• Title/Summary/Keyword: solvent analysis

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Bacillus thuringiensis Cry4A and Cry4B Mosquito-larvicidal Proteins: Homology-based 3D Model and Implications for Toxin Activity

  • Angsuthanasombat, Chanan;Uawithya, Panapat;Leetachewa, Somphob;Pornwiroon, Walairat;Ounjai, Puey;Kerdcharoen, Teerakiat;Katzenmeier, Gerd;Panyim, Sakol
    • BMB Reports
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    • v.37 no.3
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    • pp.304-313
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    • 2004
  • Three-dimensional (3D) models for the 65-kDa activated Cry4A and Cry4B $\delta$-endotoxins from Bacillus thuringiensis subsp. israelensis that are specifically toxic to mosquito-larvae were constructed by homology modeling, based on atomic coordinates of the Cry1Aa and Cry3Aa crystal structures. They were structurally similar to the known structures, both derived 3D models displayed a three-domain organization: the N-terminal domain (I) is a seven-helix bundle, while the middle and C-terminal domains are primarily comprise of anti-parallel $\beta$-sheets. Circular dichroism spectroscopy confirmed the secondary structural contents of the two homology-based Cry4 structures. A structural analysis of both Cry4 models revealed the following: (a) Residues Arg-235 and Arg-203 are located in the interhelical 5/6 loop within the domain I of Cry4A and Cry4B, respectively. Both are solvent exposed. This suggests that they are susceptible to tryptic cleavage. (b) The unique disulphide bond, together with a proline-rich region within the long loop connecting ${\alpha}4$ and ${\alpha}5$ of Cry4A, were identified. This implies their functional significance for membrane insertion. (c) Significant structural differences between both models were found within domain II that may reflect their different activity spectra. Structural insights from this molecular modeling study would therefore increase our understanding of the mechanic aspects of these two closely related mosquito-larvicidal proteins.

Development of Analytical Method for Quality Control from New Herbal Medicine(HPL-4) (새로운 생약복합제제(HPL-4)의 품질관리를 위한 분석법 개발)

  • Kim, Se-Gun;Sharma, Dipak Kumar;Lamichhane, Ramakanta;Lee, Kyung-Hee;Han, Sang-Mi;Jung, Hyun-Ju
    • Korean Journal of Pharmacognosy
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    • v.45 no.4
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    • pp.338-345
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    • 2014
  • HPL-4 is a new herbal formulation developed for the treatment of osteoarthritis. In this study, we took HPL-4 to develop a method for simultaneous determination of nine marker compounds (protocatechuic acid, chlorogenic acid, liriodendrin, nodakenin, ${\beta}$-$\small{D}$-(3-O-sinapoyl)frucofuranosyl-${\alpha}$-$\small{D}$-(6-O-sinapoyl)glucopyranoside, quercitrin, 6-gingerol, decursin and decursinol angelate) present in the formulation. The analytes were separated by UPLC system consisting of diode array detector at 205 nm and RP-amide column, and solvent system of $ACN/0.1%H_3PO_4$. Analytical method was validated to evaluate its linearity, limit of detection (LOD), limit of quantification (LOQ), precision, and accuracy. All standards showed good linearity ($R^2$ >0.99) in the rage of $0.25-400.0{\mu}g/mL$. The LOD and LOQ were within the range of 0.021-0.148 and $0.070-0.448{\mu}g/mL$, respectively. Intra- and inter-day precision was less than RSD 4.0% and the accuracy was range from 92.00-104.81% with RSD<4.2%. The results suggest that the developed UPLC method is precise, accurate and effective, and could be applied for quality control of HPL-4 formulation.

Preparation of Liquefied Cellulose and Analysis of Its Components by GC-MS Spectrometry (액화셀룰로오스의 제조 및 GC-MS에 의한 그 성분 분석)

