• Title/Summary/Keyword: HPLC Pattern

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Isolation and Characterization of a Novel Polysaccharide Producing Bacillus polymyxa A49 KCTC 4648P

  • Ahn, Sung-Gu;Suh, Hyun-Hyo;Lee, Chang-Ho;Moon, Seong-Hoon;Kim, Hee-Sik;Ahn, Keug-Hyun;Kwon, Gi-Seok;Oh, Hee-Mock;Yoon, Byung-Dae
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.171-177
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    • 1998
  • The strain A49, which produces a new type of extracellular polysaccharide was isolated from soil samples. From morphological, physiological and biochemical tests, the strain A49 was identified as a Bacillus polymyxa and named Bacillus polymyxa A49. Bacillus polymyxa A49 was found to produce a highly viscous extracellular polysaccharide when grown aerobically in a medium containing glucose as the sole source of carbon. The polysaccharide (A49 POL) showed a homogeneous pattern on gel permeation chromatography (GPC) and its molecular weight was estimated to be about 1.6 mega dalton (mDa). The FT-IR spectrum of A49-POL revealed typical characteristics of polysaccharides. As a result of investigations with HPLC and carbozole assay, A49-POL was found to consist of L-fucose, D-galactose, D-glucose, D-mannose, and D-glucuronic acid, with the molar ratio of these sugars being approximately 1:2:7:50:12. Rheological analysis of A49 POL revealed that it is pseudoplastic and has a higher apparent viscosity at dilute concentrations than does xanthan gum. The consistancy factor of A49 POL was found to be higher, and the flow index of A49 POL lower, than xanthan gum. Its apparent viscosity was comparatively unstable at various temperatures. the A49 POL showed the highest apparent viscosity at pH 3. When salts were added to A49 POL solution, the solution was compatible with up to 10% KCl, 35% NaCl, 55% $CaCl_2$, 55% $MgCl_2$, 55% $K_2HPO_4$, and 110% $Ca({NO_3})_2$, respectively.

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In vitro Dissolution and in vivo Bioequivalence Study of Controlled Release Carbamazepine Formulation (Epileptol CR® vs Tegretol CR® in Healthy Male Korean Volunteers

  • Kim, Ji-Young;Kim, Hyung-Tae;Kim, Chong-Kook
    • Journal of Pharmaceutical Investigation
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    • v.38 no.5
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    • pp.335-342
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    • 2008
  • The bioequivalence of two carbamazepine preparations was conducted. The in vivo bioequivalence study in 20 healthy male Korean volunteers was designed by using a single dose, randomized, 2-period crossover with a 3-weeks washout period between the doses. Prior to the in vivo study, an in vitro comparative dissolution test was performed by the paddle and basket method as described in the bioequivalence guidance of the Korea Food and Drug Administration (KFDA). Based on the similar dissolution pattern between two preparations in the dissolution test, the two formulations are demonstrated to be pharmaceutically equivalent. In addition, in vivo bioequivalence test was used to reconfirm the in vitro dissolution results. In the in vivo bioequivalence study, the plasma concentrations of carbamazepine up to 144 h after the administration were determined using a validated HPLC method with UV detection and the bioequivalence between the two drug products was assessed by statistical analysis of the log transformed mean ratios of $C_{max}$, $AUC_{0-t}$ and $AUC_{0-\infty}$. The mean maximum concentration ($C_{max}$) of the test and reference were found to be $1467.0{\pm}335.8\;ng/mL$ and $1465.9{\pm}310.3\;ng/mL$, respectively. The 90% confidence intervals (C.I.) of $C_{max}$ were in the range from 0.95 to 1.05. As for the $AUC_{0-t}$ and $AUC_{0-\infty}$, test values were $110027.1{\pm}27786.4\;ng/mL{\cdpt}h$, $128807.0{\pm}34563.2\;ng/mL{\cdot}h$ and $105473.6{\pm}26496.2\;ng/mL{\cdot}h$, $125448.5{\pm}35975.5\;ng/mL{\cdot}h$, respectively. The 90% C.I. of $AUC_{0-t}$ were 0.97 to 1.10 and of $AUC_{0-\infty}$, 0.99 to 1.09 and thus were within the log 0.8-log 1.25 interval proposed by the KFDA. A two-way ANOVA showed no significant difference between the two formulations. Based on these statistical analysis, it was concluded that the test formulation is bioequivalent to the reference.

