• 한방재활의학과학회지
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• 제27권2호
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• pp.29-38
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• 2017

Objectives The present study aimed to investigate the change in marker compounds, anti-oxidative and anti-inflammatory effects of salt-water processed Cortex Eucommiae. Methods To evaluate the influence of processing on anti-oxidant effect of Cortex Eucommiae, changes in total phenol, total flavonoid, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) free radical scavenging, and ferric reducing antioxidant power (FRAP) between processed and raw Cortex Eucommiae were assessed. In addition, nitrite assay was conducted to determine the influence of processing on anti-inflammatory effect of Cortex Eucommiae. Cell viability was also examined as to elucidate whether processing affects cytotoxicity of Cortex Eucommiae. Finally, high-performance liquid chromatography (HPLC) analysis was conducted to monitor changes in pinoresinol diglucoside amount of processed and raw Cortex Eucommiae. Results Salt-water processed Cortex Eucommiae showed higher total phenol and flavonoid amount, compared to raw Cortex Eucommiae. Furthermore, anti-oxidative activity of processed Cortex Eucommiae was improved as discovered in DPPH, ABTS, and FRAP assays. Anti-inflammatory effect of Cortex Eucommiae was also enhanced following salt-water processing, as evidenced in nitrite assay. HPLC analysis found that the amount of pinoresinol diglucoside, widely known as the marker compound of Cortex Eucommiae, increases through salt-water processing. All experiments were performed with non-toxic concentration of Cortex Eucommiae; processing did not affect the cytotoxicity of Cortex Eucommiae up to the currently adopted concentration. Conclusions The present results support that salt-water processing of Cortex Eucommiae is beneficial in terms of marker compound amount, anti-oxidative, and anti-inflammatory activities. Additional investigations are needed to standardize the processing method of Cortex Eucommiae.

• 생약학회지
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• 제49권4호
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• pp.312-321
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• 2018

Recently, consumer demand for functional cosmetics containing natural ingredients has been greatly expanded. To develop the natural cosmetic materials, we selected 3 plants, Chrysanthemum zawadskii Herbich (CZ), Perilla frutescens (L.) Britton var. acuta Kudo (PF), and Rosa multiflora Thunberg (RM) which showed high total flavonoid contents (TFC), total polyphenol contents (TPC), and strong DPPH radical scavenging effect. We determined astragalin, chlorogenic acid, and rosmarinic acid as a marker compound for quantitative analysis of the content of each material and standardization of the quality standards and manufacturing standards through LC/MS analysis. HPLC-DAD was used to simultaneously analyze these marker components of three natural product complexes (Mix) and to validate the analytical method through experiments such as linearity, accuracy and precision. The detection wavelengths were set at 210, 265, and 330 nm. The detected 3 compounds from extract of CZ, PF, RM showed significant linearity ($R^2${\geq_-}$0.9947). The limit of detection (LOD) of chlorogenic acid, astragalin and rosmarinic acid were $8.29{\mu}g/ml$, $2.28{\mu}g/ml$, and $27.00{\mu}g/ml$, respectively. The limit of quantification (LOQ) of chlorogenic acid, astragalin and rosmarinic acid were $25.11{\mu}g/ml$, $6.92{\mu}g/ml$, and $81.83{\mu}g/ml$, respectively. The contents of the three indicators of Mix were 19.82-24.71 mg/g of chlorogenic acid, 43.80-46.02 mg/g of astragalin, and 46.33-48.57 mg/g of rosmarinic acid.

• Bulletin of the Korean Chemical Society
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• 제34권7호
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• pp.2081-2085
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• 2013

A quantitative method for determining levels of three bioactive compounds based on pattern recognition was developed and fully validated for the quality control of Alismatis Rhizoma (AR) by HPLC. Separation conditions were optimised using an Optimapak $C_{18}$ column ($250mm{\times}4.6mm$, 5 ${\mu}m$) with a mobile phase of acetonitrile and 0.1% aqueous phosphoric acid and detection wavelengths of 205 and 245 nm. Method validation yielded acceptable linearity ($r^2$ > 0.9998) and percent recovery (98.06%-101.71%). Limits of detection ranged from 0.08 to 0.15 ${\mu}g/mL$. Levels of the three bioactive compounds, alisol C acetate, alisol B, and alisol B acetate, in AR were 0.07-0.45, 0.38-10.32, and 1.13-8.59 mg/g dried weight, respectively. Pattern analyses based on these three compounds were able to differentiate Chinese and Korean samples accurately. The results demonstrate that alisol B and its acetate may be used as marker compounds for AR quality and can be regulated to no less than 0.36 and 1.29 mg/g of dried sample, respectively. The method described here is suitable for quantitative analyses and quality control of multiple components in AR.

