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conferenceseries.com
Volume 8, Issue 5 (Suppl) J Chromatogr Sep Tech, an open access journal
ISSN: 2157-7064 Chromatography 2017
August 07-09, 2017
August 07-09, 2017 | Rome, Italy
4th World Congress on
Chromatography
J Chromatogr Sep Tech 2017, 8:5(Suppl)
DOI: 10.4172/2157-7064-C1-032
Identifying natural synergist from Pongamia pinnata using high-speed counter-current chromatography
combined with isobolographic analysis
Weihong He and Hao Yin
South China Sea Institute of Oceanology, China
For identifying the synergistic compounds from Pongamia pinnata, an approach based on high-speed counter-current
chromatography (HSCCC) combined with isobolographic analysis was designed to detect the synergistic effects in the complex
mixture. In the approach, the complex mixture was considered as the combination of two individual samples for isobolographic
analysis: the target compound and the mixture with complete removal of the target compound (subtracted residue). The two samples
were prepared by HSCCC, and were used for the calculation of the expected effect of their combination. Using this approach, three
compounds representing the major peaks in the HPLC of the brine shrimp toxic extract from P. pinnata (brine shrimp lethality
test (BST) LC50 36.5 μg/mL), pinnatin (1), 3,7-Dimethoxy-3,4'-methylenedioxy flavone (2), and karanjin (3), were prepared from
the extract, and were tested for their synergistic potency by BST. The two-phase solvent system containing n-hexane-ethyl acetate–
MeOH–water (14:7:10:10, v/v/v/v) was selected for the one-step HSCCC separation according to the partition coefficient values
(K). The extract was separated into seven fractions (Fr1–7) by HSCCC with a total mass recovery of 96.3%. Fr2, 4, and 6 were the
peak fractions corresponding to compounds 3, 2, and 1, respectively. The purities and recoveries of the target compounds after
the chromatographic analysis were 95.9%–97.5% and 92.2%–96.1%, respectively. The subtracted residue of each target compound
was performed by recombining all HSCCC fractions except the fraction containing the target compound. Isobolographic analysis
disclosed a significant synergistic effect between karanjin and its subtracted residue (potency ratio of 0.47), which gave clear evidence
that the toxicity of the extract results from synergistic interactions, and karanjin was one of the synergists participating in the
interaction. The other two compounds were excluded from the synergism because these two compounds showed additive effects with
their subtracted residues.
Analysis of free fatty acids in olive oils by UPHLC–MS
Zeid Abdullah Alothman
King Saud University, Saudi Arabia
Asimple, fast, highly efficient and direct method using ultra-performance liquid chromatography coupled to mass spectrometry
has been established for the simultaneous separation, identification and quantitation of a few saturated and unsaturated fatty
acids in olive oils from various countries. Many methods have already been found in the literature for the analysis of fatty acids. No
sample pretreatment techniques were employed such as extraction or derivatization for the analysis of target acids from oil samples,
as the oil samples were just diluted, filtered and then directly injected to the instrument. The chromatographic separations of all
target fatty acids were achieved on a Hypersil Gold C18 column of particle size 1.9 µm, 50×2.1 mm I.D, while the gradient elution
using a binary mobile phase mixture of acetonitrile and water at a flow rate of 1.5 ml/min was adopted for achieving optimum
separations. The identification and quantitation of target compounds was accomplished using selected ion reaction monitoring mode.
The recoveries of the fatty acids were obtained higher than 89% with good validation parameters; linearity (r2>0.992), detection
limit between 0.09 and 0.24 µg/ml, run to run and day to day precisions with percent relative standard deviation lower than 2.4% at
both low (1 µg/ml) and medium (10 µg/ml) concentration levels. The total content of fatty acids in each individual oils was found
in the range of 472.63–7751.20 µg/ml of olive oil, while oleic acid was found to be the major fatty acid among all analyzed oils with
the amount 3785.94 µg/ml (maximum) in Syrian olive oil. The obtained validation parameters confirm that the proposed analytical
method is rapid, sensitive, reproducible and simple and it could be applied for the successful evaluation of fatty acids in various oils
and other matrices. All the fatty acids were efficiently eluted in a time of less than 8 min with well resolved peaks by employing the
proposed method.