• Title/Summary/Keyword: HPLC analysis

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MONOMER RELEASE FROM PIT AND FISSURE SEALANT FOLLOWING BY SURFACE TREATMENT AND CURING TIME (치면열구전색제의 표면처리와 중합시간 증가에 따른 미반응 모노머 용리 평가)

  • Seo, Hyun-Woo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.1
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    • pp.122-129
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    • 2007
  • The purpose of this study was to evaluate the effects of a various light curing time on the residual monomers released from light-cured dental sealant, and to examine the effectiveness of surface treatment in reducing the oxygen-inhibited layer of light-cured dental sealant($Helioseal^{(R)}$ F, Vivadent, Liechtenstein). Specimens were cured with a halogen light curing unit(XL 3000, 3M, USA) for 20, 40, 60s. Surface treatment of a light-cured dental sealant included no treatment(control group), a 10-seconds exposure to distilled water(Group I), 10-seconds manual application using a cotton pellet wetted with 75% alcohol(Group II), and 10-seconds application of a water/pumice slurry using a rubber cup on a slow-speed handpiece The specimens were eluted in distilled water for 10 minutes. All elutes were analyzed by HPLC for identification and quantitive analysis of monomers. The results of this study can be summarized as follows. 1. None of the chromatograms of the tested sealant displayed peaks with the same retention time as that of the standard solution, except for TEGDMA. 2. The release of TEGDMA decreased with increasing curing time in conventional halogen light. 3. All surface treatment group had a decrease of monomer release in comparison with no treatment group. 4. Treatment that Group III eliminated the greatest amount of any type of residual monomers. 5. The elution of unreacted monomers from curing with halogen curing unit for 60s and Group III was less than other groups.

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Isolation of Antifungal Substances by Bacillus amyloliquefaciens IUB158-03 and Antagonistic Activity against Pathogenic Fungi (Bacillus amyloliquefaciens IUB158-03이 생산하는 항진균물질의 분리와 항균활성)

  • Kim, Hye-Young;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.96-103
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    • 2009
  • For the control of pathogenic microorganisms, Bacillus spp. were isolated from diseased pepper fruits in Korea. Among them, Bacillus sp. IUB158-03 showed high inhibitory effect on mycelial growth and spore germination of C. gloeosporioides and Botrytis cinerea. The strain was identified as B. amyloliquefaciens IUB158-03 based on its physiological, biochemical characteristics and Microlog analysis. The highest level of antifungal substances by B. amyloliquefaciens IUB158-03 were obtained when the bacterium was cultured in medium containing 2% soluble starch, 3% yeast extract, 0.5% tryptone, 0.5% $NH_4H_2PO_4$, and 1% NaCl (pH 6.0) at $25^{\circ}C$ for 72 hrs. The antifungal substances were purified by butanol extraction, silica gel column chromatography, preparative thin layer chromatography, and high performance liquid chromatography. The purified antifungal substance was confirmed $R_f$ 0.27 by TLC. This substance exhibited antifungal activity against Fusarium solani, Rhizoctonia solani, Botrytis cineria, Alternata alternaria of plant pathogenic fungi and Trichophyton mentagrophytes, Epidermophyton floccosum, Cryptococcus neoformans of human pathogenic fungi.

Comparison of Effective Constituents of Korean Paeony Roots(Paeoniae radix) Cultivated in Different Regions (한국산 작약의 산지별 유효성분 비교)