• 생약학회지
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• 제49권3호
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• pp.270-277
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• 2018

Pyungwi-san has been used to treat the digestive system diseases, physconia, nausea, anorexia, and dyspepsia in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was optimized for simultaneous determination of the 14 marker components, spinosin, liquiritirn apioside, liquiritin, narirutin, 6'''-feruloylspinosin, hesperidin, liquiritigenin, glycyrrhizin, 6-gingerol, atractylenolide III, honokiol, atractylenolide II, magnolol, and atractylenolide I in Pyungwi-san extract. All analytes were separated on a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) with maintained at $45^{\circ}C$. The mobile phase consisted of 0.1% (v/v) aqueous formic acid and acetonitrile. The MS conditions were as follows: capillary voltage 3.3 kV, extractor voltage 3.0 V, RF lens voltage 0.3 V, source temperature $120^{\circ}C$, desolvation temperature $300^{\circ}C$, desolvation gas 600 L/h, cone gas 50 L/h and collision gas 0.14 mL/min. The coefficient of determination of 14 analytes was 0.9989-1.0000. The limits of detection and quantification values of the all analytes were 0.04-2.56 and 0.13-7.69 ng/mL, respectively. As a result of the analysis using the established LC-ESI-MS/MS method, the 5 components, spinosin, 6'''-feruloylspinosin, atractylenolide III, II, and I derived from Zizyphi Fructus and Atractylodis Rhizoma, were not detected in this extract. On the other hand, the 9 components except for the 5 components were 4.15-498.87 mg/kg in lyophilized Pyungwi-san extract. Among these components, glycyrrhizin, marker compound of Glycyrrhizae Radix et Rhizoma, was detected the most amount as a 498.87 mg/kg.

• 대한본초학회지
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• 제33권6호
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• pp.29-34
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• 2018

Objectives : It is necessary to manage herbal medicines based on effectiveness by comparing the efficacy of herbal medicines by region. In this study, we compared anti-oxidative activity and marker compounds of Eucommiae Cortex by regional groups. Methods : Eucommiae Cortex grown and harvested in Gangjin, Sancheong, Yeongwol, Jangsu, and Jecheon were used. Eucommiae Cortex was extracted in distilled water at $100^{\circ}C$ for 3 hours, and filtered. filtered. Extract samples were freeze-dried at $-80^{\circ}C$. Comparison of anti-oxidant activity in Eucommiae Cortex samples from regional groups was measured in 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) free radical scavenging, and ferric reducing antioxidant power (FRAP) between regional groups of Eucommiae Cortex. In addition, high-performance liquid chromatography (HPLC) analysis was conducted to compare pinoresinol diglucoside concentration by regional groups. Results : HPLC analysis found that pinoresinol diglucoside concentration, widely known as the marker compound of Eucommiae Cortex, was the highest in Gangwon Yeongwol. There was a significant difference in anti-oxidative activity of Eucommiae Cortex between regional groups as discovered in DPPH, ABTS and FRAP assays. DPPH free radical scavenging was the highest in Jeonbuk Jangsu. ABTS free radical scavenging was the highest in Jeonbuk Jangsu and Chungbuck Jecheon. FRAP was the highest in Jeonbuk Jangsu. Conclusions : Although pinoresinol diglucoside concentration was high, anti-oxidative activity was not proportionately high. Pinoresinol diglucoside concentration was the highest in Gangwon Yeongwol. Anti-oxidative activity was the highest in Jeonbuk Jangsu.

• 대한한의학방제학회지
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• 제21권1호
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• pp.80-90
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• 2013

Objectives : The water extract of Hwangnyeonhaedok-tang (HHT), composed of the Scutellariae Radix, Coptidis Rhizoma, Phellodendri Cortex and Gardeniae Fructus has been traditionally used to treat fever, inflammation, gastritis and hypertension in east asia. However, little is known about the ameliorative effects of HHT on atopic dermatitis. Therefore, the purpose of this study was to investigate the therapeutic effect of HHT on atopic dermatitis Methods : We investigated the inhibitory effects of HHT on the production of proinflammatory cytokines in rat peritoneal mast cells (RPMCs), on the scratching behavior in ICR mice, and on atopic dermatitis symptoms in 2,4-dinitrofluorobenzene (DNFB)-induced atopic dermatitis-like model hairless mice. Results : Levels of TNF-${\alpha}$ and IL-$1{\beta}$ increased by PMA plus A23187 co-treatment were significantly inhibited by HHT in a dose-dependent manner. HHT also inhibited the histamine release from RPMCs stimulated by compound 48/80, which promotes histamine release. The oral administration of HHT reduced the scratching behavior induced by compound 48/80 and histamine in ICR mice. Furthermore, the intradermal treatment of HHT reduced the ear edema, skin lesions, and atopic molecular marker (IgE and IL-4) in DNFB-induced atopic dermatitis model mice. Conclusions : These results suggest that HHT may be used as a potential treatment for AD as a prescription for treatment of atopic dermatitis.

