• Title/Summary/Keyword: HPLC Chromatography

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Fatty Acid Combination of Major Triglyceride in Hagfish Flesh Lipids (먹장어 지질의 주된 트리글리세리드의 지방산조성)

  • LEE Eung-Ho;WADA Shun;KOIZUMI Chiaki;OHSHIMA Toshiaki;NONAKA Junsaku
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.4
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    • pp.291-298
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    • 1984
  • The extracted hagfish (Eptatretus burgeri) flesh lipid was separated into following fractions by column chromatography on Bio-beads SX-2 and Sephadex LH-20 prior to gab chromatographic analysis of their fatty acid compositions: polar lipid, triglyceride and free fatty acid. The major fatty acids of total lipid and triglyceride in hagfish were $C_{16:0},\;C_{16:1},\;and\;C_{18:1}$. The ratio of $C_{18:0}/C_{18:1}$ in the total lipid and triglyceride of hagfish was 0.1. The polar lipid of the hagfish muscle was mainly composed of phosphatidyl choline ($65.5\%$) and phosphatidyl ethanolamine ($28.0\%$). The triglyceride obtained was fractionated into four fractions by HPLC on the basis of partition numbers. Both the fatty acid composition and triglyceride composition on the basis of the total carbon number in the acyl chains of the triglyceride were analysed by the GLC. From the information obtained on triglyceride compositions based on the total carbon number by GLC and the partition number by HPLC and fatty acid composition by GLC, the combination of fatty acid in each triglycerides was estimated. A computer was used for estimation of the fatty acid combination in the triglyceride because hagfish lipid triglyceride was composed of various kinds of fatty acids. Fortyfour kinds of triglyceride were estimated. The major triglycerides in hagfish flesh lipid were found to those of ($1{\times}C_{16:0},\;2{\times}C_{18:1};\;13.5\%$), ($1{\times}C_{16:0},\;1{\times}C_{18:0},\;1{\times}C_{18:1};\;7.2\%$), ($1{\times}C_{16:1},\;2{\times}C_{18:1};\;5.4\%$), ($2{\times}C_{16:0},\;1{\times}C_{22:5};\;5.2\%$), ($1{\times}C_{14:0},\;2{\times}C_{18:1};\;4.5\%$), ($2{\times}C_{18:1},\;1{\times}C_{22:5};\;3.6\%$), ($1{\times}C_{14:0},\;1{\times}C_{18:0},\;1{\times}C_{18:1};\;2.7\%$) and ($1{\times}C_{14:0},\;1{\times}C_{16:0},\;1{\times}C_{18:2};\;2.2\%$).

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A Study on the Analytical Method of Artificial Sweeteners in Foods (식품 중 인공감미료의 분석법에 관한 연구)

  • Kim, Hee-Yun;Yoon, Hae-Jung;Hong, Ki-Hyung;Lee, Chang-Hee;Park, Sung-Kwan;Choi, Jang-Duck;Choi, Woo-Jeong;Park, Sun-Young;Kim, Ji-Hye;Lee, Chul-Won
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.14-18
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    • 2004
  • Analysis methods of artificial sweeteners, aspartame, acesulfame potassium, sodium saccharin, and sucralose isolated from foods were developed using high performance liquid chromatography, HPLC conditions for aspartame, acesulfame potassium, and sodium saccharin were: column, Symmetry $C_{18}(3.9mm\;i.d{\times}150mm,\;5{\mu}m)$; mobile phase, 0.05M sodium phosphate monobasic : acetonitrile (9 : 1, pH 3.5, containing 0.01M tetrapropylammonium hydroxide); detector, UV detector at 210 nm. HPLC condition for sucralose were : column, Symmetry $C_{18}(3.9mm\;i.d{\times}150mm,\;5{\mu}m)$; mobile phase, water:methanol (7 : 3); detector, refractive index detection (sensitivity = 16). Recoveries of artificial sweeteners in foods including soft drinks, fruit and vegetable beverages, alcoholic beverages, fermented milk beverages, soybean milk, ice cream, snacks, chewing gums, jam, honey, kimchi salted food, special dietary products, processed fish products, candies, food additive mixtures, chocolate and cocoa were 76.1-101.3%, 82.3-103.2%, 83.1-103.7%, and 80,6-99.5% for aspartame, acesulfame potassium, sodium saccharin, and sucralose, respectively.

