HPLC detection method of biotin intermediate diamino substance

A detection method, the technology of diamino substances, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that the direct detection of the reaction liquid cannot be directly applied, and the content cannot be accurately detected, so as to achieve a good reaction end point, high precision, and control The effect of the reaction endpoint

Active Publication Date: 2020-03-17
SHANGYU NHU BIOCHEM IND +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When using this method to directly detect the reaction solution, there will be a problem that the diamino compound C and the solvent peak overlap, and its content cannot be accurately detected.
Therefore, the above method cannot be directly applied to the direct detection of the reaction solution in the biotin preparation process.

Method used

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  • HPLC detection method of biotin intermediate diamino substance
  • HPLC detection method of biotin intermediate diamino substance
  • HPLC detection method of biotin intermediate diamino substance

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Experimental program
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Effect test

Embodiment 1

[0041] 1. Instruments and reagents

[0042] Instrument model: Dionne liquid chromatograph, U3000

[0043] Reagents: Acetonitrile is chromatographically pure reagents, other reagents are analytically pure reagents

[0044] 2. Preparation of standard solution

[0045] Preparation of diamino standard solution: Take an appropriate amount of amino standard and dissolve it in the mobile phase, and if necessary, sonicate to dissolve completely.

[0046] Preparation of buffer solution: Weigh 0.015moL of dipotassium hydrogen phosphate and dissolve it in 1L of deionized water, and filter it with a 0.22um microporous membrane for use.

[0047] 3. Preparation of samples to be tested

[0048] Use a capillary to take 0.05g of the biotin-synthesized reaction solution into a 15ml sampling bottle, add 10mL of mobile phase to dissolve, and ultrasonically prepare it for later use.

[0049] The preparation process of biotin in this patent is as follows: bromine on uric acid generates monobenz...

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Abstract

The invention belongs to the technical field of biotin intermediate content detection, and particularly relates to an HPLC detection method of a process intermediate diamino substance in biotin synthesis. The method comprises the following steps: dissolving a cyclization reaction solution in a biotin synthesis process by using a mobile phase, and then, carrying out detecting under the following chromatographic conditions: chromatographic columns are C18-WCX mixed columns, the detection wavelength is 195-220 nm, the test time is 16-20 min, and the mobile phase is an acetonitrile-phosphate buffer solution. According to the detection method provided by the invention, the appearance time of the diamino substance of a substance to be detected can be stabilized within 10-12min, so that the problem of peak appearance superposition of the diamino substance and a solvent peak is solved, and the monitoring is more accurate and reliable.

Description

technical field [0001] The invention belongs to the technical field of content detection of biotin intermediates, and in particular relates to an HPLC detection method of a diamino compound in the process of biotin synthesis. Background technique [0002] During the synthesis of biotin (as shown in formula a), a diamino compound (as shown in formula b) will be produced. This substance is an intermediate process product of uric acid to generate biotin. After carbonylation and post-treatment, d-biotin can be obtained. [0003] [0004] The methods in the prior art all use biotin as the target product for detection, resulting in the neglect of diamino compounds that can be converted into biotin. Because diaminos appear near the dead time, it is not easy to determine their residues, thus affecting the overall yield of biotin. At present, the detection methods of biotin mainly include microbial method, liquid chromatography, enzyme-linked immunoassay, spectroscopic method and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/86
CPCG01N30/02G01N30/8631
Inventor 吕国锋王昌泽单国红杭姣王振振李小军段文杰韦念想
Owner SHANGYU NHU BIOCHEM IND
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