• Title/Summary/Keyword: UV and fluorescence detection

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Dissolved organic matter characteristics and bacteriological changes during phosphorus removal using ladle furnace slag

  • Noh, Jin H.;Lee, Sang-Hyup;Choi, Jae-Woo;Maeng, Sung Kyu
    • Membrane and Water Treatment
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    • v.9 no.3
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    • pp.181-188
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    • 2018
  • A sidestream contains the filtrate or concentrate from the belt filter press, filter backwash and supernatant from sludge digesters. The sidestream flow, which heads back into the sewage treatment train, is about 1-3% less than the influent flow. However, the sidestream can increase the nutrient load since it contains high concentrations of phosphorus and nitrogen. In this study, the removal of PO4-P with organic matter characteristics and bacteriological changes during the sidestream treatment via ladle furnace (LF) slag was investigated. The sidestream used in this study consisted of 11-14% PO4-P and 3.2-3.6% soluble chemical oxygen demand in influent loading rates. LF slag, which had a relatively high $Ca^{2+}$ release compared to other slags, was used to remove $PO_4-P$ from the sidestream. The phosphate removal rates increased as the slag particle size decreased 19.1% (2.0-4.0 mm, 25.2% (1.0-2.0 mm) and 79.9% (0.5-1.0 mm). The removal rates of dissolved organic carbon, soluble chemical oxygen demand, color and aromatic organic matter ($UV_{254}$) were 17.6, 41.7, 90.2 and 77.3%, respectively. Fluorescence excitation-emission matrices and liquid chromatography-organic carbon detection demonstrated that the sidestream treatment via LF slag was effective in the removal of biopolymers. However, the removal of dissolved organic matter was not significant during the treatment. The intact bacterial biomass decreased from $1.64{\times}10^8cells/mL$ to $1.05{\times}10^8cells/mL$. The use of LF slag was effective for the removal of phosphate and the removal efficiency of phosphate was greater than 80% for up to 100 bed volumes.

Analysis of Selected Water-Soluble Vitamin B1, B2, B3, and B12 Contents in Namul (Wild Greens) Consumed in Korea (국내에서 섭취되는 나물류에 함유된 일부 수용성 비타민의 함량 분석)

  • Yoon, Jimin;Chung, Haejung;Kim, Younghwa
    • The Korean Journal of Food And Nutrition
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    • v.32 no.1
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    • pp.61-68
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    • 2019
  • In this study, the contents of water-soluble vitamins $B_1$ (thiamin), $B_2$ (riboflavin), $B_3$ (niacin), and $B_{12}$ (cyanocobalamin) in namuls (wild greens), such as salads and side dishes, consumed in Korea were quantified by high-performance liquid chromatography (HPLC) with UV and fluorescence detection. All samples were prepared with recipes used in Korea. All analyses were performed under the quality control of vitamin $B_1$, $B_2$, $B_3$, and $B_{12}$. The vitamin $B_1$, $B_2$, and $B_3$ contents in namuls consumed in Korea were analyzed. The highest content of vitamin $B_1$ was $3.018{\pm}0.016mg/100g$ in Putgochudoenjang-muchim. The highest contents of vitamin $B_2$, $B_3$, and $B_{12}$ were $0.279{\pm}0.003mg/100g$ in Gul-muchim, $12.241{\pm}0.040mg/100g$ in Chamchi-salad, and $8.133{\pm}0.371{\mu}g/100g$ in Pijogae-muchim, respectively. These results showed that animal-based ingredients in salads provided for good intake of vitamin $B_{12}$. These results can be used as basic data for food composition tables and improvement of the national health of Koreans.

