• The Korea Journal of Herbology
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• v.30 no.3
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• pp.19-24
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• 2015

Objectives : HPL-1, a novel herbal medicine which is composed of five herbs such as Kalopanacis Cortex, Chaenomelis Fructus, Raphani Semen, Atractylodis Rhizoma and Pulvis Aconiti Tuberis Purificatum, was developed for treatment of osteoarthritis. This study is aimed to develop analytical method for consistent quality control of HPL-1 and validate chromatographic method. Methods : Chromatographic analysis was performed using ultra-high performance liquid chromatography - diode array detector (UHPLC-DAD) equipped with RP-amide column, column oven, and auto sampler. Marker compounds [protocatechuic acid, chlorogenic acid, liriodendrin, 3,5-dicaffeoylquinic acid, ${\beta}$-D-(3-O-sinapoyl)-fructofuranosyl-$\alpha$-D-(6-O-sinapoyl)glucopyranoside and benzoylmesaconine] were separated by step gradient elution of acetonitrile and 0.1% phosphoric acid/water. The method validation was evaluated by quantitative validation parameters of linearity, accuracy, precision, limit of detection (LOD) and limit of quantification (LOQ) according to KFDA guideline.Results : An optimized method for six marker compounds in HPL-1 was established by UHPLC-DAD. The correlation coefficient (R²) with each calibration curve was greater than 0.99. The LOD and LOQ were within the range of 0.008-0.090 and $0.023-0.274{\mu}g/mL$, respectively. The relative standard deviation (RSD) of intra- and inter-day variability were less than 4.0%. The result of recovery test was range from 93.3-106.3% with RSD < 4.0%.Conclusions : These results suggest that the quantitative UHPLC method is precise, accurate, effective for quality evaluation of HPL-1. The method may also contribute to improve quality of crude drug preparations used for treatment of various diseases.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.395-402
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• 2005

Biologically active recombinant human follicle stimulating hormone (rhFSH) was produced in Chinese hamster ovary cells and purified by a series of chromatographic steps. The chromatographic steps included anion-exchange chromatography (DEAE Sepharose F/F, Q Sepharose F/F), hydrophobic interaction chromatography (Source 15 PHE), and hydroxyapatite chromatography (Macro-Prep ceramic hydroxyapatite type I). A distinctive step of the purification process developed was the use of ZnCl$_2$ for the removal of non-glycosylated or lowly-glycosylated FSH and impurities through co-precipitation with Zn$^{2+}$. Purified rhFSH was identified and characterized by several physicochemical and biological methods such as gel electrophoresis, high-performance liquid chromatography, amino acid analysis, carbohydrate analysis, and biological activity. The overall yield of the purification was ~$30\%$. The rhFSH preparation obtained showed high purity (>$99\%$) and high in vivo potency (>16,000 IU/mg). Carbohydrate analysis suggested that the purified rhFSH contained approximately $40\%$ (w/w) carbohydrate with di­or tri-antennary structure on average, which is somewhat more heavily sialylated than commercially available rhFSH. In conclusion, the results of these analyses established an identity of the purified rhFSH with natural FSH from human pituitary glands, and furthermore, the purified rhFSH preparation showed higher in vivo potency and was slightly more heavily sialylated than commercially available rhFSH.

• Journal of the Korean Chemical Society
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• v.36 no.1
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• pp.66-74
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• 1992

A method for the multiresidual simultaneous analysis of 11 thiocarbamates was studied using HPLC. Thiocarbamate in Chinese cabbage was analyzed in the order of extraction, partition, and cleanup in their optimum condition. Acetone was chosen as an extracting solvent. As a partitioning solvent, the mixture of 50% methylene chloride and petroleum ether containing extremely small water content showed good recoveries of thiocarbamate from the water layer. Partition efficiency was affected by pH of the water layer; it remained almost constant under the acidic and neutral condition while decreasing under the basic condition. The comparison done in cleanup step showed that the column chromatographic method is superior to the treatment of coagulating reagent. As an absorbent, the mixture of charcoal, magnesia, and celite with the ratio of 1 : 2 : 4 gave better recoveries and also effectively removed chlorophyll. Over the total procedure, the average recoveries for thiocarbamates in Chinese cabbage were 91% at about 2 ppm fortification level within the relative standard deviation of 8%, and the minimum detection limit (MDL) was 2.2${\sim}$9.3 ng.

• Natural Product Sciences
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• v.12 no.1
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• pp.55-61
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• 2006

A description of simple, isocratic and precise reversed phase HPLC methods is given for simultaneous quantification of pholcodine and ephedrine hydrochloride together with either carbinoxamine maleate or terfenadine in antitussive-antihistaminic oral pharmaceutical formulations. Separations were carried out on X-Terra and symmetry shield C18 column $(250\;{\times}\;4.6\;mm,\;5\;{\mu}m)$. The used isocratic elution systems were either $0.02\;M\;KH_2PO_4-acetonitrile$ in the ratio of 75 : 25 and pH adjusted to 7.70 with orthophosphoric acid or sodium hydroxide, for syrup (method A), or 0.02 octanesulphonic acid sodium salt solution-acetonitrile-acetic acid in the ratio of 75 : 25 : 0.5 for suspension (method B). The elution of both mixtures was achieved with a flow rate of 1 ml/min. Detection was carried out by UV absorbance at wavelengths of 220 and 250 nm for syrup and suspension, respectively. The quantification of the components in synthetic mixtures and actual syrup and suspension were calculated using the internal standard technique with metoclopramide HCl and codeine phosphate as internal standards (IS), respectively. The methods, for both mixtures, were validated and met all the requirements for the quality control analysis recommended by FDA and ICH.

