Quantitative detecting method for estrogen in milk
A quantitative detection method, estrogen technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low estrogen content and high requirements for detection methods, achieve high automation, improve detection sensitivity, and simple operation Effect
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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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