug analysis and detection. The present invention uses ion exchange resins to treat samples of amino acid raw materials to be tested, absorbs sugar in amino acid raw materials, and then uses high-performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) to measure five kinds of sugar impurities in amino acid raw materials. The gradient elution is carried out with water as the mobile phase, and the polar bonded phase column is used for separation. Nitrogen is used as the auxiliary gas for direct sample injection. It has been verified that the method can achieve baseline separation of each component, and the content of five sugar impurities in amino acid APIs is determined by external standard method. The method of the invention has the advantages of high sensitivity, simple operation and high accuracy, and is suitable for simultaneously determining the contents of five kinds of carbohydrate impurities in amino acid raw materials.

Description

technical field [0001] The invention relates to a method for determining trace carbohydrate impurities in amino acid raw materials, belonging to the technical field of drug analysis and detection. Background technique [0002] Amino acids are the basic units that constitute the first nutritional element in the human body - protein, and play an important role in human growth, reproduction and immunity. The production methods of amino acids mainly include extraction method, fermentation method, chemical synthesis, enzymatic method and other methods. The traditional extraction method, chemical synthesis method and enzymatic method are difficult to achieve the purpose of industrial production due to the high cost of precursors and complicated processes. At present, fermentation methods are generally used in industry. Sugar is introduced into the process as a sugar source for microorganisms in the fermentation process, and may become a trace impurity in amino acid APIs. However,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/74G01N30/88G01N30/06G01N30/14G01N30/86G01N30/12G01N30/08G01N30/04G01N30/34
CPCG01N30/02G01N30/04G01N30/06G01N30/08G01N30/12G01N30/14G01N30/34G01N30/74G01N30/8634G01N30/8679G01N30/88G01N2030/047G01N2030/126G01N2030/143G01N2030/8818G01N2030/884G01N2030/8872
Inventor 惠人杰姜峰薛灵杰杨荻
Owner JIANGNAN UNIV