한약 탕전 팩의 미생물 연구

유영법,마진열,하혜경,황대선,김복규,신광수,신현규,Yu, Young-Beob,Ma, Jin-Yeul,Ha, Hye-Kyung,Huang, Dae-Sun,Kim, Bok-Kyu,Shin, Kwang-Soo,Shin, Hyun-Kyoo 대한한의학방제학회 2007 大韓韓醫學方劑學會誌 Vol.15 No.2

Objectives: This study presents observation of microorganism such as total aerobic bacteria, total fungus, E. coli, Pseudonomas aerugjnosa, Staphylococcus aureus, and Salmonella typhimurium in herbal decoction manufactured by Korean medical clinic. Methods: We examined to observe microorganism using the requirements for the experimental methods recommended by FDA. For the identification, we observed microscopic methods and carried out polymerase chain reaction (PCR) and DNA purification. The purified DNA samples were analyzed by DNA sequencer. As compared with NCBI database. the results were identified by sequences similarity. Results and conclusion: 26 (55%) of 46 decoctions observed positive for microbial test. 12 (46 %) of 26 positive decoctions exceed requirement of microbial limit test. These microbial colony identified genus of Bacillus using microscopic and DNA sequencing methods.

Human Immunodeficiency Virus Type I에 대한 용규(龍葵) 추출물의 억제활성

유영법,Yu, Young-Beob 한국한의학연구원 2004 한국한의학연구원논문집 Vol.10 No.1

For the purpose of developing new anti-HIV agents from natural sources, the extracts of Solanum nigrum L. were tested for their inhibitory effects on HIV-1 replication and its essential enzymes as the reverse transcriptase (RT), protease and ${\alpha}$-glucosidase. In the assay of HIV-1-infected human T-cell line, water extracts inhibited the HIV- 1 -induced cytopathic effects with IC (inhibitory concentration) of 100 ug/ml. Moreover water extracts (100ug/ml) of aerial parts showed strong activity of 32.6% on anti-HIV-1 PR using the activity of the enzyme to cleave an oligopeptide. In the HIV-1 reverse transcriptase inhibition assay, aqueous extract a inhibited 17.4%, but no glucosidase inhibitory activities. We found out this result, for these samples it is possible that the inhibition of the viral replication in vitro is due to the inhibition at least one of PR and RT. It would be of great interest to identify the compounds which are responsible for this inhibition, since all therapeutically useful agent up to date are PR, RT and ${\alpha}$-glucosidase inhibitors.

능소화 잎 및 줄기 추출물의 Human Immunodeficiency Virus Type I 억제활성

유영법 한국자원식물학회 2012 한국자원식물학회지 Vol.25 No.2

적 요능소화 추출물의 HIV-1에 대한 항바이러스 효과를 복제 관련 효소에 대한 억제실험과 바이러스 복제억제 실험을 통하여 살펴보았다. 역전사효소 억제활성을 ELOSA 방법으로 실험한 결과 능소화 줄기의 물 추출물이 100 ㎍/㎖농도에서 각각 37.9%의 활성을 나타내었고, 능소화 잎과줄기의 메탄올 추출물에서 33.6% 및 31.5%의 HIV-1 protease 억제활성 나타 내었다. 그리고 HIV-1 복제 억제활성은 MT-4 세포에 대한 HIV-1 유도 세포변성억제를 광학현미경으로 관찰하여 살펴본 결과 줄기의 물 추출물이100 ㎍/㎖ 농도에서 HIV-1 바이러스 증식을 완전히 억제하였다. For the elucidation of action mechanism on anti-HIV of natural resources, the extracts of Campsis grandiflora were tested for their inhibitory effects on HIV-1 replication and its essential enzymes as the reverse transcriptase (RT),protease and α- glucosidase. In the assay of HIV-1-infected human T-cell line, water extracts of stem inhibited the HIV-1-induced cytopathic effects with IC (inhibitory concentration) of 100 ㎍/ml. Moreover water extracts (100 ㎍/ml) of stem showed strong activity of 37.9% on anti-HIV-1 RT using Enzyme Linked Oligonucleotide Sorbent Assay (ELOSA)method. In the HIV-1 protease inhibition assay, methanol extracts of stem and leaf extract showed 33.6% and 31.5%inhibition of the enzyme activity to cleave an oligopeptide resembling one of the cleavage sites in the viral polyprotein which can only be processed by HIV-1 protease, but did not exhibited glucosidase inhibitory activities. From these results,it is suggested that the inhibition of the viral replication in vitro is due to the inhibition of reverse transcriptase by water extracts of stem of Campsis grandiflora.

