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Molecularly imprinted microsphere for detecting teicoplanin and preparation method and application thereof

A technology of molecularly imprinted microspheres and teicoplanin, applied in separation methods, other chemical processes, solid adsorbent liquid separation, etc., achieving good application prospects, low detection limits, and high recovery rates

Active Publication Date: 2018-10-16
GUANGZHOU CENT FOR DISEASE CONTROL & PREVENTION (GUANGZHOU HYGIENE INSPECTION CENT GUANGZHOU CENT FOR FOOD SAFETY RISK SURVEILLANCE & ASSESSMENT INST OF PUBLIC HEALTH OF GUANGZHOU MEDICAL UNIV)
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the synthesis of molecularly imprinted polymers with good molecular recognition capabilities in aqueous matrices has been a challenge.

Method used

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  • Molecularly imprinted microsphere for detecting teicoplanin and preparation method and application thereof
  • Molecularly imprinted microsphere for detecting teicoplanin and preparation method and application thereof
  • Molecularly imprinted microsphere for detecting teicoplanin and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The reaction route of sugar-based molecularly imprinted microspheres is as follows: figure 1Shown, mainly comprise the following steps: in the round bottom flask of 250mL, 200mg methyl methacrylate, 700mg divinylbenzene and 100mg azobisisobutyronitrile are dissolved in the mixed solvent of 128mL acetonitrile and toluene (75 / 25, v / v). Stir at room temperature for 10 min under nitrogen blowing. 100 μg of template molecules and 0.242 g of 4-vinylphenylboronic acid were dissolved in 5.0 mL of phosphate buffer solution (0.01 mol / L, pH 9.5), and added dropwise to the above solution. Then ultrasonic degassing for 5 minutes, followed by nitrogen purging for 10 minutes. Then the obtained mixed solution was sealed and stirred with an electromagnetic stirrer, and then the temperature was raised to 60° C., and the polymerization reaction was carried out for 24 hours. The resulting MIMs were centrifuged at 9000 rpm for 10 min. Subsequently, the above MIMs were repeatedly washed...

Embodiment 2

[0047] MIMs and NIMs microspheres were prepared by the method of Example 1. 10 mg of MIMs (NIMs) was added to 2 mL of buffer solutions with different pH values ​​containing 100 μg / L teicoplanin, shaken horizontally at room temperature for 2 h, and then centrifuged. The concentration of teicoplanin in the supernatant was detected by UPLC-MS / MS.

[0048] Figure 6 is the adsorption capacity of MIMs and NIMs for teicoplanin at different pH values. Boric acid can covalently react with cis-diol in alkaline aqueous solution to form cyclic ester, and when the medium becomes acidic, the cyclic ester is free. Therefore, pH will affect the adsorption amount of MIMs. When the pH is 9.0, the adsorption capacity of MIMs is the largest.

Embodiment 3

[0050] MIMs microspheres were prepared by the method in Example 1. A 3 mL empty column was directly filled with 60 mg of dry MIMs, and both ends of the filler were sealed by sieve plates. The resulting column was a MISPE column. Then use 3mL methanol and 3mL 0.05mol / L phosphate buffer (pH 7.0) to activate the column, then load the sample, then wash with 2.0mL deionized water, and use the eluent to rinse the loaded MISPE column . The obtained eluate was collected, blown with nitrogen at 40° C., and then dissolved in 0.1% formic acid solution, and detected by UPLC-MS / MS.

[0051] By changing the type and volume of the eluent, the extraction conditions of the solid-phase extraction column are optimized to obtain the best eluent and elution volume.

[0052] Figure 7 and Figure 8 Respectively represent the pH value of the eluent and the impact of its volume on the recovery rate of teicoplanin, the best eluent is pH=4.0 buffer solution+methanol (1 / 1), and the best elution volum...

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Abstract

The invention discloses a molecularly imprinted microsphere for detecting teicoplanin and a preparation method and application thereof. The preparation method includes: selecting a molecule similar toa glycosyl portion in a teicoplanin molecular structure as a template molecule, 4-vinylphenylboronic acid and methyl methacrylate as functional monomer and divinyl benzene as a crosslinking agent toprepare the molecularly imprinted microsphere. When pH value is 9.0, the synthesized molecularly imprinted microsphere can be combined with target glycopeptide antibiotic; when the pH value is loweredto below 4.0, the combined glycopeptide antibiotic can be released, so that a glycosyl molecularly imprinted solid-phase extraction column prepared on this principle can intercept the glycopeptide antibiotic in the solid-phase extraction column in a slightly alkaline condition, then the glycopeptide antibiotic is eluted in an acidic condition. On the basis of the glycosyl molecularly imprinted solid-phase extraction column, teicoplanin in serum and urine samples can be detectedly effectively, and the molecularly imprinted microsphere has high recovery rate, low detection limit and good application prospect.

Description

technical field [0001] The invention relates to the field of molecularly imprinted microspheres, in particular to a molecularly imprinted microsphere for detecting teicoplanin and its preparation and application. Background technique [0002] Since the British scientist Alexander Fleming discovered penicillin in 1928, antibiotics have been widely used around the world to treat and prevent infectious diseases. With the extensive application of antibiotics, pathogenic microorganisms have developed tolerance and resistance to antibiotics and other drugs. The emergence of this kind of drug resistance makes the normal dosage of the drug no longer exert its proper bactericidal effect, and even makes the drug completely ineffective, which makes it difficult to treat the disease and easily spread the disease. Therefore, antibiotic-resistant bacterial infections have become one of the most serious public health problems. Glycopeptide antibiotics are mainly used to treat serious inf...

Claims

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Application Information

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IPC IPC(8): C08F212/36C08F220/14C08F230/06C08J9/26B01J20/26B01J20/28B01J20/30B01D15/22
CPCB01D15/22B01J20/264B01J20/28021B01J2220/4812C08F212/36C08J9/26C08J2201/0422C08F220/14C08F230/06
Inventor 谭磊陈坤才邓芬芳罗晓燕潘心红李军涛钟嶷刘于飞
Owner GUANGZHOU CENT FOR DISEASE CONTROL & PREVENTION (GUANGZHOU HYGIENE INSPECTION CENT GUANGZHOU CENT FOR FOOD SAFETY RISK SURVEILLANCE & ASSESSMENT INST OF PUBLIC HEALTH OF GUANGZHOU MEDICAL UNIV)
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