Method for quantitatively detecting lipopeptide

Abstract

The invention relates to a method for quantitatively detecting lipopeptide, comprises the following steps: (1) separating a to-be-detected sample to obtain lipopeptide; (2) reacting the lipopeptide with a fluorescence indicator to obtain fluorescently-labeled lipopeptide; (3) detecting the fluorescently-labeled lipopeptide by using high performance liquid chromatography; and (4) computing the lipopeptide content through chromatographic peak area. Compared with the prior art, the method has the advantages that reaction condition is mild, steps are simple, the detection is flexible, and trace lipopeptide can be detected.

Description

technical field [0001] The invention relates to a method for quantitatively detecting lipopeptides, in particular to a method for detecting lipopeptides using fluorescent markers. Background technique [0002] Lipopeptide is a biosurfactant with a wide variety of sources and a special structure. Lipopeptides have excellent surface and interface properties unmatched by chemical surfactants, and are environmentally friendly; lipopeptides have a wide range of biological activities, such as antibacterial, antitumor, anticoagulant, cell lysing, and changing enzyme activities. Therefore, microbial lipopeptides have broad application prospects in the field of surfactants and biomedicine. [0003] Due to the high surface activity of lipopeptides and the generally low dosage, the detection of lipopeptides in the application system has always been a difficult problem. The traditional gravimetric method is to add hydrochloric acid to the test solution to precipitate the lipopeptide. ...

AI Technical Summary

Problems solved by technology

The traditional gravimetric method is to add hydrochloric acid to the test solution to precipitate the lipopeptide. The precipitate is repeatedly extracted with an organic solvent, evaporated, dried, and weighed to quantify the lipopeptide. The main disadvantage of this detection method is that the system needs to add Hydrochloric acid precipitates the lipopeptide, and the organic solvent is repeatedly extracted. The steps are cumbersome, and the extraction process is easily interfered by other fat-soluble substances, so the pure lipopeptide cannot be obtained, and can only be roughly quantified; TLC is to extract the lipopeptide. The sample is separated by thin-layer chromatography, and the separated lipopeptide spots are hydrolyzed. After the spots are hydrolyzed, specific staining is carried out. The lipopeptides are quantified by comparing the size and color depth of the stained spots. The main disadvantages of this detection method are: separation by thin-layer chromatography The lipopeptide spots need to be hydrolyzed with concentrated hydrochloric acid at high temperature, the reaction process is complicated and the conditions are harsh. At the same time, the separation efficiency of thin-layer chromatography is low and the quantitative error is large; Separation, detection by ultraviolet detector, determination of lipopeptide peak by retention time, quantitative analysis of lipopeptide by analysis of peak area or peak height, the main disadvantage of this detection method is: complicated pretreatment is required for complex samples on site, so as not to affect The separation effect of the chromatographic column, and the detection limit is also high (suitable for the detection of samples with a lipopeptide content of about several hundred ppm), and it is impossible to detect trace lipopeptides

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