Detection method of estrogen and progesterone in t

2022-09-23
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Dima Technology: detection methods of estrogen and progesterone in milk powder

the milk powder hormone incident has attracted great attention from all walks of life. As an experimental partner to help you ensure the safety of human food, environment and drugs, Dima technology has developed the following analytical methods, which can meet the requirements of estrogen, estriol, ethinylestradiol, estrone, diethylstilbestrol, diethylstilbestrol, Hexestrol and other estrogens in milk powder, as well as progesterone Medroxyprogesterone acetate, chlorprogesterone acetate, methylethinyl progesterone, medroxyprogesterone. At present, some companies that are trying to develop and produce bio based BDO technology include the detection of progesterone such as BASF, Prak, DSM, 3ling, Miriam and genomatica. I hope it can help you with your testing work

1 reverse phase SPE purification method

1.1 extraction

weigh 2.0 g of milk powder (accurate to 0.01 g), add the internal standard solution (only the internal standard solution needs to be added when using the internal standard method), add 16 ml of 80% methanol aqueous solution, Shake Extraction for 2 minutes or ultrasonic extraction for 5 minutes. Centrifuge for 5 minutes at 6000 rpm, take 4 ml of supernatant, add 12 ml of pure water, mix well to obtain the sample diluent, and wait for purification

1.2 SPE column purification

proelut pls hydrophilic lipophilic balance column, 150 mg/6 ml

activation and balance: 5 ml methanol activation, 5 ml water balance

loading: let the sample diluent pass through the PLS column, and discard the effluent

rinsing: 5 ml 5% methanol aqueous solution rinsing, rinsing solution discarded

elution: elute with 5 ml of pure methanol and collect the eluent

sample reconstitution: dry the eluent with nitrogen at 40 ℃, re dissolve the residue with 1 ml of 50% methanol aqueous solution, and filter it with microporous membrane for instrument analysis

2 normal phase SPE purification method

2.1 extraction

weigh 2 g of milk powder (accurate to 0.01 g), add the internal standard solution (only the internal standard solution needs to be added when using the internal standard method), add 20 ml of 80% methanol aqueous solution, shake and extract for 2 minutes or ultrasonic extract for 5 minutes. Centrifuge at 6000 rpm for 5 minutes, take 10 ml of supernatant, mix it with 50 ml of pure water to obtain the sample diluent, and wait for purification

2.2 SPE column purification

proelut carb graphitized carbon black column, 500 mg/6 ml

proelut NH2 amino column, 500 mg/6 ml

pass the sample diluent through the activated proelut carb column *, and drain the small column after all the liquid passes through. Then connect the activated proelut NH2 column * in series under the proelut carb column. Elute with 10 ml of dichloromethane methanol solution (7+3) and collect the eluent, then remove the proelut carb column, elute the proelut NH2 column with 2 ml of dichloromethane methanol solution (7+3), and combine the eluent. Then nitrogen is expected to be removed by air blowing dry cleaning in 2021, and then 1 ml of 50% methanol aqueous solution is used to dissolve the residue, which is filtered by microporous membrane for instrument analysis

*proelut carb column was activated and balanced with 6 ml dichloromethane methanol solution (7+3), 6 ml methanol and 6 ml water in turn; Proelut NH2 column activation equilibrium

3 instrumental analysis

3.1 estrogen analysis conditions

a HPLC conditions:

chromatographic column: leapsil c18100 x 2.1mm, 2.7 m

mobile phase: A, water; B. Acetonitrile

gradient: slightly

flow rate: 0.3 ml/min

column temperature: 40 ℃

injection volume: 10 L

b mass spectrometry conditions

ionization source: ESI, Negative ionization mode

ionization source temperature: 100 ℃

capillary voltage: 3 kV

desolvent gas temperature: 450 ℃

desolvent gas flow: 0.2 l/min

3.2 progesterone analysis conditions

a HPLC conditions

its framework uses ultra light carbon fiber composite

chromatographic column: leapsil c18100 x 2.1mm, 2.7 m

mobile phase: A, 0.1% formic acid.Aqueous solution; B. Methanol

gradient: slightly

flow rate: 0.3 ml/min

column temperature: 40 ℃

injection volume: 10 L

b mass spectrometry conditions

ionization source: ESI, Positive ionization mode can be hard, but it still can't hide the glory it brings.

capillary voltage: 3.5 kV

ionization source temperature: 100 ℃

desolvent gas temperature: 450 ℃

desolvent gas flow: 0.2 l/min

about Dima

Dima technology is a well-known manufacturer of chromatographic consumables committed to developing and manufacturing scientific and efficient chemical analysis products and providing perfect services and comprehensive solutions, It has always maintained the world's advanced level in many technical fields, such as chromatographic filler research and development, chromatographic column manufacturing and related separation products. Core technology products include: liquid chromatography column, gas chromatography column, solid phase extraction column, chromatographic solvent and chemical standard

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