Conversion of Ginseng Saponin with the Enzyme Produced by Rhizopus sp. (Part 1) Confirmation of Conversion of Ginsenoside- Rb$_1$to Ginsenoside-Rd (Rhizopus sp.가 생산하는 효소에 의한 인삼 Saponin의 전환 (제1보) Ginsenoside-Rb$_1$에서 Ginsenoside-Rd로의 전환확인)

  • 김상달;서정훈
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.267-273
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    • 1982
  • Among 12 kinds of ginsenosides in ginseng saponin, ginsenoside-Rb$_1$was contained the most abundantly. But ginsenoside-Rd which is similar to ginsenoside-Rb$_1$in structure, was known to be superior to ginsenoside-Rb$_1$pharmaceutically. In order to convert ginsenoside-Rb$_1$into ginsenoside-Rd by microbial enzyme treatment, a Rhizopus sp. was selected among various strais of molds found in rotten ginseng roots. Enzyme was prepared from the extract of wheat bran koji culture by ammonium sulfate precipitation (1.0 sat'd) and succeeding ammonium sulfate fractionation method (0.6-0.9 sat'd). For the purpose of use as substrate, saponins were purified by the several purification steps from alcohol extract of red ginseng roots. We obtained the total saponin which was composed of 36.5% of ginsenoside Rb$_1$, 12.2% of ginsenoside-Rd and other ginsenosides. For increase of ginsenoside-Rb$_1$ component ratio, we also obtained further purified ginsenoside-Rb group saponin containing 54.5% of ginsenoside-Rb$_1$, 1.1% of ginsenoside- Rd and other ginsenosides from purified the total saponin. In the enzymatic reaction system including the total saponin or the ginsenoside-Rb group saponin, we confirmed the specific conversion of ginsenoside-Rb$_1$to ginsenoside-Rd proportionally and no change of any other ginsenoside patterns by thin layer chromatography and high performance liquid chromatography.

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Preparation and Release Properties of Oromucosal Moisture-activated Patches Containing Lidocaine or Ofloxacin (오플록사신 및 리도카인 함유 수분 감응성 구강점막 패취제의 제조 및 방출 특성)

  • Gwak, Hye-Sun;Song, Yeon-Hwa;Chun, In-Koo
    • Journal of Pharmaceutical Investigation
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    • v.35 no.6
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    • pp.417-422
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    • 2005
  • This study was aimed to design and formulate the moisture-activated patches containing ofloxacin and lidocaine for antibacterial and local anesthetic action. The solubility of lidocaine at $32^{\circ}C$ in various vehicles decreased in the rank order of PG $759.5{\pm}44.5\;mg/mL$ > PGL > IPM > PEG 300 > PEG 400 > Ethanol > PGMC > DGME > PGML > OA > $Captex^{\circledR}\;300$ > $Captex^{\circledR}\;200$ > water $(4.0{\pm}0.1\;mg/mL)$. Ofloxacin revealed very low solubility, which the highest solubility was obtained from PEG 400 $(18.7{\pm}6.3\;mg/mL)$ among the vehicles used. The addition of lactic acid increased the solubility of ofloxacin dramatically; the solubility at 5% lactic acid was $133.7{\pm}9.7\;mg/mL$. As $2-hydroxypropyl-{\beta}-cyclodextrin$ was added at the concentrations of 40, 80, 120, 160 and 200 mM, the solubilities of lidocaine and ofloxacin were enhanced up to three and two times, respectively, with concentration-dependent pattern. Gel intermediates for filmtype patches were prepared with mucoadhesive polymer, viscosity builders, lidocaine or ofloxacin at pH values from 5 to 7. Gels were cast onto a release liner and dried at room temperature. Dried patch was attached onto an adhesive backing layer, thus forming a patch system. Patches containing a single drug component were characterized by in vitro measurement of drug release rates through a cellulose barrier membrane. The release study was carried out at $37^{\circ}C$ using a Franz-type cell. Receptor solutions were isotonic phosphate buffers (pH 7.4). Samples $(100\;{\mu}L)$ were taken over 24 hours and quantitated by a verified HPLC method. The releases from all tested were proportional to the square root of time. The release rates were 0.9, 157.3 and $281.7\;{\mu}g/cm^{2}/min^{1/2}$ for the lidocaine patches and 19.8,37.2 and $50.7\;{\mu}g/cm^{2}/min^{1/2}$ for the ofloxacin patches at the concentrations of 0.3, 0.5 and 1 %, respectively. The release rates were dose dependent in both drug patches $(R^{2}\;=\;0.9077\;for\;lidocaine;\;R^{2}\;=\;0.9949\;for\;ofloxacin)$ and those were also thickness-dependent $(R^{2}\;=\;0.9246\;for\;lidocaine;\;R^{2}\;=\;0.9512\;for\;ofloxacin)$.

