• Title/Summary/Keyword: HPLC Chromatography

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The antioxidative and cytoprotective effect of Lonicerae japonicae Flos water extracts on the ultraviolet(UV)B-induced human HaCaT keratinocytes (금은화 물추출물의 항산화 효과와 Ultraviolet(UV)B로 유도된 사람 각질형성세포 손상에 대한 보호효과)

  • Seo, Seung-Hee;Bae, Gi-Sang;Choi, Sun Bok;Jo, Il-Joo;Kim, Dong-Goo;Shin, Joon-Yeon;Song, Ho-Joon;Park, Sung-Joo;Choi, Mee-Ok
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.63-71
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    • 2014
  • Objectives : Lonicerae japonicae Flos(LJF) has been reported to exhibit anti-oxidant, anti-inflammatory, anti-viral, anti-rheumatoid properties. However, it is still largely unknown whether LJF inhibits the ultraviolet(UV)B-induced oxidative damage in human HaCaT keratinocytes. Therefore in this paper, we investigated the anti-oxidative capacity and protective effect of LJF against UVB-induced oxidative demage in human HaCaT keratinocytes. Methods : To evaluate the anti-oxidative activity of LJF extracts, we measured total phenolic contents, total flavonoid contents, antioxidant capacity, and superoxide scavenging activity. To give an oxidative stress to HaCaT cells, UVB was irradiated with $200mJ/cm^2$ to HaCaT cells. To detect the protective effect of LJF against UVB, we measured cell viability, DNA fragmentation and reactive oxygen species (ROS) production. In addition, we performed high-performance liquid chromatography (HPLC) analysis to find a major component of LJF. Results : LJF contained phenolic and flavonoid contents, and showed the anti-oxidant and superoxide scavenging activity. The UVB-induced oxidative conditions led to the cell death, DNA fragmentation and reactive oxygen species (ROS) production. However, pretreatment with LJF reduced oxidative conditions, including inhibition of cell death, DNA fragmentation and ROS production. In addition, we found out chlorogenic acid as major component of LJF. Conclusions : These results could suggest that LJF contained anti-oxidative contents and exhibited protective effects against UVB on human HaCaT keratinocytes. And the effective compound of LJF which could show protective activities against UVB is chlorogenic acid. Thus, LJF and chlorogenic acid would be useful for the development of drug or cosmetics treating skin troubles.

Analysis of selenoaminoacids and selenoproteins in blood serum of sows fed by selenium fortified feed (셀레늄강화 사료를 먹인 모돈 혈청에서의 셀레노아미노산 및 셀레노단백질 분석)

  • Park, Myungsoon;Lee, Sung Hoon;Pak, Yong-Nam
    • Analytical Science and Technology
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    • v.28 no.3
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    • pp.196-203
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    • 2015
  • Selenium species (inorganic selenium, selenoaminoacids, and selenoproteins) were analyzed using anion exchange and affinity chromatography, which were connected to ICP/MS for the blood serum of sows fed by seleniumfortified feed. The Anion Exchange PRP X-100 column was used for the analysis of inorganic selenium (Se4+ and Se6+) and selenoaminoacids. The HEP column was used to separate SelP from GPx+SeAlb in selenoproteins. A quantitative analysis was performed using the post-column isotope dilution technique. The lactating sows were divided into three groups and fed by selenium fortified feed (organic 0.3 mg/kg, 0.6 mg/kg and inorganic 0.6 mg/kg) for four weeks. The test groups showed increases in selenoaminoacids compared with the control group, except the inorganic feed group. There was no significant difference between the organic feed groups. All test groups showed increases in selenoproteins. In particular, SelP showed a large increase that was 1.5 times higher than the other proteins.

Identification and Quantification of Glucosinolates in Rapeseed (Brassica napus L.) Sprouts Cultivated under Dark and Light Conditions