Quantitative Analysis for Components of Epimedium koreanum (음양곽 주성분의 정량분석)

  • Han, Yong-Nam;Hwang, Keum-Hee;Lee, Mie-Soon
    • Korean Journal of Food Science and Technology
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    • v.28 no.4
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    • pp.616-623
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    • 1996
  • Eum Yang Kwak, the aerial part of Epimedium koreanum, is widely used as a folk medicine for stimulant in man, tonic, and hypotensive purpose. The plant contains icariin (a specific flavonoid), magnoflorine (an alkaloid) and tannin, but their contents are not known until now. In this paper, a quantitative analysis method for them was developed. Determination of icariin and magnoflorine was successfully achived by high performance liquid chromatography equipped with a UV detector in the ranges of $0.1{\sim}0.4\;mg$ and $0.002{\sim}0.1\;mg\;per\;ml$ sample, respectively. Extraction of the plant was carried out with water or 50% ethanol using different decocting temperatures and times. Icariin was well extracted either by water ($100^{\circ}C$, 3hr) or 50% ethanol ($85^{\circ}C$, 1hr), and its content in the plant was measured to be 0.94%. On the other hand, magnoflorine was fully extracted by 50% ethanol ($85^{\circ}C$, 1hr), and its content was determined to be 0.16%. Therefore, decoction of the medicinal plant with water at $100^{\circ}C$ for 3hr turned out to be recommendable for the best extraction.

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Analysis of Four Pesticides, Isoproturon, Phenmedipham, Pyridate and Nitenpyram Residues by High-Performance Liquid Chromatography with Diode-Array Detector (HPLC를 이용한 Isoproturon, Phenmedipham, Pyridate 및 Nitenpyram 4종 성분의 잔류농약 분석법 개발)

  • Yang, Sung-Yong;Koo, Yun-Chang;Wang, Zeng;Heo, Kyeong;Kim, Hyeong-Kook;An, Eun-Mi;Shin, Han-Seung;Lee, Jin-Won;Lee, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1165-1170
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    • 2010
  • A method for the determination of four pesticide compounds, urea (isoproturon), bis-carbamate (phenmedipham), thiocarbamate (pyridate) and vinyllidenediamine (nitenpyram) were examined and analyzed by HPLC with C-18 column ($250\;mm{\times}4.6\;mm$, $5\;{\mu}m$ diameter particle size). Mobile phase consisted of deionized water, acetonitrile and 50 mM $KH_2PO_4$ (pH 2.5). Isoproturon and phenmedipham analytical condition was isocratic elution of the column with 50% solvent A (acetonitrile) and 50% solvent B (deionized water); pyridate was 85% solvent A (acetonitrile) and 15% solvent B (deionized water) at a flow rate of 1 mL/min; and nitenpyram analytical condition was 90% solvent A (50 mM $KH_2PO_4$, pH 2.5) and 10% solvent B (acetonitrile) at a flow rate of 1 mL/min. In results, retention times were 6.12, 8.63, 9.40 and 12.76 min for isoproturon, phenmedipham, pyridate and nitenpyram, respectively. All injection volumes were $10\;{\mu}L$ and the limit of quantitation was 0.05 mg/kg for four pesticide compounds, respectively. Recovery rate test was performed with three farm products, rice, apple and soybean. Four pesticide compounds were spiked at concentrations of 0.05, 0.1 and 0.5 mg/kg. The recovery rates were ranged from 70.18% to 118.08% and the standard deviations of all experiments were within 10%.

Development of Rapid Analytical Method of Forbidden Medicines in Dietary Supplements Using LC-ESI-Tandem MS (LC-ESI-tandem MS를 이용한 기능성표방식품 중 부정유해물질 신속검사체계 개발)

  • Kim, Hee-Yun;Jang, Young-Mi;Joo, Hyun-Jin;Jung, Young-Hyun;Lee, Myoung-Sook;Park, Jong-Seok;Lee, Kwang-Ho;Lee, Hwa-Mi
    • Korean Journal of Food Science and Technology
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    • v.39 no.4
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    • pp.372-379
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    • 2007
  • A high-performance liquid chromatography-electrospray ionization (HPLC-ESI) tandem MS was developed for the rapid and simultaneous determination of forbidden medicines in dietary supplements. Thirteen medicinal components such as PDE-5 inhibitors and their analogues, and the newly identified dimethylsildenafil and xanthoanthrafil, were included in this study. After tentative standardization of molecular ions in both polarities using thirteen references on the mass spectrometer, with ESI-continuous infusion via the syringe pump method, the relative intensity of the ions present in the resulting spectra was quantitatively compared. From the results, the ion mode was selected depending on each reference's characteristics. A HPLC method coupled with the ESI mode was developed considering the matrix effect and interference depending on the type of sample. The validation test of the developed method was followed by carrying out precision, accuracy, recovery, sensitivity and linearity, etc. The method showed sufficiently high sensitivity, reproducibility, and specificity, and produced 4 times faster results when compared with the existing HPLC/UV method for the determination of forbidden compounds in dietary supplements.

