• Journal of Environmental Health Sciences
• /
• v.41 no.5
• /
• pp.358-367
• /
• 2015

Objectives: This study was performed to evaluate the analytical method for PAH metabolites in human urine using enzyme hydrolysis and solid-phase extraction coupled with LC-(ESI)-MS/MS technique. Methods: We employed HPLC tandem mass spectrometry techniques with appropriate pre-treatment for analysis of 16 OH-PAHs in human urine. Samples were hydrolysis by ${\beta}$-flucuronidase/Aryl sulfatase, and target compounds were extracted by solid-phase extraction with a strata-x cartridge. Cross-validation was performed between Eulji University and Green Cross laboratories with 200 human urine samples. Results: The accuracies were between 90.3% and 118.8%, and precisions (relative standard deviations) were lower than 10%. The linearity obtained was satisfying for the 16 OH-PAH compounds, with a coefficient of determination ($r^2$) higher than 0.99. The results of cross-validation at the two organizations were compared by ICC (interclass correlation coefficient) values. The cross-validation results were excellent or good for all compounds. Conclusion: An analytical method was validated for low nanogram levels of 16 OH-PAHs in human urine. Also, satisfying results were obtained for method validation such as accuracy, precision and ICC of cross-validation.

• Korean Journal of Environmental Agriculture
• /
• v.25 no.4
• /
• pp.371-381
• /
• 2006

New proteomic techniques have been pioneered extensively in recent years, enabling the high-throughput and systematic analyses of cellular proteins in combination with bioinformatic tools. Furthermore, the development of such novel proteomic techniques facilitates the elucidation of the functions of proteins under stress or disease conditions, resulting in the discovery of biomarkers for responses to environmental stimuli. The ultimate objective of proteomics is targeted toward the entire proteome of life, subcellular localization biochemical activities, and the regulation thereof. Comprehensive analysis strategies of proteomics can be classified into three categories: (i) protein separation via 2-dimensional gel electrophoresis (2-DE) or liquid chromatography (LC), (ii) protein identification via either Edman sequencing or mass spectrometry (MS), and (iii) proteome quantitation. Currently, MS-based proteomics techniques have shifted from qualitative proteome analysis via 2-DE or 2D-LC coupled with off-line matrix assisted laser desorption ionization (MALDI) and on-line electrospray ionization (ESI) MS, respectively, toward quantitative proteome analysis. In vitro quantitative proteomic techniques include differential gel electrophoresis with fluorescence dyes. protein-labeling tagging with isotope-coded affinity tags, and peptide-labeling tagging with isobaric tags for relative and absolute quantitation. In addition, stable isotope-labeled amino acids can be in vivo labeled into live culture cells via metabolic incorporation. MS-based proteomics techniques extend to the detection of the phosphopeptide mapping of biologically crucial proteins, which ale associated with post-translational modification. These complementary proteomic techniques contribute to our current understanding of the manner in which life responds to differing environment.

• Bulletin of the Korean Chemical Society
• /
• v.34 no.12
• /
• pp.3629-3634
• /
• 2013

In this study, an improved method for the quantitative analysis of ginkgolic acids (GAs) in Ginkgo biloba leaf extract was developed. The samples were extracted with a mixture of chloroform and 50 % ethanol, after which the chloroform extract was dried and reconstituted in methanol. GAs with 13:0, 15:1, and 17:1 in the extract were successfully separated within 40 min and determined with high throughput performance using an online column-switching HPLC method using an SP column C8 SG80 ($4.6{\times}150mm$, $5{\mu}m$) and a Cadenza 5CD C18 column ($4.6{\times}150mm$, $3{\mu}m$). The developed HPLC method was validated for Ginkgo biloba leaf extract. The validation parameters were specificity, linearity, precision, accuracy, and limits of detection and quantitation (LODs and LOQs, respectively). It was found that all of the calibration curves showed good linearity ($r^2$ > 0.9993) within the tested ranges. The LODs and LOQs were all lower than $0.04{\mu}g/mL$. The established method was found to be simple, rapid, and high throughput for the quantitative analysis of GAs in ten commercial Ginkgo biloba leaf extract and dietary supplements. The samples were also analyzed in LC-electrospray ionization (ESI) tandem mass spectrometry (MS/MS) - multiple-ion reaction monitoring (MRM) mode to confirm the identification results that were obtained by the column switching HPLC-DAD method. The developed method is considered to be suitable for the routine quality control and safety assurance of Ginkgo biloba leaf extract.