  • Lee, Min-Ki;Arasu, Mariadhas Valan;Chun, Jin-Hyuk;Seo, Jeong Min;Lee, Ki-Teak;Hong, Soon-Taek;Kim, In Ho;Lee, Yong-Hwa;Jang, Young-Seok;Kim, Sun-Ju
    • Korean Journal of Environmental Agriculture
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    • v.32 no.4
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    • pp.315-322
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    • 2013
  • BACKGROUND: This study was performed for the identification and quantification of glucosinolate (GSL) contents in seven varieties of rapeseed (Brassica napus L.) sprouts cultivated under dark and light conditions. METHODS AND RESULTS: Crude glucosinolates (GSLs) were desulfated by treating with aryl sulfatase and purified using diethylaminoethyl sepharose (DEAE) anion exchange column. Individual GSLs were quantified using high-performance liquid chromatography (HPLC) with electrospray ionization-tandem mass spectrometry (ESI-MS/MS). Eleven GSLs including six aliphatic (progoitrin, sinigrin, glucoalyssin, gluconapoleiferin, gluconapin, and glucobrassicanapin), four indolyl (4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrassicin) and one aromatic (gluconasturtiin) were identified based on the fragmentation patterns of MS spectrum. Aliphatic GSLs were noted as the predominant group with average 85.2% of the total contents. The most abundant GSLs were progoitrin which was ranged at $8.14-118.68{\mu}mol/g$ dry weight (DW). The highest total GSL amounts were documented in 'Hanra' ($146.02{\mu}mol/g$ DW) under light condition and 'Mokpo No. 68' ($86.67{\mu}mol/g$ DW) in dark condition, whereas the lowest was in 'Tamra' (30.13 and $14.50{\mu}mol/g$ DW) in both conditions. The sum of aliphatic GSLs attributed > 80% in all varieties, except 'Tamra' (67.7% and 64.9% in dark and light conditions, respectively) in the total GSL accumulation. Indolyl GSLs were ranged $2.41-15.73{\mu}mol/g$ DW, accounted 2.78-33.6% of the total GSLs in rapeseed varieties. CONCLUSION(S): These results provide valuable information regarding potential beneficial GSL contents individually. This study attempts to contribute to knowledge of the nutritional properties of the different varieties of rapeseed plants. These results may be useful for the evaluation of dietary information.

Solid Phase Extraction(SPE) of Curcuminoids from Turmeric by Optimization Analytical Condition (최적 분석조건에 의한 강황으로부터 Curcuminoids의 고체상추출(SPE))

  • Lee, Kwang Jin;Ma, Jin Yeul;Kim, Young Jun;Kim, Young Sik
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.10
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    • pp.4927-4935
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    • 2012
  • Turmeric is a medicinal herb with various pharmacological activities. Curcumin, demethoxycurcumin(DMC) and bisdemethoxycurcumin(BDMC) were extracted from turmeric by dipping and ultrasonic wave method. And extraction efficiency was identified using solid phase extraction(SPE). The extracted sample were simultaneously separated and analyzed from three kinds of commercial $C_{18}$ reversed-phase high performance liquid chromatography using water and acetonitrile as mobile phase with isocratic elution mode. Flow rate 1.0mL/min, injection volume $10{\mu}L$ and column oven temperature $40^{\circ}C$ at 425nm wavelength has been conducted. From the experimental results, the optimum mobile phase composition of water/acetonitrile 50/50vol% using RS tech column. It is evident that the amount of curcuminoids extracted(extraction time 4h) by 100% MeOH was higher than any aqueous MeOH composition. Finally, in 100% water extraction, the amount(mAU${\times}$mim) of curcuminoids extracted by SPE was 14.3 and 24.5 times respectively higher than ultrasonic wave and dipping method. The shown results can be applied as sources for pharmaceuticals and functional material.

Inhibitory Effects of Aralia cordata Thunb Extracts on Nitric Oxide Synthesis in RAW 264.7 Macrophage Cells (독활(Aralia cordata Thunb) 추출물의 Nitric Oxide Synthesis 저해효과)

  • Kang, Chang-Ho;Koo, Ja-Ryong;So, Jae-Seong
    • Korean Journal of Food Science and Technology
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    • v.44 no.5
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    • pp.621-627
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    • 2012
  • Assessment was made of the effects of Aralia cordata Thunb (DH) on the cell proliferation, inducible nitric oxide synthase (iNOS) mRNA gene expression and nitric oxide (NO) production in RAW 264.7 macrophage cells. For the screening of anti-inflammatory activities, ethanolic extracts of 55 species of traditional herbal medicines were examined for inhibitory effects, and it was confirmed that DH possessed inhibitory effects on NO production. As a result, DH significantly decreased the production of NO and iNOS gene expression at a concentration of $250{\mu}g/mL$. The chloroformsoluble fractionates have the strongest No synthesis inhibitory effect. It is presumed that the inhibition of NO production in LPS-stimulated RAW 264.7 cells by DH components occurred via the modulation of iNOS and DH, and that the active compound from DH may be useful for therapeutic management of inflammatory-associate diseases.