In Vitro Determination of Dengue Virus Type 2 NS2B-NS3 Protease Activity with Fluorescent Peptide Substrates

  • Khumthong, Rabuesak;Angsuthanasombat, Chanan;Panyim, Sakol;Katzenmeier, Gerd
    • BMB Reports
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    • v.35 no.2
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    • pp.206-212
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    • 2002
  • The NS2B-NS3(pro) polyprotein segment from the dengue virus serotype 2 strain 16681 was purified from overexpressing E. coli by metal chelate affinity chromatography and gel filtration. Enzymatic activity of the refolded NS2B-NS3(pro) protease complex was determined in vitro with dansyl-labeled peptide substrates, based upon native dengue virus type 2 cleavage sites. The 12mer substrate peptides and the cleavage products could be separated by reversed-phase HPLC, and were identified by UV and fluorescence detection. All of the peptide substrates (representing the DEN polyprotein junction sequences at the NS2A/NS2B, NS2B/NS3, NS3/NS4A and NS4B/NS5 sites) were cleaved by the recombinant protease NS2B-NS3(pro). No cleavage was observed with an enzymatically inactive S135A mutant of the NS3 protein, or with a modified substrate peptide of the NS3/NS4A polyprotein site that contained a K2093A substitution. Enzymatic activity was dependent on the salt concentration. A 50% decrease of activity was observed in the presence of 0.1M sodium chloride. Our results show that the NS3 protease activity of the refolded NS2B-NS3(pro) protein can be assayed in vitro with high specificity by using cleavage-junction derived peptide substrates.

Determination of nadolol enantiomers in human plasma using a coupled achiral-chiral high-performance liquid chromatography method

  • Lee, Seung-Beom;Pham, Thuy-Vy;Mai, Xuan-Lan;Le, Thi-Anh-Tuyet;Nguyen, Thi-Ngoc-Van;Kang, Jong-Seong;Mar, Woongchon;Kim, Kyeong Ho
    • Analytical Science and Technology
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    • v.33 no.2
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    • pp.59-67
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    • 2020
  • Nadolol is a β-blocker drug, which effectively manages hypertension and angina pectoris. Its chemical structure allows the formation of four possible stereoisomers. A coupled column high-performance liquid chromatographic (HPLC) system with UV and fluorescence detection was investigated for simultaneously determining four nadolol enantiomers in human plasma. The plasma samples were prepared using a convenient liquid-liquid extraction process and passed through HPLC. Nadolol was initially separated from the endogenous compounds or other impurities in human plasma on a Phenomenex silica column, and its enantiomers were resolved and determined on a Chirapak AD-H column. The developed HPLC method achieved an effective chiral separation and significantly eliminated endogenous compound interference. This optimal HPLC method was validated following FDA guidelines. The results showed good selectivity, linearity, accuracy (90.50 % - 105.27 %), and precision (RSDs < 9.52 %) for each enantiomer. This method was also successfully applied to determine nadolol enantiomers in the plasma samples of a healthy male volunteer (after orally administering 80 mg racemic nadolol), proving its suitability for nadolol stereoselective pharmacokinetic studies.

An Advanced Kinetic Method for HO2·/O2-· Determination by Using Terephthalate in the Aqueous Solution

  • Kwon, Bum Gun;Kim, Jong-Oh;Kwon, Joong-Keun
    • Environmental Engineering Research
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    • v.17 no.4
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    • pp.205-210
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    • 2012
  • Hydroperoxyl radical/superoxide anion radical ($HO_2{\cdot}/O_2^-{\cdot}$, $pK_a$=4.8) as an intermediate is of considerable importance in oxidation processes. Hence, the method of detecting $HO_2{\cdot}/O_2^-{\cdot}$ with high sensitivity is necessary to be developed. To achieve this objective, this study newly employed terephthalate (TA) as a probe for the measurement of $HO_2{\cdot}/O_2^-{\cdot}$ in the kinetic method presented in our previous study. This method was based on the hydroxylation of TA to produce mainly hydroxyterephthalic acid or hydroxyterephthalate (OHTA), which was analyzed by fluorescence detection (${\lambda}_{ex}$=315nm, ${\lambda}_{ex}$=425nm). The life-time of $HO_2{\cdot}/O_2^-{\cdot}$ and its concentration formed from the photolysis technique of $H_2O_2$ were reported in this study. At range of pH 2-10, the life-time of $HO_2{\cdot}/O_2^-{\cdot}$ was 51-422 sec. In particular, an increase in the life-time with pH was observed. The sensitivities of the kinetic method by using TA were always higher with 1.7-2.5 times at pH 8.0 than those by using benzoic acid. From these results, this study can contribute to understanding the basic functions of $HO_2{\cdot}/O_2^-{\cdot}$ in oxidation processes.

