HPLC detection method of MES and NHS residues

A detection method and detection wavelength technology, which is applied in the field of HPLC detection, can solve the problems of single quantitative increase and difficulty, and achieve the effects of good linear range, high accuracy, and simple and easy operation method

Active Publication Date: 2015-03-11
ZHEJIANG JINGJIA MEDICAL TECH CO LTD
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Problems solved by technology

[0003] Two substances, 2-morpholineethanesulfonic acid (MES) and N-hydroxysuccinimide (NHS), are more polar, and their solubility in water is greater, which actually achieves a separate quantitative increase for the two substances Difficulty

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  • HPLC detection method of MES and NHS residues
  • HPLC detection method of MES and NHS residues
  • HPLC detection method of MES and NHS residues

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Embodiment Construction

[0028] 1. Device information

[0029] Chromatograph: Shimadzu high performance liquid chromatograph LC-20AB;

[0030] Chromatographic column: ZORBAX.SB-C18 4.6mm*250mm*5μm;

[0031] Injection volume: 10 μL.

[0032] 2. Investigation of MES and NHS detection methods by high performance liquid chromatography

[0033] 2.1 Preparation of MES and NHS test solution

[0034] MES test solution (300μg / mL): Accurately weigh 30.09mg of the sample, dissolve it in purified water, and dilute to 100mL.

[0035] NHS test solution (300μg / mL): Accurately weigh 30.07mg of the sample, dissolve it in purified water, and dilute to 100mL.

[0036] 2.2 Determination of detection wavelength

[0037] The test solutions of MES and NHS were taken respectively and scanned at full wavelength on the ultraviolet-visible spectrophotometer. The results show that NHS absorbs from 300nm and reaches the maximum at 200nm, MES absorbs from 220nm and reaches the maximum at 200nm, because the organic phase of t...

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Abstract

The invention discloses an HPLC detection method of MES and NHS residues. A reversed-phase C18 silica bonded chromatographic column is used, a mobile phase consists of a phosphate buffer of pH 6.5 to 6.8, and an organic solvent, and the detection wavelength is 205 to 210nm; the organic solvent is methanol or acetonitrile. According to the method, samples do not need to be specially treated, the operation method is simple and easy to implement, the accuracy is high, the needs in actual detection can be met, and two substances MES and NHS can be detected to reach microgram scale.

Description

technical field [0001] The invention relates to an HPLC detection method for MES and NHS residues. Background technique [0002] Among the relatively stable chemical cross-linking methods nowadays, the cross-linking system of carbodiimide has attracted more and more attention. The cross-linking system of carbodiimide can be applied to biological polysaccharides, polypeptides, proteins, nucleic acids and polymer modification, etc. In various fields of biology and organic synthesis, its own advantages such as low toxicity, rapid cross-linking effect, and stable results have excellently made up for the safety problems caused by the traditional use of formaldehyde and glutaraldehyde cross-linking methods. Compared with physical cross-linking methods such as thermal cross-linking, the cross-linking system of carbodiimide is more stable, and the cross-linked products are more uniform. 2-Morpholineethanesulfonic acid (MES) provides a relatively stable pH range in the cross-linking...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/74
Inventor 杨中科龚彦铭陈卫英丁祥李娟李好雨张兆利杨安顺蒋小龙彭红卫
Owner ZHEJIANG JINGJIA MEDICAL TECH CO LTD
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