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sample pretreatment method for simultaneous detection of Malachite green residual liquid chromatography -visible light and fluorescence in freshwater aquaculture water body

A malachite green and liquid chromatography technology, which is applied in the pretreatment field of liquid chromatography detection of malachite green residues in water bodies, can solve the problem of unstable recovery rate of malachite green, high requirements for pretreatment methods, and the addition of potassium borohydride High volume and other problems, to achieve the effect of less pollution, simple method and improved recovery rate

Active Publication Date: 2019-08-30
BOHAI UNIV
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  • Claims
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Problems solved by technology

However, in the actual detection process, due to the difference in raw material malachite green content, or the impact of the pH of the chromatographic buffer on potassium borohydride, in the potassium borohydride reduction step, it is often encountered that the addition of potassium borohydride is too high or the addition is too high. This is an important reason for the unstable recovery rate of malachite green; liquid chromatography-visible light and fluorescence detection method is expected to solve this problem
However, liquid chromatography-visible light and fluorescence combined detection method has high requirements for sample pretreatment methods. It is urgent to seek a simple and effective pretreatment method that can be used for water malachite green liquid chromatography-visible light and fluorescence detection. The traditional method requires a large amount of organic solvents, the steps are cumbersome, and the required amount of potassium borohydride must be accurate

Method used

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  • sample pretreatment method for simultaneous detection of Malachite green residual liquid chromatography -visible light and fluorescence in freshwater aquaculture water body
  • sample pretreatment method for simultaneous detection of Malachite green residual liquid chromatography -visible light and fluorescence in freshwater aquaculture water body
  • sample pretreatment method for simultaneous detection of Malachite green residual liquid chromatography -visible light and fluorescence in freshwater aquaculture water body

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030]Put 20mL of aquaculture fresh water in a centrifuge tube, add 10mL of 1μg / mL malachite green standard solution, filter through a 0.45μm membrane, then add 2mL of extract solution I, 20mL of extract solution II, 1) The preparation method of extract solution I: p-toluene Sulfonic acid 0.24g, hydroxylamine hydrochloride 3g, dissolve in ultrapure water, and dilute to 100mL; 2) The extract II is a mixed solution of acetonitrile and dichloromethane (1:2, v / v); vortex mixer After mixing thoroughly, add 15g of anhydrous sodium sulfate solid, centrifuge at 4500r / min for 15min, pour the supernatant in the centrifuge tube into a 100mL evaporating flask, and use a rotary evaporator at a water temperature of 35°C with a rotational speed of 50r Evaporate the solvent under reduced pressure at 1 / min, add 1mL potassium borohydride solution with a mass concentration of 1% and 1mL acetonitrile into the evaporating flask. Solution and dilute to 3mL, inject sample after 0.45μm membrane filtr...

Embodiment 2

[0044] Put 20mL of aquaculture fresh water in a centrifuge tube, add 10mL of 1.0μg / mL malachite green standard solution, filter through a 0.45μm membrane, add 2mL of extract solution I, and 20mL of extract solution II, 1) The preparation method of the extract solution I: p-toluene 0.24g of sulfonic acid, 3g of hydroxylamine hydrochloride, dissolved in ultrapure water, and dilute to 100mL; 2) The extract II is a mixed solution of acetonitrile and dichloromethane (1:2, v / v), vortex mixer After mixing thoroughly, add 15g of anhydrous sodium sulfate solid, centrifuge at 4500r / min for 15min, pour the supernatant in the centrifuge tube into a 100mL evaporating flask, use a rotary evaporator, and use a rotary evaporator to cool the mixture at a water temperature of 30°C , the solvent was evaporated to dryness under reduced pressure under the rotary evaporation speed of 70r / min, the solvent was evaporated to dryness under reduced pressure, 1mL of potassium borohydride solution with a m...

Embodiment 3

[0046] Put 20mL of aquaculture fresh water in a centrifuge tube, add 10mL of 1.0μg / mL malachite green standard solution, filter through a 0.45μm membrane, add 2mL of extract solution I, 20mL of extract solution II, 1) The preparation method of extract solution I: p-toluene 0.24g of sulfonic acid, 3g of hydroxylamine hydrochloride, dissolved in ultrapure water, and dilute to 100mL; 2) The extract II is a mixed solution of acetonitrile and dichloromethane (1:2, v / v), vortex mixer After mixing thoroughly, add 15g of anhydrous sodium sulfate solid, centrifuge at 4500r / min for 15min, pour the supernatant in the centrifuge tube into a 100mL evaporating flask, use a rotary evaporator, and use a rotary evaporator to cool the mixture at a water temperature of 45°C. , the solvent was evaporated to dryness under reduced pressure under the rotary evaporation speed of 30r / min, and the solvent was evaporated to dryness under reduced pressure, and 1mL of potassium borohydride solution with a ...

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Abstract

The present invention discloses a sample pretreatment method for simultaneous detection of malachite green residual liquid chromatography -visible light and fluorescence in freshwater aquaculture water body. The method comprises following specific steps: 20 mL of breeding fresh water is filtered through a membrane, then is placed in a centrifuge tube, then 2 mL of extracting solution I and 20 mL of extracting solution II is added, after thoroughly mixed on a vortex mixer, 15 g of anhydrous sodium sulfate solid is added, after centrifugated at 4500 r / min for 15 min, supernatant in the centrifuge tube is poured into a 100 mL evaporation flask, the solvent is evaporated under reduced pressure using a rotary evaporator, a mass concentration of 1% of potassium borohydride solution and 1 mL of acetonitrile is added to the evaporation flask, after ultrasonic vibration, the solution in the evaporation flask is transferred out, the evaporation flask is washed with 1 mL of acetonitrile, the solution is combined and the volume is adjusted to 3 mL, after filtered by a membrane, sample is injected, and the malachite green residue is simultaneously detected through the liquid chromatography-visible light and fluorescence.

Description

technical field [0001] The present invention relates to a pretreatment method for liquid chromatographic detection of residual malachite green in water bodies, in particular to a sample pretreatment method for liquid chromatography-visible light and fluorescence simultaneous detection of malachite green residual in freshwater aquaculture water . Background technique [0002] Malachite Green (C 23 h 25 N 2 , MG) The scientific name is tetramethyldiaminotriphenylmethane, also known as product green, malachite green, basic green, etc., is a class of synthetic triphenylmethane chemical substances. It can be used as dyeing of silk, leather and paper, as well as biological dyeing agent, and can also be used to prevent and treat saprolegniasis, gill mold and melon worm disease of various aquatic animals. Malachite green has special effects in the treatment of saprolegniasis, has been widely used in aquaculture, and has also been used in the transportation and storage of aquatic...

Claims

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Application Information

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IPC IPC(8): G01N30/74G01N30/14
CPCG01N30/14G01N30/74
Inventor 李秀霞王珍励建荣孙协军刘雪飞孙彤马莹莹
Owner BOHAI UNIV
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