• 제목/요약/키워드: reverse-phase high performance liquid chromatography

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Development of Analysis Method of Caffeine and Content Survey in Commercial Foods by HPLC (HPLC를 이용한 카페인의 분석법 개발 및 시판 식품중 함유량 조사)

  • Kim, Hee-Yun;Lee, Young-Ja;Hong, Ki-Hyoung;Lee, Chul-Won;Kim, Kil-Saeng;Ha, Sang-Chul
    • Korean Journal of Food Science and Technology
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    • 제31권6호
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    • pp.1471-1476
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    • 1999
  • A simple and practical method for determination of caffeine in foods was developed. The analysis of caffeine was performed by reverse phase high performance liquid chromatography using a ${\mu}-Bondapak\;C_{18}$ column at isocratic condition with methanol-acetic acid-water(20 : 1 : 79) on UV detector at 280 nm. The clean-up and extraction of caffeine in samples were based on a simple pretreatment using a Sep-Pak $C_{18}$ cartridge. Recovery rates obtained with this method for cider, candy, cookie, milk, ice cream and persimmon leaf tea were 99.23%, 99.50%, 99.17%, 99.37%, 98.93% and 99.10% respectively. And the detection limit of caffeine was $0.1\;{\mu}g/mL$. With this method, the range of caffeine contents extracted from coffee, green tea, black tea, Oolong tea(tea bag), soft drinks, ice cream, milk and commercial confectionery were $3.38{\sim}37.50\;mg/g,\;16.30{\sim}26.10\;mg/g,\;10.80{\sim}16.65\;mg/g,\;11.25\;mg/g,\;0.06{\sim}0.11\;mg/g,\;0.04{\sim}0.44\;mg/g,\;0.04{\sim}0.39\;mg/g\;and\;0.10{\sim}1.80\;mg/g$, respectively. But caffeine was not detected in the other tea such as Acanthopanax sessiliflorum tea, Angelica gigas tea, Angelica tea, Arrow root tea, Duchu'ng tea, Dunggulle tea, Ganoerma lucidum tea, Ginger tea powder, Persimmon leaf tea, Ssanghwa tea and Cocoa mix powder.

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Measurement of the Anti-oxidative Properties of Extract from Medicinal Plants Using an On-line HPLC-DPPH Assay (HPLC와 DPPH radical 소거능 측정 방법의 결합에 의한 약용 식물 추출물의 항산화 활성 비교)

  • Im, Do-Youn;Pyo, Byoung-Sik;Kim, Sun-Min;Lee, Kyoung-In
    • Journal of Life Science
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    • 제27권1호
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    • pp.44-49
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    • 2017
  • Natural anti-oxidative compounds have important disease prevention and food preservation properties, in addition to anti-bacterial, anti-inflammation, anti-cancer, and skin whitening effects. High-performance liquid chromatography (HPLC), with an ultra vilolet (UV) detector coupled to a reverse phase C18 column and an online measurement system for 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radicals, was used to search for potent antioxidative compounds in crude extracts. The online HPLC-DPPH assay was then applied to confirm antioxidative compounds in water extracts from Radix of Pueraria lobata, Rhizoma of Zingiber officinale, Fructus of Chaenomeles sinensis, Cortex of Ulmus pumila, and Radix of Astragalus membranaceus. To determine the yields of the extracts, the Brix% of each extract solution was measured using a refractometer. When the relative DPPH radical scavenging ability values of the water extracts were compared with those of a positive control (ascorbic acid), the water extracts of P. lobata, C. sinensis, and U. pumila were 7.77%, 4.71%, and 4.19%, respectively. The results suggest that this method provides a useful assay for rapid measurement of DPPH radical scavenging abilities and conformation of antioxidative compounds in natural products. Moreover, it can reduce the time spent on the separation of active compounds from natural materials, such as medicinal plants, in addition to the use of reagents for separation.

