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Kit for quantitatively detecting various beta-lactamase inhibitor residues and using method thereof

A technology for quantitative detection of lactamase, which is applied in the direction of material analysis by observing the impact on chemical indicators, and analysis by making materials undergo chemical reactions, can solve the problem of high-throughput, rapid detection, and inability to achieve 4 Simultaneous detection, high detection cost and other problems, to achieve the effect of wide screening range, low price and high detection sensitivity

Pending Publication Date: 2021-04-16
GUANGDONG UNIV OF EDUCATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the detection of β-lactamase inhibitors mainly includes high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS / MS) and other instrumental analysis methods, which have high sensitivity and accuracy. However, high-end instruments and equipment and professional technicians are required. At the same time, the requirements for sample pretreatment are high, the detection cost is high and the time is long, and it is difficult to meet the needs of high-throughput and rapid detection; and only one or two of them can be detected. , can not achieve 4 kinds of simultaneous detection

Method used

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  • Kit for quantitatively detecting various beta-lactamase inhibitor residues and using method thereof
  • Kit for quantitatively detecting various beta-lactamase inhibitor residues and using method thereof
  • Kit for quantitatively detecting various beta-lactamase inhibitor residues and using method thereof

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Effect test

Embodiment 1

[0034] Embodiment 1 Preparation method 1 of the kit of the present invention

[0035] (1) For the configuration of the β-lactamase solution, use extended-spectrum β-lactamase and dilute it with a buffer to obtain a β-lactamase solution containing 50,000 units per milliliter.

[0036] (2) The configuration of the β-lactamase inhibitor standard solution, accurately weigh the sulbactam standard, dissolve it in buffer and adjust the volume to 10.00 mg / mL.

[0037] (3) The preparation of the substrate chromogenic stock solution, weighed nitrocefamin, dissolved in high quality dimethyl sulfoxide (DMSO) and diluted to 10 mg / mL.

[0038] (4) For the configuration of the buffer, a 0.5mol / L phosphate buffer solution with a pH of 7 is used.

Embodiment 2

[0039] Embodiment 2 The preparation method 2 of the kit of the present invention

[0040] (1) For the configuration of β-lactamase solution, use extended-spectrum β-lactamase, dilute it with a buffer containing 5% bovine serum albumin, 15% trehalose, 2% polyvinylpyrrolidone, and 5% proclin300 It is a β-lactamase solution containing 50,000 units per milliliter.

[0041] (2) The configuration of the β-lactamase inhibitor standard solution, accurately weigh the standard potassium clavulanate, dissolve it in a buffer containing 2% polyvinylpyrrolidone, and adjust the volume to 10.00 mg / mL.

[0042] (3) Preparation of substrate chromogenic stock solution: weigh nitrocefatin and dissolve in high-quality dimethyl sulfoxide (DMSO) containing 10% L-cysteine, 10% polyvinylpyrrolidone, and 10% glycerin Dilute to 10mg / mL.

Embodiment 3

[0043] Embodiment 3 Preparation method 3 of the kit of the present invention

[0044] (1) For the configuration of β-lactamase solution, use extended-spectrum β-lactamase, dilute it with a buffer containing 2% bovine serum albumin, 8% trehalose, 1% polyvinylpyrrolidone, and 2% proclin300 It is a β-lactamase solution containing 50,000 units per milliliter.

[0045] (2) The configuration of the β-lactamase inhibitor standard solution, accurately weigh the tazobactam standard, dissolve it in a buffer containing 1% polyvinylpyrrolidone, and adjust the volume to 10.00 mg / mL.

[0046] (3) Preparation of substrate chromogenic stock solution, weigh nitrocefatin and dissolve in high-quality dimethyl sulfoxide (DMSO) containing 5% L-cysteine, 5% polyvinylpyrrolidone, and 5% glycerin Dilute to 10mg / mL.

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Abstract

The invention discloses a kit for quantitatively detecting various beta-lactamase inhibitor residues and a using method thereof, which belong to the technical field of food safety rapid detection. The kit comprises a beta-lactamase solution, a beta-lactamase inhibitor standard substance solution, a developing stock solution, a buffer solution and a 96-hole black lightproof elisa plate, and can be used for quantitatively detecting multiple residues of sulbactam, clavulanic acid, tazobactam and avibactam in liquid milk at the same time. The detection limits of sulbactam, clavulanic acid, tazobactam and avibactam in the milk sample are respectively 4.7 mug / kg, 95.8 mug / kg, 2.0 mug / kg and 4.6 mug / kg; the linear ranges are respectively 34.2-622.8 mug / kg, 275.7-10233.3 mug / kg, 4.9-108.8 mug / kg and 9.6-118.7 mug / kg. The kit is wide in screening range, can effectively avoid the problem of missed detection caused by the fact that lawbreakers select and avoid sulbactam which is commonly detected and use beta-lactamase inhibitors of other varieties, and is particularly suitable for high-throughput screening.

Description

technical field [0001] The invention belongs to the technical field of rapid detection of food safety, and more specifically relates to a method capable of simultaneously detecting sulbactam, clavulanic acid, tazuobatan and aviol in liquid milk A method and application for the quantitative multi-residue detection of four β-lactamase inhibitors in Avibactam. Background technique [0002] Sulbactam, clavulanic acid, tazobactam, and avibactam are four common β-lactamase inhibitors that can competitively and irreversibly inhibit β-lactamase activity. At present, there are two main reasons for the residue of β-lactamase inhibitors in food: one is endogenous factors. In the prevention and treatment of animal diseases, in order to improve antibacterial activity, β-lactamase inhibitors are often used together with The combined use of β-lactam antibiotics leads to residual β-lactamase inhibitors in animal tissues and raw milk; the second is exogenous factors, β-lactamase can decompo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 刘凤银吴晓丹李滢穆洪涛袁学文梁岳
Owner GUANGDONG UNIV OF EDUCATION
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