Method for rapidly detecting succinic acid content in fermentation broth

Abstract

The present invention provides a method for rapidly detecting a succinic acid content in fermentation broth, and belongs to the technical field of biochemical engineering. According to the method, the fermentation broth is subjected to a pretreatment, and then is directly added to a supporter (silica gel plates, cellulose acetate membrane or chromatography filter paper and the like can be adopted); after drying and immobilization, a copper sulfate-methanol solution is adopted to carry out a color reaction; after completely drying, a thin layer chromatogram scanner is adopted to analyze an absorbance value of a color spot; the succinic acid content in the fermentation broth is calculated according to a linear relation between the succinic acid content and the absorbance value. The method provided by the present invention has characteristics of is simpleness, easy operation, rapid analysis speed, high accuracy, good reproducibility and low detection cost, is applicable for meeting the requirements of the experimental study of the succinic acid fermenting, and the requirements of the analysis and the detection in the practical production, and is a good analysis and detection method for the succinic acid.

Description

technical field [0001] The invention relates to a detection method in the technical field of succinic acid fermentation, in particular to a method for rapidly detecting the content of succinic acid in a fermentation broth based on the color reaction between succinic acid and copper sulfate. Background technique [0002] Succinic acid (Succinic acid) is an important organic acid product, which is mainly used in the following four fields: one is as a surfactant, detergent additive and foaming agent; the other is as an ion chelating agent for the electroplating industry to prevent metal corrosion and pitting; the third is used in the food industry as acidifiers, pH modifiers, flavor substances and antibacterial agents; the fourth is the production of pharmaceuticals, antibiotics, amino acids and vitamins. The sales volume of succinic acid in these four fields exceeds 400 million US dollars per year. In recent years, the National Engineering Research Center for Engineering Plas...

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Problems solved by technology

Among them, liquid chromatography (National Standard of the People's Republic of China GB / T 5009.157-2003; Cao Jianlei et al. Construction and fermentation performance of succinic acid-producing recombinant Escherichia coli. Journal of Applied and Environmental Biology, 2010, 16(6): 851-857. ) has the advantages of low detection limit, good reproducibility and high precision, and is a common method for the accurate quantitative analysis of succinic acid in fermented liquid at present, but HPLC needs to use an expensive liquid chromatograph, which has high cost and long detection time. Moreover, the various requirements for the system are also very high; method (Zhao Dayun et al. Gas chromatography analysis of organic acid components in potherb mustard pickle marinade. Journal of Shanghai Jiaotong University (Agricultural Science Edition), 2003, 21(3): 220-225, 245.) both have expensive equipment and complicated operations. cumbersome, time-consuming detection and analysis, and gas chromatography also needs to derivatize succinic acid, therefore, these two methods are rarely used in the detection and analysis of succinic acid in fermentation broth; some reports use thin-layer chromatography Method (He Lingyun et al. Determination of succinic acid content in pinellia medicinal materials in different production areas by TLC scanning method. Chinese patent medicine, 2002, 24 (4): 290-292.; Wang Guangzhong et al. Determination of earthworms by TLC scanning method The content of succinic acid in medicinal materials. Journal of Pharmaceutical Analysis, 1997, 17 (4): 274-275.) to analyze the content of succinic acid in traditional Chinese medicine ingredients, but the detection and analysis process requires a long time for chromatographic development, due to the use of acid-base indicators Chromogenic reagent staining, quantitative analysis is easily interfered by other organic acids in the fermentation broth

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