In Vivo Measurement of Extracellular Monoamines and Their Metabolites in the Rat Posterior Hypothalamus Using Microdialysis Technique (미세투석법을 이용하여 흰쥐 후 사상하부에서 세포외액의 모노아민과 대사체들의 생체내 측정)

  • Sung, Ki-Wug;Kim, Seong-Yun;Cho, Young-Jin;Lee, Kweon-Haeng;Lee, Sang-Bok
    • The Korean Journal of Pharmacology
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    • 제28권1호
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    • pp.1-9
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    • 1992
  • Catecholamines, serotonin and their metabolites were measured in the posterior hypothalamus of urethane-anesthetized normotensive Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) using brain microdialysis which is a recently developed experimental method to measure the release of neurotransmitters and their metabolites at the localized brain area in vivo. Microdialysis probe was implanted stereotaxically to the rat posterior hypothalamus and perfused by Ringer's solution. Monoamines and their metabolites were quantified by reverse phase high performance liquid chromatography with electrochemical detection. In vitro recovery test of microdialysis showed that there exist inverse relationship between the perfusion flow rate and the relative recovery of neurochemical compounds. The estimated extracellular concentration of dopamine was about 32 nM, of norepinephrine 50 nM, of epinephrine 50 nM, of serotonin 73 nM, of 3, 4-dihydroxyphenylacetic acid (DOPAC) 281 nM, of homovanillic acid (HVA) 181 nM, and of 5-hydroxyindoleacetic acid (5HIAA) 3767 nM in the hypothalamic perfusate of the normotensive rat. There was no difference in the basal level of monoamines between the SHR and the WKY. In contrast, the level of DOPAC, HVA and 5HIAA in SHR was higher than that in the WKY, This study demonstrated that the microdialysis technique should be an applicable tool for in vivo measurement of central neurochemical substances.

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Bioequivalence of Boryung Torsemide Tablet to Torem Tablet (Torasemide 10 mg) by High Performance Liquid Chromatography/UV Detector

  • Cho, Hea-Young;Kang, Hyun-Ah;Park, Chan-Ho;Kim, Se-Mi;Kim, Dong-Ho;Park, Sun-Ae;Kim, Kyung-Ran;Hur, Hyeon;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • 제35권5호
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    • pp.323-328
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    • 2005
  • The purpose of the present study was to evaluate the bioequivalence of two torasemide tablets, Torem tablet (Roche Korea Co., Ltd., Korea, reference drug) and Boryung Torsemide tablet (Boryung Pharmaceutical Co., Ltd., Korea, test drug), according to the guidelines of Korea Food and Drug Administration (KFDA). After adding an internal standard (furosemide) to human serum, serum samples were extracted using 5 mL of ethyl acetate. Compounds were analyzed by reverse-phase HPLC method with UV detection. This method showed linear response over the concentration range of 0.05 ug/mL with correlation coefficient of 0.999. The lower limit of quantitation using 0.5 mL of serum was 0.05 ug/mL which was sensitive enough for pharmacokinetic studies. Twenty-eight healthy male Korean volunteers received each medicine at the torasemide dose of 20 mg in a $2{\times}2$ crossover study. There was a one-week washout period between the doses. Serum concentrations of torasemide were monitored by an HPLC-UV for over a period of 12 hr after the administration. $AUC_{t}$(the area under the serum concentration-time curve from time zero to 12 hr) was calculated by the linear trapezoidal rule method. $C_{max}$ (maximum serum drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the serum concentration-time data. Analysis of variance was carried out using logarithmically transformed $AUC_{t}$ and $C_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the crossover design was properly performed. The 90% confidence intervals of the $AUC_{t}$ ratio and the $C_{max}$ ratio for Boryung Torsemide/Torem were log 0.97-10g 1.03 and log 0.93log 1.12, respectively. These values were within the acceptable bioequivalence intervals of log 0.80-log 1.25. Thus, the criteria of the KFDA guidelines for the bioequivalence was satisfied, indicating Boryung Torsemide tablet and Torem tablet are bioequivalent.