Pharmacokinetic Modeling of Reversible Interconversion between Prednisolone and Prednisone (가역적상호대사과정 모델을 이용한 Prednisolone과 Prednisone의 약동학적 분석)

  • Shin, Jae-Gook;Yoon, Young-Ran;Cha, In-June;Jang, In-Jin;Lee, Kyung-Hoon;Shin, Sang-Goo
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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    • pp.269-281
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    • 1996
  • Pharmacokinetics of prednisolone and prednisone undergoing reversible interconversion were analyzed from the model including this metabolic process. Blood samples were drawn serially upto 12 hours after I,V. bolus injection of 1 mg/kg prednisolone sodium phosphate and prednisone into 8 dogs as a crossover manner. Plasma concentrations of those two steroids were simultaneously measured with the method of HPLC. After injection, plasma concentrations of administered prednisolone and prednisone were declined with a biexponential pattern and their metabolic partner was rapidly formed. Plasma concentrations of those metaboite were decayed in parallel with their parent steroids throught the elimination phase. Apparent clearances of prednisolone and prednisone were $11.1{\pm}2.0\;ml/min/kg$ and $45.9{\pm}6.4\;ml/min/kg$, and they were underestimated by 29.4% and 33.6% compared to their real clearances$(15.7{\pm}4.4\;and\;69.2{\pm}17.7\;ml/min/kg)$ estimated using reversible interconversion model. Apparent volume of distribution of prednisolone$(1.32{\pm}0.43\;L/kg)$ and prednisone$(4.81{\pm}2.75\;L/kg)$ were overestimated by 53.5 and 52.7% and were compared to the real volumes $(0.86{\pm}0.30\;and\;3.15{\pm}2.13\;L/kg)$. Mean residence time of prednisolone$(2.0{\pm}0.61\;h)$ and prednisone$(1.74{\pm}0.74\;h)$ were much longer than the real sojourn time$(0.93{\pm}0.26\;and\;0.88{\pm}0.54\;h)$. Essential clearances In the reversible interconversion were greater as following orders: $Cl_{21}$(44.3 ml/min/kg) > $Cl_{20}$(24.2 ml/min/kg) > $Cl_{12}$ (7.9 ml/min/kg) > $Cl_{10}$(7.8 ml/min/kg). Estimated mean values of RF, EE, $%X^1_{ss}$ and $RHO^2_1$ were $0.31{\pm}0.10$, $1.49{\pm}0.23$, $69.3{\pm}16.7%$ and $0.65{\pm}0.10$, respectively. These results suggested that true pharmacokinetic parameters estimated from the model including reversible interconversion were significantly different from the apparent parameters estimated from the conventional mamillary model, and disposition of these two steroids seemed to be well explained by the model including reversible interconversion.

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Mobility of Pesticides from Soil in Different Slope by Simulated Rainfall Under Field Conditions (포장에서 인공강우를 이용한 경사도별 농약의 이동 특성 연구)

  • Kim, Seong-Soo;Kim, Tae-Han;Lee, Sang-Min;Park, Hong-Ryeol;Park, Dong-Sik;Lim, Chun-Keun;Hur, Jang-Hyun
    • The Korean Journal of Pesticide Science
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    • v.12 no.1
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    • pp.24-33
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    • 2008
  • In this study, the mobility of seven pesticides from soil in different slopes was investigated by simulated rainfall under field conditions. Simulated rainfall subjected to $22\;mm\;hr^{-1}$ was treated using rainfall simulator after 12 hr of pesticide treatment. Amounts of the pesticides were measured in run-off water samples. The soil samples collected before and after rainfall from upper, middle and lower parts and three different depths of sloped-plot were also analyzed. At result, the order of the amount of pesticide residues was $0{\sim}15$ > $15{\sim}30$ > $30{\sim}45\;cm$ of soil depth and no pattern was shown in upper, middle and lower, and different slopes in soil samples. all pesticides from the run-off water samples collected from soils were detected maximum 96% within 60 minutes after first collection except carbendazim and cypermethrin which have the lowest water solubilities. These results revealed that mobility of pesticides can be dependant mainly on soil textures and physicochemical properties of pesticides. Therefore, it can be suggested that selection of pesticides should be considered for properties of pesticide in the alpine and sloped-land.

