• Microbiology and Biotechnology Letters
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• v.4 no.4
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• pp.131-137
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• 1976

Aspergillus niger S-1 was proved to be a good hesperidinase producer which have been selected for naringinase utilization. Enzyme of this strain had good characteristics and purified relative high degree with good recovery by ammonium sulfate or aceton treatment. Results obtained were summarized as follows (1) The enzyme was most active at 60$^{\circ}C$, when the reaction was performed in the pH 4.0 for 30min. Optimum pH for enzyme activity was 5.0 and activity was retained 78％ at pH value 3.5. (2) Hesperidinase activity retained 95％ of its full activity after treatment at 60$^{\circ}C$ for 30min at pH value 4.0., 70％ at 70$^{\circ}C$ and 65％ at 80$^{\circ}C$. Most stable pH of this enzyme was showed 5.0 after treatment for 24hr at 4$^{\circ}C$ (3) Only Magnesium ion activated enzyme reaction, while other metallic ions, Cu$\^$＋＋/, Mn$\^$＋＋/, Pb$\^$＋＋/, Mo$\^$＋＋/, Ag$\^$＋＋/, Hg$\^$＋＋/ inhibited. (4) Eleven fold purification with 35％ recovery was obtained in the case of 60％ aceton treatment and 10-fold purification with 5.6％ recovery was showed with 40％ aceton comparing to the crude extract Enzyme. (5) Crude enzyme precipitated with 0.4-0.6 saturated ammonium sulfate contained 13f6 of the original enzyme activity with 48-fold increase in specific activity and enzyme has been purified 25 fold with a yield 19％ by 0.6-5.8 saturation. (6) Hesperidinase formation was noticeably increased by addition of small amount of orange-peel extraction on the wheat bran medium.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.285-292
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• 2016

In the present study, we evaluated the in vitro wound-healing properties of two types of rice cell extracts (RCEs; prepared using ethanol and pressurized hot water extraction methods), using human dermal fibroblasts and keratinocytes. The effects of the RCEs (at 25–100 μg/ml) on cytotoxicity and cell migration were assessed. Both RCEs were not cytotoxic to the two cell types, instead increasing their proliferation by up to 25% in a dose-dependent manner compared with the controls. Furthermore, both RCEs significantly enhanced the migratory ability of the two cell types (fibroblast, 230–450%; keratinocyte, 170–350%). Additionally, we examined the effect of the RCEs on type I collagen synthesis, which is important in the wound reconstruction process. The RCEs significantly increased collagen type I mRNA and protein levels to a degree comparable to that induced by vitamin C. These results suggest the RCEs to be candidate materials for use in promoting wound healing, through their actions of increasing cell migration and accelerating wound re-epithelialization.

• Journal of Pharmaceutical Investigation
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• v.39 no.1
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• pp.73-78
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• 2009

A simple LC-MS/MS method of rabeprazole in human plasma was developed and validated. Rabeprazole and Internal standard (I.S), omeprazole, were extracted from human plasma by liquid liquid extraction, chromatographic separation of rabaprazole in plasma was achieved at $45^{\circ}C$ with a Shiseido UG120 $C_{18}$ column and methanol-10 mM ammonium acetate buffer (pH 9.42 with ammonium water), as mobile phase. Rabeprazole produced a protonated precursor ion [$(M+H)^+$] at m/z 360.10 and corresponding product ion at m/z 242.21. Internal standard produced a protonated precursor ion [$(M+H)^+$] at 346.09 and corresponding product ion at m/z 198.09. This method showed linear response over the concentration range of $1{\sim}500\;ng/mL$ with correalation coefficient greater than 0.99. The lower limit of quantitation (LLOQ) using 0.2 mL plasma was 1 ng/mL, which was sensitive enough for pharmacokinetics studies. The method was specific and validated with a limit of quantitation of 1 ng/mL. The intra-day and inter-day precision and accuracy were acceptable for all samples including the LLOQ. The applicability of the method was demonstrated by analysis of plasma after administration of a single 10 mg dose to 36 healthy subject. From the plasma rabeprazole concentration versus time curves, the mean $AUC_t$ (The area under the plasma concentration-time curve from time 0 to 12 hr ) was $691.36{\pm}321.88\;ng{\cdot}hr/mL$, $C_{max}$ (maximum plasma drug concentration) of $353.21{\pm}131.52\;ng/mL$ reached $3.4{\pm}1.1\;hr$ after adiministration. The mean biological half-life of rabeprazole was $1.37{\pm}0.75\;hr$. Based on the results, this simple method could readily be used in pharmacokinetics studies.

• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.134-140
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• 2004

Optimal extraction method for antioxidant compounds from coat of grape dreg was established. Extracts were prepared with ethanol solutions containing phosphoric, formic, acetic, HCl, TFA, and citric acids. Antioxidant compounds of grape coat were determined by HPLC analysis and evaluated for antioxidizing effects using in vitro model system. Peonidin-3-glucoside content was the highest in 0.1% HCl-added ethanol extract. The extract prepared from pure ethanol without organic acids showed the highest content of cyanidin-3-glucoside among samples tested. Resveratrol and quercetin contents, the most important antioxidants, were highest in 0,1% HCl-added extract. Electron-donating ability was high in 0.1% acetic acid-added extract. SOD-like activities were 95.08% and 94.39% in 0.1% formic and 0.1% citric acid extracts, respectively. Inhibitory effects on peroxidation of egg yolk lecithin were observed in phosphoric (60.51%), citric (59,27%), and formic acid (56,77%) extracts. Hydrogen radical-scavenging activity was 59.47% in 0,1% HCl extract. Results suggest addition of 0.1% HCl in ethanol solution affords the highest yield in antioxidant compounds and antioxidant activities.

• Journal of Pharmaceutical Investigation
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• v.35 no.6
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• pp.437-443
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• 2005

A rapid, selective and sensitive reversed-phase HPLC method for the determination of a major metabolite of terfenadine, fexofenadine, in human serum was developed, validated, and applied to the pharmacokinetic study of terfenadine. Fexofenadine and internal standard, haloperidol were extracted from human serum by liquid-liquid extraction with acetonitrile and analyzed on a $Symmetry^{TM}$ C8 column with the mobile phase of 1% triethylamine phosphate (pH 3.7)-acetonitrile (67:33, v/v, adjusted to pH 5.6 with triethylamine). Detection wavelength of 230 nm for excitation, 280 nm for emission and flow rate of 1.0 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed fexofenadine concentration (50 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 10-500 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of serum was 10 ng/mL, which was sensitive enough for the pharmacokinetic studies of terfenadine. The overall accuracy of the quality control samples ranged from 95.70 to 114.58% for fexofenadine with overall precision (% C.V.) being 3.53-14.39%. The relative mean recovery of fexofenadine for human serum was 90.17%. Stability studies (freeze-thaw, short-term, extracted serum sample and stock solution) showed that fexofenadine was stable during storage, or during the assay procedure in human serum. However, the storage at $-70^{\circ}C$ for 4 weeks showed that fexofenadine was not stable. The peak area and retention time of fexofenadine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of fexofenadine in human serum samples for the pharmacokinetic studies of orally administered Tafedine tablet (60 mg as terfenadine) at three different laboratories, demonstrating the suitability of the method.

• Journal of Pharmaceutical Investigation
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• v.35 no.4
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• pp.265-271
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• 2005

A rapid, selective and sensitive reversed-phase HPLC method for the determination of etodolac in human serum was developed, validated, and applied to the pharmacokinetic study of etodolac. Etodolac and internal standard, ibuprofen were extracted from human serum by liquid-liquid extraction with hexane/isopropanol (95:5, v/v) and analyzed on a Luna C18(2) column with the mobile phase of 1% aqueous acetic acid-acetonitrile (4:6, v/v). Detection wavelength of 227 nm and flow rate of 1.0 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^3$ factorial design using a fixed etodolac concentration $(1\;{\mu}g/mL)$ with respect to its peak area and retention time. And also, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of $0.05-40\;{\mu}g/mL$ with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of serum was 0.05 ${\mu}g/mL$, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 92.00 to 110.00% for etodolac with overall precision (% C.V.) being 1.08-10.11%. The percent recovery for human serum was in the range of 76.73-115.30%. Stability studies showed that etodolac was stable during storage, or during the assay procedure in human serum. The peak area and retention time of etodolac were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of etodolac in human serum samples for the pharmacokinetic studies of orally administered Lodin XL tablet (400 mg as etodolac) at three different laboratories, demonstrating the suitability of the method.