  • Sung, Won-Yong;Yoon, Gwang-Ro;Jang, Sang-Moon
    • Food Science and Preservation
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    • v.7 no.3
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    • pp.297-302
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    • 2000
  • To compare the contents of well-known effective constituents, paeoniflorin, albiflorin and benzoic acid, of Korean paeony roots cultivated at different regions, this study was to provide the basic information for the quality evaluation of paeony roots. HPLC analysis showed that the paeoniflorin contents of white paeony roots of 3 years grown at Eui-seong gun, Kyungpook was the highest of 2.06% and red paeony roots of 3 years grown at Im-sil gun, Junpook was the lowest of 1.33%. The highest level of albiflorin contents was 2.04% of red paeony root grown at Ul-jin gun, Kyungpook, while the lowest levels were 0.05% of Eui-seong gun, Kyungpook and 0.06% of Im-shil gun, Junpook. The highest level of benzoic acid contents was 0.12% of red paony roots grown at Choung-do gun, Kyungpook and that grown at Ul-jin gun, Kyungpook was the lowest. Based on the average of three available constituents, the paeony roots cultivated at Kyungpook province was the highest contents of 2.03%, while 1.47% at Junpook province. These results suggest that Kyungpook province is the best region for the cultivation of white and red paeony roots, the combining of three available constituents would be more accurate than use of paeoniflorin only.

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The chemical constituents, antioxidant activity and anti-inflammatory activities of Solanum nigrum Linne by different harvest time (수확시기별 까마중 전초의 성분, 항산화활성 및 항염증 활성)

  • Choi, Seongkyu;Seo, Kyoung-Sun;Kim, Yong-Doo;Yun, Kyeong-Won;Choi, Bong-suk;Jin, Seong-Woo;Kang, Kyeong-Yun;Cho, In-Kyung;Kim, Kyung-Je
    • Food Science and Preservation
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    • v.23 no.6
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    • pp.866-875
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    • 2016
  • In this study, we investigated the variation in free sugars, amino acids, antioxidant activity, and anti-inflammatory activity of Solanum nigrum Linne based on harvest time. Major amino acids identified by HPLC analysis were proline, histidine, and serine. The highest content of total amino acids were found in S. nigrum aerial parts and roots harvested on July 10th and August 10th. Four kinds of free sugars (fructose, glucose, sucrose, maltose) were detected in S. nigrum, and the free sugar content varied significantly with harvest time. The fructose content of S. nigrum decreased with as harvest time increased. The total polyphenol content of S. nigrum was highest in those harvested on August 30th. The antioxidant activity of ethanol extract from S. nigrum collected at different harvest times were measured by DPPH and ABTS radical scavenging assays. The anti-inflammatory activity of these extracts were assayed via nitric oxide suppression in C6 glioma cells with a lipopolysaccharide (LPS)-induced inflammatory response. The anti-inflammatory activity and antioxidant effects were the highest in the extract from S. nigrum collected on August 30th. Good correlations were observed between antioxidant and anti-inflammatory activities in ethanol extract of S. nigrum roots harvested on August 30th.

In Vitro Pharmacodynamics of CKD-602 in HT-29 Cells

  • Park, In-Sook;Ahn, Mee-Ryung;Suh, Soo-Kyung;Choi, Hong-Serck;Sohn, Soo-Jung;Yang, Ji-Sun;Yoo, Tae-Moo;Kuh, Hyo-Jeong
    • Archives of Pharmacal Research
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    • v.25 no.5
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    • pp.718-723
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    • 2002
  • CKD-602 (7-[2-(N-isopropylamino)ethyl]-(20S)-camptothecin) is a recently-developed synthetic camptothecin analogue and currently under clinical development by Chong Kun Dang Pharm (Seoul, Korea). CKD-602 showed potent topoisomerase inhibitory activity in vitro and broad antitumor activity against various human tumor cells in vitro and in vivo in animal models. This study describes the pharmacodynamics of the immediate and delayed cytotoxicity induced by CKD-602 in a human colorectal adenocarcinoma cell line, HT-29, and its intracellular drug accumulation by HPLC. The present study was designed to address whether the higher activity of CKD-602 with prolonged exposure is due to delayed exhibition of cytotoxicity and/or an accumulation of anti proliferative effect on continuous drug exposure. The drug uptake study was performed to determine whether the delayed cytotoxicity is due to a slow drug accumulation in cells. CKD-602 produced a cytotoxicity that was exhibited immediately after treatment (immediate effect) and after treatment had been terminated (delayed effect). Both the immediate and delayed effects of CKD-602 showed a time dependent decrease in 4IC_{50}$ values. Drug uptake was biphasic and the second equilibrium level was obtained as early as at 24hr, indicating that the cumulative and delayed antitumor effects of CKD-602 were not due to slow drug uptake. On the other hand, CKD-602 treatment was sufficient to induce delayed cytotoxicity after 4hr, however, longer treatment (>24hr) enhanced its cytotoxicity due to the intracellular accumulation of the drug, which requires 24hr to reach maximum equilibrium concentration. In addition, $C^n$$\times$T=h analysis (n=0.481) indicated that increased exposure times may contribute more to the overall antitumor activity of CKD-602 than drug concentration. Additional studies to determine the details of the intracellular uptake kinetics (e.g., concentration dependency and retention studies) are needed in order to identify the optimal treatment schedules for the successful clinical development of CKD-602.

