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Isolation of Alliin in Garlic and Its Quantitative Determination by High Performance Liquid Chromatography and Studies on the Antimicrobial Efforts of Alliin and Ethanol Extracts from Korean Garlic(Alliium sativum L.) (마늘 중 고속 액체 크로마토그래피에 의한 알린의 분리 및 정량과 Alliin과 에탄올 추출물의 항균효과에 관한 연구)

  • 위성언
    • The Korean Journal of Food And Nutrition
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    • v.16 no.4
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    • pp.296-302
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    • 2003
  • First. the purification and analysis of alliin in garlic from different origins by alliin-HPLC determination method were studied. Allinase in garlic was inactivated by heating in boiling water followed by extraction of alliin in garlic with 80% methanol. To remove free amino acids and alliin homologs in garlic, garlic extract was separated by cation exchange column which was packed with amberlite CG-120 resin using 40L d-water as eluent. Alliin in garlic extract was crystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=70:29:1 and then recrystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=75:24:1. Obtained alliin was identified by melting point. TLC, microscope observation and mass spectrometry. High performance liquid chromatography (HPLC) following pre-column derivatization of cystein derivatives with o-phthaldialdehyde/2-mercaptoethanol has succeessfully been applied to the analysis of various garlics. Each alliic of standard solution and garlic extract was derivatized to isoindole derivative by o-phthaldialdehyde /2-mercaptoethanol and then analyzed by HPLC. Six point calibration was done by using alliin peak area. Lineality was observed at 0 ∼ 1.0mg/ml of alliin concentration. Weighted regression line function was Y=6254X - 256077. By this function, alliin contents in various garlics were 0.34 ∼ 0.73% fresh weight. Second study was designed to evaluate the effects of garlic extracts of various concentrations on the growth of various pathogenes (Eubacterium limonsum, Bacteroides fragilis, Salmonella typhimurium, Salmonella typhi, Shigella sonnei, Kiebsiella pneumoniae, Enterobacter cloacae, Pserdomonas aeruginosa, Escherichia coli). For antimicrobial effects against microorganism, totally minimal inhibition concentrations (MIC) of alliin were from 5,000 to 20,000ppm. MIC of ethanol extract were 1,250 to 10,000ppm.

Determination of Synthetic Food Colours by HPLC with Photodiode Array Detector (HPLC를 이용한 타르색소의 분리정량)

  • Yang, Ho-Chul;Heo, Nam-Chil
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.30-35
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    • 1999
  • A simple, rapid, efficient method is for extraction of 13 synthetic water-soluble food colours (Tartrazine, Amarnth, Indigo carmine, New coccine, Sunset yellow FCF, Allura red AC, Eosine, Fast Green FCF, Brilliant Blue FCF, Erythrosine, Acid red, phloxine, Rose Bengal) by polyamide resin and for their quantitative by high performance liquid chromatography (HPLC). Colours (coal-tar dyes) were extracted with polyamide resin and then determinated by HPLC. The HPLC conditions using a reverse phase partition type column $(Nova-pak\;C_{18})$, photodiode array (PDA) detector and 1% Ammonium acetate / 60% acetonitrile in water as eluent, were acceptable for various kinds of colorants. By the use of the proposed method, a survey of coal-tar dyes was carried out on 20 samples and that were detected $4.76{\sim}133.47\;ppm$.

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Determination of 3-phenoxybenzoic Acid in Urine and Exposure Assessment of Pyrethroid Insecticides to Human Being (요중 3-phenoxybenzoic acid 미량 분석 및 pyrethroid계 살포자 노출 평가)

