• Title/Summary/Keyword: Sep-pak $C_{18}$

Search Result 87, Processing Time 0.023 seconds

New Methods for Isolation of Sesquiterpene from Panax ginseng (인삼 Sesquiterpene의 새로운 분리방법)

  • 위재준;신지영
    • Journal of Ginseng Research
    • /
    • v.21 no.3
    • /
    • pp.214-218
    • /
    • 1997
  • New simple methods for the Isolation of sesquiterpenes from Panax ginseng were developed. First, volatile compounds were isolated by simultaneous distillation and extraction (SDE) with 30% methanol and $\alpha$-hexane instead of water and ethyl ether/pentane (1:1). Secondly, head space volatiles in U-shaped tube at 7$0^{\circ}C$ were passed through C18 Sep-Pak by nitrogen gas streaming and the adsorbed volatiles were fluted by $\alpha$-hexane. TLC analysis showed that the volatile concentrates consisted mainly of terpenes when colored by vanillin-sulfuric and. GC/MS data revealed that approximately 30 sesquiterpenes of molecular weight 204 occupied 81.1% or more of the volatile concentrates isolated by those two newly developed methods. Among these, alloaromadendrene, germacrene B, isocaryophyllene, $\alpha$-neoclovene, ${\gamma}$-muurolene, $\beta$-panaslnsene, and $\alpha$-humulene were identified as being major sesqulterpenes by authentic samples or literatme search Key words : Panax ginseng, volatile compound, sesquiterpene, isolation, new method, GC/MS.

  • PDF

Determination of Ursodeoxycholic Acid in Crude Drug Formulations by HPLC and SPE Using Selective Pre-column Derivatization with 2-Bromoacetyl-6-methoxynaphthalene (2-Bromoacetyl-6-methoxynaphthalene을 형광유도체화제로 HPLC와 SPE를 이용한 생약제제 중 Ursodeoxycholic acid의 정량)

  • 진창화;임수희;이기진;심형섭;조의환;염정록
    • YAKHAK HOEJI
    • /
    • v.46 no.6
    • /
    • pp.392-397
    • /
    • 2002
  • A simple and sensitive high performance liquid chromatographic method to quantitate ursodeoxycholic acid in crude drug pharmaceuticals was investigated. Ursodeoxycholic acid react with 2-bromoacetyl-6-methoxynaphthalene (Br-AMN) in the presence of triethylamine to form highly fluorescent derivative. The derivatization procedure was performed at 7$0^{\circ}C$ and completed within 30 min. The optimal wavelength of the fluorescence detector are λ$_{ex}$=300 nm and λ$_{em}$ = 460 nm. The LOD of the ursodeoxycholic acid was 25 ng/mι based on the S/N =3, and the LOQ was 80 ng/mι based on S/N = 10. Crude drug pharmaceuticals pretreated by solid phase extraction (Sep-pak $C_{18}$ cartridge) which were shown very good separation and recovery values for the compound.d.

A Study on the Determination of Residual Antibiotics and synthetic antibacterial agents in Meat(I) -Microbiological Assay- (식육중의 잔류 항생.항균제의 검정에 관한 연구(I) -생물학적 검정법-)

  • 류재천;송윤선;박종세;장준식;신보승언
    • Journal of Food Hygiene and Safety
    • /
    • v.8 no.1
    • /
    • pp.1-8
    • /
    • 1993
  • 육류중에 잔류하는 항생물질 및 항균제를 검출하기 위해 본 실험에서는 3종의 균주 Bacillus subtilis ATCC 6633, Micrococcus luteus ATCC 9341, BAcillus cereus var. mycoides ATCC 11778을 사용하여 실험하였다. 시료의 clean-up은 항생·항균제의 물리화학적 성질을 고려하여 우선 McIlvaine buffer를 가하여 homogenize하고 hexane으로 defatting 시킨 후, chloroform으로 추출한 액과 Sep-Pak C18과 Bakerbond SPE carboxylic acid column에 흡착시킨 후 추출한 액을 각각 시험용액 A, B, C,로 하였다. 각 test solution을 paper disk를 사용하여 함균 배지에 올려놓고 overnight culture 후 inhibition pattern을 통해 여러 종류의 항생·항균제를 계통적으로 검출하였다. 본 실험에서 macrolide계와 tetracycline계 등은 0.1ppm 이하의 detection limit을 보였으며, penicillinrP는 0.001ppm이하의 높은 detection limit을 나타내므로서 시료중에 잔류하는 극미량까지도 검출할 수 있었다. 본 방법은 식육중의 잔류 항생·항균제를 동시에 간단하게 계통적으로 분류하는데 있어 좋은 방법이라고 생각되며 항생·항균제의 체계적인 1차 screening 수단으로서 유용한 방법이라 사료된다.

