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A detection method of form mercury in animal tissue cells

A detection method and animal tissue technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of high detection limit, poor selectivity, unfavorable trace form mercury detection, etc., and achieve stability enhancement and mutual conversion prevention. Effect

Active Publication Date: 2020-06-09
ZUNYI INST OF PROD QUALITY INSPECTION & TESTING +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the cumbersome pretreatment of gas chromatography, many interference factors, poor selectivity of ECD, and high detection limit, it is not conducive to the detection of trace forms of mercury in cells; gas chromatography-atomic absorption method has low sensitivity; high performance liquid chromatography- The pretreatment process of atomic fluorescence spectrometry is complicated, and the form of mercury in the cells is easy to change during the pretreatment process, which affects the accuracy of the measurement results

Method used

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Effect test

Embodiment 1

[0024] The detection method of form mercury in animal tissue cells of the present invention comprises the following steps:

[0025] (1) Sample pretreatment: After treating the cell sample with 0.3% triton for 15 minutes, sonicate for 60 minutes, shake and shake, take 100 μL of sample solution, add 500 μL of extractant, shake and shake well, centrifuge at 12,000 rpm for 15 minutes at 4°C, and take the supernatant The liquid was analyzed by UPLC-ICP-MS after passing through a 0.22μm filter membrane;

[0026] (2) Chromatographic conditions: Chromatographic column: Althena C18, 120A, 4.6×250mm, 5μm; mobile phase: 10mmol / L ammonium acetate, 0.05% L-cysteine, 5% methanol, adjust pH to 2.0- with concentrated hydrochloric acid 2.5, dilute to 1L with primary water; flow rate: 1.1mL / min; injection volume: 20μL;

[0027] (3) Mass spectrometry conditions: ICP-MS working parameters: optimize the instrument conditions with 1.0μg / L tuning solution before measurement, among them, ICP-MS of T...

Embodiment 2

[0030]Standard music: Inorganic mercury, methylmercury, and ethylmercury standard solutions were purchased from the China Institute of Metrology, diluted step by step to 1000μg / L and then mixed to prepare a mixed standard solution containing 100μg / L of mercury, and then extracted solution and add a certain amount of cell culture medium to dilute inorganic mercury, methylmercury and ethylmercury step by step. Supernatant detection; on-machine detection obtained r2=0.9998 for inorganic mercury, f(x)=16373.8678*x+161.7391; r2=0.9998 for methylmercury, f(x)=14744.4700*x+214.245; r2=0.9997f(x)=14601.0805*x+69.5239; linear range: 0~5μg / L.

Embodiment 3

[0032] Recovery rate: After adding a certain amount of three forms of mercury mixture to the cultured cells, extract and detect the data of group A; then culture 4 copies of the same tissue cells under the same conditions as the data of group A, and select two of them to directly extract the form Mercury, get the data of group B, add three kinds of mercury mixed solution according to the ratio of 1:1 of the data of group A, and extract the inorganic mercury under the same conditions after shaking well, get the data of group C, get the recovery rate after calculation, the mercury is 91.6 %; methylmercury is 96.7%, ethylmercury is 97.8%.

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Abstract

The invention discloses a method for detecting mercury species in animal tissue cells. The method includes a step (1) of preparing standardized product solution; a step (2) of pre-treating a sample: after cell wall cracking, using an extractant to perform extraction and centrifugalization, taking supernatant, filtering the supernatant and then analyzing the supernatant through an instrument; a step (3) of adopting ultra performance liquid chromatography - inductively coupled plasma mass spectrometry to perform determination, wherein the instrument conditions are as follows: a mobile phase of ultra-high performance liquid chromatography is ammonium acetate, L-cysteine and methanol system, and the pH is adjusted to 2.0-2.5. According to the method for detecting mercury species in the animaltissue cells, the interference of sodium ions in cell culture solution can be eliminated, and the stability and accuracy of the method are greatly improved.

Description

technical field [0001] The invention belongs to the field of trace metal detection, and in particular relates to an ultra-high performance liquid chromatography-inductively coupled plasma mass spectrometry method for determining form mercury in animal tissue cells. Background technique [0002] Hg-containing traditional Chinese medicine has a long history in traditional Chinese medicine in my country. Cinnabar is one of the mercury-containing traditional Chinese medicines. It has the effects of clearing the heart, calming the nerves, improving eyesight, and detoxifying. Wind, dim vision, aphthous sore throat, sore swollen toxin and other syndromes are listed as the first tranquilizers, and its main component is HgS. [0003] Mercury-containing substances are toxic substances, so there is a risk of poisoning when taking cinnabar, and improper use may even lead to death. At present, there is no report on the form of mercury in cells after the main component of cinnabar, HgS, i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 周绍均陆远富罗砚文向丽萍鲁艳柳王奥李露刘桂岚
Owner ZUNYI INST OF PROD QUALITY INSPECTION & TESTING
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