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감마선 조사 당귀(Angelica gigas Nakai)의 유효성분 안정성 및 유전독성학적 안전성 연구
유영법(Young-Beob Yu),조성기(Sung-Kee Jo) 한국식품영양과학회 2000 한국식품영양과학회지 Vol.29 No.2
건조 생약재 당귀의 감마선 조사 위생화의 가능성을 검토하기 위하여 감마선 조사후 유효성분 안정성 및 유전독성학적 안전성을 평가하고자 하였다. 감마선 조사 당귀의 유효성분 안정성을 검증하기 위해 당귀의 유효성분인 decursin과 decursinol angelate를 분리, 정제하고 이를 표준품으로 하여 HPLC법으로 정량분석을 실시하였다. 구조 이성체인 decursin과 decursinol angelate는 normal phase column에서 좋은 분리능을 보여주었으며, 역상 column에서는 column의 길이와 내경이 분리능에 영향을 미침을 확인할 수 있었다. 감마선 조사(10 kGy) 당귀와 비조사 당귀(0 kGy)의 유효성분 정량결과, 조사 당귀에서 decursin의 함량이 0.27% 정도 증가하였으나 유의성은 관찰되지 않았으며, decursinol angelate는 조사 및 비조사당귀의 비교 결과 함량변화가 인정되지 않았다. Chromatogram의 pattern에서도 감마선 조사에 따른 변화가 관찰되지 않았다. 이는 방사선 조사 건조 생약재의 유효성분의 안정성을 확보하는데 중요한 결과로 생각된다. 또한 10 kGy 감마선 조사 당귀의 열수 추출물은 Salmonella thyphimurium 균주를 이용한 복귀돌연변이 실험에서 복귀돌연변이 수를 증가시키지 않았으며, CHO 세포를 이용한 소핵유발성 검증에서도 소핵빈도를 증가시키지 않았다. 이로써 감마선 조사된 당귀의 추출물이 돌연변이원으로 작용하지 않으며, 세포 핵분열중에 독성을 나타내지 않음을 알 수 있었다. 이상의 결과로 보아 생약재 당귀를 감마선으로 조사한 후에도 당귀내 유효성분이 그대로 보전되는 것으로 사료되며, 감마선 조사 당귀 추출물의 독성학적 안전성은 앞으로 생체내 유전독성시험, 만성독성 및 생식독성시험 등을 통하여 확인될 수 있을 것으로 사료된다. In the present studies, we assessed the stability of active components and toxicological safety of irradiated Angelica gigas Nakai (Danggui). In order to confirm the stability of active components in the γ-irradiated roots of Danggui, the quantitative analysis of decursin and decursinol angelate of γ-irradiated sample was carried out by high performance liquid chromatographic (HPLC) methods using reverse phase columns and normal phase columns. From the root of Danggui, decursin and decursinol angelate were isolated by a silica gel column chromatography (toluene : ether (1 : 1), Hexane : EtOAc (15 : 1)). And then the struc- tures were confirmed in the ¹H and ¹³C-NMR analysis. The HPLC chromatograms of decursin and decursinol angelate in γ-irradiated Danggui were similar with those of non-irradiated sample. In the examination of in vitro genotoxicity of the water extract from γ-irradiated Danggui using Salmonella reversion assay (Ames test) and micronucleus test in Chinese hamster ovary (CHO) cells, mutagenicity was not exhibited in the two assays with or without metabolic activation. These results suggest that active components in the γ-irradiated Danggui should be stable and that the safety of γ-irradiated Danggui could be revealed in further tests in vivo.
HPLC로 표준화한 가미삼황산(加味三黃散) 분획물(SH-21-B)의 랫드에 대한 단회경구투여독성시험
유영법(Young-Beob Yu),김선형(Seon Hyeong Kim),윤유식(Yoo-Sik Yoon) 한국독성학회 2005 Toxicological Research Vol.21 No.3
Gami-Samhwang-San, a herbal prescription for obesity treatment, is composed of seven crude herbs such as Rehmanniae Radix Preparata, Ephedrae Herba, Scutellariae Radix, Acori Gramineri Rhizoma, Polygalae Radix, Typhae Pollen, Armeniacae Semen, Menthae Herba. In this study, marker substances in n-butanol fraction (SH-21-B) from Gami-Samhwang-San were analyzed by high performance liquid chromatography (HPLC) and acute toxicity of standardized SH-21-B was evaluated by good laboratory practices (GLP) guideline of Korea Food and Drug Administration. Therefore, we confirmed that there were baicalin of 15.92%, amygdalin of 6.57% and ephedrine of 2.49% in SH-21-B. SH-21-B was administered in rats at dose of 0 ㎎/㎏, 2,000 ㎎/㎏, and 5,000㎎/㎏. Clinical signs of both sexes of rats were observed daily for 14 days after single oral administration. Two female rats, one administered at 2,000 ㎎/㎏ and the other administered at 5,000 ㎎/㎏, died, but no dead animal was observed among male rats. Therefore LD_(50) in the female rat is observed to be 8,710 ㎎/㎏, and MLD (Minimun Lethal Dose) of the male rat is observed to be more than 5,000 ㎎/㎏.
