Experimental determination of hormone residues in foods of animal origin
Experimental determination of hormone residues in foods of animal origin
Hormone residues in foods of animal origin may jeopardize the health of consumers, with sex and growth-promoting hormones, which are currently the most used, posing the greatest risk to human health. Source: Food Safety Monitoring Technology (Chemical Industry Press)
Operation method
HPLC-MS/MS method
Materials and Instruments
Animal tissue Move 1. Pre-treatment Weigh 10 g of sample, accurately add 0.5 mL of internal standard solution (13C-hexenestrol, 13C-testosterone, 13C-estradiol) (20 ng/mL), add 10 mL of sodium acetate-acetic acid buffer solution at pH 5.2, homogenize, and then add 10 uL of β-glucosidase-sulfate esterase solution, and then perform enzyme digestion by oscillation at (37±1)°C overnight. After removal and cooling, 36 mL of methanol was added and extracted by shaking for 30 min, then transferred to a centrifuge tube and centrifuged at 2000 g for 10 min. 40 mL of hexane was added to the supernatant and extracted twice, and the hexane layer was discarded. In the water-methanol layer, 0.5 mL of n-propanol was added, and the methanol was removed by rotary evaporation at 50 ℃. The HLB solid phase extraction column (6 mL, 500 mg) was activated with 6 mL methanol, 6 mL water. 50 mL of water was added to the solution after rotary evaporation and passed through the column at 2-3 mL/min. After washing Pan with 6 mL of water, the extraction column was blown dry with nitrogen. The column was eluted with 10 mL of 5 mmol/L triethylamine methanol solution, the eluate was blown dry with nitrogen, 0.3 mL of trichloromethane was added to dissolve the residue, and 3 mL of n-hexane was added for the next purification step. The cinnamon gum solid phase extraction column (6 mL, 500 mg) was activated with 6 mL of hexane, the solution was passed through the column and washed with 5 mL of hexane. The column was eluted with 6 mL of water-saturated ethyl acetate, and the eluate was nitrogen-dried and dissolved in 2 mL of methanol-ethyl acetate (40 + 60) solution for passing through the amino column. Amino column (6 mL, 500 mg) was washed with 3 mL methanol-ethyl acetate (40 + 60) solution, 3 mL water-saturated ethyl acetate activation, the previous step solution over the column and receive, then add 3 mL methanol - ethyl acetate (40 + 60) solution elution and receive, nitrogen gas blowing dry, add 0.5 mL of mobile phase dissolved on the machine. 2. Determination (1) androgen determination of liquid chromatographic conditions column: phenyl column (2.0 × 250 mm, 5 um); mobile phase: methanol - water, gradient elution, the initial conditions of 65% water - 35% methanol, maintained for 3 min, 27 min linear gradient so that the proportion of methanol 100%, maintained for 5 min, 1 min the proportion of methanol down to 35%, maintained for 20 min to the next injection. Flow rate: 0.2 mL/min. (2) Mass spectrometry reference conditions for androgen determination Ionization source: electrospray positive ion mode; capillary voltage: 3.5 KV; conical hole voltage: 70 V; RF lens 1 voltage: 30 V; RF lens 2 voltage: 0.5 V; source temperature: 100 ℃; desolvent gas temperature: 300 ℃; desolvent gas flow rate: 400 L/h; electron multiplication voltage: 650 V; injection chamber pressure: 2.8 × 10-3 mbar; androgen determination mass spectrometry reference conditions. 10-3 mbar; (3) Estrogen determination of liquid chromatographic conditions column: phenyl column (2.0 mm × 250 mm, 5 um); mobile phase: acetonitrile - water, gradient elution, the initial conditions of 65% water - 35% acetonitrile, keep 3 min, 27 min linear gradient so that the proportion of acetonitrile 100%, keep 5 min, 1 min the proportion of acetonitrile down to 35%, keep 20 min to the next injection. Flow rate: 0.2 mL/min (4) Mass spectrometry conditions for estrogen determination Ionization source: electrospray negative ion mode; capillary voltage: 3.1 KV; dimensional pore voltage: 80 V; RF lens 1 voltage: 40 V; RF lens 2 voltage: 0.5 V; source temperature: 100 ℃; desolvation gas temperature: 300 ℃; desolvation gas flow rate: 400 L/h; electron multiplication voltage: 650 V; injection chamber pressure: 2.8 × 10 -3 mbar. For more product details, please visit Aladdin Scientific website.
Sodium acetate-acetic acid buffer solution β-glucosidase-sulfatase solution Methanol Hexane Water saturated ethyl acetate
Centrifuge tube HLB solid phase extraction column Liquid chromatography conditioned columns