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Method for determining content of pyrroloquinoline quinone in biological sample by magnetic solid-phase extraction-capillary zone electrophoresis

A biological sample and capillary technology, applied in the field of biological analysis, can solve the problems of cumbersome steps, heavy workload, and expensive C18 column, and achieve the effect of good symmetry and improved separation efficiency

Inactive Publication Date: 2013-07-10
JIANGSU INST OF NUCLEAR MEDICINE
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Problems solved by technology

The PQQ capillary electrophoresis detection method reported in the literature is to add β-alanine to the buffer system, and use reverse-phase C18 small column solid-phase extraction to separate and enrich PQQ in the cell culture medium. The detection limit is 0.1-0.2 μM, and the PQQ peak type There is tailing phenomenon, the C18 small column used in solid phase extraction is expensive, and the steps are cumbersome and the workload is heavy

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  • Method for determining content of pyrroloquinoline quinone in biological sample by magnetic solid-phase extraction-capillary zone electrophoresis
  • Method for determining content of pyrroloquinoline quinone in biological sample by magnetic solid-phase extraction-capillary zone electrophoresis
  • Method for determining content of pyrroloquinoline quinone in biological sample by magnetic solid-phase extraction-capillary zone electrophoresis

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Embodiment 1

[0033] (1) Capillary electrophoresis analysis conditions:

[0034] Uncoated quartz capillary column (41.5cm×50μm, Hebei Yongnian County Ruifeng Chromatography Device Co., Ltd.), the new capillary was washed with methanol for 10min, distilled water for 5min, 1mol / L NaOH solution for 30min, distilled water for 5min, and finally Rinse with running buffer for 10-15min. Both the running buffer and the sample solution were filtered with a 0.22 μm microporous syringe filter. Before each injection, rinse with distilled water and running buffer for 2 minutes. The detection wavelength is 249nm, the injection method is pressure injection (0.5psi, 10.0 seconds), the separation voltage is 25kV, and the capillary column temperature is 25°C. The running buffer is: 10mmol / L borax-10mmol / Lβ-cyclodextrin mixture, pH=8.8, containing 0.1% trifluoroacetic acid.

[0035] (2) Selection of buffer system

[0036] The buffer system and concentration directly affect the migration and separation of i...

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Abstract

The invention provides a method for determining the content of pyrroloquinoline quinone in a biological sample by magnetic solid-phase extraction-capillary zone electrophoresis and belongs to the field of a life analyzing technology. According to the method disclosed by the invention, SiO2 and adsorbents with different polarities are mixed; and the adsorbents have different selective properties on compounds with different hydrophobic properties, so that the effective separation can be carried out. A capillary electrophoresis method adopted by the invention adds a modifying agent beta-cyclodextrin; trifluoroacetic acid is firstly added into a capillary electrophoresis buffering system to solve the trailing problem of a peak in a capillary electrophoresis picture; and the symmetrical property of the peak is obviously better than that of a document report. The method disclosed by the invention is rapid and convenient, and can be used for determining the content of the PQQ in milk by utilizing a magnetic solid-phase extraction technology being combined with capillary electrophoresis firstly. Compared with a document method, the method disclosed by the invention improves the separation efficiency by 6 times.

Description

technical field [0001] The invention discloses a method for measuring pyrroloquinone quinone content in biological samples by magnetic solid-phase extraction-capillary zone electrophoresis, belonging to the technical field of bioanalysis. Background technique [0002] Pyrroloquinoline quinine PQQ is a new prosthetic group of oxidoreductases, mainly distributed in prokaryotes, some plants and mammals, and has a high content in milk, especially breast milk, and is considered essential in the body One of the vitamins. [0003] The analysis methods of PQQ in biological samples include gas chromatography-mass spectrometry, high performance liquid chromatography, capillary electrophoresis, etc. Chromatography B, 739 (2000) 101–107.]. Capillary electrophoresis is a rapid analysis method developed in the 1980s, which has the advantages of fast analysis speed, high separation efficiency, and less consumption of samples and reagents. Compared with high performance liquid chromatogr...

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Application Information

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IPC IPC(8): G01N27/447G01N1/34
Inventor 周杏琴毛师师张建康曹国宪钦晓峰
Owner JIANGSU INST OF NUCLEAR MEDICINE
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