• Toxicological Research
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• v.24 no.3
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• pp.207-212
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• 2008

Pesticide residues play several key roles as environmental and food pollutants and it is crucial to develop a method for the rapid determination of pesticide residues in environments. In this study, a simple, effective, and sensitive method has been developed for the quantitative analysis of methoxyfenozide in water and soil when kept under laboratory conditions. The content of methoxyfenozide in water and soil was analyzed by first purifying the compound through liquid-liquid extraction and partitioning followed by florisil gel filtration. Upon the completion of the purification step the residual levels were monitored through high performance liquid chromatography(HPLC) using a UV absorbance detector. The average recoveries of methoxyfenozide from three replicates spiked at two different concentrations and were ranged from 83.5% to 110.3% and from 98.1% to 102.8% in water and soil, respectively. The limits of detection(LODs) and limits of quantitation(LOQs) were 0.004 vs. 0.012 ppm and 0.008 vs. 0.024 ppm, respectively. The method was successfully applied to evaluate the behavioral fate of a 21% wettable powder(WP) methoxyfenozide throughout the course of 14 days. A first-order model was found to accurately fit the dissipation of methoxyfenozide in water with and a $DT_{50}$ value of 3.03 days was calculated from the fit. This result indicates that methoxyfenozide dissipates rapidly and does not accumulate in water.

• BMB Reports
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• v.49 no.11
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• pp.585-586
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• 2016

Extracellular vesicles (EVs) are natural carriers of biomolecules that play central roles in cell-to-cell communications. Based on this, there have been various attempts to use EVs as therapeutic drug carriers. From chemical reagents to nucleic acids, various macromolecules were successfully loaded into EVs; however, loading of proteins with high molecular weight has been huddled with several problems. Purification of recombinant proteins is expensive and time consuming, and easily results in modification of proteins due to physical or chemical forces. Also, the loading efficiency of conventional methods is too low for most proteins. We have recently proposed a new method, the so-called exosomes for protein loading via optically reversible protein-protein interaction (EXPLORs), to overcome the limitations. Since EXPLORs are produced by actively loading of intracellular proteins into EVs using blue light without protein purification steps, we demonstrated that the EXPLOR technique significantly improves the loading and delivery efficiency of therapeutic proteins. In further in vitro and in vivo experiments, we demonstrate the potential of EXPLOR technology as a novel platform for biopharmaceuticals, by successful delivery of several functional proteins such as Cre recombinase, into the target cells.

• Korean Journal of Environmental Agriculture
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• v.36 no.1
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• pp.63-66
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• 2017

BACKGROUND: The phenol concentrations in water samples were determined using gas chromatography after derivatization of the analyte to phenyl acetate followed by extraction using a large volume of solvent. However, this procedure requires an additional purification step and is not analytically efficient. METHODS AND RESULTS: In this study, phenol was first extracted from an acidified water sample using ethyl acetate and then acetylated using acetic anhydride in the presence of a small amount of water and $K_2CO_3$. The derivative was extracted using 1mL of n-butyl acetate. One microliter of the extract was analyzed by GC-MS without further purification. The calibration curve showed good linearity with the $r^2$ value of 0.9968. The method detection limit and the limit of quantitation were estimated to be $0.18{\mu}g/L$ and $0.56{\mu}g/L$, respectively. Repeatability (RSD, n=3) and recovery (n=3) were 9.1%-4.3% and 90.6%-110.5%, respectively. The concentrations of phenol in a few samples of stream water were distributed in the range of $2.51-7.51{\mu}g/L$. CONCLUSION: This method is simpler and faster to implement than those currently utilized and shows high analytical reliability. It can be applied to the quantitative determination of phenol concentrations in surface water and groundwater samples.

• Bulletin of the Korean Chemical Society
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• v.34 no.2
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• pp.489-494
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• 2013

An improved method for the preparation of monomethine cyanine dyes with quinoline nucleus by one-pot three-component using 1-methyl-2-quinolinethione, quaternized 2- or 4-methylheterocyclic compounds and methyl p-toluenesulfonate as starting materials was described. Compared with the traditional methods, the new synthetic method reduced the reaction steps, shortened the reaction time, avoided the separation and purification of the intermediate and reduced cost. The dyes absorbed in the region 478.0-563.0 nm and had molar extinction coefficients of $1.3{\times}10^4-9.4 {\times}10^4L\;mol^{-1}\;cm^{-1}$. Their fluorescence maxima and Stokes shifts were in the range of 525.2-594.4 nm and 16.2-80.6 nm in different solvents, respectively. From the spectral properties of the dyes in different solvents, it could be found that the ${\lambda}_{max}$ of the dyes were shorter in protonic solvents, and showed hypsochromic shifts with the increase of polarity of the solvents.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1018-1023
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• 2001

Oleamide (cis-9-octadecenamide) is endogenous primary amide of fatty acid that is produced in small amounts in animal brains. It is known to induce sleep and to lower temperature by destroying the lipid plasma membrane structure of cells, thereby disclosing gap junction channels. To develop a new biological production method for oleamide, a screening program was conducted to isolate a microorganism producing oleamide. Among 1,500 soil microorganisms tested, KK90378 exhibited a potent positive reaction with Dragendoff`s reagent, used to detect the primary amide of oleamide. KK90378 was identified as a Streptomyces species based on cultural and morpohological characteristics, the presence of diaminopimelic acid in the cell wall, and the sugar patterns for the whole-cell extrat. Streptomyces sp. KK90378 produced oleamide 3 days after culture at $28^{\circ}C$, pH 7.2 A series of purification steps, including hexane extraction, silica gel column, and preparative thin layer chromatographies, were performed for the purification of oleamide. A spectrophotometric analysis using $^1H$, $^13C$-NMR, and GC-MS confirmed that the chemical structure of the purified oleamide was identical to that of authentic oleamide.

