• Journal of Life Science
• /
• v.18 no.8
• /
• pp.1159-1163
• /
• 2008

The purpose of this study was to establish a high-throughput assay method capable of estimating specific lipase activity at oil-water interface. The method is based on the fact that fluorescence intensity of fluorescein is affected by pH. The pH-dependence might be used to monitor pH change caused by the release of fatty acid through the action of lipase. Assay was performed by incubating a reaction mixture containing oil emulsion, fluorescein and enzyme and by monitoring fluorescence intensity periodically. Fluorescence intensity decreased linearly with a rate proportional to the enzyme amount. Linear relationship was observed between enzyme amount and reaction rate which was calculated from a graph of fluorescence change against time. The assay was possible at different pH conditions in the range of pH 6.0-8.0.

• Korean Chemical Engineering Research
• /
• v.56 no.6
• /
• pp.826-831
• /
• 2018

This study presents fabrication of bi-compartmental particles labeled by multiple fluorescence. To compartmentalize fluorescent expression at the particle, two fluorescent dyes with less overlap of the excitation and emission spectra are selected. To ensure the fluorescence stability, the fluorescent dyes contain acrylate functional groups in the molecules so that they can be cross-linked together with monomers constituting the particle. Strong fluorescent expression and compartmentalization were observed at the particle fabricated using the selected fluorescent dyes through confocal microscopy. Furthermore, long-term fluorescence stability was verified by measuring fluorescent expression and intensity for 4 weeks. We anticipate that the bi-compartmental particles labeled by multiple fluorescence can be widely used for multi-target drug delivery system, analysis of 3 dimensional Brownian motion, and investigation of 3 dimensional complex self-assembled morphologies.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.10
• /
• pp.1689-1694
• /
• 2008

The ability to detect Salmonella spp. is essential in the prevention of foodborne illness. This study examined a Salmonella spp. detection method involving the application of immunomagnetic separation and immunoliposomes (IMS/IL) encapsulating sulforhodamine B (SRB), a fluorescent dye. A quantitative assay was conducted by measuring the fluorescence intensity of SRB that was produced from an immunomagnetic bead-Salmonella spp.-immunoliposome complex. The results indicated detection limits of $2.7{\times}10^{5}$ and $5.2{\times}10^{3}$ CFU/ml for Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterka subsp. enterka serovar Typhimurium (S. Typhimurium), respectivley. The signal/noise ratio was improved by using 4% skim milk as a wash solution rather than 2% BSA. In addition, higher fluorescence intensity was obtained by increasing the liposome size. Compared with the conventional plating method, which takes 3-4 days for the isolation and identification of Salmonella spp., the total assay time of to h only including 6 h of culture enrichment was necessary for the Salmonella detection by IMS/IL. These results indicate that the IMS/ IL has great potential as an alternative rapid method for Salmonella detection.

• Bulletin of the Korean Chemical Society
• /
• v.31 no.7
• /
• pp.1875-1880
• /
• 2010

A dicycle pyrazoline derivative, 1-phenyl-5-(p-fluorophenyl)-3,4-($\alpha$-p-fluoro-tolylenecyclohexano) pyrazoline, was synthesized and characterized by elemental analysis, IR, UV-vis, fluorescence spectra and X-ray single crystal diffraction. Density function theory (DFT) calculations were performed by using B3LYP method with 6-$311G^{**}$ basis set. The optimized geometry can well simulate the molecular structure. Vibrational frequencies were predicted, assigned and compared with the experimental values, which suggest that B3LYP/6-$311G^{**}$ method can well predict the IR spectra. Both the experimental electronic absorption spectra and the predicted ones by B3LYP/6-$311G^{**}$ method reveal three electron-transition bands, with the theoretical ones having some red shifts compared with the experimental data. Natural bond orbital analyses indicate that the absorption bands are mainly derived from the contribution of n $\rightarrow\pi^*$ and $\pi\rightarrow\pi^*$ transitions. Fluorescence spectra determination shows that the title compound can emit blue-light at about 478 nm. On the basis of vibrational analysis, the thermodynamic properties of title compound at different temperature have been calculated, revealing the correlations between $C^0_{p,m}$, $S^0_m$, $H^0_m$ and temperature.

• Food Science and Biotechnology
• /
• v.18 no.4
• /
• pp.978-984
• /
• 2009

An analytical method for the simultaneous determination of 9 quinolones (QNs) in porcine, chicken, and bovine muscles was developed and validated using liquid chromatography-fluorescence detector (LC-FLD). The samples were extracted using a liquid-liquid extraction (LLE) process. Chromatographic separation was achieved on a reverse phase $C_8$ column with a gradient elution using a mobile phase of 200 mM ammonium acetate buffer (pH 4.5) and acetonitrile (ACN). The proposed method was validated according to the Food and Drug Administration (FDA) guideline for bioanalytical assay procedures. Recoveries of QNs were 83.1-111.9% with relative standard deviations (RSDs) below 15%. Linearity within a range of 30-500 ${\mu}g/kg$ was obtained with the correlation coefficient ($R^2$) of 0.9967-0.9999. The limits of detection (LOD) were 1-16 ${\mu}g/kg$. These values were lower than the maximum residues limits (MRLs) established by the European Union (EU). The present method was successfully applied to determine QNs in edible muscles.

