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Quantitative detecting method for estrogen in milk

A quantitative detection method and estrogen technology, which can be applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of extremely high detection method requirements and low estrogen content, and achieve improved detection sensitivity, high degree of automation, and simple and easy operation. row effect

Active Publication Date: 2014-03-12
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a high-sensitivity, high-precision, online rapid sample extraction analysis technology for the problems of complex matrix of milk samples, low estrogen content under normal circumstances, and extremely high requirements for detection methods, so as to realize the detection of traces in milk samples. Quantitative rapid automated determination of estrogen

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  • Quantitative detecting method for estrogen in milk
  • Quantitative detecting method for estrogen in milk
  • Quantitative detecting method for estrogen in milk

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 detects the pretreatment of sample

[0036] Take 3mL uniformly mixed milk sample, add three deuterium-labeled estrogen internal standards, and vortex mix. The final concentration of internal standard is 50ppt. Next, 100 μL of 30% trichloroacetic acid was added to the sample, and mixed. Add 8 mL of ethyl acetate and n-hexane (1:1) solvent, shake and extract for 10 min, then centrifuge at 10,000 rpm for 10 min. The supernatant was transferred and blown dry with nitrogen in a water bath at 50°C. Repeat the extraction twice. Dissolve the nitrogen-dried residue in 1 mL of acetonitrile, incubate at 50°C for 5 minutes, vortex mix for 30 seconds, let stand at room temperature for 5 minutes, and freeze at -20°C for 30 minutes. The samples were centrifuged at 10,000 rpm for 5 min at 4°C, the supernatant was transferred to another test tube, and dried with nitrogen in a water bath at 50°C. The residue was dissolved by swirling with 150 μL of sodium bicarbonate bu...

Embodiment 2

[0037] Example 2 Analysis of estrogen concentration by using on-line solid phase extraction-two-dimensional liquid chromatography tandem mass spectrometry

[0038] Online solid phase extraction-two-dimensional liquid chromatography tandem mass spectrometry selection conditions: 2 Agilent 1200SL liquid chromatography binary pumps, G1367C automatic sampler, G1316B column thermostat with Valco six-way valve. The mass spectrometer is equipped with an electrospray ionization source (ESI) and Analyst 1.4.2 data processing software for API 5000,

[0039] Strata-X online solid-phase extraction column (20mm×2.0mm×20μm, purchased from Phenomenex) was used for sample purification. Eclipse Plus phenylhexyl column (2.1 mm×100 mm×3.5 μm, purchased from Agilent) was used for sample analysis. Ion source: electrospray ionization source (ESI), positive ion detection; ion source voltage: +5500V; temperature: 500°C; source gas 1 (GS1, N 2 ) pressure 65psi; gas 2 (GS2, N 2 ) pressure 55psi; air c...

Embodiment 3

[0041] The drawing of embodiment 3 standard curve

[0042] Quantitative determination was carried out by matrix addition internal standard method. The drawing of the standard curve of the internal standard method includes: preparing a blank milk sample, weighing 8 parts, each 3mL. The internal standard of the same concentration was added respectively, and the final concentration of the internal standard was 50ng / L. Add different concentrations of estrogen standard substances in turn, so that the final concentrations of the standard points of the standard curve are 0ng / L, 2.0ng / L, 5.0ng / L, 10.0ng / L, 20.0ng / L, 40.0ng / L, 80.0 ng / L and 100ng / L. The above-mentioned matrix addition standard is extracted and determined by liquid chromatography-tandem mass spectrometry in the same way as the sample, and the ratio of the peak area of ​​the measured component in the added sample to the peak area of ​​the internal standard is the ordinate, and the concentration of the measured componen...

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Abstract

The invention provides a quantitative detecting method for estrogen in milk. The quantitative detecting method includes steps of 1) performing derivatization pretreatment for a milk sample; and 2) measuring the content of the estrogen in the milk sample by the aid of online solid-phase extraction and two-dimensional liquid chromatogram tandem mass spectrum analysis. Owning to sample derivatization measurement, detection sensitivity is greatly improved. A two-dimensional liquid chromatogram tandem mass spectrum system is used for detecting, accordingly, online automatic purification of the sample is realized, and the detection efficiency of the sample is greatly improved. In addition, the analysis method is fine in recovery rate, high in automation degree and simple and easy in operation, and can fast and synchronously analyze the content of various types of trace estrogen in the sample.

Description

technical field [0001] The invention relates to the field of food inspection, in particular to a quantitative detection method for estrogen in milk. Background technique [0002] In the past decade, estrogen, as a typical endocrine disruptor, has received extensive attention. Estrogen-like endocrine disruptors (EDCs) are a class of compounds that can bind to estrogen receptors, interfere with normal hormone secretion in the body, cause the quality of germ cells to decline, and cause serious harm such as carcinogenic, teratogenic, and mutagenic. Estrogen can lead to human endocrine imbalance, especially low-dose long-term exposure has a serious impact on specific groups of people (such as fetuses, newborn babies, and prepubertal children), such as precocious puberty, obesity, etc., which have evolved into social problems. Another major hazard of estrogen is its carcinogenic effect. Foreign epidemiological surveys show that the incidence of testicular cancer and uterine canc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
Inventor 陈刚郑晓春李思聪杜一楠米晓霞方美英
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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