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Capillary electrophoretic analysis method for trace detection of phenazine-1-carboxylic acid

A technology of capillary electrophoresis and analysis method, which is applied in the field of capillary electrophoresis analysis for micro-detection of phenazine-1-carboxylic acid, which can solve the problems of few pesticide residues and achieve the effects of lower detection limit, higher precision, and accurate and simple method

Inactive Publication Date: 2009-08-12
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently rarely used in the analysis of pesticide residues

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] (1) Equipment and raw materials

[0017] Beijing Binda 1229—high-efficiency capillary electrophoresis analyzer (with UV monitor), Nanjing Qianpu HW-2000 chromatographic workstation, quartz capillary column with an inner diameter of 50-100 μm and a length of 30-70 cm (Hebei Yongnian Optical Fiber Factory).

[0018] Methanol (HPLC grade), triple distilled water (conductivity 0.082μS / cm)

[0019] (2) Preparation of phenazine-1-carboxylic acid crystals

[0020] 1. Acidify the fermentation broth fermented by Pseudomonas M18 with 1.0M HCl to pH=2.0, mix the treated fermentation broth with 3 times the volume of chloroform, shake and extract, centrifuge at 3000×g for 20 minutes, and take the organic The phase was evaporated to dryness to obtain the crude extract of phenazine-1-carboxylic acid.

[0021] 2. The crude extract of phenazine-1-carboxylic acid is subjected to silica gel column chromatography with chloroform as the eluent, and a bottle is collected every 30 minutes, ...

Embodiment 2

[0033] Example 2: Determination of phenazine-1-carboxylic acid in fermentation broth.

[0034] Take 180 μL of Pseudomonas M18 fermentation broth after 10 h of fermentation, and add 1M HCl to adjust the pH value to 2.0. Mix the treated fermentation broth with 540 μL of chloroform, shake and extract, centrifuge at 12000×g for 5 minutes, take out 500 μL of organic phase and vacuum-dry it, and finally dissolve it in 500 μL of methanol solution containing 200 μg / mL phenazine, and mix well.

[0035] Under the following conditions: separation voltage 20kV, detection wavelength 254nm buffer, column temperature: 25°C, 10mM pH6.6 phosphate buffer, 20cm height differential pressure injection for 75s, capillary inner diameter 75μm, length 59cm, capillary electrophoresis analysis, measured The concentration of phenazine-1-carboxylic acid in the 10h Pseudomonas M18 fermentation broth was 26.81 μg / mL.

Embodiment 3

[0036] Example 3: Determination of phenazine-1-carboxylic acid residues in soil.

[0037] Evenly apply 1 mL of phenazine-1-carboxylic acid methanol solution with a concentration of 1.5 μg / mL in 25 g of soil, that is, the application concentration is 0.30 mg / kg, and the treated soil is ultrasonically extracted with 100 mL of dichloromethane, and the extract After performing solid phase extraction, collect the solid phase extraction eluate, concentrate to dryness, and obtain the sample to be tested after vacuum drying.

[0038] The sample to be tested was dissolved in 5mL methanol, and filtered through a 46 μm filter membrane, and the filtrate was analyzed under the electrophoresis conditions in Example 1, and the concentration of phenazine-1-carboxylic acid in the extract could be measured to be 1.385 μg / mL, the recovery rate of this extraction was 92.35%.

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Abstract

The invention relates to an analysis method for micro-detecting the capillary electrophoresis of phenazine-1-carboxylic acid, which comprises the quantitative analysis of an internal standard method and an external reference method. Electrophoresis conditions are that: a quartz capillary column with an inner diameter of between 50 and 100mu m is adopted, and the length is between 30 and 70cm; the separation voltage is between 15 and 25kV; the detection wavelength is between 230 and 260nm; the column temperature is between 23 and 28 DEG C; buffer solution is phosphorous acid buffer solution with a concentration of between 5 and 20mM, and the pH value is between 6.0 and 7.0; and a sample loading mode is to load for 60 to 80s at a height pressure difference of between 10 and 20cm, and a sample is dissolved into methanol or methanol solution containing an internal standard substance with certain concentration. The method has good repetitiveness, is simple, convenient and feasible, performs on-line enrichment on the phenazine-1-carboxylic acid by electrostacking enrichment technology during the detection, greatly reduces the limit of detection of the method, and has better linear relation when the phenazine-1-carboxylic acid is between 0.075 and 30mu g / mL.

Description

technical field [0001] The present invention relates to a capillary electrophoresis analysis method for trace detection of phenazine-1-carboxylic acid, in particular to a method for detecting trace amounts of phenazine-1-carboxylic acid by electro-stacking non-aqueous phase capillary enrichment electrophoresis, including internal standard method Quantitative analysis with external standard method. Background technique [0002] Phenazine-1-carboxylic acid is an antibiotic substance that has a strong inhibitory effect on pathogenic bacteria such as wheat take-all, watermelon anthracnose and pepper blight. The good biological control effect of phenazine-1-carboxylic acid has led people to devote themselves to Research and develop new phenazine-1-carboxylic acid biopesticides to replace some heavily polluted chemical pesticides. Shenzimycin is a microbial-derived pesticide that is being widely promoted and applied in China. Its main active ingredient is phenazine-1-carboxylic a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447G01N1/28
Inventor 高彬张雪洪彭华松王威
Owner SHANGHAI JIAO TONG UNIV
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