• Title/Summary/Keyword: fluorometric HPLC

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Validation of chlorophyll algorithm in Ulleung Basin, East/Japan Sea

  • Yoo, Sin-Jae;Kim, Hyun-Cheol;Lee, Jeong-ah;Park, Mi-Ok
    • Korean Journal of Remote Sensing
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    • v.18 no.1
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    • pp.35-42
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    • 2002
  • The results of our observation in May 2000 indicated that the SeaWiFS algorithm (O'Reilley et al., 1998), which was adopted for OSMI data processing, overestimated the actual chlorophyll values. This was rather unexpected in that there were good reasons to expect that the bio-optical properties of East/Japan Sea belonged to Case 1 water and in such case, the OC2 algorithm would give unbiased estimates of actual chlorophyll a values. In November 2000, a cruise conducted bio-optical surveys in the same area. This time we added HPLC (High Performance Liquid Chromatography) method for measuring chlorophyll a concentration to the standard fluorometric method, which we hale been using during the past Fluorometric method with acidification is known to result in under/overestimation of chlorophyll values in many parts of the world oceans, while it is easier and cheaper than HPLC method. To our surprise, the comparison of HPLC chlorophyll and fluorometric chlorophyll values show that fluorometric values gave an underestimation up to 50%. This error was due to the presence of accessory pigments such as chlorophyll b. Considering this error, our precious result of May 2000(Yoo et al., 2000) might have to be reinterpreted. Calculation of reflectance at 490 and 555nm, however, indicated that this is not still enough to explain the discrepancies.

Measurement of δ-Aminolevulinic Acid in Urine by Fluorometric HPLC and Colorimetric Methods (비색법과 HPLC 법에 의한 요중 δ-Aminolevulinic acid의 측정치 비교)

  • Ahn, Kyu-Dong;Yeon, You-Yong;Lee, Byung-Kook
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.4 no.1
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    • pp.17-24
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    • 1994
  • The urinary excretion of ${\delta}$-aminolevulinic acid has been widely used as a measure of the biological effect of lead in lead exposed workers. It is usually measured by colorimetric method based on the color reaction of ALA-pyrrole with Ehrlich's reagent. But the results of ${\delta}$-ALA in urine by this method are somewhat artificially higher than expected due to the urinary ALA-like compound such as aminoacetone. On the other hand, the recently developed fluorometric HPLC method is very sensitive and specific for the measuring urinary ALA. In order to compare the data obtained by two methods and to investigate the interrelation between two methods, 117 lead workers with different lead exposure were checked urinary ${\delta}$-ALA, blood lead and other lead exposure related indices. The results obtained are as follows; 1. Urinary excretion of ${\delta}$-ALA by colorimetric method is 2.15 times higher than HPLC method in overall, revealing 2.47 times in workers of blood lead less than $20{\mu}g/dl$, 2.53 times in workers of blood lead $21-40{\mu}g/dl$ and 1.86 times in workers of blood lead over $41{\mu}g/dl$, respectively. 2. While the correlation coefficients of ${\delta}$-ALA measured by colorimetric method with blood lead and blood ZPP was 0.571 and 0.629, those of ${\delta}$-ALA measured by HPLC with blood lead and blood ZPP were 0.6l0 and 0.637. All the correlation coefficients were statistically significant, but there was no statistical difference of correlation coefficients between two methods. 3. The correlation coefficient of urinary excretion of ${\delta}$-ALA between two method was 0.838 without any correction, but it was 0.852 with the correction of specific gravity 1.024. 4. Simple linear regression of ${\delta}$-ALA measured by HPLC method on ${\delta}$-ALA measured by colorimetric method was (ALA-UPH)=-0.245+0.536 (ALA-UCO) without any correction and it was (SP ALA)=-0.525+0.598 (SP ALA-UCO) with the correction of specific gravity 1.024. With above results, it is recommended that the diagnostic criteria of ${\delta}$-ALA for lead poisoning needed to be revised if ${\delta}$-ALA is measured by HPLC rather than colorimetric method.

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Composition and Distribution of Phytoplankton with Size Fraction Results at Southwestern East/Japan Sea