유영법(Young-Beob Yu),Mi-Jung Kim(김미정),Do-Young Yum(염도영),Bon-Taq Koo(구본탁),Hye-Kyeong Ha(하혜경),Hyun-Kyu Shin(신현규),Jin-Yeul Ma(마진열),Jong-Cheol Park 한국한의학연구원 2006 한국한의학연구원 연구보고서 Vol.- No.-
The purpose of this study was to examine the effects of the Hominis placenta extracts and Astragali radix extracts on improvement of senile osteoporosis (Type 2) in SAM P6. Placenta ex-tracts (Beuronel<SUP>®</SUP>) produced by INBIONET Corp, Daejeon, Korea is hydrolysate of dried human placenta that contains interferon, prostaglandin E1, lysozyme, glycosaminoglycans (GAG) and various amino acid such as leusine, alanine and glutamic acid. In traditional oriental medicine, Placenta (Jahageo) is indicated for insufficiency of blood and tidal fever due to steaming bone disorder. At 10 weeks after birth, Placenta ex. and Astrali Radix ex. was given 100μL and 200μL/kg/day (i.p.), 3times a week for 4 weeks in SAM P6 mice. We measured complete blood cells (CBC) such as RBC, HGB, Hct, PLT, MPB and MCHC. And we analyzed the plasma concentrations of blood urea nitrogen, creatinine, inorganic phosphate and total iron. In addition, we tested bone mineral density (BMD) using the soft X-ray (Lunar Pixirnus). As a result, in SAM P6, injection of Placenta ex. (200μL) increased in RBC, HB and PLT, in comparison with control group. It was also found that the inorganic phosphate levels increased significantly in the injection groups of the Placenta ex. from that of the control group, but blood urea nitrogen was no significant. Moreover, Placenta ex. and Astrali Radix ex. were showed a trend of increase in bone mineral density (BMD) of the vertebrae lumbales, femur and tibia in P6 mice (Fig. 1). These findings suggest that Placenta ex. and Astragali Radix ex. are effective in preventing bone loss in SAMP6. In conclusion, these extracts help on improvement of osteoporosis in SAMs through probably hematogenous functions. <그림참조>
HPLC로 표준화한 가미홍화탕(KH-19)의 랫드와 Beagle견에 대한 단회경구투여독성시험
유영법(Young-Beob Yu),성현제(Hyun Jea Sung),윤유식(Yoo-Sik Yoon) 한국독성학회 2005 Toxicological Research Vol.21 No.1
Gami-Honghwa-Tang (KH-19), a herbal prescription for reducing the side effect of radiotherapy, is composed of eight crude herbs such as Rehmanniae Radix Preparata, Angelicae Gigantis Radix, Cnidii Rhizoma, Paeoniae Radix, Corni Fructus, Moutan Cortex Radicis, Lycii Fructus, Carthami Flos, and Glycyrrhizae Radix. In this study, marker substances in KH-19 were analyzed by high performance liquid chromatography-diode array detector (HPLC-DAD) and safety evaluation of standardized KH-19 was evaluated by good laboratory practices (GLP) guideline of Korea Food and Drug Administration. HPLC-DAD was employed to determine the quantities and the qualities of several marker substances such as 5-hydroxymethyl-2-furaldehyde (5-HMF), paeonol, loganin, paeoniflorin, glycyrrhizin, and decursin in the KH-19. In acute oral toxicity study on rat, transient inhibition of body weight was shown, but change in general symptom was not detected. No dead animal was observed up to 5,000 mg/kg in both male and female animals. In acute oral toxicity study on Beagle dog, transient vomiting, diarrhea, anorexia, and body weight reduction were observed, However, no dead animal was observed up to 2,000 mg/kg in both male and female animals.
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.