After a pilot scale production of a rhamnolipidic biosurfactant, it is purified using two methods, chemical separation and chromatography. We use UPLC-MS/MS for the characterization of the chemical species.

1. Insert your information hereIntroduction Results
References
Biousurfactants are surface-active agents, whose potential application
range includes the use in bioremediation of hydrocarbon contaminated
soils. (Behrens et al. 2016). Rhamnolipids (RLs), the most studied type
of biosurfactants, makes hydrocarbons soluble for degradation through
cell activity. RL diversity arises from the heterogeneous congener
mixtures synthesized which can vary in alkyl chain length ranging from
C8 to C14 and their isomers for mono or di-rhamnolipids, raising more
than 60 different congeners. The aim of this work was to know the
diversity of the RL congeners produced by the Pseudomonas
aeruginosa 6K-11 strain and detected using ultra pressure liquid
chromatography tandem mass spectrometry (UPLC-MS/MS) method.
Purification and determination of Rhamnolipid congeners through ultra-performance liquid
chromatography tandem mass spectrometry (UPLC-MS/MS), employing corn oil as carbon source
M.V.G. Alván,K.K.Valladares,D.G.Martinez,F.Merino,S.Gutierrez.
U.N.M.S.M.-LaboratoryofMicrobiologyand MicrobialBiotechnology.Lima-PE
mgracial13@gmail.com
Methods
Results
MONO-
RHAMNOLIPID
CONGENER
TRANSITION
% ABUNDANCE
SEP-SN AMB-SN PA-T
Rha-C8-C10 475 - 305 0 0 1.90
Rha-C10-C8 475 - 333 0 0.00 2.03
Rha-C10-C10 503 - 333 0 82.87 80.61
Rha-C10-C12:1 529 - 333 0 8.57 8.26
Rha-C121:-C10 529 - 359 0 0 0.20
Rha-C10-C12 531 - 333 0 8.55 5.18
Rha-C12-C10 531 - 361 0 0 1.81
Total 0 100% 100%
DI-RHAMNOLIPID
CONGENER
Rha-Rha-C8-C10 621 - 451 0 1.26 1.15
Rha-Rha-C10-C8 621 - 479 59.04 1.99 1.36
Rha-Rha-C10-C10 649 - 479 40.96 68.44 74.42
Rha-Rha-C10-C12:1 675 - 479 0 9.73 6.54
Rha-Rha-C12:1-C10 675 - 505 0 0 0.22
Rha-Rha-C10-C12 677 - 479 0 14.76 12.01
Rha-Rha-C12-C10 677 - 507 0 3.83 4.30
Total 100% 100% 100%
Conclusion
Table 1. % Relative abundance was calculated based on the individual congener peak area divided by the sum total area for all RLs for either
mono- or di- RL fractions as per current practice for published literature in this area.
This study showed that the congener produced and the nitrogen source are
not related. Also, the best method to purify rhamnolipids depends on the
purpose of the research, since the purification through acid precipitation is
cost effective, fast and the 14 congeners are obtained, but there is still
contaminating molecules. On the other hand, adsorption and ion exchange
chromatography reduce the abundance of contaminating molecules and, in
the case of using both techniques, 2 rhamnolipids congeners are obtained.
0 10 20 30 40 50 60 70
Rha-Rha-C10-C8
Rha-Rha-C8-C10
Rha-C10-C8
Rha-C8-C10
Rha-Rha-C10-C10
Rha-C10-C10
Rha-Rha-C10-C12:1
Rha-Rha-C12:1-C10
Rha-C10-C12:1
Rha-C121:-C10
Rha-Rha-C10-C12
Rha-Rha-C12-C10
Rha-C12-C10
Rha-C10-C12
% ABUNDANCE
RLCONGENER
RL Congeners detected using UPLC MS/MS
PA-T AMB-SN SEP-SN
Graphic 1. Abundance percentage of RL congener detected in different samples. PA-T: tryptone as nitrogen source and purification with acid
precipitation method. AMB-SN: Sodium Nitrate as nitrogen source, and purified trough adsorption chromatography. SEP-SN: Sodium Nitrate as nitrogen
source, and purified trough adsorption and ion exchange chromatography.Rudden, M.2015. Applied microbiology and biotechnology.
Behrens, B.2016.Analytical and bioanalytical chemistry,.
Reiling, H 1986 Applied and environmental microbiology Acknowledges: Supported by Proyecto FINCYT under Contract Nº238-IA-2013
Methods
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