  • Park, Mi-Ok
    • Ocean Science Journal
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    • v.41 no.4
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    • pp.301-313
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    • 2006
  • Abundance and distribution of phytoplankton in seawater at southwestern East/Japan Sea near Gampo were investigated by HPLC analysis of photosynthetic pigments during summer of 1999. Detected photosynthetic pigments were chlorophyll a, b, $c_{1+2}$ (Chl a, Chl b, Chl $c_{1+2}$), fucoxanthin (Fuco), prasinoxanthin (Pras), zeaxanthin (Zea), 19'-butanoyloxyfucoxanthin (But-fuco) and beta-carotene (B-Car). Major carotenoid was fucoxanthin (bacillariophyte) and minor carotenoids were Pras (prasinophyte), Zea (cyanophyte) and But-fuco (chrysophyte). Chl a concentrations were in the range of $0.16-8.3\;{\mu}g/land$ subsurface chlorophyll maxima were observed at 0-10m at inshore and 30-50 m at offshore. Thermocline and nutricline tilted to the offshore direction showed a mild upwelling condition. Results from size-fraction showed that contribution from nano+picoplankton at Chl a maximum layer was increased from 18% at inshore to 69% at offshore on average. The maximum contribution from nano+picoplankton was found as 87% at St. E4. It was noteworthy that contribution from nano+picoplanktonic crysophytes and green algae to total biomass of phytoplankton was significant at offshore. Satellite images of sea surface temperature indicated that an extensive area of the East/Japan Sea showed lower temperature ($<18\;^{\circ}C$) but the enhanced Chi a patch was confined to a narrow coastal region in summer, 1999. Exceptionally high flux of low saline water from the Korea/Tsushima Strait seemed to make upwelling weak in summer of 1999 in the study area. Results of comparisons among Chi a from SeaWiFS, HPLC and fluorometric analysis showed that presence of Chi b cause underestimation of Chi a about 30% by fluorometric analysis but overestimation by satellite data about 30-75% compared to HPLC data.

Detection of Aflatoxins in Soybean Food by HPLC (고속액체 크로마토 그라피에 의한 대두식품중 아플라톡신의 검출)

  • Kim, Young-Kook;Roh, Jung-Koo
    • Korean Journal of Food Science and Technology
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    • v.17 no.4
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    • pp.295-303
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    • 1985
  • Aflatoxin $B_1$, $B_2$, $G_1$, and $G_2$were quantitatively detected by the high pressure liquid chromatography on a Micropak-CN column, with Hexane-THF-IPA-water, using a Lichrosorbpacked flowceil in the fluorometric detector. Under those conditions, the minimum detectable amount of aflatoxin $B_1$ was 0.2 ng. HPLC was used in determining amount of aflatoxins in the commercially manufactured soybean food and home-made Meju. Aflatoxin producing abilities of strains used in the industrially fermented soybean food were also studied with the HPLC technique. Although aflatoxin-like substances were detected in a few samples on TLC, they were not identified with the HPLC retention times of standard aflatoxins. The commercial fungal strains used in Korea had no aflatoxin producing abilities.

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High Performance Liquid Chromatographic Assay of Ofloxacin in Plasma (혈장중 Ofloxacin의 HPLG분석)

  • Baek, Chae Sun;Kim, Young Su
    • Korean Journal of Clinical Pharmacy
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    • v.10 no.1
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    • pp.38-41
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    • 2000
  • A high-performance liquid chromatographic method with fluorometric detection was evaluated for analysis of ofloxacin in plasma. Biological fluids (plasma, $200\;{\mu}L$) were prepared for assay by protein precipitation with chlorofurm. The detection of ofloxacin and triamterene as an internal standard were performed at 358 nm for excitation and 495 nm for emission. The HPLC separation was carried out on Ultrasphere ODS column (4.6 mm${\times}25\;cm,\;5\;{\mu} m$) with acetonitrile $(45\%)$-phosphoric acid $(1.5\%)\;containing\;0.3\%$ sodium laurylsulfate as the mobile phase. The flow-rate was 1.0 mL/min. The calibration graphs were linear from 3.0 to 80 ng/mL with r=0.998. The minimal detectable concentration in plasma was 1.5 ng/mL. The proposed technique is reproducible, selective, reliable and sensitive.

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Application of FMOC-Cl for the Quantitative Determination of N-linked Oligosaccharides (FMOC 표식에 의한 Sugar Chain의 분석)

  • Kim, Dong-Hyun;Hwangbo, Sik;Chung, Gu-Yong
    • Korean Journal of Food Science and Technology
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    • v.29 no.4
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    • pp.630-634
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    • 1997
  • A fluorescence tagging agent, FMOC-Cl (9-fluorenylmethyl chloroformate) was used for the determination of 1-amino-oligosaccharide intermediates generated from glycoproteins by peptide-N $(N-acetyl-{\beta}-D-glucosaminyl)$ asparagine amidase (N-Glycanase, PNGase F). The derivatives were separated on an Amido 80 column by HPLC using a gradient system with 25 to 51% aqueous acetonitrile and monitored by a fluorometric detector. The detection limit of FMOC-amino-oligosaccharides was $0.05{\sim}1.5$ pmol with fluorometric detection at 278 nm.

