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Method for detecting various mushroom toxins in blood and urine

A urine and blood technology, applied in the field of detection of various mushroom toxins in blood and urine, to achieve the effect of simple pretreatment, high extraction efficiency and good separation effect

Inactive Publication Date: 2019-05-03
刘思洁
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the problem that conventional detection methods cannot detect the toxin concentration in the patient's urine and blood due to the low concentration of toxins in the occurrence of poisonous mushroom poisoning, and adopts a fast, simple and effective pretreatment technology to provide a It can quickly, effectively and accurately detect at least α-amanitin, β-amanitin, γ-amanitin, dihydroxyphallotoxin, and carboxydihydroxyphallotoxin in blood or urine Simultaneous rapid qualitative and accurate quantitative detection of 6 kinds of mushroom toxins including peptide and carboxytrihydroxy phalloidin

Method used

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  • Method for detecting various mushroom toxins in blood and urine
  • Method for detecting various mushroom toxins in blood and urine
  • Method for detecting various mushroom toxins in blood and urine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The method for detecting multiple mushroom toxins in blood and urine comprises the following steps:

[0052] 1) Sample pretreatment

[0053] 1. Blood sample: take 5.00mL of blood, centrifuge at 1500r / min for 5min, absorb 1.00mL of upper plasma into a 10mL centrifuge tube, add 1mL of acetonitrile solution containing 0.5% (V / V) acetic acid, vortex extract for 2min, and Centrifuge at 5000r / min for 5min, take 2.0mL of the supernatant in a 10mL test tube, dry it with nitrogen in a water bath at 55°C, add 250μL of the initial mobile phase (2mmol / L ammonium acetate aqueous solution with a mass ratio of 70-60:30-40 and 2mmol / L of ammonium acetate acetonitrile solution) to constant volume, vortexed and shaken for 1 min, filtered through a 0.22 μm filter membrane, and then determined.

[0054] 2. Urine sample: Take an appropriate amount of urine and filter it through a 0.22μm filter membrane for UPLC-MS / MS analysis.

[0055] 2) Preparation of standard solution

[0056] Mixed ...

Embodiment 2

[0083] The standard series of 6 kinds of mushroom toxins with different concentrations were prepared with blank matrix solution, analyzed by UPLC-MS / MS, and the linear range, regression equation, correlation coefficient (r) and detection limit were obtained. Sample processing is carried out according to the method of the present invention, 3 different levels of mixed standard solutions of 6 kinds of mushroom toxin contents are added to the matrix sample, and each added concentration is measured in parallel 6 times to determine the precision and accuracy.

[0084] 1) Linear range, regression equation, detection limit

[0085] The concentration of 6 kinds of mushroom toxins in blood samples was linear in the range of 10μg / L-100μg / L. The detection limits of the six mushroom toxins were 0.2 μg / L for α-amanitin, 0.2 μg / L for β-amanitin, 0.2 μg / L for γ-amanitin, and 0.1 μg / L for dihydroxyphallotoxin. μg / L, carboxydihydroxy phallotoxin 0.2μg / L, carboxytrihydroxy phallotoxin 0.2μg / L. ...

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Abstract

The invention discloses a method for detecting various mushroom toxins in blood and urine. Through the optimization on experimental conditions, the method for determining alpha-amanitin, beta-amanitin, gamma-amanitin, phalloidin, phallacidin, phallisacin contents in the blood and urine of poisoning patients by using ultra-high-performance liquid chromatography-mass spectrometry / mass spectrometry can be established for the first time. Thus, the scientific detection method can be provided for the disposal of emergency public health events.

Description

technical field [0001] The invention relates to the technical field of emergency detection. More specifically, it relates to a method for detecting various mushroom toxins in blood and urine. Background technique [0002] There are about 500 kinds of poisonous mushrooms in my country, of which about 421 kinds contain less toxins or can be eaten after treatment, more than 30 kinds contain strong toxicity that can kill, and at least 16 kinds are extremely toxic. One poisonous mushroom may contain multiple toxins, and one toxin can be present in many kinds of poisonous mushrooms. Jilin Province is rich in mushroom resources. With the development and utilization of mushroom resources, accidents of accidentally eating poisonous mushrooms often occur. This has all caused great threat to people's health and even life. In 2015, a total of 169 reports of food poisoning public health emergencies (hereinafter referred to as food poisoning incidents) were received through the public ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 刘思洁吴永宁方赤光崔勇石矛马杰
Owner 刘思洁
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