  • Seo, Jong-Chul;Song, Jae-Seok;Choi, Hong-Soon
    • The Korean Journal of Pesticide Science
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    • v.11 no.2
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    • pp.87-94
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    • 2007
  • Pyrethroid insecticide have widely been used for agricultural sector and residential environments. To assess the exposure of insecticide which is absorbed through skin the analysis of urinary metabolite is essential. At present, the urinary 3-PBA was analyzed using liquid-phase extraction. But LPE have many limitations, such as long pre-treatment time and low recovery. So, this study was conducted to determine the optimum conditions for analysing 3-PBA in urine using solid phase extraction. Furthermore, this study intend to investigate the relation of concentrations of pyrethroid, deltamethrin in air and 3-PBA in urine. The optimum condition for hydrolysis was found to be done with hydrochloric acid for one hour. The recovery rates of 3-PBA were $84.6%{\pm}1.2%$, $54.8{\pm}0.9%$, $99.8{\pm}1.2%$ with XAD-2, XAD-7, XAD-16 using as the aborbents and acetone as eluents respectively. But acetonitrle and methanol gave low recovery rate and methyl cellosolve could not elute the compound. The amount of acetone for elution were 6mL, 9mL, 3mL for XAD-2, XAD-7, XAD-16 as absorbents respectively. The non-absorbed rates was $0.8{\pm}0.5%$, and $0.7{\pm}0.3%$ under XAD-16, mesh size 140-200, amount of resin 1.4g and the flow rate of eluent was 0.1mL/min. In the concentration process, we obtained 11 times higher concentration of material. The amounts of urinary 3-PBA were. The LODs of 3-PBA and deltamethrin were 0.004 mg/L, 0.038 mg/L, respectively. The further research of minute monitoring which include spray pattern, environmental condition is needed And more research about the relation between total pyrethroid exposure and urinary various metabolite are also necessary.

Developmemt of Rice Husk Pellets as Bio-filter Media of Bio Scrubber Odor Removal System (왕겨펠렛 생물담체 개발 및 이를 이용한 bio scrubber형 악취제거 시스템 성능평가)

  • Bae, Jiyeol;Han, Sangjong;Park, Ki Ho;Kim, Kwang-Soo
    • Journal of Korean Society for Atmospheric Environment
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    • v.34 no.4
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    • pp.554-566
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    • 2018
  • The rice husk contains nutrients which can be easily utilized by microorganisms, and also has a water retaining ability, which played a crucial part in enabling it to become a biofilter media. In this study, we evaluated the applicability of rice husk pellet bio-scrubber as a microbiological carrier. The pelletization experiment of rice husk as a biological media was performed using PVA and EVA binder. Also, the feasibility tests of rice husk as a biological media for odor removal were carried out in order to know whether rice-husk contains useful components as a media for microbiological growth or not. Lastly, a combined test for odor gas absorption and biological oxidation was conducted using a lab scale bio-filter set-up packed with rice-husk pellets as wet-scrubber. The major components of the rice husk were carbon, hydrogen, nitrogen, and oxygen, while carbon acted as the main ingredient which comprised up to 23.00%. The C : N : P ratio was calculated as 45 : 1 : 2. Oxygen uptake rate, yield and decay rate of the rice husk eluent was calculated to be $0.0049mgO_2/L/sec$, 0.24 mgSS/mgCOD and 0.004 respectively. The most stable form of rice husk pellets was produced when the weight of the rice husk, EVAc, PVAc, and distilled water was 10 : 2 : 0.2 : 10. The prepared rice husk pellets had an apparent density of 368 g/L and a porosity of 59.00% upon filling. Dry rice husks showed high adsorption capacity for ammonia gas but low adsorption capacity for hydrogen sulfide. The bio-filter odor removal column filled with rice husk pellets showed more than 99.50% removal efficiency for NH3 and H2S gas. Through the analysis of circulation water, the prime removal mechanism is assumed to be the dissolution by water, microbial nitrification, and sulfation. Finally, it was confirmed that the microorganisms could survive well on the rice husk pellets, which provided them a stable supply of nutrients for their activity in this long-term experiment. This adequate supply of nutrients from the rice husk enabled high removal efficiency by the microorganisms.