Evaluation of Tar Dyes Used in Commercial Foods (식품 중 합성첨가물 사용실태 조사 연구 -타르색소 중심으로-)

  • 윤미혜;김국주;김정임;황선일;문수경;정은주;김재관
    • Journal of Food Hygiene and Safety
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    • 제15권2호
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    • pp.108-113
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    • 2000
  • This study was carried out to propose a simple method for the extraction of seven tar dyes such as tartrazine, sunset yellow FCF, amaranth, erythrosine, allura red, brilliant blue FCF and indigo carmine using aminopropyl amine cartridge and to determine the content of the dyes in candies, soft drinks, ice bars and okchuns produced in Korea. The tar dyes were simultaneously analyzed by reverse phase high performance liquid chromatography(HPLC). The recovery rates of the dyes ranged from 65.8% to 99.6%. The contents of the dyes in candies, soft drinks, ice bars and octhuns were N.D.∼50.1 mg/kg, N.D.∼49.9 mg/kg, N.D.∼56.0 mg/kg and N.D.∼867.3 mg/kg, respectively. The types of the dyes used most frequently for candies, soft drinks and ice bars were tartrazine, brilliant blue and amaranth, respectively. Of the samples, tartrazine was used frequently, and indigo carmine was not used at all.

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Hypoallergenic and Physicochemical Properties of the A2 β-Casein Fractionof Goat Milk

  • Jung, Tae-Hwan;Hwang, Hyo-Jeong;Yun, Sung-Seob;Lee, Won-Jae;Kim, Jin-Wook;Ahn, Ji-Yun;Jeon, Woo-Min;Han, Kyoung-Sik
    • Food Science of Animal Resources
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    • 제37권6호
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    • pp.940-947
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    • 2017
  • Goat milk has a protein composition similar to that of breast milk and contains abundant nutrients, but its use in functional foods is rather limited in comparison to milk from other sources. The aim of this study was to prepare a goat A2 ${\beta}$-casein fraction with improved digestibility and hypoallergenic properties. We investigated the optimal conditions for the separation of A2 ${\beta}$-casein fraction from goat milk by pH adjustment to pH 4.4 and treating the casein suspension with calcium chloride (0.05 M for 1 h at $25^{\circ}C$). Selective reduction of ${\beta}$- lactoglobulin and ${\alpha}_s$-casein was confirmed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis and reverse-phase high-performance liquid chromatography. The hypoallergenic property of A2 ${\beta}$-casein fraction was examined by measuring the release of histamine and tumor necrosis factor alpha from HMC-1 human mast cells exposed to different proteins, including A2 ${\beta}$-casein fraction. There was no significant difference in levels of both indicators between A2 ${\beta}$-casein treatment and the control (no protein treatment). The A2 ${\beta}$-casein fraction is abundant in essential amino acids, especially, branched-chain amino acids (leucine, valine, and isoleucine). The physicochemical properties of A2 ${\beta}$-casein fraction, including protein solubility and viscosity, are similar to those of bovine whole casein which is widely used as a protein source in various foods. Therefore, the goat A2 ${\beta}$-casein fraction may be useful as a food material with good digestibility and hypoallergenic properties for infants, the elderly, and people with metabolic disorders.

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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    • 제31권1호
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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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Enzymatic Synthesis and Characterization of Structured Lipids from Docosahexaenoic Acid - Enriched Fish oil and Soybean oil (어유와 대두유로부터 기능성 유지의 효소적 합성)

  • Kim, Yu-Mi;Jeon, Mi-Sun;Lee, Jeung-Hee;Lee, Ki-Teak
    • Food Science and Preservation
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    • 제15권3호
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    • pp.437-444
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    • 2008
  • Structured lipids(SLs) were synthesized by enzymatic interesterification with DHA-enriched fish oil(containing 27% docosahexaenoic acid) and soybean oil in the hatch-type reactor. The interesterification was performed for 24 hr at $55^{\circ}C$ and TLIM(immobilized lipase from Thermonyces lanuginosa, 10% by weight of total substrates) was mixed with 180 rpm of shaking. The fish oil and soybean oil were interesterifed with several weight ratio(fish oil : soybean oil, 2:8, 3:7, 4:6, 5:5, w:w), Reverse-phase high performance liquid chromatography with an evaporative light-scattering detector separated the triglyceride species of SLs. The products contained the newly synthesized peaks. Especially, one of peaks was distinctively increased with the increasing weight ratio from 2:8 to 5:5 while the peak of trilinolein (LLL) decreased vice versa. The effect of antioxidants such as catechin, BHT(Butylated hydroxytoluene), and their combinations on the oxidative stability in SL were investigated. Oxidative stability was carried out under oven test at $60^{\circ}C$ over 72 hr thereafter SLs were analyzed for total fatty acid content, rancimat, peroxide value, electronic nose and TBARS value. Among all combinations of antioxidant, the highest stability was obtained from 200 ppm of catechin. Besides, total tocopherol ($\alpha$, $\gamma$, and $\delta$-tocopherol), iodine and saponification value were analyzed in which iodine and saponification value of SLs were 151.19 and 182.35.