• Journal of Food Hygiene and Safety
• /
• v.13 no.1
• /
• pp.41-46
• /
• 1998

A postcolumn derivatization method was tried for the simultaneous determination of four major aflatoxins ($B_1,\;B_2,\;G_1,\;and\;G_2$) by high-performance liquid chromatography with fluorescence detection. As compared with a previous precolumn derivatization method, it was found that the postcolumn derivatization combined with an electrochemical cell (Kobra cell) was less time-consuming, safer, improved the sensitivity and selectivity, and provided good recoveries for aflatoxin $B_1$ (88.9%) and $G_1$ (100.5%). This method showed linearity from 10~100 ppb for aflatoxin $B_1\;and\;G_1$, and from 3~30 ppb for aflatoxin $B_2\;and\;G_2$. However, aflatoxin Bz and Gz were not detected satisfactorily although they showed good resolution.

• Bulletin of the Korean Chemical Society
• /
• v.21 no.5
• /
• pp.483-486
• /
• 2000

A method is described for the fluorimetric determination of nickel, based on the formation of $Ni(II)-\alpha-(2-Benzimidazolyl)-\alpha'$, $\alpha''$ -(N-5-Nitro-2-Pyridylhydrazone)-toluene complex in the presence of a non-ionic surfactant. The complex has practically no fluorescence in the absence of surfactant, but the addition of Triton X-100 makes possible the fluorimetric determination of low concentrations of Ni(II) as it enhances the fluorescenceintensity of the complex by up to about 5-fold. This method is very sensitive and selectrive for the direct determination of nickel ion. The optimum conditions are a Triton X-100 concentration of 2.0 mL(5.0%, v/v) and pH $9.0\pm0.2(ammonium$ chloride-ammonia buffer). The fluorescence is measured at 337 nm of emission wavelength under 300 nm of excitation wavelength. The fluorescence intensity is a linear function of the concentration of Ni(II) in the range 5-70 ng/mL, and the detection limit is 2.0 nm/mL. The proposed method has been successfully applied to the determination of trace amounts of Ni(II) in food and human hair samples.

• Analytical Science and Technology
• /
• v.27 no.4
• /
• pp.187-195
• /
• 2014

It is very important to minimize the damage of latent fingerprints at enhancing the contrast. This study proves the enhancement effects of cyanoacrylate-fumed latent fingerprints using p-dimethylaminobenzealdehyde (DMAB) on semi-porous surfaces and the influence factors. The latent fingerprints in experiment were developed for cyanoacrylate treatment in a vacuum chamber and used after drying at room temperature for 24 hours. For fluorescence staining, the cyanoacrylate-developed marks using DMAB were sublimated during 48 hours under the different conditions of surface area, temperature, atmospheric pressure. First experiment showed how surface area effects on the sublimation rate and fluorescence intensity by DMAB of particle size and container size. In addition, the fluorescence staining using DMAB with solvent-free contact method had the greatest fluorescence intensity after 36 hours and a low fluorescence intensity over a certain size of surface area. Second experiment showed that the evaporation of DMAB solid crystals got a satisfying result in a temperature of $20^{\circ}C$ and reduced time to get the greatest fluorescence intensity. It took a long time to get a optimum level of fluorescence intensity at $30^{\circ}C$ or more and it was less effective in fluorescence intensity. Third experiment on the pressure indicated that the fluorescence intensity of vacuum was weaker than nonvacuum but it was inapplicable to very high variations in pressure.

• Journal of Life Science
• /
• v.23 no.2
• /
• pp.306-310
• /
• 2013

We compared the efficacy of the competitive ELISA method for measuring the level of urinary aflatoxin M1 (AFM1) with that of the HPLC-fluorescence detector (HPLC-FLD) method. The recovery rate of AFM1 with the ELISA method was 105% (73-124%), and the coefficient of variation of the analysis was 6.85%. The ELISA method showed a 0.20 pg/ml and 0.62 pg/ml limit of detection and limit of quantitation, respectively. In correlation analysis, the two methods showed a very strong and statistically significant correlation (R=0.96, p<0.01). However, in spite of the strong correlation, the ELISA method tended to overestimate the urinary AFM1 concentration compared to the HPLC-FLD method. These results suggest that the competitive ELISA method may be a useful technique for measuring the AFM1 level in high-throughput urine samples, but it needs to be corrected with a regression equation from regression analysis with the HPLC-FLD method.

• Bulletin of the Korean Chemical Society
• /
• v.33 no.1
• /
• pp.105-110
• /
• 2012

A europium (III)-sensitized, spectrofluorimetric (FL) method is presented for the determination of sparfloxacin (SPAR) using an anionic surfactant, sodium dodecyl benzene sulphonate (SDBS). The method is based on the strong fluorescence (FL) enhancement of SPAR after the addition of $Eu^{3+}$ ions as fluorescence probes. The experimental results indicated that the FL intensity of the SPAR-$Eu^{3+}$ system was enhanced markedly by SDBS. The maximum FL emission signal was obtained at about 615 nm when excited at 372 nm. The experimental conditions that affected the FL intensity of the SPAR-$Eu^{3+}$-SDBS system were optimized systematically. The enhanced FL intensity of the system exhibited a good linear relationship with the SPAR concentration over the range of $1.5{\times}10^{-9}-1.2{\times}10^{-7}mol\;L^{-1}$ with a correlation coefficient (r) of 0.9987. The limit of detection ($3{\delta}$) was $4.15{\times}10^{-10}mol\;L^{-1}$ with a relative standard deviation (RSD) of 1.65%. This method was successfully applied for the determination of SPAR in pharmaceuticals, and human serum and urine samples with higher sensitivity, wide dynamic range and better stability. The possible interaction mechanism of the system is also discussed in detail by ultraviolet absorption spectra and FL spectra.