Method for detecting polyoxins B by high performance liquid chromatography method

A high-performance liquid chromatography and polydamicin technology, which is applied in the field of high-performance liquid chromatography detection of polydamicin B, can solve the problems of affecting the accuracy of analysis, inability to achieve separation, and long detection time, and achieve high accuracy , good separation, short running time effect

Active Publication Date: 2014-05-07
陕西麦可罗生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Such as Chinese patent application 200810039564.0 Quantitative analysis and detection method of polyoxin B, reported the Oxford cup bioassay method of polyoxin B, the detection method disadvantages: poor reliability, long detection time
In the existing HPLC detection method, better separation between polyoxin B and some homologues cannot be achieved, which affects the accuracy of analysis

Method used

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  • Method for detecting polyoxins B by high performance liquid chromatography method
  • Method for detecting polyoxins B by high performance liquid chromatography method
  • Method for detecting polyoxins B by high performance liquid chromatography method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Test equipment and conditions:

[0033] Waters 1525 high performance liquid chromatograph, Waters 2487 ultraviolet detector;

[0034] Chromatographic column: C18 embedded with polar groups (250×4.6mm, 5μm);

[0035] Mobile phase: 0.1% trifluoroacetic acid aqueous solution;

[0036] Flow rate: 1.0 mL / min;

[0037] Detection wavelength: 260nm;

[0038] Column temperature: 25°C;

[0039] Injection volume: 10 μL.

[0040] experiment procedure:

[0041] Weigh 0.02g of polyoxin B standard substance, dissolve in a 50mL volumetric flask with pure water, constant volume, shake well, pass through a 0.45μm membrane filter, and prepare for the test standard solution.

[0042] Weigh a sample containing 0.02g polyamycin B, dissolve it in a 50mL volumetric flask with pure water, constant volume, shake well, pass through a 0.45μm filter membrane, and prepare for the test sample solution.

[0043] Get the above-mentioned test standard product and sample solution, inject high-perf...

Embodiment 2

[0045] Test equipment and conditions:

[0046] Waters 1525 high performance liquid chromatograph, Waters 2487 ultraviolet detector;

[0047] Chromatographic column: C18 embedded with polar groups (250×4.6mm, 5μm);

[0048] Mobile phase: 0.05% trifluoroacetic acid aqueous solution;

[0049] Flow rate: 0.8 mL / min;

[0050] Detection wavelength: 260nm;

[0051] Column temperature: 25°C;

[0052] Injection volume: 10μL;

[0053] experiment procedure:

[0054] Weigh 0.02g of polyoxin B standard substance, dissolve in a 50mL volumetric flask with pure water, constant volume, shake well, pass through a 0.45μm membrane filter, and prepare for the test standard solution.

[0055] Weigh a sample containing 0.02g polyamycin B, dissolve it in a 50mL volumetric flask with pure water, constant volume, shake well, pass through a 0.45μm filter membrane, and prepare for the test sample solution.

[0056] Get above-mentioned test sample solution, inject high performance liquid chromatogr...

Embodiment 3

[0058] Test equipment and conditions:

[0059] Waters 1525 high performance liquid chromatograph, Waters 2487 ultraviolet detector;

[0060] Chromatographic column: C18 embedded with polar groups (250×4.6mm, 5μm);

[0061] Mobile phase: methanol: 0.13% trifluoroacetic acid aqueous solution = 3%: 97%;

[0062] Flow rate: 0.8 mL / min;

[0063] Detection wavelength: 260nm;

[0064] Column temperature: 25°C;

[0065] Injection volume: 10μL;

[0066] experiment procedure:

[0067]Weigh 0.02g of polyoxin B standard substance, dissolve in a 50mL volumetric flask with pure water, constant volume, shake well, pass through a 0.45μm membrane filter, and prepare for the test standard solution.

[0068] Weigh a sample containing 0.02g polyamycin B, dissolve it in a 50mL volumetric flask with pure water, constant volume, shake well, pass through a 0.45μm filter membrane, and prepare for the test sample solution.

[0069] Get the above-mentioned test standard product and sample solutio...

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Abstract

The invention relates to a method for detecting polyoxins B by high performance liquid chromatography. Following chromatographic conditions are adopted: a chromatographic column takes alkylsilane bonded silica gel embedded into a polar group as a fixed phase and takes organic phase-water phase as a mobile phase; the flow speed is 0.5-1.5mL/min; detection wavelength is 240-290nm; the column temperature is 10-40 DEG C; sample size is 5-20 microliters; an ultraviolet detector or a diode array detector is adopted for the detector. Compared with a bioassay method and an HPLC (high performance liquid chromatography) method in the prior art, the method according to the invention has the advantages that the separation degree is favorable and the accuracy is high by the cooperation of the chromatographic column and the mobile phase; by the detection method, the polyoxins B and homolog with the similar structure are separated well; the detection accuracy of the polyoxins B is improved; the running time of the detection method is short; the appearance time of the polyoxins B main peak is 8 minutes.

Description

technical field [0001] The invention belongs to the technical field of pesticide detection and analysis, and in particular relates to a high performance liquid chromatography detection method for polyoxin B. Background technique [0002] Polyoxin, also known as polyoxin, is a nucleoside antibiotic isolated from S.cacaoi var.asoensis in Japan. In my country, it is a metabolite of Streptomyces aureus (S.aurea chromogenes 4896). [0003] Polyoxin is a mixture of 14 different homologues from A to N. The main active ingredient is polyoxin B, which belongs to the polyoxypyrimidine nucleoside agricultural antibiotic fungicides and can effectively inhibit the fungal cell wall skeleton Components - the synthesis of chitin, which prevents the biosynthesis of the cell wall of the bacteria and leads to the death of the bacteria. Polyoxin B is a safe pesticide with high efficiency, low toxicity and no environmental pollution, so it is widely used in the control of apple leaf spot, toba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 颜瑞莉杨宏勃李蒲民
Owner 陕西麦可罗生物科技有限公司
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