• Journal of Plant Biotechnology
• /
• v.37 no.2
• /
• pp.196-204
• /
• 2010

With the completion of genome sequencing of several organisms, attention has been focused to determine the function and functional network of proteins by proteome analysis. The recent techniques of proteomics have been advanced quickly so that the high-throughput and systematic analyses of cellular proteins are enabled in combination with bioinformatics tools. Furthermore, the development of proteomic techniques helps to elucidate the functions of proteins under stress or diseased condition, resulting in the discovery of biomarkers responsible for the biological stimuli. Ultimate goal of proteomics orients toward the entire proteome of life, subcellular localization, biochemical activities, and their regulation. Comprehensive analysis strategies of proteomics can be classified as three categories: (i) protein separation by 2-dimensional gel electrophoresis (2-DE) or liquid chromatography (LC), (ii) protein identification by either Edman sequencing or mass spectrometry (MS), and (iii) quanitation of proteome. Currently MS-based proteomics turns shiftly from qualitative proteome analysis by 2-DE or 2D-LC coupled with off-line matrix assisted laser desorption ionization (MALDI) and on-line electrospray ionization (ESI) MS, respectively, to quantitative proteome analysis. Some new techniques which include top-down mass spectrometry and tandem affinity purification have emerged. The in vitro quantitative proteomic techniques include differential gel electrophoresis with fluorescence dyes, protein-labeling tagging with isotope-coded affinity tag, and peptide-labeling tagging with isobaric tags for relative and absolute quantitation. In addition, stable isotope labeled amino acid can be in vivo labeled into live culture cells through metabolic incorporation. MS-based proteomics extends to detect the phosphopeptide mapping of biologically crucial protein known as one of post-translational modification. These complementary proteomic techniques contribute to not only the understanding of basic biological function but also the application to the applied sciences for industry.

• Journal of Food Hygiene and Safety
• /
• v.18 no.4
• /
• pp.195-201
• /
• 2003

An easily available, simultaneous identification/determination procedure for sildenafil, homosildenafil, tadalafil, vardenafil in adulterated health related foods was established by using a combination of three different analytical methods; thin layer chromatography(TLC), liquied chromatography-mass spectrometry (LC/MS) and high-performance liquied chromatography (HPLC)/photo-diode-array detector. The sample solution for TLC was applied to silica gel 60 $F_{254}$ plates with ethylacetate/acetonitrile/25%ammonia (90:10:5) as a developing solvent. Spots were located under UV radiation at 254 nm and dragendolfs reagent. Mass spectra of the compounds by LC/MS were investigated with electrospray ionization (ESI) interface, under positive ion mode. The HPLC analysis was performed on a column of capcell pack $C_{18}$ (UG120, 4.6${\times}$250mm I.D. 5 ${\mu}$m)with 0.1% sodium 1-hexansulfonate (in 0.1% phosphoric acid)/acetnitrile (73:27) as a mobile phase, and effluent was minitored with a photo-diode-applied to commercial foods, Sildenafil content was inthe range of 0.4mg/g~360.9 mg/g from 7 out of 35 samples. Homosildenafil content was in the range of 2.2 mg/g~336.0 mg/g from 7 out of 35 samples. Tadalafil content was 429.3 mg/g, 9.6 mg/500 mg from 2 out of 35 samples. The procedure described here is available for the screening of sildenafil, homosildenafil, tadalafil, vardenafil.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.36 no.4
• /
• pp.303-314
• /
• 2010

In this study, the antibacterial, antioxidative effect and component analysis of Pinus koraiensis leaf extracts were investigated. MIC values of the ethyl acetate fraction from P. koraiensis leaf extracts on P. acnes, S. aureus, P. ovale, and E. coli were 0.06 %, 0.25 %, 0.13 % and 0.50 %, respectively. The results showed that the antibacterial activity of the ethyl acetate fraction on P. acnes, P. ovale. and S. aureus was more prominent. The aglycone fraction of P. koraiensis leaf extracts ($22.93\;{\mu}g/mL$) showed more higher free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of P. koraiensis leaf extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The 50 % ethanol extract ($0.70\;{\mu}g/mL$) showed the most prominent ROS scavenging activity. Also the ethyl acetate ($1.04\;{\mu}g/mL$) and the aglycone fraction ($1.43\;{\mu}g/mL$) showed very high antoxidant activity. The protective effects of extract/fractions of P. koraiensis leaf extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The P. koraiensis leaf extracts showed cellular membrane protective effects in a concentration dependent manner ($5{\sim}50\;{\mu}g/mL$). TLC and HPLC chromatogram of the ethyl acetate fraction obtained from hydrolysis of P. koraiensis leaf extracts revealed 2 main bands (PK-4, PK-6) and peaks (peak 1, peak 2), which were identified as kaempferol-3-O-glucoside (PK-6, peak 1) and kaempferol-3-O-arabinoside (PK-4, peak 2) by LC/ESI-MS/MS, respectively. These results indicate that extract/fractions of P. koraiensis can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging ROS, and protect cellular membrane against ROS. Extract/fractions of P. koraiensis can be applicable to new cosmeceuticals for antioxidant, antiaging, and antibacterial activity.

