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HPLC method for quick teicoplanin fermentation liquor unit detection

A technology of teicoplanin and detection method, applied in the detection field, can solve the problems of affecting the accuracy and sensitivity of analysis, increasing the pressure of the chromatographic column, damaging the chromatographic column, etc., so as to achieve the detection method that is simple, fast, strong in specificity, good accuracy

Inactive Publication Date: 2014-05-07
鲁南新时代生物技术有限公司
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Problems solved by technology

The disadvantages of this method are: the analysis time of each sample is 60 minutes, which is not convenient for rapid detection; the flow rate is high, and long-term use will cause the column pressure to increase, increase pump loss, damage the chromatographic column, and affect the accuracy and sensitivity of the analysis.

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  • HPLC method for quick teicoplanin fermentation liquor unit detection
  • HPLC method for quick teicoplanin fermentation liquor unit detection

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Embodiment Construction

[0020] The present invention will be further described below in conjunction with specific experiments, but the present invention is not limited.

[0021] 1. Sample processing method

[0022] Take a certain amount of fermentation broth, adjust the pH to 10 with 10% NaOH solution, put it in a centrifuge tube after ultrasonication for 30min, centrifuge at 10000rpm for 10min, take the supernatant and put it in another 10mL centrifuge tube, and put it in a centrifuge tube at 10000rpm After centrifugation for 10 min, the supernatant was taken, passed through a 0.22 μm filter membrane, and tested on the machine.

[0023] 2. Chromatographic conditions

[0024] (1) Chromatographic column: reversed-phase C18 column with octadecylsilane bonded silica gel as filler;

[0025] (2) detector: ultraviolet detector;

[0026] (3) Detection wavelength: λ=277nm;

[0027] (4) Mobile phase: A: 0.01mol / L ammonium acetate solution, B: 90% acetonitrile aqueous solution, used after filtration and ul...

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Abstract

The invention discloses an HPLC method for quick teicoplanin fermentation liquor unit detection. The method comprises the following steps: processing teicoplanin fermentation liquor with alkali, performing ultrasonography, performing centrifugation for two times, enabling clear liquor to pass a filtration film at one step, and then placing the filtrated liquor on a machine to be detected and analyzed, wherein a high-performance liquid chromatograph (HPLP) is mainly equipped with a reversed-phase C18 column; a detector is an ultraviolet detector; a mobile phase is an ammonium acetate solution and an acetonitrile buffer solution for gradient elution, and data is collected and analyzed at a condition of 277 nm. The detection method provided by the invention is simple, convenient and fast, and is high in sensitivity and good in accuracy.

Description

technical field [0001] The invention belongs to the technical field of detection methods, and in particular relates to a rapid HPLC detection method of a teicoplanin fermentation broth unit. Background technique [0002] Teicoplanin belongs to glycopeptide antibiotics and has strong antibacterial activity against Gram-positive bacteria. It is often used clinically for serious Gram-positive bacterial infections, and its effect on Staphylococcus aureus is often better than the glycopeptide antibiotic Vancomycin Therefore, the production and research and development of teicoplanin have been paid more and more attention. The earliest teicoplanin was fermented by Parenti et al. from a strain of swimming actinomycete A.teichomyceticus nov.sp., which is now commonly used ATCC31121; domestic research mainly includes rare actinomycetes SIIA92-363, A .teichomyceticus 98-1-227 and its SIIA 05-03-136, teicoplanin can be isolated from the fermentation liquid of these three strains. The...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 赵志全王军李娜
Owner 鲁南新时代生物技术有限公司
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