• Title/Summary/Keyword: open column chromatography

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Biological Activities and Bioactive Compounds in the Extract of Acer tegmentosum Maxim. Stem (산겨릅나무 줄기추출물의 생리활성 및 유효성분 분리)

  • Hong, Bo-Kyong;Eom, Seok-Hyun;Lee, Chan-Ok;Lee, Ji-Won;Jeong, Jong-Hyun;Kim, Jae-Kwang;Cho, Dong-Ha;Yu, Chang-Yeon;Kwon, Yong-Soo;Kim, Myong-Jo
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.4
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    • pp.296-303
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    • 2007
  • Acer tegmentosum (Acereaceae) has been used a source of traditional medicines for the treatment of hepatic disorders in Korea. This research was conducted to determine biofunctional activities of A. tegmentosum stem extract and to identify its bioactive components. Methanolic extract from A. tegmentosum stem was partitioned by using organic solvents, including n-hexane, ethyl acetate, n-butanol, and water. Two compounds were isolated by using an ODS column chromatography from ethyl acetate soluble fraction shown to the strongest antioxidant activity ($RC_{50}=3.15\;{\mu}g/m{\ell}$) among the fractions. The isolated compounds were analyzed by $^1H$ and $^{13}C$ NMR, IR, UV/VIS, MS spectrum data and identified as catechin, ${\rho}-Hydroxyphenethyl$ alcohol $1-O-{\beta}-_D-(6'-O-galloyl)-glucopyranoside$. The compounds have shown strong antioxidant activity, with similar activity to BHA ($RC_{50}=2\;{\mu}g/m{\ell}$). Especially, ${\rho}-Hydroxyphenethyl$ alcohol 1-O-{\beta}-_D-(6'-O-galloyl)-glucopyranoside$ was shown strong anti-lipid peroxidative activity. However, the compounds were not shown antimicrobial activities. In antimicrobial activity assays, ethyl acetate soluble fraction was effective to bacterial inhibition, such as Escherichia coli and Klebsiella pneumonia, with minimum inhibitory concentrations in $125\;{\mu}g/m{\ell}$. Otherwise, antifungal activity against Candida albicans was shown in n-hexane soluble fraction exhibiting $63\;{\mu}g/m{\ell}$ of minimum inhibitory concentration. In anticomplementary activity assays, water soluble fraction was the most effective exhibiting 24% inhibitory activity.

Method Validation and Quantification of Lutein and Zeaxanthin from Green Leafy Vegetables using the UPLC System (UPLC를 이용한 lutein과 zeaxanthin의 분석법 검증 및 엽채류에서의 정량적 평가)

  • Kim, Suna;Kim, Ji-Sun
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.686-691
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    • 2012
  • The objective of this research is to present method development and validation for the simultaneous determination of lutein and zeaxanthin using ultra performance liquid chromatography (UPLC). Also, rapid quantification was performed on six green leafy vegetables (Allium tuberosum, Aster scaber, Hemerocallis fulva, Pimpinella brachycarpa, Sedum sarmentosum and Spinacia oleracea) that are commonly consumed in Korea. Separation and quantification were successfully achieved with a Waters Acquity BEH C18 ($50{\times}2.1mm$, $1.7{\mu}m$) column by 85% methanol within 5 min. Two compounds showed good linearity ($r^2$ > 0.9968) in $1-150{\mu}g/mL$. Limit of detection (LOD) and quantification (LOQ) for lutein and zeaxanthin were 1.7 and 5.1 g/mL and 2.1 and 6.3 g/mL, respectively. The RSD for intra- and inter-day precision of each compound was less than 10.69%. The recovery of each compound was in the range of 91.75-105.13%. Aster scaber and Spinacia oleracea contained significantly higher amounts of lutein ($4.06{\pm}0.24$ and $3.97{\pm}0.10mg$/100 g of fresh weight), respectively.

Mytilin B, an Antimicrobial Peptide from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus : Isolation, Purification, and Characterization (참담치(Mytilus coruscus) 혈구(hemocyte) 유래 항균 펩타이드 mytilin B의 정제 및 특성 분석)

