Rhamnolipids have extensive potential applications and are the most promising biosurfactants for commercialization. The efficient and accurate identification and analysis of these are important to their production, application and commercialization. Accordingly, significant efforts have been made to identify and analyse rhamnolipids during screening of producing strains, fermentation and application processes. Cationic cetyltrimethylammonium bromide-methylene blue (CTAB-MB) test combines a series of indirect assays to efficiently assist in the primary screening of rhamnolipids-producing strains, while the secretion of rhamnolipids by these strains can be identified through TLC, FTIR, NMR, electrospray ionization mass spectrometry (ESI-MS) and HPLC-MS analysis. Rhamnolipids can be quantified by colorimetric methods requiring the use of concentrated acid, and this approach has the advantages of reliability, simplicity, low-cost and excellent reproducibility with very low technological requirements. HPLC-MS can also be employed as required as a more accurate quantification method. In addition, HPLC-ELSD has been established as the internationally acceptable measure of rhamnolipids for commercial purposes. The preparation of well-accepted rhamnolipids standards and modifications of analysis operations are essential to further enhance the accuracy and improve the simplicity of rhamnolipid analysis.Key points• Current status of R&D works on determination of rhamnolipids is listed• Advantages and disadvantages of various types analysis are summarized• Limitations of current rhamnolipid quantification are discussed Graphical abstract.
Keywords: Analysis; Colorimetric methods; Identification; Rhamnolipids.