Extraction and Preprocessing Methods for Ginsenosides Analysis of Panax ginseng C.A. Mayer (인삼의 진세노사이드 분석을 위한 추출 및 전처리법)

  • Kim, Geum-Soog;Hyun, Dong-Yun;Kim, Young-Ock;Lee, Sung-Woo;Kim, Young-Chang;Lee, Seung-Eun;Son, Yeong-Deck;Lee, Min-Jeong;Park, Chung-Berm;Park, Ho-Ki;Cha, Seon-Woo;Song, Kyung-Sik
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.6
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    • pp.446-454
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    • 2008
  • An advanced extraction method by ultrasonic extraction with applied solid phase extraction (SPE) has been developed for the determination of simultaneous eight major ginsenosides, namely ginsenosides Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, and Rd in the root of Panax ginseng. Four extraction methods including n-BuOH reflux extraction (Method A), 70% EtOH reflux extraction (Method B), 50% MeOH reflux extraction with SPE (Method C), and 50% MeOH ultrasonication with SPE clean-up process (Method D) were investigated for the determination of eight major ginsenosides. Total contents of ginsenosides were highest by extraction of Method C as $2.408{\pm}0.011%$. However, Method D was evaluated as relatively simpler and more efficient method due to short extraction time, small solvent consumption and less expensive, compared to conservative reflux method. Ginsenosides were also satisfactorily separated with good resolution and the accuracy range was between 1.05 and 4.06% as relative standard deviation (RSD) by Method D. SPE condition and HPLC condition were further optimized for determination of eight major ginsenosides by the ultrasonic extraction method. Conclusively, ultrasonic extraction of 2 g sample of ginseng using ultrasonic bath and 1 loading for SPE was evaluated as proper condition for extraction of ginseng.

Structures and some Properties of the Antimicrobial Compounds in the Red Alga, Symphyocladia latiuscula (참보라색우무에서 추출한 항균물질의 구조 및 특성)

  • LIM Chi-Won;LEE Jong-Soo;CHO Young-Je
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.4
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    • pp.280-287
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    • 2000
  • Three antimicrobial compounds (SL-l, SL-2 and SL-3) were isolated and identified from the marine red alga, Symphyocladia latiuscula. In addition, their biological functionalities such as cytotoxicity and desmutagenic activity were investigated. From the cryophyllized S. JatiuscuJa, SL-l, SL-2 and SL-3 were purified by solvent extractions and HPLC.SL-2 was crystallized in benzene-diethyl ether solvent. On the EI-MS spectra, it was found that they had three bromines in their structure which showed typical signal strength ratios at $M^+, [M+2]^+, [M+4]^+, [M+6]^+ (13: 38: 37: 12)$. $SL-l$ was identified as 2,3,6-tribromo-4,5-dihydroxybenzyl alcohol ($C_8H_7Br_3O_3, MW=374$) by NMR and MS spectra. SL-2 was assigned as 2,3,6-tribromo-4,5-dihydroxybenzyl methyl ether ($C_8H_7Br_3O_3, MW=388$) and confirmed by X-ray crystallographic analysis. SL-3 was presumed as an isomer of SL-2. Methanol extract of the S. latiuscula showed antimicrobial activities against all strains tested (bacteria, 15 strains; yeasts, 17 strains; fungi, 4 strains), much or less. The strongest inhibition activity of the methanol extract was to the Vibrio mimicus ($50 {\mu}g/ml$) and V. vulnificus ($50 {\mu}g/ml$). The mice injected intraperitoneally with 3 mg of SL-l and 5 mg of 5L-2 showed no acute toxicity response. SL-2 showed higher desmutagenic activity than SL-l against PhIP and MeIQx.