The Study on the Separation of the Subsidiary Elements in Iron and Steel by Using Ion Exchangers (II). The Separation of Anions (이온 교환수지에 의한 철 및 강의 분석에 관한 연구 (제2보). 음이온 성분의 분리)

  • Byoung-Cho Lee;Myon-Yong Park;Kee-Chae Park
    • Journal of the Korean Chemical Society
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    • v.17 no.6
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    • pp.428-433
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    • 1973
  • The quantitative separations of a mixture containing equal amount of each anion such as Si(IV), As(V), P(V), S(VI), W(VI) and Cr(VI) are carried out by the elution through 20${\times}3.14cm^2$ column of anion exchange resin, Dowex 1${\times}$8. The eluents are a mixture of 0.07 M hydrochloric acid and 0.03 M sodium chloride (pH = 1.30) for Si(IV), As(V) and P(V) species, a mixture of 0.6 M sodium chloride and 0.3 M sodium hydroxide for S(VI), W(VI) and Cr(VI) species, and 0.1 N sodium sulfite (pH = 3.48) for P(V) and As(V) species. The subsidiary anions in a standard mixture such as Si(IV), As(V), S(VI), P(V) and W(VI) are separated together from large amount of Fe(III) by the elution through 30cm${\times}3.14cm^2$ column of the resin, Dowex${\times}$50w${\times}$12, using a mixture of 0.1 M sodium nitrate and 2 percent dimethylsulfoxide aqueous solution as an eluent. Si(IV), As(V), S(VI), P(V) and W(VI) eluted together are separated quantitatively under the same conditions as in the separations of the anion mixture. By the conditions obtained in the separations of the standard mixture, Fe(III) and all of the subsidiary anions in steel are quantitatively separated.

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Separation Characteristics of Mandelic Acid in Kromasil HPLC Column (Kromasil HPLC 칼럼에서 Mandelic Acid의 분리특성)

  • Kim, Byung Lip;Kim, Jong Min;Kim, Woo Sik;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.46 no.4
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    • pp.681-685
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    • 2008
  • Chiral separation of racemic mandelic acid was achieved on a Kromasil KR100-5CHI-TBB column. Some chromatographic parameters (resolution, number of theoretical plates, capacity factor) are calculated under different separation conditions such as changes of mobile phase compositions (hexane/t-BME = 85/15 - 10/90) as well as formic acid concentrations for adjusting pH (0.1, 0.5, 1.0 v/v%). Flow rate versus number of theoretical plates was compared to evaluate column efficiency. To determine the adsorption isotherms, PIM (Pulse Input Method) was carried out. At the concentrations of racemic mandelic acid between 0.1 and 0.3 mg/ml, L- and D-mandelic acids have the same retention times of 8.8 and 9.4 min respectively. Mandelic acid isotherms show a linear form under the concentrations of 0.3 mg/ml with eluent (hexane/t-BME = 75/25). As the concentrations of mandelic acids increase, nonlinear Langmuir isotherms were observed as $C_{S,L}=3.358C_{M,L}/(1+0.0897C_{M,L})$ for L-mandelic acid and, $C_{S,D}=3.692C_{M,D}/(1+0.1457C_{M,D})$ for D-mandelic acid.

Analysis of THMs and PCBs by Indirect Photometric Method in High Performance Liquid Chromatography (고성능 액체 크로마토그래피의 간접 분광광도법에 의한 THM과 PCB의 분석)

  • Chung, Yongsoon;Lee, Kangwoo
    • Journal of the Korean Chemical Society
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    • v.39 no.12
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    • pp.918-924
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    • 1995
  • THMs and PCBs were separated and analysed with elution on Novapak ODS or $\mu-Bondapak$ phenyl column by an eluent containing p-nitrophenol (p-NP). THMs studied were CHCl3, CHBrCl2, CHBr2Cl, and CHBr3, and PCBs used were Aroclor 1221, 1242, 1248, $\alpha-$ and $\beta-BHC.$ It was thought that the retention on the stationary phase and sensitivities of the samples are related to the interaction between the sample and stationary phase or p-NP. THMs were separated completely on the ODS column by elution with MeOH-water (30 : 70) containing $1.0{\times}10^{-4}$ M p-NP and some of PCBs were separated on the phenyl column by elution with $CH_3CN$-water(50 : 50) containing $1.0{\times}10^{-4}$ M p-NP. Detection limits of THMs were from $1.0{\times}10^{-4}$ g to $1.0{\times}10^{-6}$ g. Aroclors were $2{\times}10^{-6}$ g, and $\alpha-$ and $\beta-BHC$ were $2{\times}10^{-4}$ g and $1.0{\times}10^{-4}$ g respectively.

