A method for high-throughput detection of multiple different types of environmental pollutants in urine using LC-MS-MS

A LC-MS-MS and environmental pollutant technology, applied in the field of environmental analysis, can solve the problems of low sensitivity of mass spectrometry of a single compound, insufficient range of pollutant types, and difficult separation of various compounds, so as to save manpower and material costs and simplify pretreatment Fast, stable and reproducible results

Active Publication Date: 2021-11-19
XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, the GC-MS method sample pretreatment requires complex steps such as acidification, liquid-liquid extraction, solid-phase extraction, and derivatization, which are time-consuming and laborious. Moreover, GC is difficult to apply to compounds whose structures are easily destroyed by heat, and the application has certain limitations.
The types of pollutants involved in the LC-MS method are not extensive enough, and multiple detections are required to detect many different types of pollutants, resulting in a relatively large sample volume and a relatively long time-consuming
Therefore, the existing technology is still facing challenges in the comprehensive evaluation of pollutant exposure, and it is urgent to develop a sensitive and stable detection method for detecting multiple different types of pollutants with a small amount of samples at one time.
[0005] For liquid phase analysis, the difficulties in detecting multiple different types of compounds at one time are: it is difficult to completely separate different types of compounds in liquid phase, and the sensitivity of mass spectrometry of a single compound is reduced.
It is still difficult to develop a sensitive and stable detection method for detecting many different types of pollutants with a small number of samples at once.

Method used

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  • A method for high-throughput detection of multiple different types of environmental pollutants in urine using LC-MS-MS
  • A method for high-throughput detection of multiple different types of environmental pollutants in urine using LC-MS-MS
  • A method for high-throughput detection of multiple different types of environmental pollutants in urine using LC-MS-MS

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] The sample pretreatment steps are: 100ul urine sample is added to the internal standard solution, then 50ul 1mol / L ammonium acetate aqueous solution (adjusted to pH 5.0), then β-glucuronyl esterase is added, mixed and incubated at 37°C for 3h. After the incubation, take out and add 500ul of acetonitrile containing 1% formic acid, vortex fully for 15s, centrifuge to get the supernatant, concentrate the supernatant to 100ul by vacuum centrifugation, and test it on the machine, the injection volume is 20ul. The method is simple to operate and requires less sample volume, which overcomes the characteristics of tedious, complicated, time-consuming and sample volume-consuming pretreatment in other methods.

[0071] Liquid phase conditions: Chromatographic column specifications: C18 reverse chromatographic column, 3.0mm*100mm; the aqueous phase is 5mmol / L ammonium acetate aqueous solution, the organic phase is pure methanol, and the initial ratio of mobile phase is 95% aqueous ...

Embodiment 2

[0080] Embodiment 2 method specificity

[0081] Method specificity: Use the method of the present invention to measure blank double-distilled water and urine spiked samples to see if there is an interference peak at the peak position of the target compound, so as to judge the specificity of the method for each target compound. Experimental results such as Figure 5 and Image 6 shown.

[0082] Compared Figure 5 and Image 6 It can be seen that the specificity of the method of the present invention is good, and the peak shape of each target compound is good. The blank water sample measured by this method has no interference peak at the peak position of the target compound, and the specificity of the method is high.

Embodiment 3

[0083] Embodiment three matrix effect

[0084] Matrix effect detection: the urine sample was spiked directly for detection on the machine, the urine sample was spiked with the method to remove the matrix and then tested on the machine, and double-distilled water without matrix was used to prepare a certain concentration of the sample and tested on the machine respectively. Obtain the response value of each target compound, see the impact of the matrix on the response of each target compound, and the removal effect of this method on the matrix. The test results are shown in Table 4.

[0085] Table 4 Matrix effects of 22 target compounds

[0086]

[0087]

[0088] The experimental results show that the matrix inhibition rate of the target compound most affected by the matrix is ​​as high as 92.5% when the urine sample is directly tested on the machine, and the response value becomes significantly larger after the sample pretreatment method in this method. Among them, the...

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Abstract

The invention discloses a method for using LC-MS / MS high-throughput detection of various types of environmental pollutants in urine, which includes the following steps: (1) sample pretreatment: the urine after enzymolysis is treated with formic acid Fully vortexed after the acetonitrile treatment, centrifuged to get the supernatant, the supernatant was concentrated by vacuum centrifugation to obtain the injection sample detected on the machine; (2) adopt LC-MS / MS injection to detect the injection sample: liquid phase The conditions are: the chromatographic column adopts a reversed-phase C18 chromatographic column; the mobile phase A is 4-6 mmol / L ammonium acetate aqueous solution; the mobile phase B is methanol; gradient elution is adopted; Multiple ion detection mode. The method of the invention has the characteristics of rapid detection, high efficiency, high throughput, strong versatility, high sensitivity, good stability and reproducibility, and is suitable for large-scale detection of biological samples in epidemiological research.

Description

technical field [0001] The invention belongs to the field of environmental analysis, and in particular relates to the development, verification and application of a method for detecting various types of environmental pollutants in urine by using LC-MS / MS high-throughput. Background technique [0002] At the same time as China's rapid economic development, the resulting environmental pollution, especially the impact on health, has attracted much attention. In recent years, I have been engaged in the detection of environmental pollutants in human samples and found that different types of environmental pollutants can be detected in the urine of the general population, but they are based on different detection methods. Since people are inevitably exposed to the cumulative exposure of diverse pollutants rather than a single pollutant, how to comprehensively evaluate the exposure level of multiple pollutants in the human body is the premise and basis for studying the impact of env...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N33/50
CPCG01N30/02G01N30/06G01N33/50G01N2030/027G01N2030/062
Inventor 田英赵莎莎王伟业张军
Owner XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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