  • Lee, Min Jeong;Oh, Ryunkyoung;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;Park, Jung Youn;Seo, Jung-Kil;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1301-1315
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    • 2018
  • We purified an antimicrobial peptide from the acidified hemocyte extract of Mytilus coruscus by $C_{18}$ reversed-phase high-performance liquid chromatography (RP-HPLC). The peptide was 4041.866 Da based on matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS) and the 25 amino acids of the N-terminus sequence were identified. Comparison of this sequence of the purified peptide with the N-terminus sequences of other antimicrobial peptides revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We also identified a 312 bp open-reading frame (ORF) encoding 103 amino acids based on the obtained amino acid residues. The nucleotide sequence of this ORF and the amino acid sequence also revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We synthesized two antimicrobial peptides with an alanine residue in the C-terminus, and designated them mytilin B1 and B2. These two antimicrobial peptides showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus and Streptococcus parauberis (minimal effective concentration, MECs $41.6-89.7{\mu}g/ml$), gram-negative bacteria, including Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, and Vibrio ichthyoenteri (MECs $7.4-39.5{\mu}g/ml$), and the fungus Candida albicans (MECs $26.0-31.8{\mu}g/ml$). This antimicrobial activity was stable under heat and salt conditions. Furthermore, the peptides did not exhibit significant hemolytic activity or cytotoxic effects. These results suggest that mytilin B could be applied as alternative antibiotic agent, and they add to the understanding of the innate immunity of hard-shelled mussels.

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.

Urinary Mutagenicity, Physical Symptoms and Stress of Nurses Handling Anticancer Drugs (항암제 취급 간호사의 소변중 돌연변이 유발능과 자각증상 및 스트레스)

  • 김봉임
    • Journal of Korean Academy of Nursing
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    • v.26 no.4
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    • pp.963-975
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    • 1996
  • The purpose of this study was to call attention to the mental, physical and occupational hazards of the anticancer-drug-handling nurses by examining the possible urinary mutagenicity and measuring physical symptoms and stress level of the nurses exposed to anticancer drugs. The experimental group of the urinary mutagenicity assay was 14 nurses handling anticancer drugs at the medical wards of a hospital located in J city ; the control group was 12 psychiatric nurses of the same hospital. The test material was the nurses' 24hrs urine, which was concentrated by XAD-2 column chromatography. Tester strains were TA98(±S9 mix), TA100(±S9 mix), TA1535(±S9 mix) and TA1537(±S9 mix) ; Salmonella mammalian-microsomal test(Ames test) was employed for the urinary mutagenicity assay. The physical symptoms of which the nurses experienced were investigated through self-reports on open-questionnaires. The stress levels of the experimental group were measured by a stress measuring instrument developed by this author. Reliability of this instrument was found to be adequate (Cronbach's Alpha=0.9079). To ascertain the urinary mutagenicity of the experimental group, the mean and the standard deviation of the colonies of Tester strains appearing on the minimal plates were taken and compared differences between two groups. T-test was employed for the significance test of two groups. The physical symptoms were compared between the two groups through the analysis of the nurse' self-reports. The mean and standard deviation of the stress levels of the experimental group were also calculated and were examined through t-test. The results were summarized as follows : 1. The experimental group revealed significantly higher urinary mutagenicity both in the activation method test and the non-activation method test of the tester strains TA98, TA100 and TA1535. In the case of TA1537, two groups showed no difference in the non-activation method test, but the activation method revealed difference. 2. The physical symptoms were also much more frequently reported in the experimental group. 79.3% of the experimental group reported more than 1 kind of physical symptoms. On the other hand, 33.2% of the control group complained of 1 kind of physical symptom. The items with high symptom frequency were 'headache', 'itching sensation', 'corneal congestion', 'skin allergy' 3. The mean score of stress in the experimental group was 2.41(range 1-4). The experimental group showed the stress level above 2.0 in the 14 of 15 items in all. The highest stress level were recorded in the following items in the order quoted, 'I fear that anticancer drug may touch any part of body while handling it.', 'I feel concerned there is no protective countermeasure against anticancer drug handling.', 'I am afraid the anticancer drug handling may produce a fetal loss in the future'.

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Toxicokinetics of rifapentine in beagle dogs (Beagle dog에 있어서 rifapentine의 독성동태연구)

  • Shin, Ho-chul;Lee, Hye-suk;Cha, Shin-woo;Han, Sang-seop;Roh, Jung-ku;Kim, Jin-suk;Lee, Won-chang
    • Korean Journal of Veterinary Research
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    • v.35 no.4
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    • pp.815-822
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    • 1995
  • The toxicokinetics of rifapentine was studied after an oral administration to beagle dogs. High-performance liquid chromatography(HPLC) using column-switching technique was performed to determine the serum concentrations of rifapentine. The pharmacokinetic profiles of rifapentine were analysed using one-compartment open model. Following a single oral administration of 10mg/kg, pharmacokinetic parameters were determined as follows: maximum serum concentration($C_{max}$), $28.90{\mu}g/ml$; maximum concentration time($T_{max}$), 3.7hr; elimination half-life($t_{1/2}$, 4.7hr; area under the curve(AUC), $339.0{\mu}g{\cdot}hr/ml$; volume of disiribution/bioavailability (Vd/F), 0.21 l/kg; lag time, 24min; absorption rate constant($k_a$), $0.445hr^{-1}$; elimination rate constant($k_{el}$), $0.148hr^{-1}$. After 6 month multiple oral doses of 10mg/kg/day, parameters were as follows: $C_{max}$, $34.40{\mu}g/ml$; $T_{max}$, 2.6hr; $t_{1/2}$, 6.7hr; AUC, $391.3{\mu}g{\cdot}hr/ml$; Vd/F, 0.291/kg; $k_a$, $0.976hr^{-1}$; $k_{el}$, $0.104hr^{-1}$. The consistant kinetic parameters after a single and multiple oral administration show that there was no accumulation of rifapentine after 6 month oral administration. We also simulated the concentration of rifapentine after oral multiple administration of 10 and 50mg/kg/ day, based on the parameters obtained form the single administration. The measured serum concentrations of rifapentine were well fitted to the simulated results. The simulated results show that rifapentine readily reaches to steady-state after about 3 doses and the steady-state serum concentrations($C_{ss}$) are fluctuated in between $2.2{\sim}25.2{\mu}g/ml$, and $10.6{\sim}125.2{\mu}g/ml$ at the doses of 10 and 50mg/kg/day, respectively.