• Journal of Pharmaceutical Investigation
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• v.35 no.6
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• pp.423-429
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• 2005

A selective and sensitive reversed-phase HPLC method for the determination of fenoprofen in human serum was developed, validated, and applied to the pharmacokinetic study of fenoprofen calcium. Fenoprofen and internal standard, ketoprofen, were extracted from human serum by liquid-liquid extraction with diethyl ether and analyzed on a Luna C18(2) column with the mobile phase of acetonitrile-3 mM potassium dihydrogen phosphate (32:68, v/v, adjusted to pH 6.6 with phosphoric acid). Detection wavelength of 272 nm and flow rate of 0.25 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed fenoprofen concentration $(2\;{\mu}g/mL)$ with respect to its peak area and retention time. And also, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of $0.05-100\;{\mu}g/mL$ with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was $0.05\;{\mu}g/mL$, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 92.27 to 109.20% for fenoprofen with overall precision (% C.V.) being 5.51-11.71 %. The relative mean recovery of fenoprofen for human serum was 81.7%. Stability (freeze-thaw, short and long-term) studies showed that fenoprofen was not stable during storage. But, extracted serum sample and stock solution were allowed to stand at ambient temperature for 12 hr prior to injection without affecting the quantification. The peak area and retention time of fenoprofen were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of fenoprofen in human serum samples for the pharmacokinetic studies of orally administered Fenopron tablet (600 mg as fenoprofen) at three different laboratories, demonstrating the suitability of the method.

• Applied Biological Chemistry
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• v.41 no.1
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• pp.60-66
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• 1998

During the screening for anti-complementary activity from 10 kinds of edible mushrooms, an alkali extract of Agaricus bisporus showed the highest activity through the complement fixation test. The crude anti-complementary material(AB-0) from Agaricus bisporus was obtained by the alkali extraction using 1 N NaOH containing 5% urea$(65^{\circ}C)$, followed by methanol reflux, dialysis and lyophilization. The fraction AB-O showed potent anit-complementary and anti-tumor activity against sarcoma-180 injected mice. The fraction AB-0 was divided into 5 fractions(AB-20, AB-40, AB-60, AB-80, AB-A) by gradual acetone precipitation. Among them fraction AB-20 having the highest activity and yield was found to contain 39% carbohydrate and 46% protein. The anti-complementary protein-bound polysaccharide AB-20 consisted of glucose, arabinose, xylose, galactose and mannose in a molar ratio of 6.49 : 1.98 : 1.24 : 1.00 : 0.71, respectively and its main component amino acids were alanine(20.59%), isoleucine(16.85%), glutamine+glutamic acid(14.12%) and leucine (13.83%). The anti-complementary activity of AB-20 was decreased greatly by periodate oxidation, but decreased slightly by pronase digestion. This indicates that polysaccharide moiety is corelated with the anti-complementary activity and that protein is also involved in the activity.

• Journal of Pharmaceutical Investigation
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• v.36 no.2
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• pp.89-95
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• 2006

A selective and sensitive reversed-phase HPLC method for the determination of pentoxifylline in human serum was developed, validated, and applied to the pharmacokinetic study of pentoxifylline. Pentoxifylline and internal standard, chloramphenicol, were extracted from the serum by liquid-liquid extraction with dichloromethane and analyzed on a Luna CI8(2) column with the mobile phase of acetonitrile-0.034 M phosphoric acid (25:75, v/v, adjusted to pH 4.0 with 10 M NaOH). Detection wavelength of 273 nm and flow rate of 0.8 mL/min were used. This method showed linear response over the concentration range of 10-500 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of the serum was 10 ng/mL, which was sensitive enough for pharmacokinetic studies of pentoxifylline. The overall accuracy of the quality control samples ranged from 89.3 to 92.7% for pentoxifylline with overall precision (% C.V.) being 4.1-9.2%. The relative mean recovery of pentoxifylline for human serum was 105.8%. Stability (stock solution, short and long-term) studies showed that pentoxifylline was not stable during storage. But three freeze-thaw cycles and extracted serum samples were stable. This method showed good ruggedness (within 15% C.V.) and was successfully applied for the analysis of pentoxifylline in human serum samples for the pharmacokinetic studies of orally administered $Trental^{\circledR}$ tablet (400 mg pentoxifylline), demonstrating the suitability of the method.

• Applied Biological Chemistry
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• v.21 no.1
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• pp.63-67
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• 1978

The incorporation of activity in plasma lipid was determined after injection of $u-C^{14}-glucose$ to normal laying hen. The extraction of total lipid from plasma was carried out by the method of Floch et al. with some modifications. The counting of carbon-14 activity was conducted by a Beckman Liquid Scintillation System, The concentration of plasma lipid was estimated as much as 3,070mg per 100ml of plasma taken at 5 minutes after injection of tracer amount of $u-C^{14}-glucose$. The specific activity per gram of plasma lipid carbon atom per injected dose per kg body weight increased gradually following the time after injection until 120 minutes, whereas the glucose activity decreased logarithmically. The partial incorporation quotient of activity in plasma lipid from that of glucose was 0.73% until 120 minutes after the injection.