가미삼황산(加味三黃散) 분획물(SH-21-B)의 지표성분 정량과 구조활성상관(QSAR) 예측

유영법,Yu, Young-Beob 대한암한의학회 2006 大韓癌韓醫學會誌 Vol.11 No.1

Objective: Gami-Samhwang-San, a herbal prescription for obesity treatment, is composed of seven crude herbs such as Ephedrae Herba, Scutellariae Radix, Acori Gramineri Rhizoma, Polygalae Radix, Typhae Pollen, Armeniacae Semen, Nelumbo Folium. This study was aimed to evaluate marker substances in n-butanol fraction (SH-21-B) from Gami-Samhwang-San by high performance liquid chromatography (HPLC). And we predicted inhibition activity of major compounds of Gami-Samhwang-San using Quantitative Structure Activity Relationships (QSAR) Methods: The separation was performed on a YMC J,sphere-H80 CI8(250${\times}$4.6 mm I.D) column by gradient elution with $H_3PO_4$ buffers in acetonitrile as the moblie phase at a flow-rate of 1.0ml/min. Results: HPLC was employed to determine the quantities and the qualities of several marker substances such as ephedrine, pseudoephedirne, baicalin, ${\beta}-asarone$, tenuifoliside, naringenin, amygdalin and hyperoside in the SH-21-B. Conclusion: We suggest this results could be a useful evidence for quality control of SH-21-B.

Analysis of Scutellaria baicaleinsis Georgi (Scutellariae Radix) by LC-DAD and LC-ESI/MS

유영법,최필선,구성태,장수환 한국자원식물학회 2018 한국자원식물학회지 Vol.31 No.6

In this study, baicalin, as a marker substance of Scutellariae Radix, was quantitatively analyzed by a highperformance liquid chromatography-photodiode array detector (HPLC-DAD). We identified wogonoside, baicalein, andwogonin in the Scutellariae Radix by a high performance liquid chromatography-electrospray ionization-mass spectrometer(HPLC-ESI-MS). The baicalin was separated on a Xterra C18 column (5 ㎛, 4.6 x 250 ㎜) using mobile phase consisting of38% acetonitrile in 0.68% phosphoric acid. The baicalin spectrum in the Scutellariae Radix extracts was coincided bycomparing with UV-visible spectrum (200-550 ㎚) of baicalin standard in the library. The amount of baicalin in ScutellariaeRadix was 10.46%, which is higher than KFDA’s guideline. The marker substances of Scutellariae Radix showed a strongbase peak [M]+ in the positive detection mode following as; baicalin (m/z; 271 [MH+-sugar]+, 447 [M+H]+), wogonoside(m/z; 285 [MH+-sugar]+, 461 [M+H]+), baicalein (m/z; 271 [M+H]+), wogonin (m/z; 285 [M+H]+). These results areconsistent with the fragment pattern and molecular weight of standard components from literature.

Effects of Triterpenoids and Flavonoids Isolated from Alnus firma on HIV-1 Viral Enzymes

유영법,Hirotsugu Miyashiro,Norio Nakamura,Masao Hattori,박종철 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.7

Triterpenoids and flavonoids isolated from Alnus firma S. Z. were found to inhibit HIV-1 virus replication and controlled its essential enzymes. In this study, the inhibition of HIV-1 viral replication and its essential enzymes, such as reverse transcriptase, protease and α-glucosidase, were observed using 18 Korean plant extracts. Among the extracts, the methanol extract of Alnus firma leaves showed potent inhibition against the HIV-1 induced cytopathic effect (CPE) in MT-4 cells on microscopic observation (the minimum concentration for complete inhibition of HIV-1 induced CPE, IC=50 µg/mL). Thus, 14 compounds were isolated and identified from the methanol extract of Alnus firma leaves. Of these compounds, the alnustic acid methyl ester exhibited inhibition against HIV-1 protease, with an IC50 of 15.8 µM, and quercetin, quercitrin and myricetin 3-O-β-D-galactopyranoside displayed inhibition against HIV-1 reverse transcriptase, all with IC50 values of 60 µM. Based on these results, the viral replication inhibition of the methanol extract of Alnus firma leaves was adjudged to be acutely related to the protease inhibition activation of the alnustic acid methyl ester as well as the reverse transcriptase inhibition activation of flavonoids.

Angelica keiskei 지상부의 화학성분

유영법,이종호,최명락,옥광대,박종철,Yu, Young-Beob,Lee, Jong-Ho,Choi, Myeong-Rak,Ok, Kwang-Dae,Park, Jong-Cheol 한국생약학회 1996 생약학회지 Vol.27 No.1

Nucleoside, flavonol glycoside and disaccharide have been isolated from the aerial part of Angelica keiskei Koidz and identified by means of spectral analysis as adenosine, hyperoside and sucrose, respectively.