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The Antiproliferative Effects of Compounds Isolated from Schisandra chinensis (오미자로부터 분리된 화합물의 암세포 증식 억제 효과)

  • Suh, Won-Se;Park, So Yeon;Min, Byung Sun;Kim, Sea Hyun;Song, Jeong Ho;Shim, Sang Hee
    • Korean Journal of Food Science and Technology
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    • v.46 no.6
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    • pp.665-670
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    • 2014
  • We isolated twelve lignans and three terpenoids were isolated from the n-hexane fraction of Schisandra chinensis extract. Using spectroscopic data and comparison with available literature, the following compounds were identified: (1) wuweizisu C, (2) gomisin N, (3) deoxyschisandrin, (4) gomisin A, (5) schisandrin, (6) chamigrenal, (7) schisanlactone D, (8) methylgomisin O, (9) angeloylgomisin O, (10) (-)-gomisin $L_2$, (11) schisandronic acid, (12) (-)-gomisin $L_1$, (13) (+)-gomisin $K_3$, (14) gomisin J, and (15) tigloylgomisin H. Notably, this was the first finding that compound (8) was isolated from this plant. Each compound was evaluated for its in vitro cytotoxic activities toward HL-60 (human leukemia), HeLa (human cervical carcinoma), and MCF-7 (breast cancer) cell lines. Compounds (7), (8), and (9) exhibited strong cytotoxic effects on HL-60 ($IC_{50}$ 7.37, 6.60, and $8.00{\mu}M$, respectively), whereas compound (6) exhibited weak cytotoxicity towards MCF-7 ($IC_{50}$ $30.50{\mu}M$). In addition, compound (8) showed the strongest activity towards HeLa cells ($IC_{50}$ $1.46{\mu}M$).

Inhibitory Effects of Carex humilis Extract on Elastase Activity and Matrix Metalloproteinase-1 Expression (산거울 추출물의 Elastase 활성 저해 및 Matrix Metalloproteinase-1 발현 억제 효과)

  • Park, Sun-Hee;Lee, Kang-Hyuk;Han, Chang-Sung;Kim, Ki-Ho;Kim, Young-Heui
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.2
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    • pp.129-136
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    • 2010
  • In order to evaluate anti-wrinkle activity of Carex humilis extract, free radical scavenging activity, elastase inhibitory activity and reduction of expression Matrix metalloproteinase-1 (MMP-1) mRNA and MMP-1 protein were investigated. The roots of Carex humilis were extracted with 95 % ethanol and successively partitioned with organic solvents with increasing polarity of the solvents. Each fraction of organic solvent were investigated by using free radical scavenging activity and elastase inhibitory activity test. Among them, EtOAc fraction showed antioxidant activity ($SC_{50}$=4.89 ${\mu}g/mL$) and elastase inhibitory activity ($IC_{50}$=23.5 ${\mu}g/mL$). EtOAc fraction was developed on silica gel by open-column chromatography and consecutively re-developed on C18 resin by prep-HPLC to give ${\alpha}$-viniferin as a major component, which was confirmed by spectrometric analysis. In the assay on expression of MMP-1 mRNA by RT-PCR and protein by western-blot, EtOAc layer (10 ~ 100 ${\mu}g/mL$) was reduced about 50 ~ 60 %, 50 ~ 65 % respectively and ${\alpha}$-viniferin (0.5 ~ 2 ${\mu}g/mL$) was inhibited about 60 ~ 75 %, 55 ~ 65 % respectively in human fibroblast. Therefore, our findings suggest that EtOAc layer of Carex humilis containing ${\alpha}$-viniferin can be useful as an active ingredient for cosmeceuticals of anti-wrinkle effects.