Validated HPLC Method for the Pharmacokinetic Study of Atenolol and Chlorthalidone Combination Therapy in Korean Subjects

  • Kang, Hyun-Ah;Kim, Hwan-Ho;Kim, Se-Mi;Yoon, Hwa;Cho, Hea-Young;Oh, Seaung-Youl;Choi, Hoo-Kyun;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.5
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    • pp.331-338
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    • 2006
  • A rapid, selective and sensitive reverse-phase HPLC methods for the determination of atenolol and chlorthalidone in human serum and whole blood were validated, and applied to the pharmacokinetic study of atenolol and chlorthalidone combination therapy. Atenolol and an internal standard, pindolol, were extracted from human serum by liquid-liquid extraction, and analyzed on a $\mu$-Bondapak C18 $10-{\mu}$ column in a mobile phase of methanol-0.01 M potassium dihydrogenphosphate(30:70, v/v, adjusted to pH 3.5) and fluorescence detection(emission: 300 nm, excitation: 224 nm). Chlorthalidone and an internal standard, probenecid, were extracted form human whole blood by liquid-liquid extraction, and analyzed on a Luna C18 $5-{\mu}$ column in a mobile phase of acetonitrile containing 77% 0.01 M sodium acetate and UV detection at 214 nm. These analysis were performed at three different laboratories using the same quality control(QC) samples. The chromatograms showed good resolution, sensitivity, and no interference by human serum and whole blood, respectively. The methods showed linear responses over a concentration range of 10-1,000 ng/mL for atenolol and 0.05-20 ${\mu}g/mL$ for chlorthalidone, with correlation coefficients of greater than 0.999 at all the three laboratories. Intra- and inter-day assay precision and accuracy fulfilled international requirements. Stability studies(freeze-thaw, short-, long-term, extracted sample and stock solution) showed that atenolol and chlorthalidone were stable. The lower limit of quantitation of atenolol and chlorthalidone were 10 ng/mL and 0.05 ${\mu}g/mL$, respectively, which was sensitive enough for pharmacokinetic studies. These methods were applied to the pharmacokinetic study of atenolol and chlorthalidone in human volunteers following a single oral administration of Hyundai $Tenoretic^{\circledR}$ tablet(atenolol 50 mg and chlorthalidone 12.5 mg) at three different laboratories.

Co-occurrence of Deoxynivalenol and Zearalenone in Cereals and their Products (곡류와 그 가공품에서 Deoxynivalenol과 Zearalenone의 분석)

  • Ok, Hyun-Ee;Chang, Hyun-Joo;Choi, Sung-Wook;Lee, Na-Ri;Kim, Hyun-Jung;Koo, Min-Sun;Chun, Hyang-Sook
    • Journal of Food Hygiene and Safety
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    • v.22 no.4
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    • pp.375-381
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    • 2007
  • Deoxynivalenol (DON) and zearalenone (ZEN) are naturally occurring toxins produced by Fusarium species, which may grow on cereals. The aims of this study were to determine the incidence and contamination levels of DON and ZEN in cereal products. Seventy samples of cereal products were randomly selected from retail outlets during 2005 and 2006. DON and ZEN were analyzed by using high performance liquid chromatography with fluorescence and UV-detector, respectively. Detection limits were $4.4{\mu}g\;kg^{-1}$ for DON and $3.4{\mu}g\;kg^{-1}$ for ZEN. DON and ZEN were detected in 37 and 17, respectively, of the 70 samples, but the levels found were very low. In particular, out of 70 samples, 12 samples of corn and barley were co-contaminated with DON and ZEN, with levels ranging from 5.6 to $1842.3{\mu}g\;kg^{-1}$ for DON and 12.1 to $174.9{\mu}g\;kg^{-1}$ for ZEN, respectively. However, DON and ZEN were not detected in breakfast cereals and wheat flour. The highest level was found in dried corn kernel samples that confirmed by LC-MS. This study show that DON and ZEN co-contaminate with low levels in cereal products.