• 한국식품영양과학회지
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• 제44권1호
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• pp.85-88
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• 2015

차조기 열수 추출물을 개별인정형 건강기능식품 기능성 원료로 개발하기 위하여 지표성분 표준화를 위한 rosmarinic acid의 분석법 설정과 분석법에 대한 밸리데이션을 실시하였다. 1% acetic acid가 첨가된 water와 methanol을 이동상으로 하고 Zorbax extended-C18($4.6{\times}150mm$, $5{\mu}m$) 칼럼을 사용하여 기울기 용리(gradient elution) 방법으로 분석하였다. 본 연구에서는 분석법을 확립하고 분석법에 대하여 특이성, 직선성, 정확성과 정밀성 그리고 회수율에 대하여 확인하였다. Rosmarinic acid의 검량선은 $R^2$=0.9995로 좋은 선형성을 보였으며 정량한계와 검출한계는 각각 $1.2{\mu}g/mL$와 $0.36{\mu}g/mL$였다. 일내와 일간 분석에서 상대표준편차(RSD)는 각각 3.21%와 1.43% 미만으로 나왔다. 정밀성 측정 결과에서는 97.04~98.98%로 나왔고, RSD는 0.25~1.97%였다. 그러므로 HPLC를 이용한 rosmarinic acid의 분석법이 차조기 열수 추출물 기능성원료 표준화를 위한 적합한 시험법임이 검증되었다. 본 시험법에 따라 분석한 차조기 열수 추출물 내의 rosmarinic acid의 함량은 세 lot를 3회 분석하였을 때 평균 0.03%로 나왔다. 따라서 본 연구를 통하여 확립된 rosmarinic acid의 분석법이 차조기열수 추출물 개별인정형 건강기능식품 기능성 원료 개발을 위한 기초자료로 활용될 것으로 사료된다.

• 한방재활의학과학회지
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• 제27권1호
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• pp.19-25
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• 2017

Objectives Eucommiae cortex is one of the frequently used herbs for musculoskeletal disorders, with well-known anti-inflammatory effect. Meanwhile, powdered form of herbal drugs is more advantageous than decoction form, in that storing and taking is more convenient. Thus, the authors aimed to investigate whether the drying method affects the index compound level and anti-inflammatory effect of eucommiae cortex water extract. Methods Eucommiae cortex was extracted in distilled water at $100^{\circ}C$ for 3 hours, filtered, and then evaporated under vacuum. One half of the sample was freeze-dried at $-80^{\circ}C$. Another half of the sample was added with dextrin and then spray-dried at $180^{\circ}C$. To assess the possible change in index compound content, pinoresinol diglucoside was selected as the index marker. The content level of the index compound in various extract sample was quantified through high performance liquid chromatography. In addition, iNOS assay in LPS-induced RAW 264.7 cells was adopted to determine the anti-inflammatory effect of various extract samples. Furthermore, MTT assay was performed to confirm that the result of iNOS assay was not due to cytotoxicity. Results There was no significance difference in index compound content between extract samples obtained through two different drying methods. Anti-inflammatory activity of extract samples were similar at the matching concentration, regardless of the drying methods. Extract samples did not show any significant cytotoxicity. Conclusions Extract samples of eucommiae cortex were obtained through freeze-drying and spray-drying. Neither change in index compound content nor difference in anti-inflammatory activity was observed between drying methods.

• 대한본초학회지
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• 제29권3호
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• pp.35-42
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• 2014

Objectives : Since single extract powders was released at 1987, the insurance fee has not been changed, but the price of raw material has been increasing. According to this, Pharmaceutical company couldn't invest on quality of the product, so, the quality of single extracts went down and lost the consumer's trust. We checked the contents of marker ingredients in single extract thereby to recover the reliability of insurance-covered herbal preparations. Methods : we bought total twelve products of eight different single extracts of two main pharmaceutical companies among total 65 kinds of single extract powder stipulated in KHP (The Korean Herbal Pharmacopoeia) monograph III at Jan. 2013. Assays of selected single extract powders are performed by KHP regulation. And we surveyed price fluctuation of relevant herb raw materials from 2005 to 2012. Results : Among twelve products, eight single extract powders were suitable by the KHP regulation. But four products didn't reach the content amount of KHP. Marker contents in the single extracts product of Pueraria Root, Licorice, Peony root and Scutellaria Root of A company were 70%, 1%, 23.7% and 75.1%, respectively. Conclusions : We can acertain whether there's a quality problem in the insurance-covered single extract powders. But, A company is no longer producing these improper single extract powders. As a medicine, single extract powders needs to be strictly quality controlled by the company, and regularly monitored by the KFDA.

• 한국표면공학회지
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• 제35권4호
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• pp.193-198
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• 2002

Alloys of XD45 (Ti45A12Nb2Mn-0.8vol%TiB$_2$) and XD47 (Ti47A12Nb2Mn-0.8vol%TiB$_2$) were oxidized between 800 and $1000^{\circ}C$ in air, and their oxidation characteristics were studied. The oxide scales consisted primarily of an outer $TiO_2$ layer, an intermediate $Al_2$$O_3$-rich layer, and an inner mixed layer of ($TiO_2$+$Al_2$$O_3$). Nb tended to present at the lower part of the oxide scale, whereas Mn at the upper part of the oxide scale. The Pt marker tests indicated that the outer oxide layer grew primarily by the outward diffusion of Ti and Mn, and the inner mixed layer by the inward transport of oxygen.