Anti-oxidant and α-Glucosidase Inhibition Activity of Extracts or Fractions from Diospyros lotus L. Leaves and Quantitative Analysis of Their Flavonoid Compounds (고욤잎 추출물과 분획물의 항산화 및 alpha-glucosdiase 저해 활성 및 플라보노이드 화합물의 정량)

  • Kim, Seon-Young;Kim, Sang Jun;Kim, Ji-Ae;Kim, Da Hye;Kwak, Seol Hwa;Chung, Chang Ho;Jeon, In Hwa;Jang, Seon Il;Jeong, Seung-Il
    • Journal of Life Science
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    • v.24 no.9
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    • pp.935-945
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    • 2014
  • Persimmon leaves were commonly consumed as beverages, but were also used as popular folk medicine in Asia. The purpose of this work was to assess the biological activities of Diospyros Lotus L. extracts (DLLE). Various solvent extracts, including n-Hexnae, $CHCl_3$, EtOAc, and n-BuOH fractions, were obtained from the methanol extract of Diospyros Lotus L. leaves. The increasing interest in the powerful biological activity of plant phenolics and flavonoids outlined the necessity for determining their content in medicinal herbs. In this study, the total polyphenol and flavonoid contents (TPC and TFC) in the EA fraction were higher than those of other fractions. The biological activities of DLLE were tested using the 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity assay, as well as superoxide dismutase (SOD) activity as an anti-oxidant effect and ${\alpha}$-glucosidase inhibitory activity as an anti-diabetic effect. The EA fraction with high TPC and TFC values showed the highest anti-oxidant effect and high ${\alpha}$-glucosidase inhibition. The EA fractions were further purified into eight fractions using open column chromatography. Higher anti-oxidant and anti-${\alpha}$-glucosidase activity were observed in polar fractions. The content of the flavonoids, including quercein-3-O-rutinoside, kaempferol-3-O-glucoside, myricetin, luteolin, and kaempferol, were analyzed in effective fractions using high-performance liquid chromatography (HPLC). The results suggest that DLLE have anti-oxidative and anti-diabetic effects and thus, have the potential as anti-diabetic materials and as a source for natural health products.

Isolation and Identification of Flavonoids from Corn Silk (옥수수수염에 함유된 Flavonoids의 분리 및 동정)

  • Kim, Sun-Lim;Kim, Mi-Jung;Lee, Yu-Young;Jung, Gun-Ho;Son, Beom-Young;Lee, Jin-Seok;Kwon, Young-Up;Park, Yong-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.4
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    • pp.435-444
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    • 2014
  • This study was carried out to isolate and characterize the flavonoids present in corn silks. Maysin content in the unpollinated corn silks (Kwangpyeongok) showed its highest level at 3 days after silking, and decreased thereafter, while the content of open pollinated silks were consistently decreased after silking. This result indicates that the maysin content is considerably affected by the pollination of corn silk. Unpollinated corn silks were collected with excising, and ethanol employed to extract flavonoids at common temperature for 9 days. After extraction, chlorophyll, lipids etc. were removed with methylene chloride, then submitted to flash column cartridge ($150{\times}40mm$ i.d.) packed with a preparative $RP-C_{18}$ bulk packing material ($125{\AA}$, $55-105{\mu}m$), and monitored at 352 nm. Four fractions, fraction-I, -II, -III, and -IV, were isolated from ethanolic extract of corn silks. Absorption spectrum of fraction I showed its maximum intensity (${\lambda}_{max}$) at 327 nm and 239 nm, fraction-II showed its maximum intensity at 339 nm and 274 nm, fraction-III showed its maximum intensity at 345 nm and 277 nm, and fraction-IV showed its maximum intensity at 352 nm, 270 nm, 257 nm, respectively. On the baisis of ESI micro-TOF analysis, fraction-I was identified as chlorogenic acid (m/z 355, 3-(3,4-dihydroxycinnamoyl) quinic acid, $C_{16}H_{18}O_9$), fraction-II identified as a mixture of chlorogenic acid and luteolin 3'-methyl ether 7-glucuronosyl-($1{\rightarrow}2$)-glucuronide (m/z 653, $C_{28}H_{28}O_{18}$), fraction-III identified as a mixture of chlorogenic acid luteolin 7-O-neohesperidoside (m/z 595, $C_{27}H_{30}O_{15}$), and luteolin 3'-methyl ether 7-glucuronosyl-($1{\rightarrow}2$)-glucuronide, and fraction-IV identified as maysin (m/z 577, 2"-O-${\alpha}$-L-rhamnosyl-6-C-(6-deoxy-xylohexose-4-ulosyl)luteolin, $C_{27}H_{28}O_{14}$), respectively. From the ethanolic extract of corn silks, fraction-I was obtained about 35 mg/100 g F.W., fraction-II was about 48 mg/100 g F.W., fraction-III was about 46 mg/100 g F.W., and fraction-IV was about 138 mg/100 g F.W., respectively.