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A kind of hplc detection method of biotin intermediate diamino

A detection method, the technology of diamino substances, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to accurately detect the content, inability to directly apply the reaction solution to direct detection, etc.

Active Publication Date: 2020-10-20
SHANGYU NHU BIOCHEM IND +1
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  • Summary
  • 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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  • A kind of hplc detection method of biotin intermediate diamino
  • A kind of hplc detection method of biotin intermediate diamino
  • A kind of hplc detection method of biotin intermediate diamino

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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 in particular relates to an HPLC detection method for a diamino compound in the process of biotin synthesis. The method comprises the steps of: dissolving the cyclization reaction solution in the biotin synthesis process with mobile phase and then detecting under the following chromatographic conditions: chromatographic column C18-WCX mixed column; detection wavelength is 195-220nm, and test time is 16-20min , and the mobile phase was acetonitrile-phosphate buffered saline. The detection method provided by the invention can stabilize the peak eluting time of the diamino substance to be tested at 10-12 minutes, solves the problem that the peaks of the diamino compound and the solvent peak overlap, and makes the monitoring 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 Patents(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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