• Korean Journal of Environmental Agriculture
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• v.23 no.4
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• pp.251-257
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• 2004

Analytical methods were developed to determine cyclosulfamuron residues in soil, water, rice grain and straw using high-performance liquid chromatography (HPLC) with ultraviolet absorption detection. In these methods, cyclosulfamuron was extracted with aqueous $Na_2HPO_4$/acetone and acetone/methanol mixture from soil and rice samples respectively. Liquid-liquid partition coupled with ion-associated technique, Florisil column chromatography, and solid-phase extraction (SPE) were used to separate cyclosulfamuron from interfering co-extractives prior to HPLC analysis. For water sample, the residue was enriched in $C_{18}$-SPE cartridge, cleaned up in situ, and directly subjected to HPLC. Reverse-phase HPLC under ion-suppression was successfully applied to determine cyclo-sulfamuron in sample extracts with the detection at its ${\lambda}_{max}$ (254 nm). Recoveries from fortified samples averaged $87.8{\pm}7.1%$ (n=12), $97.3{\pm}7.2%$ (n=12), $90.8{\pm}6.6%$ (n=6), and $78.5{\pm}6.7%$ (n=6) for soil, water, rice grain and straw, respectively. Detection limits of the methods were 0.004 mg/kg, 0.001 mg/L, 0.01 mg/kg and 0.02 mg/kg for soil, water, rice grain and straw samples, respectively.

• Journal of Food Hygiene and Safety
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• v.12 no.4
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• pp.288-293
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• 1997

Analytical method using capillary GC/ECD was developed to determine trace residues of chlofluazuron, 1-[3, 5-dichloro-5-trifluoromethyl-2-pyridyloxy)phenyl]-3-(2, 6-difiuorobenzoyl), in meat, and applied to analyze the residues in domestic and imported meats. The analytical scheme developed does not require column chromatographic cleanup; chlorfiuazuron was extracted with diethyl ether and petroleum ether (50: 50), partitioned against acetonitrile, cleaned up with silica Sep-Pak cartridge, identified GC/ECD, and comfirmed by GC/MS. The mean recoveries of the pesticide in meat fortified with standard solution 0.1, 0.5, 0.1 mg/kg were ranged from 82 to 95%. The limit of detection and limit of quantitation were 0.001 and 0.005 mg/kg, respectively. Chlorfluazuron residues were not found in domestic samples, but found in imported Australian beef ranging from 0.02 to 0.17 mg/kg, detected by 18% among the samples.

• Natural Product Sciences
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• v.14 no.3
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• pp.171-176
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• 2008

Column chromatographic separation of 70% EtOH extract of the roots of Korean cultivated-wild ginseng led to the isolation of ten ginsenosides (1 - 10). The isolated compounds were identified as ginsenoside $Rg_1$ (1), ginsenoside Re (2), ginsenoside Rc (3), ginsenoside $Rb_1$ (4), ginsenoside $Rb_2$ (5), ginsenoside Rd (6), ginsenoside $Rg_3$ (7), ginsenoside $F_2$ (8), ginsenoside $Rb_3$ (9), and ginsenoside $Rd_2$ (10) by physicochemical and spectroscopic methods. The compounds (1 - 10) were for the first time isolated from the roots of Korean cultivated-wild ginseng.

• Korean Journal of Pharmacognosy
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• v.27 no.4
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• pp.366-370
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• 1996

Fruits of Rubus coreanum (Korean name: Bog-bun-ja) have been used in oriental traditional medicine as the remedies for impotence, pollution, premature ejaculation and frequency of urination etc. It is known to have phenolic compounds as an astringent. By means of chromatographic methods, four hydrolyzable tannins were isolated from the fruits of R. coreanum. The structures of these compounds were established as gallic acid, 2,3(S)-HHDP-D-glucopyranose, sanguiin H-4 and sanguiin H-6 on the basis of physicochemical and spectroscopic evidences.

• Natural Product Sciences
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• v.6 no.1
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• pp.11-15
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• 2000

Two soyasaponins were isolated from the aerial parts of Astragalus tribuloides Del. Their structures were established on the basis of spectroscopic and chemical methods. In addition, ursolic acid, ${\beta}-sitosterol\;{\beta}-D-glucoside$ and isorhamnetin 3-O-glucoside were isolated and identified by comparing their mp, spectral and chromatographic data with those of authentic samples. This is the first report of screening and isolation of the chemical constituents of this species of genus Astragalus.

• Natural Product Sciences
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• v.10 no.6
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• pp.262-267
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• 2004

A reversed-phase liquid chromatographic method for decursin and decursinol angelate of Angelicae gigantis Radix, an important crude drug in Korean traditional medicine, was developed and validated. Decursin and decursinol angelate, the structure isomer (pyranocoumarin) each other, are the main organic constituents in Angelicae gigantis Radix. This method was developed using a RP-18 column, UV detector at 280 nm and 50% acetonitrile solution containing 0.01 M sodium dodecyl sulfate and 25 mM sodium dihydrogen phosphate (pH 5.0) as the mobile phase. Various validation parameters were included and evaluated satisfactorily. Linearity was established in range 2-75 mg/ml of decursin and decursinol angelate (correlation coefficient = 0.9997 and 0.9995, respectively). This analytical method showed good accuracy (98.1% and 99.5%, respectively). Precision (repeatability) revealed a relative standard deviation value of 1.71% (decursin) and 3.19% (decursinol angelate). For intermediate precision measure the considered variables were equipment and days. A robustness test showing the influence of deferent counter-ion concentration in mobile phase was also performed.