Comparison of Chlorophyll Contents and Patterns of Korean and Japanese Powder Green Tea (한일 말차(抹茶)의 클로로필 함량 및 패턴 비교)

  • 김기선;고즈쿠에노부유키;한재숙
    • Journal of the East Asian Society of Dietary Life
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    • v.13 no.6
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    • pp.586-592
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    • 2003
  • This research used High Performance Liquid Chromatography(HPLC) to analyze and compare the content and the pattern of chlorophyll to find the difference in quality, water temperature, and the stirring time between the three different types of Korean and Japanese powder green tea, which were currently on the market. Chlorophyll contents of Japanese-made medium quality tea (S5: 8.34mg/g), and low quality tea (S6: 8.23mg/g) were relatively high, and that of Korean-made medium quality tea (S2 ; 4.87mg/g) was lower. The water temperature of 85$^{\circ}C$ and 95$^{\circ}C$ were high except in Korean-made medium and low quality powder green tea (S2 and S3). Japanese-made powder tea required 60 seconds of stirring time and Korean-made tea required 15 seconds, but 30 seconds seem to be the ideal stirring time for both of them. Chlorophyll a, b, pheophytin a, b, and pheophorbide were confirmed from the chlorophyll pattern. There were 1.5∼2 times more of chlorophyll a in Japanese-made one compared to that of Korean-made one; also 1.5∼2 times more of pheophytin a in Korean-made one than Japanese-made one. It might be concluded that Japanese-made powder green tea was higher in quality.

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Cirsium japonicum Extracts Show Antioxidant Activity and PC12 Cell Protection against Oxidative Stress (좁은잎 엉겅퀴 추출물의 산화방지 활성 및 산화적 스트레스에 대한 PC12 세포 보호효과)

  • Jang, Miran;Kim, Gun-Hee
    • Korean Journal of Food Science and Technology
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    • v.48 no.2
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    • pp.172-177
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    • 2016
  • The phenolic compounds, antioxidant activity and neuronal cell protective effect of Cirsium japonicum extract were evaluated in this study. High performance liquid chromatography mass analysis showed that C. japonicum was composed of chlorogenic acid, linarin, and pectolinarin. C. japonicum extract showed its antioxidant activity with half-maximal inhibitory concentrations of 567 and $130{\mu}g/mL$ by DPPH and ABTS radical scavenging activity, respectively. The total antioxidant capacities of C. japonicum via DPPH, ABTS, and FRAP assays were 11.32, 100.15, and $12.76{\mu}g/mL$ trolox equivalents, respectively. In addition, the neuroprotective effect of C. japonicum extract was investigated by measuring cell viability via MTT, LDH and DCF-DA assay using $H_2O_2-damaged$ PC12 cells. C. japonicum extract showed neuronal cell protective effects in a dose-dependent manner. These results indicated that C. japonicum extract has potent antioxidant and neuronal protective effects. Therefore, C. japonicum can be regarded as an effective and safe functional food resource as natural antioxidants, and may decrease the risk of neurodegenerative disorders.

Comparison of the Biochemical Activities of Commercial Yogurts and Lactobacillus acidophilus-containing Yogurt (시판용 요구르트와 Lactobacillus acidophilus 요구르트의 생화학적 활성의 비교)

  • Ryu, Jae-Ki;Lee, Hyeong-Seon;Koo, Bon-Kyung;Kim, Hyun-Kyung
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.2
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    • pp.59-64
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    • 2015
  • Lactic acid-producing bacteria such as Lactobacillus spp. function to ferment carbohydrates and produce ATP. Such Lactobacillus spp. are used for the production of commercial yogurts. Lactobacillus spp. are beneficial to the intestinal tract, and Lactobacillus acidophilus-containing yogurts have received considerable attention because of their preventive effects against early-stage cancer of the large intestine. In this study, lactic acid-producing bacteria were cultured from three different groups: commercial solid yogurt (for eating), commercial liquid yogurt (for drinking), and Lactobacillus acidophilus-containing yogurt. We first determined the optimum culture conditions for Lactobacillus spp. and then analyzed turbidity and pH in order to compare the growth abilities and lactic acid-production capacities among the groups. Finally, high-performance liquid chromatography was used to measure the lactic acid content in the culture supernatants, and the antibacterial activities against Staphylococcus aureus and Escherichia coli were compared among the three groups. The optimum culture conditions for Lactobacillus spp. were MRS medium at $25^{\circ}C$, for 24 h. The highest turbidity was found in L. acidophilus-containing yogurt, followed by liquid yogurt and solid yogurt. Similarly, the highest lactic acid production ability was found in L. acidophilus-containing yogurt, followed by liquid yogurt and solid yogurt. Culture supernatants from the three groups did not show any antibacterial activity towards S. aureus; however, supernatants derived from L. acidophilus-containing yogurt resulted in a 1.8 mm inhibitory zone against E. coli in a paper disk diffusion test. These results revealed the high level of lactic acid-production capacity and antibacterial activity in L. acidophilus-containing yogurt.