  • PDF

Antimicrobial Activity and Characteristics of Amblytropis pauciflora Kitagawa Extract (털새동부 추출물의 항균효과 및 특성)

  • 김현수
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.27 no.5
    • /
    • pp.993-999
    • /
    • 1998
  • Methanol extract of roots of Amblytropis pauciflora Kitagawa showed the antimicrobial activity to three test strains. Antimicrobial spectra of various extracts of Amblytropis pauciflora Kitagawa were tested against 24 strains of bacteria and fungi. The crude methanol extract inhibited the growth of 12 strains of bacteria and Asp. fumigatus with the exception of yeasts. The properties of the antimicrobial substance were very stable under heat(at 12$0^{\circ}C$), acid(pH 3.0) and alkali(pH 11.0) treatment. Only the root harvested in spring showed the antimicrobial activity. Among the components extracted by butanol, ginseng saponin Rg1 and various saponin-like materials were detected by TLC analysis using a plate of silica gel 60F254. The antimicrobial compound was purified by methanol extraction, activated charcoal column chromatography, Sep-pak(C18) pretreatment and reverse phase HPLC. The purified compound was detected at 13.520 min as a single peak(about 98% purity) through the HPLC analysis.

  • PDF

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
    • /
    • v.31 no.6
    • /
    • pp.1471-1476
    • /
    • 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.

  • PDF

Purification of Antibacterial Peptide from the Skin of the Catfish Silurus asotus (메기의 껍질로부터 항균성 펩타이드의 정제)

  • Sohn, Hee-Young;Go, Hye-Jin;Park, Nam Gyu
    • Journal of Life Science
    • /
    • v.26 no.3
    • /
    • pp.296-301
    • /
    • 2016
  • An antibacterial peptide from skin extract of the catfish Silurus asotus was purified and characterized. The acidified skin extract was put through a Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient and divided into flow-through (F.T.), 10% methanol-elute (RM10), 60% methanolelute (RM60), and 100% methanol-elute (RM100) fractions. RM10, RM60, and RM 100 showed antimicrobial activity against Escherichia coli D31. On the other hand, the F.T. fraction did not show antimicrobial activity. Among the various fractions, RM 60 had the highest activity. RM 60 was partially purified on a cation exchange column (CM52) by a stepwise gradient. The ammonium acetate (pH 5.15) 0.02 M – 0.8 M fraction showed antimicrobial activity. Then an antimicrobial peptide was purified using a 0.6M fraction with strong antibacterial activity through a series of five C18 reversed-phase HPLC columns. For the characterization of the purified peptide, the molecular weight and amino acid sequence were analyzed by MALDI-TOF MS and Edman degradation. The molecular weight of this peptide was about 4182.1 [M+H]+. The amino acid sequence of this peptide was partially determined as follows: PALXXKARREAKVKF. These findings suggest that this peptide plays a significant role in the innate defense system of catfish skin.

Purification of an Antibacterial Peptide from the Gills of the Pufferfish Takifugu pardalis (졸복의 아가미로부터 항균성 펩타이드의 정제)

  • Kim, Tae Young;Go, Hye-Jin;Park, Nam Gyu
    • Journal of Life Science
    • /
    • v.27 no.1
    • /
    • pp.50-56
    • /
    • 2017
  • An antibacterial peptide was purified from an acidified gill extract of the pufferfish Takifugu pardalis. The acidified gill extract was put through a Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient and divided into a flow-through (F.T.) and 60% methanol fraction (RM 60). Among the eluents, RM 60 had potent antibacterial activity against Bacillus subtilis KCTC 1021. RM 60 was partially purified on a cationic-exchange column (SP-5PW) by a linear gradient, and the antibacterial peptide was then further purified, using a series of cationic-exchange and $C_{18}$ reversed-phase HPLC columns. For characterization of the purified peptide, its molecular weight and amino acid sequence were analyzed by MALDI-TOF MS and Edman degradation. The molecular weight of the peptide was about 1171.6 Da. The amino acid sequence of the peptide was partially determined as: STKEKAPRKQ. A comparison of the N-terminal amino acid sequence of the purified peptide with that of other known polypeptides revealed high homology with the N-terminus of the histone H3 protein, which belongs to the histone H3 family. Thus, this peptide was designated as a puffer fish gill (PFG)-related antimicrobial peptide. This is the report to describe an antimicrobial function for the N-terminus of histone H3 of an animal species. The findings suggest that this peptide plays a significant role in the innate defense system of the pufferfish.