Downregulation of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in human keratinocytes by melanogenic inhibitors

  • Ahn, Kwang-Seok;Lee, Jinseon;Kim, Yeong-Shik
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.780-803
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    • 2003
  • Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.

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A Study of Current Status on Pesticide Residues in Commercial Dried Agricultural Products (서울시 유통 건조농산물 중의 농약잔류 실태 연구)

  • Kim, Bog-Soon;Park, Seoung-Gyu;Kim, Mi-Sun;Cho, Tae-Hee;Han, Chang-Ho;Jo, Han-Bin;Choi, Byung-Hyun;Kim, Sung-Dan
    • Korean Journal of Food Science and Technology
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    • v.39 no.2
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    • pp.114-121
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    • 2007
  • The purpose of this study was to investigate 253 kinds of pesticide residues in 58 commercial dried agricultural products in Seoul. The determinations of the pesticide residues were performed using multiresidue methods and were carried out by a gas chromatography-nitrogen phosphorus detector (GC-NPD), an electron capture detector ($GC-{\mu}ECD$), a mass spectrometry detector (GC-MSD) and high performance liquid chromatography-ultraviolet detector (HPLC-UV), and a fluorescence detector (HPLC-FLD). The pesticide residue detection rate in the commercial dried agricultural products was 24.1% (14 of 58 samples). Twelve pesticide residues without maximum residue limits (MRLs) were detected. In the vegetable groups, the frequency of pesticide residues was found to be in the increasing order of dried fruiting vegetables > dried leafy vegetables > dried stalk and stem vegetables. The pesticides used on dried red pepper in the dried fruiting vegetables were varied (7 kinds) and numerous (4 of 8 samples). The pesticide types detected in the commercial dried agricultural products were in the order of pyrethroid > organochloride > organophosphorus and insecticide > fungicide > herbicide ${\cdot}$ nematicide. The primary pyrethroid pesticide detected was cypermethrin. According to the producing areas of products, large numbers of pesticide residues were found in the order of Korea, China, North Korea, USA, and Vietnam.

Analytical Methods of Hydroxyl Radical Produced by TiO2 Photo-catalytic Oxidation (TiO2 광촉매 산화 반응에서 생성된 수산기 라디칼 분석 방법)

  • Kim, Seong Hee;Lee, Sang-Woo;Kim, Jeong Jin;Kim, Soon-Oh
    • Journal of the Mineralogical Society of Korea
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    • v.28 no.3
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    • pp.245-253
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    • 2015
  • The performance of $TiO_2$ photo-catalytic oxidation process is significantly dependent on the amount of hydroxyl radicals produced during the process, and it is an essential prerequisite to quantify its production. However, precise and accurate methods for quantification of hydroxyl radicals have not been developed so far. For this reason, this study was initiated to compare existing methods for analysis of hydroxyl radicals produced by $TiO_2$ photo-catalytic oxidation and to propose a new method to overcome the limitation of established methods. To simulate $TiO_2$ photo-catalytic oxidation process, Degussa P25 which has been widely used as a standard $TiO_2$ photo-catalyst was used with the dose of 0.05 g/L. The light source of process was UVC mercury low-pressure lamp (11 W, $2,975mW/cm^2$). The results indicate that both potassium iodide (KI)/UV-vis spectrometer and terephthalic acid (TPA)/fluorescence spectrometer methods could be applied to qualitatively measure hydroxyl radicals via detection of triiodide ion ($I_3{^-}$) and 2-hydroxyterephthalic acid which are produced by reactions of iodine ion ($I^-$) and TPA with hydroxyl radicals, respectively. However, it was possible to quantitatively measure hydroxyl radicals using TPA method coupled with high-performance liquid chromatograph (HPLC). The analytical results using TPA/HPLC method show that hydroxyl radical of 0.013 M was produced after 8 hours operation of photo-catalytic oxidation under specific experimental conditions of this study. The proposed method is expected to contribute to precise the evaluation of the performance of photo-catalytic oxidation process.