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A pilot study on measuring the human cortisol using an electronic device (전자소자를 이용한 인간 감성지표 코티졸 검출에 관한 기초연구)

  • Lee, Hee-Jo;Lee, Jung-Hyun;Hwang, Yoo-Sun;Jung, Hyo-Il
    • Science of Emotion and Sensibility
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    • v.14 no.4
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    • pp.531-536
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    • 2011
  • The precise measurement of human emotion is of pivotal importance in the field of emotion science. Based on the perspective of psychoneuroimmunoendocrinology, human emotion is now considered to be measurable by body fluid. The biological molecule cortisol, which is known for the stress hormone, has been widely investigated to help researchers to estimate the stress loaded on human body. Until now, several measurement techniques such as High Performance Liquid Chromatography (HPLC), fluorometric assay, and reverse phase chromatography have been developed. However, since these measurements are expensive, take relatively long time for an operation, and they are not portable, they are not appropriate for POCT (point of care testing). In this paper we demonstrate the performance of a miniaturized-microwave resonant device in the measurement of cortisol. Our method has many advantages in that it requires a small volume of sample, has fast response time, is easy to operate and needs no labeling process. Besides, it will shed a light on the measurement techniques for emotion science.

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Studied on the Antibacterial, Antifungal Components in Some Korean Marine Sponges (한국산 해면류중의 항균, 항곰팡이 물질에 관한 연구)

  • LEE Jong-Soo;KIM In-Soo;MOON Soo-Kyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.3
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    • pp.193-202
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    • 1991
  • Antimicrobial substances were screened by paper disk plate method in marine sponges, Halichondria okadai, Halichendria sp., H iaponica and Haliclona Pemollis, collected from the south coast of Korea. Antibacterial components were detected in two species, H okadai and Halichondria sp.. Three components such as benzoic acid, okadaic acid(OA) and dinophysistoxin-1(DTX1) were identified from these sponges as the antimicrobial compounds by MS and NMR spectral data. OA$(550{\~}600{\mu}g/kg)$ and $(400{\~}490{\mu}g/kg)$ were determined from the wet H okadai and Halichondria sp., respectively, by using fluorometric HPLC analysis with 9-anthryldiazomethane(ADAM) as fluorescent labelling reagent.

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Paralytic Shellfish Toxins in the Mussel, Mytilus edulis, Caused the Shellfish Poisoning Accident at Geoje, Korea, in 1996 (96년 거제에서 패류 중독 사고를 유발한 진주담치의 마비성 독소)

  • LEE Jong-Soo;SHIN Il-Shik;KIM Young-Man;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.1
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    • pp.158-160
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    • 1997
  • Food poisoning accident occurred on May, 1996 at Oepo, Geoje County, Kyeongnam Province, Korea, and two persons were died within a few hours after ingestion of the soup prepared with wild mussel, Mytilus edulis, harvested on the sea rock. Paralytic shellfish poisons (PSP) were elucidated as the responsible toxins for the food poisons accident because the wild mussels caught after three days at the near place from the accident contained high toxicity of PSP ranged $650\~1000MU/g$ of edible meat by mouse bioassay. Gonyautoxin-1+4 $(42.7\%)$ and C1+C2 $(40.0\%)$ were detected as the major toxins in the mussels by fluorometric HPLC method. Although, the poison extracted out with drip during freezing and thawing, and the toxicity gradually decreased by boiling for 20 minutes, over 30 MU/g of toxins remained in the soup and meat, which indicated that they could be able to make food poisoning.

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Identification of an Essential Tryptophan Residue Residue in Alliinase from Garlic (Allium sativum) by Chemical Modification

  • Jin, Yeong Nam;Choe, Yong Hun;Yang, Cheol Hak
    • Bulletin of the Korean Chemical Society
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    • v.22 no.1
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    • pp.68-76
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    • 2001
  • We have employed chemical modification to identify amino acids essential for the catalytic activity of alliinase (EC 4.4.1.4) from garlic (Allium sativum). Alliinase degrades S-alkyl-L cysteine sulfoxides, causing the characteristic odor of garlic. The activity of alliinase was rapidly and completely inactivated by N-bromosuccinimide(NBS) and slightly decreased by succinic anhydride and N-acetylimidazole. These results indicate that tryptophanyl, lysyl, and tyrosyl residues play an important role in enzyme catalysis. The reaction of alliinase with NBA yielded a characteristic decrease in both the absorbance at 280 nm and the intrinsic fluorescence at 332 nm with increasing reagent concentration of NBS, consistent with the oxidation of tryptophan residues. Kinetic analysis, fluorometric titration of tryptophans and correlation to residual alliinase activity showed that modification of only one residue present on alliinase led to complete inhibition of alliinase activity. To identify this essential tryptophan residue, we employed chemical modification by NBS in the presence and absence of the protecting substrate analogue, S-ethyl-L-cysteine (SEC) and N-terminal sequence analysis of peptide fragment isolated by reverse phase-HPLC. A fragment containing residues 179-188 was isolated. We conclude that Trp182 is essential for alliinase activity.