꽃치자나무 추출물의 HIV-1 효소 억제 활성과 QSAR에 의한 활성인자 예측

유영법 한국자원식물학회 2014 한국자원식물학회지 Vol.27 No.1

In this study, we conducted the anti- HIV-1 enzymes assay in vitro and its active components were predicted by QSAR in silico for the elucidation of action mechanism on anti-HIV of natural resources. The extracts of Gardenia jasminoides var. radicans Makino were tested for their inhibitory effects on the reverse transcriptase (RT), protease and α- glucosidase. In the enzyme inhibition assay, the methanol extracts of Gardenia jasminoides var. radicans Makino stem showed a strong activity of 32.5% on the enzyme activity to cleave an oligopeptide, resembling one of the cleavage sites in the viral polyprotein which can only be processed by HIV-1 protease. Moreover the methanol extracts of stem exhibited alpha-glucosidase inhibitory activities of 26.1%. The methanol extracts (100 μ g/ml) of stem showed a weak activity of 13.4% on anti-HIV-1 RT using Enzyme Linked Oligonucleotide Sorbent Assay (ELOSA) method. However, all extracts of leaf and stem didn’t exhibit the HIV-1-induced cytopathic effects with IC (inhibitory concentration) of 100 μ g/ml in HIV-1-infected human T-cell line. From these results, it is suggested that Gardenia jasminoides var. radicans Makino extracts may possibly be involved in the inhibition of reverse transcriptase, protease and alpha-glucosidase but can’t vitally concerned with the viral replication in vitro. 꽃치자나무 추출물의 HIV-1 reverse transcriptase, protease 및 alpha-glucosidase 에 대한 억제활성실험을 실시하였다 . ELOSA 방법으로 실험한 역전사효소 억제활성 실험에서는 꽃치자나무 잎 MeOH 추출물 100 ㎍ /ml 농도에서 13.4% 의 약한 억제활성이 관찰되었고 , 줄기의 메탄올추출물에서는 32.5% 의높은 HIV-1 protease 억제활성과 줄기의 메탄올 추출물에서는26.1% 의 alpha-glucosidase 억제활성이 관찰되었다 . 그리고 HIV-1 복제 억제활성은 MT-4 세포에 대한 HIV-1 유도 세포변성억제를 광학현미경으로 관찰하여 살펴본 결과 잎과 줄기의 모든 추출물에서 HIV-1 바이러스 증식억제에 억제활성이 관찰되지 않았다 . In silico 실험결과 주요성분인 crocetin 이 81.8% 의 높은 역전사 효소활성이 예측되었으며, genipin 이 55.39%, geniposidic acid aglycone이 64.5% 역전사효소 활성이 예측되었다 . 주로 aglycone 의 활성이 높게 예측되었으며 배당체의 경우 활성이 현저하게 저하되는 것으로 나타났다.

Quantitative and Qualitative Analysis of Alkaloids in Coptis chinensis (Coptidis Rhizoma) by LC-DAD and LC-ESI/MS

유영법,배창휴 한국자원식물학회 2017 한국자원식물학회지 Vol.30 No.6

The quality control of natural products is principal key to guarantee the Good Manufacturing Practices (GMP) and Good Clinical Practices (GCP) for the functional food, pharmaceuticals and cosmeceuticals in the industry. In this study, we examined the quantitative analysis of berberine as marker substance of Coptidis Rhizoma by high performance liquid chromatography-photodiode array detector (HPLC-DAD). The HPLC method was validated and met all the requirements for the quality control analysis recommended by FDA and ICH. The berberine was separated on a Xterra C18 column (5 ㎛, 4.6 × 250 ㎜) using mobile phase consisting of distilled water and acetonitrile with KH2PO4 (3.4 g) and Na2SO4 (1.7 g). Calibration curve of berberine has been estimated (y = 42293.47x-41589 with the correlation coefficient 0.9999). The amount of berberine was calculated as 4.25%. And berberastine, palmatine, columbamine, jatrorrhizine, epiberberine, berberine and coptisine in the Coptidis Rhizoma were identified by high performance liquid chromatography - electrospray ionization-mass spectrometer (HPLC-ESI-MS) method.

사방오리 잎의 Triterpenoid 및 Flavonoid 화합물

유영법,Norio Nakamura,Hirotsugu Miyashiro,Masao Hattori,박종철 한국생약학회 2007 생약학회지 Vol.38 No.1

Abstract − In this study, three triterpenoids, two steroids and nine flavonoids were isolated from the leaves of Alnus firma Sieb. et Zucc. On the basis of spectroscopic evidences, the structures of these compounds were established as β-amyrin acetate, β- amyrin, β-sitosterol, alnustic acid methyl ester, β-sitosterol glucoside, pinocembrin, alnustinol, quercetin, quercetin-3-O-α-Larabinofuranoside, quercetin-3-O-α-L-rhamnopyranoside, quercetin-3-O-β-D-glucopyranoside, myricetin-3-O-β-D-galactopyranoside, (+)-catechin and (−)-epicatechin.