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Dual-wavelength ultraviolet spectrophotometry method for measuring content of natamycin in fermentation liquid

A technique of spectrophotometry and natamycin, applied in the field of analytical chemistry, can solve problems such as difficult to eliminate, interference of measurement results, complex HPLC system, etc., and achieve the effect of eliminating interference, detection limit and quantitative limit evaluation

Inactive Publication Date: 2014-04-02
WUHAN SUNHY BIOLOGICAL
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Problems solved by technology

The HPLC method can separate some related substances, but due to the complexity of the HPLC system, high operating costs and personnel requirements, especially the strict requirements for sample pretreatment, it is not suitable for the detection of a large number of biological fermentation samples
[0004] The determination of the content of natamycin finished product in the United States Pharmacopoeia adopts high-performance liquid chromatography, and there are also reports of using this method to determine natamycin in China, but this method is not suitable for large-scale breeding and fermentation of natamycin high-yielding bacteria. The online and fast offline detection of the process has obvious disadvantages, such as a long measurement period, high requirements for sample processing and instruments, and is not suitable for large batches of samples, etc.
[0005] There are also domestic reports on the rapid determination of natamycin in fermented liquid by ultraviolet spectrophotometry, but since natamycin fermented liquid is obtained by biological fermentation, many organic substances are used in the fermentation medium, and these substances contain other Substances with ultraviolet absorption will cause certain interference to the measurement results when measured by ultraviolet spectrophotometry. It is difficult to eliminate the interference of various impurities in the fermentation broth on the measurement results by single-wavelength ultraviolet spectrophotometry

Method used

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  • Dual-wavelength ultraviolet spectrophotometry method for measuring content of natamycin in fermentation liquid
  • Dual-wavelength ultraviolet spectrophotometry method for measuring content of natamycin in fermentation liquid
  • Dual-wavelength ultraviolet spectrophotometry method for measuring content of natamycin in fermentation liquid

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Embodiment 1

[0015] 1. Materials and equipment

[0016] Electronic balance (1 / 10,000), UV spectrophotometer, ultrasonic cleaner, natamycin reference substance (United States Pharmacopoeia Commission), natamycin fermentation broth sample.

[0017] 2. Fermentation conditions

[0018] Incline medium (%): peptone 0.05, malt extract 0.3, glucose 1.0, agar 2.0, pH7.0;

[0019] Seed medium (%): peptone 0.6, corn steep liquor 0.6, glucose 2.0, sodium chloride 1.0, pH7.0;

[0020] Fermentation medium (%): corn starch 2.0, yeast extract powder 0.5, peptone 0.5, glucose 4.0, pH 7.0.

[0021] Dig out an appropriate amount of the slant culture cultivated at 28°C and inoculate it into a seed bottle containing 25ml of seed medium, cultivate it at 28°C and 200rpm for 24-36 hours, and inoculate it in a 30L automatic fermenter according to the inoculation amount of 10%. °C, cultured at 20 L / min for 144 hours, and samples were taken every 8 hours from 72 hours to obtain natamycin fermentation broth sample...

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Abstract

The invention relates to the field of analytical chemistry, in particular to a dual-wavelength ultraviolet spectrophotometry method for measuring the content of natamycin in fermentation liquid. The method comprises the following steps: (1) measuring light absorption values of a processed sample to be measured at the wavelengths of 303 nm and 290 nm respectively by taking a 70% methanol-water solution as a blank; (2) calculating the content of the natamycin according to the following formula: C sample=(k*delta A+b)*N. The precision degree, the accuracy inspection, the detection limit and the quantitation limit of the method are effectively evaluated, so that the method is guaranteed to be scientific and accurate.

Description

technical field [0001] The invention relates to the field of analytical chemistry, in particular to a dual-wavelength ultraviolet spectrophotometric method for determining the content of natamycin in fermentation broth. Background technique [0002] Natamycin is a twenty-six-membered ring polyene macrolide antifungal agent, which specifically inhibits yeast and mold, and is widely used as a natural biological food preservative and antibacterial additive. Natamycin, also known as Pimaricin, is mainly composed of Streptomyces natalensis, Streptomyces chattanovgensis, and Phaeus Streptomyces gilvosporeus and other producing bacteria are refined through fermentation and refining. [0003] During the biological fermentation process, a large amount of organic substances, such as protein and some metabolites, will be produced in the fermentation broth, which will interfere with the determination of the sample. The HPLC method can separate some related substances, but due to the c...

Claims

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Application Information

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IPC IPC(8): G01N21/33
Inventor 詹志春王冠徐丽
Owner WUHAN SUNHY BIOLOGICAL
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