Recombinant Production and Antimicrobial Activity of an Antimicrobial Model Peptide (Uu-ilys-CF) Derived from Spoon Worm Lysozyme, Uu-ilys (개불 라이소자임 유래 항균성 모델 펩타이드(Uu-ilys-CF)의 재조합 단백질 생산 및 항균 활성)

  • Oh, Hye Young;Go, Hye-Jin;Park, Nam Gyu
    • Journal of Life Science
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    • 제31권1호
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    • pp.83-89
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    • 2021
  • Uu-ilys, an i-type lysozyme from spoon worm (Urechis unicinctus), is an innate immune factor that plays an important role in the defense against pathogens. It also possesses non-enzymatic antibacterial activity. Thus, there is a possibility to develop an antimicrobial model peptide from Uu-ilys. In this study, we report the design, production, and antibacterial activity of an Uu-ilys analog that exhibits antibacterial activity. The Uu-ilys structure was fragmented according to its secondary structures to predict the regions with antimicrobial activity using antimicrobial peptide (AMP) prediction tools from different AMP databases. A peptide containing the C-terminal fragment was predicted to exert antimicrobial activity. The chosen fragment was designated as an Uu-ilys analog containing the C-terminal fragment, Uu-ilys-CF. To examine the possibility of developing an AMP using the sequence of Uu-ilys-CF, recombinant fusion protein (TrxA-Uu-ilys-CF) was produced in an expression system that was heterologous. The produced fusion protein was cleaved after methionine leaving Uu-ilys-CF free from the fusion protein. This was then isolated through high performance liquid chromatography and reverse phase column, CapCell-Pak C18. The antibacterial activity of Uu-ilys-CF against different microbial strains (four gram-positive, six gram-negative, and one fungal strain) were assessed through the ultrasensitive radial diffusion assay (URDA). Among the bacterial strains tested, Salmonella enterica was the most susceptible. While the fungal strain tested was not susceptible to Uu-ilys-CF, broad spectrum antibacterial activity was observed.

Antimicrobial effect of Enterococcus faecalis BMSE-HMP005 isolated from human breast milk against multidrug-resistant bacteria (모유 유래 유산균 Enterococcus faecalis BMSE-HMP005의 다제내성 균에 대한 항균효과)

  • Lee, Jeong-Eun;Kim, Soo-bin;Yu, Du-na;Jo, So-Yeon;Kim, Ae-Jung;Kook, Moochang
    • Korean Journal of Food Science and Technology
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    • 제54권2호
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    • pp.209-217
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    • 2022
  • In this study, Enterococcus faecalis BMSE-HMP005 isolated from human breast milk demonstrated antimicrobial effects against multidrug-resistant (MDR) bacterial strains. The bacteriocin produced by E. faecalis BMSE-HMP005 was fractionated using reverse-phase high-performance liquid chromatography. This fraction showed antimicrobial effects against both gram-positive and gram-negative MDR bacteria. No hemolytic reactions were observed. E. faecalis BMSEHMP005 was resistant to vancomycin; however, kanamycin, ampicillin, and erythromycin showed minimum inhibitory concentrations that were lower than the acceptable range provided by the European Food Safety Authority. For artificial gastric juice and bile acid, the survival rates were 98.67% and 95.70%, respectively. These results show the potential utility of E. faecalis BMSE-HMP005 as a probiotic with remarkable antimicrobial effects against MDR bacteria.