Dataset Information

Monitoring the chemical production of citrus-derived bioactive 5-demethylnobiletin using surface-enhanced Raman spectroscopy.

ABSTRACT: To develop an accurate and convenient method for monitoring the production of citrus-derived bioactive 5-demethylnobiletin from the demethylation reaction of nobiletin, we compared surface-enhanced Raman spectroscopy (SERS) methods with a conventional high-performance liquid chromatography (HPLC) method. Our results show that both the substrate- and solution-based SERS methods correlated with the HPLC method very well. The solution method produced lower root-mean-square error of calibration and higher correlation coefficient than the substrate method. The solution method used an "affinity chromatography"-like procedure to separate the reactant nobiletin from the product 5-demthylnobiletin based on their different binding affinities to the silver dendrites. The substrate method was found simpler and faster to collect the SERS "fingerprint" spectra of the samples because no incubation between samples and silver was needed and only a trace amount of samples was required. Our results demonstrated that the SERS methods were superior to the HPLC method in conveniently and rapidly characterizing and quantifying 5-demethylnobiletin production.

SUBMITTER: Zheng J

PROVIDER: S-EPMC3786449 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature

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Monitoring the chemical production of citrus-derived bioactive 5-demethylnobiletin using surface-enhanced Raman spectroscopy.

Zheng Jinkai J Fang Xiang X Cao Yong Y Xiao Hang H He Lili L

Journal of agricultural and food chemistry 20130820 34

To develop an accurate and convenient method for monitoring the production of citrus-derived bioactive 5-demethylnobiletin from the demethylation reaction of nobiletin, we compared surface-enhanced Raman spectroscopy (SERS) methods with a conventional high-performance liquid chromatography (HPLC) method. Our results show that both the substrate- and solution-based SERS methods correlated with the HPLC method very well. The solution method produced lower root-mean-square error of calibration and ...[more]

PMID: 23885986

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Project description:Metabolomics is an emerging field of cell biology that aims at the comprehensive identification of metabolite levels in biological fluids or cells in a specific functional state. Currently, the major tools for determining metabolite concentrations are mass spectrometry coupled with chromatographic techniques and nuclear magnetic resonance, which are expensive, time consuming and destructive for the samples. Here, we report a time resolved approach to monitor metabolite dynamics in cell cultures, based on Surface Enhanced Raman Scattering (SERS). This method is label-free, easy to use and provides the opportunity to simultaneously study a broad range of molecules, without the need to process the biological samples. As proof of concept, NIH/3T3 cells were cultured in vitro, and the extracellular medium was collected at different time points to be analyzed with our engineered SERS substrates. By identifying individual peaks of the Raman spectra, we showed the simultaneous detection of several components of the conditioned medium, such as L-tyrosine, L-tryptophan, glycine, L-phenylalanine, L-histidine and fetal bovine serum proteins, as well as their intensity changes during time. Furthermore, analyzing the whole Raman data set with the Principal Component Analysis (PCA), we demonstrated that the Raman spectra collected at different days of culture and clustered by similarity, described a well-defined trajectory in the principal component plot. This approach was then utilized to determine indirectly the functional state of the macrophage cell line Raw 264.7, stimulated with the lipopolysaccharide (LPS) for 24 hours. The collected spectra at different time points, clustered by the PCA analysis, followed a well-defined trajectory, corresponding to the functional change of cells toward the activated pro-inflammatory state induced by the LPS. This study suggests that our engineered SERS surfaces can be used as a versatile tool both for the characterization of cell culture conditions and the functional state of cells over time.

Dataset Information

Monitoring the chemical production of citrus-derived bioactive 5-demethylnobiletin using surface-enhanced Raman spectroscopy.

Publications

Monitoring the chemical production of citrus-derived bioactive 5-demethylnobiletin using surface-enhanced Raman spectroscopy.

Similar Datasets

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