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Simultaneous Determination of Titanium, Zirconium and Niobium by Reactor Neutron Activation (원자로 중성자에 의한 티탄, 지르코늄 및 니오브의 동시 정량)

  • Chul Lee;Yung Chang Yim;Koo Soon Chung
    • Journal of the Korean Chemical Society
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    • v.18 no.1
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    • pp.40-46
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    • 1974
  • The fission neutron reactions of $^{47}Ti(n.p)^{47}Sc$ and $^{93}Nb(n,{\alpha})^{90m}Y$, along with epicadmium neutron reaction of $^{96}Zr(n,{\gamma})^{97}Zr$ were used for the simultaneous determination of Ti, Nb and Zr in synthetic mixture. Prior to neutron irradiation, Ti, Zr and Nb in the mixture were separated together in one group through the cation exchange column of Dowex $50{\times}8$ resin using 0.5 M ${\alpha}$-hydroxy-iso-butyric acid as the eluent. After irradiation of the eluate the product nuclides, $^{97}Zr$, ^{47}Sc$ and ^{90m}Y$, were eluted sequentially through the same column with 0.5 M ${\alpha}$-HIBA, 0.5 M ${\alpha}$-HIBA-1 N HNO_3 and 0.5 M ${\alpha}$-HIBA-2 N HNO_3$ solution, respectively. The gamma-ray spectrometry was used for the measurement of the gamma-ray activities of the eluted nuclides. The detection limits of Nb, Ti and Zr were found to be 0.2 %, 0.01 % and 0.002 %, respectively.

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Structural Analysis of Natural Indigo Colorants Extracted from Polygonum tinctorium (천연인디고 색소의 구조분석)

  • Chung, In-Mo;Lee, Kwang-Gill;Sung, Gyou-Byung;Kim, Hyun-Bok;Nam, Sung-Hee;Hong, In-Pyo
    • Journal of Sericultural and Entomological Science
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    • v.49 no.1
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    • pp.8-13
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    • 2007
  • Natural indigo colorants were prepared by extraction of Polygonum tinctorium which was harvested just in the blooming season(in the late of July). The components were analyzed by TLC and HPLC, and its structures were analyzed by FT-IR, EI-mass. The results obtained are summarized as follows; The natural indigo powder was dissolved in DMSO and developed in eluent, $CHCI_3/CH_3CN$(8.5:1.5 v/v) by means of TLC for its quality analysis. It was segregated into indirubin as a red colour and indigo as a blue colour. In case of HPLC analysis,. FT-IR spectrum of indirubin showed a peak for NH residue between 3200 and $3300cm^{-1}$. $^1H-NMR$ spectrum for indigo displayed AA'BB' spin system caused by indole structure between 6.5 and 7.7ppm of H4, 5, 6 and 7, and -NH proton for indirubin showed an singlet between 10.88 and 11.0ppm. EI-mass spectrum of indigo and indirubin both disclosed their molecular size as 262 and it implies that these two substances are isomer.

Determination of Free 4-hydroxyproline with Dansylchloride by HPLC in Human Urine (소변 중 4-hydroxyproline 분석에 관한 연구)

  • Lee, Keou-Weon;Cho, Young-Bong;Lee, Kyung-Jong
    • Journal of Preventive Medicine and Public Health
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    • v.35 no.4
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    • pp.282-286
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    • 2002
  • Objectives : The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. Methods : The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined.4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the a-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 mmol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). Results : The detection limit for the urinary free 4-Hyp was $0.364{\mu}mol/l$. The recovery rate of 4-Hyp was 99.7%, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and $0.32-56.27{\mu}mol/l$. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers ($11.82{\pm}6.73\;nmmol/mg\;Cre$) were 2-fold higher than in pre-shift workers ($5.36{\pm}5.53\;nmol;/mg\;Cre$) and controls ($4.91{\pm}4.89\;nmol;/mg\;Cre$). Conclusions : This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.