A method for rapid detection of vancomycin by liquid chromatography-mass spectrometry

A vancomycin and LC/MS technology, applied in the field of chromatographic detection, can solve the problems of short analysis time, large matrix effect, low sensitivity, etc., and achieve the effects of good sensitivity, promotion of dissolution, and good purification effect.

Active Publication Date: 2020-11-24
周迎春
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to overcome the disadvantages and deficiencies such as large matrix effect and low sensitivity of the vancomycin detection method in the prior art, the object of the present invention is to provide a method for rapidly detecting vancomycin by liquid mass spectrometry, which is suitable for detecting fat content High vancomycin content in food of animal origin, with the advantages of simple pretreatment, short analysis time, low cost, high accuracy, low detection limit, good stability, etc.

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  • A method for rapid detection of vancomycin by liquid chromatography-mass spectrometry
  • A method for rapid detection of vancomycin by liquid chromatography-mass spectrometry
  • A method for rapid detection of vancomycin by liquid chromatography-mass spectrometry

Examples

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Embodiment 1

[0055] In this example, the vancomycin standard was used to explore and determine the best chromatographic conditions and mass spectrometry conditions; the determined optimal chromatographic conditions and mass spectrometry conditions were used to measure the backfat of imported frozen pigs to determine whether it contained vancomycin Add vancomycin standard substance in negative imported frozen pig backfat, determine the recovery rate, precision and minimum detection limit of the method, the specific experimental method is as follows:

[0056] (1) Preparation of standard solution: Accurately weigh 10.63±0.01mg of 94.1% standard vancomycin, dissolve it in pure water and dilute to 100mL, make a standard stock solution with a concentration of 100μg / mL, and store at 4°C .

[0057] (2) Prepare vancomycin standard solutions with six concentrations of 2.0ng / mL, 5.0ng / mL, 10.0ng / mL, 20.0ng / mL, 50.0ng / mL, and 100.0ng / mL, using 6470 ultra-high performance liquid phase Chromatography-t...

Embodiment 2

[0082] This embodiment uses the method determined in Example 1 to measure the imported frozen pork boneless foreleg to determine whether it contains vancomycin; add vancomycin standard substance to the negative imported frozen pork boneless foreleg, and the determination method recovery, precision, and detection limit. The specific experimental method is as follows:

[0083] (1) Preparation of standard solution: Accurately weigh 10.63±0.01mg of 94.1% standard vancomycin, dissolve it in pure water and dilute to 100mL, make a standard stock solution with a concentration of 100μg / mL, and store at 4°C ;

[0084] (2) Sample preparation: Randomly select 500 g of edible part of imported frozen pig boneless foreleg meat, divide it into small pieces, grind it thoroughly with a meat grinder, and set aside;

[0085] (3) Preparation of spiked samples: Accurately weigh 2.00±0.01g of crushed imported frozen pork boneless front leg meat into a polypropylene centrifuge tube, and add the con...

Embodiment 3

[0100]In this embodiment, the method determined in Example 1 is used to measure the imported frozen pig boneless front leg skin to determine whether it contains vancomycin; add vancomycin standard substance to the negative imported frozen pig boneless front leg skin, and the determination method recovery, precision, and detection limit. The specific experimental method is as follows:

[0101] (1) Preparation of standard solution: Accurately weigh 10.63±0.01mg of 94.1% standard vancomycin, dissolve it in pure water and dilute to 100mL, make a standard stock solution with a concentration of 100μg / mL, and store at 4°C ;

[0102] (2) Sample preparation: 500 g of the edible part of the boneless front leg skin of imported frozen pigs was randomly selected, chopped with a kitchen knife (due to the high toughness of the meat skin, it cannot be completely crushed with the current grinder such as a meat grinder), and set aside;

[0103] (3) Preparation of spiked samples: Accurately we...

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Abstract

The invention belongs to the technical field of chromatographic detection, and specifically relates to a method for quickly detecting vancomycin by utilizing LC-MS (Liquid Chromatography-Mass Spectrometry). The method comprises the following steps: smashing a to-be-detected sample, adding an extracting reagent, and uniformly mixing; then carrying out ultrasonic extraction, and centrifuging, thus obtaining supernatant; extracting the supernatant by adopting water saturated normal hexane, thus obtaining a to-be-purified solution; activating through an MCX solid-phase extraction column, taking and sampling the to-be-purified solution, carrying out drip washing and impurity removing by using methanol, then eluting by using amonium methanol, and collecting an eluent; concentrating, forming a constant volume by using a constant volume solution, and filtering, thus obtaining a purified solution; detecting the purified solution by adopting an HPLC-MS (High Performance Liquid Chromatograph-MassSpectrometer). The method disclosed by the invention has the advantages that the problems of large matrix effects, low sensitivity and the like existing in a process of detecting animal derived foodand particularly the animal derived food with a high fat content in the prior art are well solved, ideal recovery and precision degree are obtained, the minimum detection limit of the method is 0.3 mug / kg, and repeatability and reproducibility of an analysis result are effectively ensured.

Description

technical field [0001] The invention belongs to the technical field of chromatographic detection, and in particular relates to a method for rapidly detecting vancomycin by liquid chromatography-mass spectrometry. Background technique [0002] Vancomycin (Molecular formula: C 66 h 75 Cl 2 N 9 o 24 ) is a tricyclic glycopeptide antibiotic against Gram-positive bacteria. Neither effective serious bacterial infection. But vancomycin has certain side effects, it can cause ear, kidney toxicity, skin allergic reaction, nervous system toxicity and organ dysfunction. If vancomycin is illegally used in animal breeding, it is likely to affect the safety of animal food, public health, and import and export trade due to drug resistance. If people eat food with vancomycin residues for a long time, it will not only endanger human health , and increases the risk of bacteria developing resistance, leading to more severe antibiotic resistance among consumers. [0003] At present, the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 周迎春刘少博韩海涛彭新然
Owner 周迎春
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