  • 조국란;황병호;공영토;도금현
    • Journal of Korea Foresty Energy
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    • v.19 no.2
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    • pp.86-92
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    • 2000
  • The liquefactions of $\alpha$-cellulose(Sigma Chemical, C-8002, 47H0383) was prepared in the presence of phenol using sulfuric acid as a catalyst under $N_2$ gas protection at $180^{\circ}C$ for 60minutes to examine its components. The ratio of $\alpha$-cellulose to phenol was 1: 6.2(w/w), and that to sulfuric acid was 1: 0.05(g/$m\ell$). The yields of liquefaction were calculated after the liquefied mixtures were passed through 1G4 glass filter. The luquefied product of $\alpha$-cellulose was analyzed using GC-MS Spectormeter. The 12 compounds identified by GC-MS Spectrometer, of which peak area covers 54% as 2,4-dimethyl phenol, p-isopropyl phenol, 1-ethyl-3,5-dimethyl benzene, o-isopropyl phenol, (E)-2,4\` dihydroxy-stilbene, 2,2\`-methylene-bisphenol, 4,4\`-methylenebisphenol, 3-methyl-2-hydroxyphenyl-(E)-2-hydroxyl-4\`-methoxy-stilbene, 1-phyenyl-1-(4\`hydroxyphenyl)methanol phenol derivatives. From this results, the reaction pathways of the liquefaction of cellulose were proposed through electrophilic substitution reaction. Phenol as a solvent might react with the reaction intermediates as well in the cellulose liquefaction.

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Optical and Electrical Properties with Various Post-Heating Temperatures in the Al-Doped ZnO Thin Films by Sol-Gel Process (졸-겔법에 의해 제조된 Al-Doped ZnO 박막의 후열처리 온도에 따른 전기 및 광학적 특성)

  • Ko, Seok-Bae;Choi, Moon-Sun;Ko, Hyungduk;Lee, Chung-Sun;Tai, Weon-Pil;Suh, Su-Jeong;Kim, Young-Sung
    • Journal of the Korean Ceramic Society
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    • v.41 no.10 s.269
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    • pp.742-748
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    • 2004
  • Isopropanol of low boiling point was used as a solvent to prepare Al-doped ZnO(AZO) thin films. A homogeneous and stable sol was made from Zn acetate a solute whose mole concentration was 0.7mol/$\iota$ and Al chloride as a dopant. Al-doped ZnO thin films were prepared by sol-gel method as a function of post-heating temperature from 500 to $700^{\circ}C$ and the optical and electrical properties were investigated. The c-axis orientation along (002) plane was enhanced with the increasing of post-heating temperature and the surface morphology of the films showed a homogeneous and nano-sized microstructure. The optical transmittance of the films post-heated below $650^{\circ}C$ was over $86\%$, but decreased at $700^{\circ}C$. The electrical resistivity of the thin films decreased from 73 to 22 $\Omega$-cm as the post-heating temperature increased up to $650^{\circ}C$, but increased greatly to 580 $\Omega$-cm at $700^{\circ}C$. XPS analysis indicated that the deterioration of electrical and optical properties was attributed to the precipitation of $Al_2O_3$ phase on the surface of AZO thin film. This result suggests that the optimum post-heating temperature to improve electrical and optical properties is $600^{\circ}C$.

Identification of the Plant Part of Gleditsia sinensis that Activates Nrf2, an Anti-oxidative Transcription Factor (조협의 부위에 따른 항산화 전사인자 Nrf2 활성 효과)

  • Choi, Jiyeon;Kim, Kyun Ha;Choi, Jun Yong;Han, Chang Woo;Ha, Ki Tae;Jeong, Han-Sol;Joo, Myungsoo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.3
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    • pp.303-309
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    • 2014
  • The fruit of Gleditsia sinensis has been extensively used as a key ingredient of an herbal remedy for the treatment of various inflammatory diseases in traditional Korean Medicine. However, the reason of using the fruit of G. sinensis for the remedy is unclear. Since Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is a key anti-inflammatory transcription factor, which is activated by the fruit of G. sinesis, we examined whether other plant parts of G. sinensis are also capable of suppressing inflammatory responses by activating Nrf2. Water extracts of various parts of G. sinensis were prepared and tested for Nrf2 activation by reporter assay and western blot analysis. Our results show that the hull of G. sinensis is the most potent in activating Nrf2. Sequential organic solvent extraction of the hull show that all the fractions had a higher potency in activating Nrf2 than the water extract, albeit differential degrees. The hull originated from Korea in general activated Nrf2 strongly compared to that of China. Chloroform fraction of the hull was further examined, showing that the fraction induced nuclear localization of Nrf2, indicative of activated Nrf2, and Nrf2-dependent gene expression including NAD(P)H dehydrogenase quinone 1 (NQO-1), glutamate-cysteine ligase catalytic subunit (GCLC), and heme oxygenase - 1 (HO-1). Therefore, our results show that, among other plant parts examined in this study, the hull of G. sinensis is the most potent, providing the experimental basis for the use of the hull of G. sinensis as an active ingredient for an anti-inflammatory remedy.

