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Method for testing tetracycline antibiotic residue in cattle pulmonary surfactant extract

A surface-active, tetracycline-based technology, used in measuring devices, instruments, scientific instruments, etc., can solve problems such as inability to truly reflect antibiotic residues, microbial growth interference, and narrow application scope.

Active Publication Date: 2018-02-16
BEIJING INST FOR DRUG CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is only used for the determination of tetracycline residues, and cannot be used for other commonly used tetracycline antibiotics, such as oxytetracycline, aureomycin, etc., and its application range is narrow
At the same time, the bovine lung surfactant extract is a nutrient substance, which may interfere with the growth of microorganisms, so that the method of comparing the size of the inhibition zone between the control substance and the test substance cannot truly reflect the amount of antibiotic residues

Method used

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  • Method for testing tetracycline antibiotic residue in cattle pulmonary surfactant extract
  • Method for testing tetracycline antibiotic residue in cattle pulmonary surfactant extract
  • Method for testing tetracycline antibiotic residue in cattle pulmonary surfactant extract

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to a method for testing tetracycline antibiotic residue in cattle pulmonary surfactant extract. The method adopts ultrahigh-performance liquid chromatography-tandem mass spectrometry for testing and includes following steps: providing an ultrahigh-performance liquid chromatograph, a triple quadrupole mass spectrometer, the cattle pulmonary surfactant extract and a reference sample; determining chromatographic conditions and mass spectrometric conditions; performing liquid preparation, testing and result processing. By using the method, 11 antibiotics including tetracycline, chlorotetracycline, terramycin, doxycycline, metacycline, minocycline, beta-doxycycline, epitetracycline, epichlorotetracycline, anhydrotetracycline and epianhydrotetracycline and known impuritiescan be tested at the same time. Linear range, sensitivity, precision and recovery rate of the method all meet analysis requirements.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a method for determining residual antibiotics in bovine lung surface-active extracts, in particular to measuring tetracycline in bovine lung surface-active extracts using ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS) Residues of tetracycline antibiotics such as tetracycline, chlortetracycline, oxytetracycline, doxycycline, metacycline, minocycline, β-doxycycline, epitetracycline, epitaxy There are as many as 11 kinds of mycins, anhydrotetracyclines and anhydrotetracyclines. Background technique [0002] Bovine lung surfactant extract is a lung surfactant isolated from healthy newborn calf lungs. The main components include triglycerides, phospholipids, free fatty acids, cholesterol and a small amount of active protein [SHU L. Study on composition analysis and quality control of Medicinal calf pulponarysurfactant[D].Beijing:Univ.of Chinese Acad...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 李珉侯金凤王成刚齐麟寇晋萍刘海涛王光裕李文东车宝泉王俊秋
Owner BEIJING INST FOR DRUG CONTROL
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