Cognitive Ability Enhancement Effects in Rats by B. mori Fibroin Enzymatic Hydrolysate (견 피브로인 효소 가수분해물의 동물 인지기능 향상 효과)

  • Yeo Joo-Hong;Lee Kwang-Gill;Kweon HaeYong;Woo Soon-Ok;Han Sang-Mi;Lee Yong-Woo;Kim Jin-Il;Kim Sung-Su;Demura Makoto
    • Journal of Sericultural and Entomological Science
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    • v.46 no.1
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    • pp.23-27
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    • 2004
  • We were investigated the cognitive ability enhancement in rats using B. mori fibroin molecular controlled hydrolysate by preparative recycle HPLC system. Also, some of its physicochemical properties and free amino acid components were investigated, too. Nuclear magnetic resonance (NMR) spectrum are measured different pattern between high (HF) and low (LF) molecular controlled B. mori fibroin samples, respectively. The in vitro test by neuron primary cell culture are showed dependent on molecular weight (Mw) of B. mori fibroin in the order of LF > MF > HF, too. However, the memorial effects in rats by in vivo test are large dependent molecular weight on that maximum 50% than control. Also, the memorial effect was higher than the enzymatic relative and acid hydrolysate, which was 50.0$\pm$2.1 and 25.9$\pm$0.32%, respectively.

Synthesis of O-(3-[18F]Fluoropropyl)-L-tyrosine (L-[18F]FPT) and Its Biological Evaluation in 9L Tumor Bearing Rat

  • Moon, Byung-Seok;Kim, Sang-Wook;Lee, Tae-Sup;Ahn, Soon-Hyuk;Lee, Kyo-Chul;An, Gwang-Il;Yang, Seung-Dae;Chi, Dae-Yoon;Choi, Chang-Woon;Lim, Sang-Moo;Chun, Kwon-Soo
    • Bulletin of the Korean Chemical Society
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    • v.26 no.1
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    • pp.91-96
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    • 2005
  • O-(3-[$^{18}$F]Fluoropropyl)-L-tyrosine (L-[$^{18}$F]FPT) was synthesized by nucleophilic radiofluorination followed by acidic hydrolysis of protective groups and evaluated with 9 L tumor bearing rat. L-[$^{18}$F]FPT is an homologue of O-(2-[$^{18}$F]fluoroethyl)-L-tyrosine (L-[$^{18}$F]FET) which recently is studied as a tracer for tumor imaging using positron emission tomography (PET). [$^{18}$F]FPT was directly prepared from the precursor of O-(3-ptoluenesulfonyloxypropyl)- N-(tert-butoxycarbonyl)-L-tyrosine methyl ester. FPT-PET image was obtained at 60 min in 9 L tumor bearing rats. The radiochemical yield of [$^{18}$F]FPT was 0-45% (decay corrected) and the radiochemical purity was more than 95% after HPLC purification. The total time elapsed for the synthesis of [$^{18}$F]FPT was 100 min from EOB (End-of-bombardment). A comparison of uptake studies between [$^{18}$F]FPT and [$^{18}$F]FET was performed. In biodistribution, [$^{18}$F]FPT showed similar pattern with [$^{18}$F]FET in various tissues, but [$^{18}$F]FPT showed low uptake in brain. Furthermore, [$^{18}$F]FPT showed higher tumor-to-brain ratio than [$^{18}$F]FET. In conclusion, [$^{18}$F]FPT seems to be more useful amino acid tracer than [$^{18}$F]FET for brain tumors imaging with PET.