• Applied Chemistry for Engineering
• /
• v.24 no.2
• /
• pp.177-183
• /
• 2013

In this study, the antioxidative activities and component analysis of Broussonetia kazinoki SIEB (B. kazinoki). extracts were investigated. B. kazinoki extract showed the effective free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}=8.53{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of the ethyl acetate fraction of B. kazinoki. extracts in the luminol-dependent $Fe^{3+}-EDTA/H_2O_2$ system was $1.69{\mu}g/mL$. The ethyl acetate fraction of B. kazinoki. extracts also exhibited more prominent cellular protective effects (${\tau}_{50}$, 183.3 min at $10{\mu}g/mL$) than that of typical antioxidant $\alpha$-tocopherol (${\tau}_{50}$ = 38.00 min) in the $^1O_2$-induced photohemolysis of human erythrocytes. Components of the ethyl acetate fraction obtained from B. kazinoki extracts were analyzed by TLC, HPLC chromatogram, LC/ESI-MS/MS and $^1H$-NMR. Consequently, Components, components were identified as the kazinol J of kazinol series and luteolin (2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4-chromenone) of flavonoid series having antioxidant activities. These results indicate that extract/ fraction of B. kazinoki can be used as antioxidants in biological systems, particularly skins exposed to UV radiation by quenching and/or scavenging $^1O_2$ and other ROS, and protecting cellular membranes against ROS. Thus, the extract/fraction of B. kazinoki could be applicable to new cosmeceuticals.

• The Korean Journal of Pesticide Science
• /
• v.20 no.3
• /
• pp.236-246
• /
• 2016

The aim of this study was to develop an analytical method for the determination of pyribencarb and its metabolite KIE-9749 in agricultural commodities. The experiment was performed with a range of concentrations $0.05{\sim}2.5{\mu}g/g$ in apple, green pepper, potato, hulled rice, soybean, pear, peach, grape and cucumber. Each samples were extracted with acetone and cleaned by dichloromethane/saline water partition and purified with Florisil solid phase extraction (SPE) cartridge and aminopropyl SPE cartridge. Pyribencarb and KIE-9749 were separated and quantified by HPLC/UVD at 265nm using acetonitrile and water as mobile phase. The recoveries of pyribencarb and KIE-9749 were within 78.3~108.4% and 73.9~113.7% with RSD below 12.2% and 15.0%, respectively. The limits of quantification (LOQ) were both $0.05{\mu}g/g$. LC/ESI-MS/MS was optimized for confirmation of residue identity.

• Microbiology and Biotechnology Letters
• /
• v.42 no.1
• /
• pp.51-57
• /
• 2014

In this study, the antioxidative effects and component analysis of Polygonum aviculare (P. aviculare) extracts were investigated. The ethyl acetate and the aglycone fraction from P. aviculare extracts were more active than (+)-${\alpha}$-tocopherol and $\small{L}$-ascorbic acid, which are known as strong antioxidants for their antioxidative activity by the DPPH method and chemiluminescence assay. The cellular protective effects of fractions of P. aviculare on the rose-bengal sensitized photohemolysis of human erythrocytes, increased in a concentration dependent manner ($1-10{\mu}l$). In particular, the ethyl acetate fraction at a concentration of $10{\mu}l$ showed the most prominent protective effect among all the extracts (${\tau}_{50}$, 314.70 min). TLC and HPLC chromatogram of the ethyl acetate fraction of P. aviculare extracts revealed 3 main bands (PA8, PA5, PA6) and peaks (peak 1, peak 2, peak 3), which were identified as myricetin-3-O-rhamnoside (myricitrin, PA8, peak 1), quercetin-3-${\alpha}$-rhamnoside (quercitrin, PA6, peak 3) by LC/ESI-MS/MS and $^1H$-NMR respectively. These results indicate that fractions from P. aviculare could be applicable to new functional cosmetics as antioxidants.

• Journal of the Korean Chemical Society
• /
• v.68 no.3
• /
• pp.139-145
• /
• 2024

This study aims to elucidate the mechanism involved in the hydrolysis of the hexacyanoferrate(III) complex ion (Fe(CN)₆^3-) and the mechanism leading to the formation of Prussian blue (Fe^III₄[Fe_II(CN)₆]₃·xH₂O, PB) in acidic aqueous solutions at moderately elevated temperatures. Hydrolysis constitutes a crucial step in generating PB through the widely used single-source or precursor method. Recent PB syntheses predominantly rely on the single-source method, where hexacyanoferrate(II/III) is the exclusive reactant, as opposed to the co-precipitation method employing bare metal ions and hexacyanometalate ions. Despite the widespread adoption of the single-source method, mechanistic exploration remains largely unexplored and speculative. Utilizing UV-vis spectrophotometry, negative-ion mode liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS), and a devised reaction, this study identifies crucial intermediates, including aqueous Fe^2+/3+ ions and hydrocyanic acid (HCN) in the solution. These two intermediates eventually combine to form thermodynamically stable PB. The findings presented in this research significantly contribute to understanding the fundamental mechanism underlying the acid-catalyzed hydrolysis of the hexacyanoferrate(III) complex ion and the subsequent formation of PB, as proposed in the sequential mechanism introduced herein. This finding might contribute to the cost-effective synthesis of PB by incorporating diverse metal ions and potassium cyanide.