Antimicrobial Effects and Antioxidative Activity of Baek-bu-ja(Aconiti koreani Rhizoma) by Extraction Solvent Ratio (추출용매 비에 따른 백부자(Aconiti koreani Rhizoma) 추출물의 항균효과 및 항산화효과)

  • Yoon, So-Jung;Kim, Jeung-Hoan;Lee, Kyoung-Hwan;Kwon, Hyo-Jung;Chun, Sung-Sook;Cho, Young-Je
    • Applied Biological Chemistry
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    • v.48 no.3
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    • pp.258-262
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    • 2005
  • For the purpose of developing natural antioxidant, the antioxidative activity and antimicrobial of phenolics isolated from Baek-bu-ja (Aconiti koreani Rhizoma) were determined. Optimum extracting condition for phenolics were water extracts. HPLC analysis showed that the four major phenolic metabolites were rosemarinic, protocatechuic, caffeic and chlorogenic acids. The water extracts of Baekbuja did not have antimicrobial activity against H. pylori; however, the ethanol extracts revealed higher antimicrobial activity. Electron donation ability on DPPH of Baekbuja ethanol extract was 20% higher than other ethanol extracts. The 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid radical decolorization (ABTS) and antioxidant protection factor (PF) were determined with extracts from Aconiti koreani Rhizoma. 94% inhibition and 1.14 PF were shown on ABTS and antioxidant protection factor with 60% ethanol extracts. Also, TBARS (thiobarbituric acid reactive substances) showed $0.19\;{\mu}M$ in the control and $0.07\;{\mu}M$ in the 80% ethanol extracts. The result suggests that Baekbuja extract may be useful as potential sources of anti Helicobacter pylori, antioxidant.

Simultaneous Identification/Determination System for Sildenafil, Homosildenafil, Vardenafil and Tadalafil as Adulterants in Foods (식품중 불법함유된 발기부전치료제 성분 검출을 위한 동시분석법 연구)

  • Jang, Jae-Hee;Park, Kun-Sang;Park, Hye-Kyung;Koo, Yong-Eui;Choi, Youn-Ju;Hwang, In-Kyung;Kim, Dai-Byung
    • Journal of Food Hygiene and Safety
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    • v.18 no.4
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    • pp.195-201
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    • 2003
  • An easily available, simultaneous identification/determination procedure for sildenafil, homosildenafil, tadalafil, vardenafil in adulterated health related foods was established by using a combination of three different analytical methods; thin layer chromatography(TLC), liquied chromatography-mass spectrometry (LC/MS) and high-performance liquied chromatography (HPLC)/photo-diode-array detector. The sample solution for TLC was applied to silica gel 60 $F_{254}$ plates with ethylacetate/acetonitrile/25%ammonia (90:10:5) as a developing solvent. Spots were located under UV radiation at 254 nm and dragendolfs reagent. Mass spectra of the compounds by LC/MS were investigated with electrospray ionization (ESI) interface, under positive ion mode. The HPLC analysis was performed on a column of capcell pack $C_{18}$ (UG120, 4.6${\times}$250mm I.D. 5 ${\mu}$m)with 0.1% sodium 1-hexansulfonate (in 0.1% phosphoric acid)/acetnitrile (73:27) as a mobile phase, and effluent was minitored with a photo-diode-applied to commercial foods, Sildenafil content was inthe range of 0.4mg/g~360.9 mg/g from 7 out of 35 samples. Homosildenafil content was in the range of 2.2 mg/g~336.0 mg/g from 7 out of 35 samples. Tadalafil content was 429.3 mg/g, 9.6 mg/500 mg from 2 out of 35 samples. The procedure described here is available for the screening of sildenafil, homosildenafil, tadalafil, vardenafil.

Studies for Component Analysis, Antioxidative Activity and ${\alpha}-glucosidase$ Inhibitory Activity from Equisetum arvense (쇠뜨기(Equisetum arvense) 추출물의 항산화 활성 분석 및 ${\alpha}-glucosidase$ 저해활성)

  • Gua, Jia;Jin, Ying-Shan;Han, Woong;Shim, Tae-Heum;Sa, Jae-Hoon;Wang, Myeong-Hyeon
    • Applied Biological Chemistry
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    • v.49 no.1
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    • pp.77-81
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    • 2006
  • This study was carried out to investigate the chemical components, and antioxidative and $anti-{\alpha}-glucosidase$ activities of Equisetum arvense extracts. In Equisetum arvense extracts were composed of 53.20% of crude fiber, 20.42% of crude ash, 15.32% of crude protein and 2.21% of crude fat. Potassium was the most predominant mineral and followed by phosphorus, calcium, magnesium, and sodium. The contents of the unsaturated fatty acids, such as linolenic acid, linoleic acid, and palmitic acid, were higher than those of saturated fatty acids. Seventy percent ethanol extract exhibited antioxidative activity with $IC_{50}$ of $168.1\;{\mu}g/ml$. The Seventy percent methanol extract showed higher ${\alpha}-glucosidase$ inhibitory activity than other solvent extracts.

