摘 要：鼠李糖脂具有亲水基团，由1～2个分子的鼠李糖构成，是一种含有鼠李糖结构的脂类物质。作为一种生物表面活性剂，鼠李糖脂具有表面活性高、毒性低、可自然降解等特征，在石油工业、食品、医药、化妆品、绿色农业和生态环境等领域具有极大的潜在应用价值。本研究采用苯酚-硫酸法、蒽酮-硫酸法、3, 5-二硝基水杨酸法，以酶标仪替代传统的分光光度计，对鼠李糖脂的质量浓度进行测定，进而开发出微量快速检测鼠李糖脂质量浓度的方法，旨在实现快速、准确、灵敏的鼠李糖脂质量浓度测定。本试验确定了苯酚-硫酸法为鼠李糖脂质量浓度的最佳检测方法，成功建立了苯酚-硫酸法微量检测体系，并优化了检测条件。利用酶标仪检测发现，在479 nm处的吸光度值及标准曲线斜率均较高，可快速检测鼠李糖脂的质量浓度，节约时间，节省危化品，减少废弃物的产生，提高检测效率。苯酚-硫酸法微量检测体系的成功应用可实现大量样本的快速筛选，操作简单，减少了时间及人力成本，减少了对昂贵设备的依赖，可快速检测转化液中鼠李糖脂的质量浓度，为筛选高转化菌株奠定了基础。

关键词：鼠李糖脂；苯酚-硫酸法；蒽酮-硫酸法；3, 5-二硝基水杨酸法；高通量筛选
中图分类号：O652.3             文献标志码：A              DOI：10.3969/j.issn.1007-7146.2025.05.010

Abstract: Rhamnolipids have hydrophilic groups and are composed of 1～2 molecules of rhamnose. They are a type of lipid substance containing rhamnose structure. As a biological surfactant, it has the characteristics of high surface activity, low toxicity, and natural degradation, and has great potential application value in the fields of petroleum industry, food, medicine, cosmetics, green agriculture, and ecological environment. This study used phenol-sulfuric acid method, anthrone-sulfuric acid method, and 3, 5-dinitrosalicylic acid method, replacing traditional spectrophotometers with enzyme-linked immunosorbent assay, to determine the mass concentration of rhamnolipids. Subsequently, a micro rapid method for detecting the mass concentration of rhamnolipids was developed, aiming to achieve rapid, accurate, and sensitive determination of rhamnolipids. The phenol-sulfuric acid method was chosen as the optimal detection method for the mass concentration of rhamnolipids, we successfully established a phenol-sulfuric acid micro detection system, and optimized the detection conditions. Using an enzyme-linked immunosorbent assay reader, it was found that the absorbance value at 479 nm and the slope of the standard curve were both high. This allows for rapid detection of the mass concentration of rhamnolipids, saving time, hazardous chemicals, reducing the generation of waste gases, and improving detection efficiency. With the successful application of the phenol-sulfuric acid micro detection system, we can achieve rapid screening of a large number of samples, with simple operation, reduced time and labor costs, and reduced dependence on expensive equipment. The system can quickly detect the mass concentration of rhamnolipids in the transformation solution, laying the foundation for screening high transformation strains.
Key words: rhamnolipids; phenol-sulfuric acid method; anthrone-sulfuric method; 3, 5-dinitrosalicylic acid method; high-throughput
（Acta Laser Biology Sinica, 2025, 34(5): 466-472）