Method for Determination of Tetracycline Antibiotic Residues in Bovine Lung Surfactant Extract
A technology of surface activity and tetracyclines, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as narrow application range, inability to truly reflect antibiotic residues, microbial growth interference, etc.
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Embodiment 1
[0049] Example 1: Determination of Tetracycline Antibiotic Residues in Bovine Lung Surfactant Extract
[0050] 1. Instruments and materials
[0051] 1.1. Instrument
[0052] Thermo Ultimate 3000 ultra-high performance liquid chromatography (Thermo Fisher, USA), Thermo TSQ Quantiva triple quadrupole mass spectrometer (Thermo Fisher, U.S.), XA205 1 / 100,000 balance (Mette Le Toledo Company, Switzerland), 20-position vacuum solid-phase extraction device (Waters Company, the United States), Milli-Q pure water machine (Merck Millipore, Germany), ZY-300 Ⅳ inhibition zone measuring instrument ( Pioneer Weifeng Co., Ltd., China), DHP-0162 electric heating constant temperature incubator (Shanghai Yiheng Scientific Instrument Co., Ltd., China).
[0053] 1.2. Materials
[0054] 1.2.1. Standards and reference substances
[0055] Tetracycline Standard (Lot No. 130306-201419), Oxytetracycline Standard (Lot No. 130305-201321), Tetracycline Hydrochloride Reference Substance (Lot No. 130488...
Embodiment 2
[0125] Example 2: Determination of Tetracycline Antibiotic Residues in Bovine Lung Surfactant Extract
[0126]Carry out with reference to Example 1, the only difference is that in "2.1, sample pretreatment method" change and process as follows: Accurately weigh 50 mg of bovine lung surfactant extract, put it in a 50 ml centrifuge tube, add McIlvaine buffer 8 ml, Vortex to mix, sonicate for 5 minutes, add 20ml of n-hexane, vortex for 5 minutes, centrifuge at 5000rpm for 5 minutes, discard the lower layer, and the supernatant is used for loading; 1. After 6ml of water activation, take the sample and carry out solid-phase extraction (combined with the use of vacuum solid-phase extraction device), successively wash with 4ml of 6% methanol, then elute with 2ml of methanol, and collect the methanol eluate; Blow dry with N2, redissolve the residue with 500 μl methanol, and take 1 μl for chromatographic injection. Record the chromatogram obtained from the test, and combine the method...
Embodiment 3
[0127] Example 3: Determination of Tetracycline Antibiotic Residues in Bovine Lung Surfactant Extract
[0128] Carry out with reference to Example 1, the only difference is that "2.1, sample pretreatment method" is changed and processed as follows: accurately weigh 50 mg of bovine lung surfactant extract, put it in a 50 ml centrifuge tube, add McIlvaine buffer 4 ml, Vortex to mix, sonicate for 15 minutes, add 15ml of n-hexane, vortex for 15 minutes, centrifuge at 4000rpm for 15 minutes, discard the lower layer, and use the supernatant for sample loading; use 150mg of Oasis HLB solid-phase extraction cartridges, wash through 6ml of methanol , 4ml of water after activation, take the sample and carry out solid-phase extraction (combined with the use of a vacuum solid-phase extraction device), successively wash with 6ml of 4% methanol, then elute with 4ml of methanol, and collect the methanol eluate; Blow dry with N2, redissolve the residue with 500 μl methanol, and take 1 μl for ...
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