Exposure Assessment of PCDD/Fs and Monitoring of Health Effects on Workers and Resident near the Waste Incinerators in Korea (국내 일부 소각장 근로자와 주변지역주민들의 PCDDs/Fs 노출과 건강 영향 평가)

  • Hong, Yun-Chul;Lee, Kwan-Hee;Kwon, Ho-Jang;Jang, Jae-Yeon;Leem, Jong-Han
    • Journal of Preventive Medicine and Public Health
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    • v.36 no.4
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    • pp.314-322
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    • 2003
  • Objectives : In this study, the exposure status of the hazardous substances from incinerators, such as polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), were studied , and the relationship between the exposure of these hazardous substances and their heath effects on the workers and residents near municipal solid waste (MSW) incinerators and an industrial incinerator investigated. Methods : Between July 2001 and Jure 2002, 13 workers at two MSW incinerators, 16 residents from the area around the two MSW incinerators, 6 residents from the control area, and further 10 residents near an industrial incinerator, estimated to emit higher levels of hazardous substances, were interviewed. Information, including sociodemographic information, personal habits, and work history, detailed gynecologic and other medical history were collected through interviews. Blood samples were also collected from 45 subjects, and analyzed for PCDD/DFs, by high resolution gas chromatography -high resolution mass spectrometry, using the US EPA 1613 method. In addition to the questionnaire survey, urinary concentrations of 8-hydroxydeoxyguanosine (8-OH-dG) and malondialdehyde (MDA) were measured as oxidative injury biomarkers. The urinary concentrations of 8-OH-dG were determined by in vitro ELISA, and the MDA by HPLC, using u adduct with thiobarbituric acid. Results : The PCDD/DFs concentrations in the residents near the industrial incinerator were higher than those in the controls, workers and residents near the MSW incinerators. The average TEQ (Toxic Equivalencies) concentrations of the PCDD/DFs in residents near the industrial incinerator were 53.4pg I-TEQs/g lipid. The estimated daily intakes were within the tolerable daily intake range (1-4 pg I-TEQ/Kg bw/day) suggested by WHO (1997) in only 30% to the people near the industrial incinerator. Animal studies have already shown that even a low body border of PCDD/DFs, such as 10 ng TEQ/kg bw, can cause oxidative damage in laboratory animals. Our study also showed that the same body burden of PCDD/DFs can cause oxidative damage to humans. Conclusions : The exposures to PCDD/DFs and the oxidative stress of residents near the industrial incinerator, were higher than those in the controls, workers and residents near the MSW incinerators. Proper protection strategies against these hazardous chemicals are needed. Because a lower body burden of PCDD/Fs, such as 10ng TEQ/kg bw, can cause oxidative damage, the tolerable daily intake range should be restrictedly limited to 1pg I-TEQ/kg bw/day.

A Comparative Study of High-Performance Liquid Chromatographic Method for Analysis of Ginseng Saponin (인삼(人蔘)사포닌분석(分析)을 위한 고속액체(高速液體)크로마토그래피법(法)의 비교연구(比較硏究))

  • Choi, Jin-Ho;Park, Kil-Dong;Han, Kang-Wan;Oh, Sung-Ki
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.11 no.3
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    • pp.81-86
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    • 1982
  • Ginseng saponins separated from Panax ginseng C.A. Meyer were analyzed by high-performance liquid chromatography using a carbohydrate analysis column. The effect on the resolution and retention time of each ginsenosides, as well as the addition effect of n-butanol on a acetonitrile/water system, was examined using various proportions of acetonitrile/water system (80/20-94/6) which have been used with typical solvent of carbohydrate analysis column. The retention time of each ginsenosides was greatly affected by the compositions and mixture proportions of the mobile phase and also markedly increased as the proprotion of acetonitrile in mobile phase increased. It was proved that acetonitrile/water system (80/20) and acetonitrile/water/n-butanol system (86/14/10) were very effective mobile phases for diol and triol sapoin analyses, respectively. According to the result obtained by this method, the PT/PD ratios of white and red ginseng saponins were 0.401 % and 0.561 %, respectively. Red ginseng increased PT/PD ratio, compared with white ginseng. This is attributed to the change of saponin pattern by processing method and the change of PT/PD ratio would greatly influence on the biochemical and pharmacological effects of ginseng and its products.

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