A Comparative Study on Quantifying Uncertainty of Vitamin A Determination in Infant Formula by HPLC (HPLC에 의한 조제분유 중 비타민 A 함량 분석의 측정불확도 비교산정)

  • Lee, Hong-Min;Kwak, Byung-Man;Ahn, Jang-Hyuk;Jeon, Tae-Hong
    • Korean Journal of Food Science and Technology
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    • v.40 no.2
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    • pp.152-159
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    • 2008
  • The purpose of this study was to determine the accurate quantification of vitamin A in infant formula by comparing two different standard stock solutions as well as various sample weights using high performance liquid chromatography. The sources of uncertainty in measurement, such as sample weight, final smaple vloume, and the instrumental results, were identified and used as parameters to determine the combined standard uncertainty based on GUM(guide to the expression of uncertainty in measurement) and the Draft EURACHEM/CITAC Guide. The uncertainty components in measuring were identified as standard weight, purity, molecular weight, dilution of the standard solution, calibration curve, recovery, reproducibility, sample weight, and final sample volume. Each uncertainty component was evaluated for type A and type B and included to calculate the combined uncertainty. The analytical results and combined standard uncertainties of vitamin A according to the two different methods of stock solution preparation were 627 ${\pm}$ 33 ${\mu}$g R.E./100 g for 1,000 mg/L of stock solution, and 627 ${\pm}$ 49 ${\mu}$g R.E./100 g for 100 mg/L of stock solution. The analytical results and combined standard uncertainties of vitamin A according to the various sample weighs were 622 ${\pm}$ 48 ${\mu}$g R.E./100 g, 627 ${\pm}$ 33 ${\mu}$g R.E./100 g, and 491 ${\pm}$ 23 ${\mu}$g R.E./100 g for 1 g, 2 g, and 5 g of sampling, respectively. These data indicate that the preparation method of standard stock solution and the smaple amount were main sources of uncertainty in the analysis results for vitamin A. Preparing 1,000 mg/L of stock solution for standard material sampling rather than 100 mg, and sampling not more than 2 g of infant formula, would be effective for reducing differences in the results as well as uncertainty.

Isolation and Characterization of Elastase Inhibitor from Areca catechu (빈랑으로부터 Elastase 저해물질의 분리 및 특성조사)

  • 조중제;이건국;조병기;최정도
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.26 no.1
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    • pp.163-186
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    • 2000
  • We have previously screened 150 medicinal plants on the inhibition of elastase and found a significant inhibitory effects of the extracts of Areca catechu L. on the aging and inflammation against the skin tissues. To isolate and identify the compounds having biological activity, we was further purified by each of the solvent fractions, silica gel column chromatography, preparative TLC and reversed-Phase HPLC. Peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as Phenolic substance using various colorimetric methods, UV, and IR. $IC_{50}$/ values of phenolic substance purified from Areca catechu were 26.9 $\mu\textrm{g}$/$m\ell$ for porcine pancreatic elastase (PPE) and 60.8 $\mu\textrm{g}$/$m\ell$ for human neutrophil elastase (HNE). This Phenolic substance showed more potent activity than those of reference compounds, oleanolic acid (76.5 $\mu\textrm{g}$/$m\ell$ for PPE, 219.2 $\mu\textrm{g}$/$m\ell$ for HNE) and ursolic acid (31.0 $\mu\textrm{g}$/$m\ell$ for PPE, 118.6 $\mu\textrm{g}$/$m\ell$ for HNE). According to the Lineweaver-Burk Plots, the inhibition against both PPE and HNE by this phenolic substance was competitive with substrate. Phenolic substance from Areca catechu exhibited high free radical scavenging effect ($SC_{50}$/ : 6 $\mu\textrm{g}$/$m\ell$) and inhibited effectively hyaluronidase activity ($IC_{50}$/: 210 $\mu\textrm{g}$/$m\ell$). These results suggest that the Phenolic substance Purified from Areca catechu showed anti-aging effect by protecting connective tissue proteins.

