caa a22 4500 605505454 CHVBK 20210128100622.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6837-1 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6837-1 Development and validation of an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the quantitative determination of rhamnolipid congeners [Elektronische Daten] [Michelle Rudden, Konstantina Tsauosi, Roger Marchant, Ibrahim Banat, Thomas Smyth] Rhamnolipids (RLs) are synthesised as a complex mixture of congeners comprising either one or two molecules of rhamnose glycosidically linked to a dimer of 3-hydroxy fatty acids varying in chain length and degree of saturation. Currently, HPLC-MS/MS is the most precise and accurate method for RL determination, while accurate quantification is limited. In this study, a rapid ultra pressure liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the rapid and quantification of individual RL congeners. Increased RLs specificity was achieved using tandem mass spectrometry in multiple reaction monitoring (MRM) mode which was used to quantify RL isomer pairs such as Rha-Rha-C8-C10/Rha-Rha-C10-C8 which are difficult to resolve chromatographically. UPLC showed an 18-fold reduction in retention time for Rha-Rha-C10-C10 (1.07 min) and a 17-fold reduction for Rha-C10-C10 (1.36), the major rhamnolipids present, compared to HPLC, with a total run time less than 2.2min. The results show that the linear range for the main RL congeners (Rha-C10-C10 and Rha-Rha-C10-C10) is 0.1 to 100μg/mL. The LOD and LOQ for Rha-C10-C10 is 0.05 and 0.1 μg/mL and for Rha-Rha-C10-C10 is 0.1 and 0.5 μg/mL, respectively. The method was validated for linearity, intra- and inter-day precision and accuracy in accordance with FDA guidelines. The method was applied for the quantification of 14 individual RL congeners produced by Pseudomonas aeruginosa ST5 and comparison of RLs composition on four different carbon sources. Quantification of the individual congeners showed a conserved congener distribution irrespective of carbon source with a preferential selection for C10 β-hydroxyacids as the lipid component of RLs. The only statistically significant differences detected were between actual RL yields on the various carbon sources. Springer-Verlag Berlin Heidelberg, 2015 Rhamnolipids nationallicence UPLC-MS/MS nationallicence Quantification nationallicence Congeners nationallicence Triple quadrupole nationallicence Rudden Michelle School of Biomedical Science, Ulster University, Coleraine, Northern Ireland, UK aut Tsauosi Konstantina School of Biomedical Science, Ulster University, Coleraine, Northern Ireland, UK aut Marchant Roger School of Biomedical Science, Ulster University, Coleraine, Northern Ireland, UK aut Banat Ibrahim School of Biomedical Science, Ulster University, Coleraine, Northern Ireland, UK aut Smyth Thomas Department of Life Sciences, Institute of Technology Sligo, Co., Sligo, Republic of Ireland aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 9177-9187 0175-7598 99:21<9177 2015 99 253 https://doi.org/10.1007/s00253-015-6837-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6837-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Rudden Michelle School of Biomedical Science, Ulster University, Coleraine, Northern Ireland, UK aut NATIONALLICENCE 700 1- Tsauosi Konstantina School of Biomedical Science, Ulster University, Coleraine, Northern Ireland, UK aut NATIONALLICENCE 700 1- Marchant Roger School of Biomedical Science, Ulster University, Coleraine, Northern Ireland, UK aut NATIONALLICENCE 700 1- Banat Ibrahim School of Biomedical Science, Ulster University, Coleraine, Northern Ireland, UK aut NATIONALLICENCE 700 1- Smyth Thomas Department of Life Sciences, Institute of Technology Sligo, Co., Sligo, Republic of Ireland aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 9177-9187 0175-7598 99:21<9177 2015 99 253