Cellular activity and guided bone regenerative effect of drug-loaded biodegradable membranes (약물함유 생체분해성 차폐막의 생채활성도 및 골조직 유도재생 효과)

  • Kim, Won-Kyeong;Choi, Sang-Mook;Han, Soo-Boo;Kwon, Young-Hyuk;Chung, Chong-Pyoung;Lee, Seung-Jin
    • Journal of Periodontal and Implant Science
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    • v.27 no.1
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    • pp.129-150
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    • 1997
  • The purpose of this study was to evaluate the effects of tetracycline(TC}, flurbiprofen, and PDGF-BB loaded biodegradable membranes on the cell-attachment, the activity of loaded PDGF-BB, in vivo release kinetics, and guided bone regenerative potentials. To evaluate the cell attachment to membranes, the number of gingival fibroblasts attached to each membrane(10% TC, 10% flurbiprofen, $200ng/cm^2$ PDGF-BB loaded membranes, drug-unloaded membrane) was counted by coulter counter and the morphologic pattern of attached cells was examined under SEM. To determine whether the activity of loaded PDGF-BB is sustained, the cellular growth and survival rate of gingival fibroblasts was used for both standard PDGF-BB and loaded PDGF-BB. For evaluation of in vivo release kinetics, drug-loaded membranes were implanted on the dorsal skin of the rats. On 1, 3, 7, 10, 14, 21, and 28 days after implantation, the amount of remaining drugs were measured by HPLC assay for TC and flurbiprofen, and by ${\gamma}-scintillation$ counter for $PDGF-BB^{1125}$. For evaluation of guided regenerative potential, the amount of new bone in the calvarial defect(5mm in diameter) of the rat was measured by histomorphometry 1 and 2 weeks after implantation of membranes. The number of cells attached to the PDGF-BB loaded membrane was largest as compared with the other mernbranes.(p< 0.05) The activity of loaded PDGF-BB was not significantly different from the activity of standard PDGF-BB.(p<0.05) After initial burst release of drug during the first 24 hours, drugs were gradually released for 4 weeks. Especially the release rate of PDGF-BB was nearly constant during 4 weeks. PDGF-BB loaded membranes(200, $400ng/cm^2$) were effective in guided bone regeneration as compared with drug-unloaded membrane. These results implicate that drug-loaded biodegradable membranes might be a useful for guided bone regeneration.

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Flavonoids and Antimicrobial Activity of the Ethanol Extract of Korean Cherry (Prunus tomentosa Thunberg) (앵두 Ethanol 추출물의 항균력과 Flavonoid)

  • 황호선;김중만;전예정;송영애;박효숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.6
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    • pp.833-839
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    • 2003
  • Quantitaties of flavonoids were determined by HPLC in three different ethanol extract (50, 70 and 95%, v/v) from Korean cherry. Antimicrobial activity of the ethanol extract and its solvent fraction (ethyl acetate and n-butanol fractions) against Staphylococcus aureus (ATCC 6538) and Staphylococcus epidermidis (ATCC 12228) was evaluated using MIC and disk test. Additionally, UV protective effect was also determined. The highest flavonoid quantity was found in 70% ethanol extract of Korean cherry. The quantities of quercitrin were 12.7 mg% and 2.1 mg% in ethyl acetate fraction and in buthanol fraction, respectively. In addition, the flavonoid in 70% (v/v) ethanol extract of Korean cherry included 14.9 mg% of quercitrin, 6.0 mg% of tannic acid and 5.2 mg% of catechin. Therefore, it was confirmed that main flavonoid of Korean cherry was quercitrin. For antimicrobial activity of ethanol extract of Korean cherry against Staphylococcus aureus (ATCC 6538) and Staphylococcus epidemidis (ATCC 12228), MIC and disk test using ethyl acetate fraction and butanol fraction were carried out. As a result, ethyl acetate fraction with higher quantity of flavonoid showed higher antimicrobial activity. Therefore, it was confirmed that the higher antimicrobial activity was brought about by flavonoid in Korean cherry. For UV protective effect of Korean cherry, ethanol extract showed the absorption zone of ultraviolet rays in 210∼380 nm including 290∼320 nm that is the wave length of UV-B harmful to the skin of human body. The pattern is the same as the absorption zone of 220∼300 nm in catechin and 210∼400 nm in quercitrin. Then, the functionalities of catechin and quercitrin were anticipated. It is expected that quercitrin in Korean cherry has the antimicrobial effect and UV protective effect related to the skin.