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Modification and Validation of an Analytical Method for Dieckol in Ecklonia Stolonifera Extract (곰피추출물의 지표성분 Dieckol의 분석법 개선 및 검증)

  • Han, Xionggao;Choi, Sun-Il;Men, Xiao;Lee, Se-jeong;Oh, Geon;Jin, Heegu;Oh, Hyun-Ji;Kim, Eunjin;Kim, Jongwook;Lee, Boo-Yong;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.37 no.3
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    • pp.143-148
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    • 2022
  • This study was to investigate an analytical method for determining dieckol content in Ecklonia stolonifera extract. According to the guidelines of International Conference on Harmonization. Method validation was performed by measuring the specificity, linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) of dieckol using high-performance liquid chromatography-photodiode array. The results showed that the correlation coefficient of calibration curve (R2) for dieckol was 0.9997. The LOD and LOQ for dieckol were 0.18 and 0.56 ㎍/mL, respectively. The intra- and inter-day precision values of dieckol were approximately 1.58-4.39% and 1.37-4.64%, respectively. Moreover, intra- and inter-day accuracies of dieckol were approximately 96.91-102.33% and 98.41-105.71%, respectively. Thus, we successfully validated the analytical method for estimating dieckol content in E. stolonifera extract.

Evaluation of HbA1c Levels Via the Latex Immunoturbidimetric Method by Using Chemistry Autoanalyzer (자동화학분석기에서의 라텍스 면역비탁법의 Autolab HbA1c 평가)

  • Jo, Yongjun;Lee, So-young;Park, Hae-il;Kim, YeongSic;Lee, Jehoon;Kim, Yonggoo;Han, Kyungja
    • Laboratory Medicine Online
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    • v.2 no.1
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    • pp.10-14
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    • 2012
  • Background: Measurement of HbA1c levels is widely used to diagnose diabetes mellitus and to evaluate and monitor plasma-glucose concentrations over 6-8 weeks. In this study, we evaluated the diagnostic performance of the newly developed latex immunoturbidimetric method by using Autolab HbA1c. Methods: We analyzed and compared the diagnostic performance of Autolab HbA1c with that of Toshiba 200FR between April 2009 and July 2009. According to guidelines (EP5-A2, EP6-P, EP9-A2) of the clinical and laboratory standards institute (CLSI), we compared linearity, precision and correlation of Autolab HbA1c with those of G7 (Tosoh Corp., Kyoto, Japan) by using high-performance liquid chromatography (HPLC) method. Results: Data obtained using Autolab HbA1c showed good linearity in mixtures of samples with low (3.1%) and high (15.1%) levels of HbA1c (r2=0.9997). In the analysis of within-run precision of the samples with HbA1c levels of 5.1% and 12.1%, the SDs were 0.04 and 0.06 and covariances of these samples were 0.8% and 0.5%, respectively. In the Deming regression model, the regression equation was as follows: Autolab HbA1c=1.0859×Tosoh HPLC-0.6957. Conclusions: In this study, Autolab HbA1c method showed better performance characteristics than Tosoh G7 did. In reference review, there was no interference of variant hemoglobin. The data acquisition time of Autolab HbA1c was lower than that of Tosoh G7. The advantages of Autolab HbA1c are that it can be used as an autoanlyzer in routine chemical analysis, it does not require pre-analytical treatment, and the samples are automatically treated with distilled water for hemolysis.

Effects of Salinity Stress on the Composition of Free Amino Acids of the Pacific abalone Haliotis discus discus (염분 스트레스가 둥근전복 Haliotis discus discus의 유리아미노산 조성에 미치는 영향)

  • Yang, Hyun-Sung;Park, Kyung-Il;Hong, Choong-Hee;Choi, Kwang-Sik
    • Journal of Aquaculture
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    • v.21 no.4
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    • pp.218-225
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    • 2008
  • Changes in 13 free amino acids (FAA) of Haliotis discus discus exposed to various salinities were investigated using a high performance liquid chromatography (HPLC). Taurine, glycine and alanine are three major contributors to the total FAA in the gill tissues of H. discus discus. Concentration of taurine was 114 imol/g dry tissue weights accounting for 76.64% of total FAA in H. discus discus. Levels of most FAAs in H. discus discus exposed to low and high salinities for 24 h decreased dramatically. Taurine concentration was slightly increased in the samples exposed to 20 psu and 25 psu for 48 h, and greatly increased after 120 h exposure. After 48 hrs exposure to 20 psu, 25 psu, 30 psu and 40 psu, methionine in H. discus discus was not detected; no methionine was detected in the sample exposed to 20 psu, 25 psu, 30 psu and 40 psu after 120 h of exposure. Taurine:glycine ratio increased depending upon hyper-hypoosmotic condition as well as period of osmotic stress. These data indicate that taurine, glycine and methionine play important role in regulating osmotic stress in H. discus discus. This study suggesting that FAA analysis is a useful tool to diagnose osmotic stress to H. discus discus.