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Preparation and Release Behavior of Atorvastatin Calcuim - Encapsulated Polyoxalate Microspheres (아토르바스타틴 칼슘을 함유한 폴리옥살레이트 미립구의 제조 및 방출거동)

  • Lee, Cheon Jung;Kim, Su Young;Lee, Hyun Gu;Yang, Jaewon;Park, Jin Young;Cha, Se Rom;Lim, Dong-Kwon;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.5
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    • pp.656-663
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    • 2014
  • Atorvastatin calcium-loaded polyoxalate (POX) microspheres were prepared by an emulsion solvent-evaporation/ extraction method of oil-in-oil-in-water ($O_1/O_2/W$) for sustained release. We investigated the release behavior according to initial drug ratio, molecular weight ($M_w$) and concentration of POX and concentration of emulsifier. The microsphere was characterized on the surface, the cross-section morphology and the behavior of atorvastatin calcium release for 10 days by scanning electron microscopy (SEM) and high performance liquid chromatography (HPLC). The analysis of crystallization was analyzed to use X-ray diffraction (XRD), differential scanning calorimeter (DSC) and Fourier transform infrared (FTIR). These results showed that the release behaviors can be controlled by preparation conditions.

Identification of a Neolignan Glycoside from the Pine Tree, Pinus densiflora Showed Antithrombotic Activity (Pinus densiflora 유래의 항트롬빈 활성을 나타내는 Neolignan Glycoside의 동정)

  • Seo, Min-Jeong;Kang, Byoung-Won;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.24 no.8
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    • pp.873-879
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    • 2014
  • The constituents from the needles of the pine tree, Pinus densiflora, were purified and investigated for antithrombotic activity. The needles were initially extracted three times with 70% ethanol, and the extract was sequentially fractionated with chloroform and n-butanol. The aqueous layer formed after n-butanol fractionation was subjected to purification by medium pressure and high pressure liquid chromatography. The two neolignans, 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl -3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside (a neolignan glycoside) and 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$) were identified by $^1H$ and $^{13}C$ NMR spectra. The effect of the purified compounds, the neolignan glycoside and icariside $E_4$ on thrombin inhibition were investigated by measuring thrombin clotting time in plasma. As a result, the clotting of the neolignan glycoside was delayed four times compared to that of icariside $E_4$. In addition, an analysis of the inhibition effect by changing the concentration showed that the clotting time was delayed in accordance with an increase in the concentration of the neolignan glycoside. Furthermore, we examined the interaction of thrombin and fibrinogen to clarify the action mechanism. As a result, the delay of clotting time in the response of thrombin and pure fibrinogen may indicate that neolignan glycosides inhibit the thrombin action in a direct manner, leading to the suppression of fibrin generation.

Real-Time Monitoring of Mitochondrial ATP Synthesis and Hydrolysis by Surface Infrared Spectroscopy

  • Yamaguchi, Ryo-Taro;Hirano-Iwata, Ayumi;Aonuma, Yuki;Yoshimura, Yuya;Shinohara, Yasuo;Kimura, Yasuo;Niwano, Michio
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.108-109
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    • 2013
  • Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

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