Quantitation of Mevinolinic Acid in Human Plasma by HPLC (고속액체크로마토그래피를 이용한 사람 혈장중 메비놀린 산의 정량)

  • Oh, Han-Suk;Park, Dong-Young;Seo, Sung-Hoon;Kim, Young-Gwan;Hong, Seon-Pyo;Choi, Young-Wook;Lee, Kyung-Tae
    • Journal of Pharmaceutical Investigation
    • /
    • v.30 no.4
    • /
    • pp.279-282
    • /
    • 2000
  • Simple and precise high-performance liquid chromatographic (HPLC) assay was developed and validated for the determination of a HMG-CoA reductase inhibitor, $lovastatin^{TM}$ and its active metabolite (mevinolinic acid) in human plasma. The method involved solid phase extraction of mevinolinic acid and internal standard using Sep-Pak Cartridge. Samples were analyzed by reversed-phase HPLC using $Capcell-Pak\;C_{18}$ column with ultraviolet detection at 238 nm. The quantitation limit of mevinolinic acid was 2 ng/ml and the calibration curve was linear over the range of 2-50 ng/ml $(r^2>0.999)$ with human plasma. The analyses of quality control samples indicated that the normal values could be predicted with an accuracy >97%. The intra- and inter-day coefficients of variation for the analyses were <10%. The average recoveries were similar (79%) for mevinolinic acid and methylmevinolinic acid. The method described has been successfully applied to the quantification of mevinolinic acid in about 1,000 human plasma samples over six-month period.

  • PDF

Screening for Components to Inhibit N-Nitrosodimethylamine Formation from Tomato (토마토로부터 N-nitrosodimethylamine 생성을 억제시키는 유효성분의 검색)

  • Choi, Sun-Young;Lee, In-Sook;Lee, Soo-Jung;Shon, Mi-Yae;Shin, Jung-Hye;Seo, Jong-Kwon;Kang, Min-Jung;Sung, Nak-Ju
    • Journal of Life Science
    • /
    • v.16 no.5
    • /
    • pp.794-798
    • /
    • 2006
  • A tomato (Lycopersicon esculentum cv. Naomi F1) juice was separated into ascorbate and phenolic portions using a Sep-pak $C_{18}$ cartridge, and its each portion was tested for inhibition of N-nitrosodimethylamine (NDMA) formation. Ascorbate and phenolic portions of tomato juice inhibited NDMA formation by $81.37{\pm}0.25%$ and $72.03{\pm}0.25%$, respectively. The phenolic portion was further fractionated by prep-HPLC and inhibitory effects of NDMA formation by 4 fractions $(1{\sim}4)$ from tomato juice was tested under the different pH conditions (pH 1.2 and 4.2). Fraction 2 inhibited NDMA formation by $50.10{\pm}0.46%$ (pH 1.2) and $64.30{\pm}0.20%$ (pH 4.2), respectively. Fraction 2 was further separated into 4 subfractions $(2a{\sim}2d)$. Subfraction 2b especially inhibited NDMA formation by $70.62{\pm}0.45%$ (pH 1.2) and $75.30{\pm}0.45%$ (pH 4.2). This subfraction was confirmed o-coumaric acid through the analysis of GC-Mass spectrum, $^1H-NMR$ and $^{13}C-NMR$.

Heptelidic Acid, a Sesquiterpene Lactone, Inhibits Etoposide-Induced Apoptosis in Human Leukemia U937 Cells

  • Kim, Jin-Hee;Lee, Choong-Hwan
    • Journal of Microbiology and Biotechnology
    • /
    • v.19 no.8
    • /
    • pp.787-791
    • /
    • 2009
  • In the course of screening for substances that inhibit etoposide (10 ${\mu}g$/ml)-induced apoptosis in human leukemia U937 cells, fungal strain F000120, which exhibits potent inhibitory activity, was selected. The active compound was purified from an ethyl acetate extract of the microorganism by Sep-pak $C_{18}$ column chromatography and HPLC, and was identified as heptelidic acid (koningic acid) by spectroscopic methods. This compound inhibited caspase-3 induction in U937 cells with an $IC_{50}$ value of 40 ${\mu}M$ after 8 h of etoposide treatment. Fluorescent dye staining with acridine orange and ethidium bromide showed that heptelidic acid inhibited apoptosis. Furthermore, it was found that DNA fragmentation and caspase-3 activation, the biological hallmarks of apoptosis, were inhibited by the compound in a dose-dependent manner, suggesting that heptelidic acid inhibits etoposide-induced apoptosis via downregulation of caspases.