• Journal of Life Science
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• v.19 no.8
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• pp.1039-1046
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• 2009

The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.

• BMB Reports
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• v.28 no.2
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• pp.129-137
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• 1995

The ryanodine-receptor $Ca^{2+}$ release channel protein in the sarcoplasmic reticulum membrane of rabbit skeletal muscle plays an important role in muscle exitation-contraction (E-C) coupling. Various types of detergents were tested, including Chaps, cholate, octylglucoside, Zwittergents, Mega-9, Lubrol PX, and Triton X-100 for solubilization of this protein. Among these, Chaps and Triton X-100 were found to optionally solubilize the channel complex. Optimum conditions for this solubilization were pH 7.4 with a salt concentration of 1 M. The addition of phospholipid in the solubilization step helped in stabilizing the protein. The purification of the receptor was performed using sucrose density gradient centrifugation. Various methods [dilution, freeze-thaw, adsorption (Biobeads), and dialysis] were investigated to incorporate the Chaps-solubilized and purified $Ca^{2+}$ release channel protein into liposomes made from different types of phospholipids. Of these, a combined method consisting of a dialysis, freeze-thaw and sonication steps yielded the best results. Reconstituted vesicles produced by this method with 95% phosphatidylcholine (from soybean extract) had good function.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.103-106
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• 2006

Anthocyanins are water-soluble glycosides and acylglycosides of anthocyanidins, having different color variations due to its substitution patterns. Anthocyanins, present in various fruits, vegetables and crops as natural colorant, have been well characterized for its bioactive properties, anti-oxidant, anti-cancer, anti-proliferative and anti-inflammatory properties. During extraction and purification, the factors, such as pH, temperature, oxygen, light, enzymes, nucleophilic agents, sugar derivatives and co-pigments, have affected on anthocyanin stability. For this reason, the extraction method should be thoroughly checked for the qualitative/quantitative analysis of anthocyanin in particular plant material. To identify the optimum extraction method of cyanidin-3-glucoside, major anthocyanin of dark purple-colored grains, Oryza sativa cv. Heugjinjubyeo, Phaselous vulgaris, Phynchosia gngularis, Sesamum indium, Rhynchosia nulubilis and Lablab purpureus, reversed-phase HPLC analysis using solvent system of acetonitrile, methanol and water were accomplished.

• Korean Journal of Soil Science and Fertilizer
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• v.44 no.2
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• pp.278-284
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• 2011

To obtain optimum configuration method and livestock wastewater loading in small-scale livestock wastewater apparatus by natural purification method for treating livestock wastewater, the small-scale livestock wastewater apparatuses were constructed with 9 kinds of combined systems such as aerobic bed (ae)-anaerobic bed (an), ae-ae, ae-anoxic bed (ox), an-an, an-ae, an-ox, ox-ae, ox-an and ox-ox livestock wastewater treatment apparatuses. Under different configuration methods, the removal rate of COD in Ae-Ae and Ae-An livestock wastewater treatment apparatus was higher than that in other configuration methods. The removal rate of T-N in Ae-An livestock wastewater treatment apparatus was higher than that in other configuration methods. Removal rates of SS and T-P were not different regardless of configureation methods in small-scale livestock wastewater treatment apparatus. Under different livestock wastewater loading, the removal rates of pollutants were higher in the order of $50L\;m^{-2}\;day^{-1}{\fallingdotseq}100L\;m^{-2}\;day^{-1}\;>\;200L\;m^{-2}\;day^{-1}$. Therefore, optimum configuration method was Ae-An livestock wastewater treatment apparatus, and optimum livestock wastewater loding was $100L\;m^{-2}\;day^{-1}$ in small-scale livestock wastewater treatment apparatus.

• Korean Journal of Environmental Agriculture
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• v.25 no.3
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• pp.202-210
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• 2006

To develop small-scale sewage treatment apparatus for detached house of agricultural village, a small-scale sewage treatment apparatus by natural purification method that consisted of packaged form of aerobic, anoxic and anaerobic bed was constructed. The efficiency of sewage treatment according to the sewage treatment method, sewage loading, and the injection method of sewage were investigated for small-scale sewage treatment apparatus of packaged form of aerobic, anoxic and anaerobic bed. The removal rate of pollutants according to the sewage treatment method for small-scale sewage treatment apparatus was high in the order of aerobic-anoxic-anoxic bed < aerobic-anoxic-anaerobic bed. The optimum filter media in small-scale sewage treatment apparatus was a broken stone. The removal rate of pollutants according to sewage loading in small-scale sewage treatment apparatus was high in the order of $1,200L/m^2{\cdot}day\fallingdotseq900L/m^2{\cdot}day\fallingdotseq600L/m^2{\cdot}day$. The removal rate of pollutants according to injection method of sewage in small-scale sewage treatment apparatus was high in the order of continuous injection $\fallingdotseq$ intermittent injection. When loaded under the optimum conditions, removal rate of BOD, COD, SS, T-N and T-P were 99, 95, 99, 83 and 96%, respectively, through this 3-stepped small-scale treatment apparatus arrayed with the order of aerobic, anoxic and anaerobic bed.