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Production and Quantitative Analysis of Trehalose Lipid Biosurfactants Using High‐Performance Liquid Chromatography
Author(s) -
Patil Harshada I.,
Pratap Amit P.
Publication year - 2018
Publication title -
journal of surfactants and detergents
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.349
H-Index - 48
eISSN - 1558-9293
pISSN - 1097-3958
DOI - 10.1002/jsde.12158
Subject(s) - chemistry , chromatography , high performance liquid chromatography , trehalose , calibration curve , column chromatography , analytical chemistry (journal) , detection limit , organic chemistry
Trehalose lipids (THL) are glycolipid biosurfactants having a wide range of biomedical and environmental applications. Low yield, high purification cost, and the absence of a valid analytical method hinders their application. Hence, in the present study a simple, rapid, and reliable isocratic high‐performance liquid chromatography (LC) method was developed for the identification and quantification of trehalose lipid biosurfactants from Rhodococcus erythropolis . THL having a minimum surface tension of 24 mN m −1 and a critical micellar concentration of 25 mg L −1 were produced using hexadecane as a substrate. A standard was developed from the crude THL mixture using thin‐layer chromatography and column chromatography and its structure was confirmed using infrared spectroscopy, mass spectroscopy, and 1 H NMR. A high performance liquid chromatography (HPLC) method for quantitation was developed using a C18 column with water/acetonitrile (80:20) as the mobile phase at a 1 mL min −1 flow rate and UV detection at 208 nm. This method was validated according to International Conference on Harmonization guidelines for linearity, precision, accuracy, robustness, LOD, and LOQ. This method was found to be linear over the range 10‐50 μg m L −1 ( r 2 =0.99801), precise, accurate, and robust. This method can detect minimum 3.2 μg mL −1 and quantify minimum 9.2 μg mL −1 of THL. Standards were developed from R. erythropolis , broth and purified standard trehalose 6,6′‐dimycolate from Mycobacterium bovis , having the same retention time of 2.0 min. The yield was calculated from the calibration curve and was found to be 25 g L −1 .