Counter anion effects in anion exchange membrane-fabricated non-aqueous vanadium redox flow battery

  • Son, Pyeong Soo;Oh, Min-Seok;Ye, Jun-Hee;Choi, Seong-Ho
    • Analytical Science and Technology
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    • v.28 no.5
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    • pp.341-346
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    • 2015
  • In order to understand the counter anionic effects in a non-aqueous vanadium redox flow battery (VRFB), we synthesized four types of electrolyte salts (1-ethyltriethamine tertafluoroborate, [E-TEDA]+[BF4], 1-ethyltriethamine hexafluorophosphate, [E-TEDA]+[PF6], 1-butyltriethylamine tertafluoroborate, [B-TEDA]+[BF4], and 1-buthyltriethamine hexafluorophosphate [B-TEDA]+[PF6]) by counter anion exchange reaction after the SN2 reaction. We confirmed the successful synthesis of the electrolyte salts [E-TEDA]+[Br] and [B-TEDA]+[Br] via 1H-NMR spectroscopy and GC-mass analysis before the counter anion exchange reaction. The electric potential of the vanadium acetylacetonate, V(acac)3, as an energy storage chemical was shown to be 2.2 V in the acetonitrile solvent with each of the [E-TEDA]+[BF4], [E-TEDA]+[PF6], [B-TEDA]+[BF4], and [B-TEDA]+[PF6] electrolyte salts. In a non-aqueous VRFB with a commercial Neosepta AFN membrane, the maximum voltages reached 1.0 V and 1.5 V under a fixed current value of 0.1 mA in acetonitrile with the [E-TEDA]+[BF4] and [E-TEDA]+[PF6] electrolyte salts, respectively. The maximum voltage was 0.8 V and 1.1 V under a fixed current value of 0.1 mA in acetonitrile with the [B-TEDA]+[BF4] and [B-TEDA]+[PF6] electrolyte salts, respectively. From these results, we concluded that in the non-aqueous VRFB more of the [PF6] counter anion than the [BF4] counter anion was transported onto the commercial Neosepta AFN anion exchange membrane.

Enhancement of cyanoacrylate-developed marks using p-dimethylaminobenzaldehyde (DMAB) on semi-porous surfaces and analysis of the influence factors on fluorescence intensity (반다공성 재질에 유류된 지문의 CA 훈증 후 p-dimethylaminobenzealdehyde(DMAB) 형광시약 적용 시 표면적과 주변 온도, 기압이 형광착색에 미치는 효과에 관한 연구)

  • Yu, Je-Sul;Kim, Ju-Hah
    • Analytical Science and Technology
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    • v.27 no.4
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    • pp.187-195
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    • 2014
  • It is very important to minimize the damage of latent fingerprints at enhancing the contrast. This study proves the enhancement effects of cyanoacrylate-fumed latent fingerprints using p-dimethylaminobenzealdehyde (DMAB) on semi-porous surfaces and the influence factors. The latent fingerprints in experiment were developed for cyanoacrylate treatment in a vacuum chamber and used after drying at room temperature for 24 hours. For fluorescence staining, the cyanoacrylate-developed marks using DMAB were sublimated during 48 hours under the different conditions of surface area, temperature, atmospheric pressure. First experiment showed how surface area effects on the sublimation rate and fluorescence intensity by DMAB of particle size and container size. In addition, the fluorescence staining using DMAB with solvent-free contact method had the greatest fluorescence intensity after 36 hours and a low fluorescence intensity over a certain size of surface area. Second experiment showed that the evaporation of DMAB solid crystals got a satisfying result in a temperature of $20^{\circ}C$ and reduced time to get the greatest fluorescence intensity. It took a long time to get a optimum level of fluorescence intensity at $30^{\circ}C$ or more and it was less effective in fluorescence intensity. Third experiment on the pressure indicated that the fluorescence intensity of vacuum was weaker than nonvacuum but it was inapplicable to very high variations in pressure.