현삼 (Scrophularia buergeriana)에서 분리한 화합물의 함량분석 및 간세포 보호 효과

나현선,오선민,신우철,황보전,김형근,윤다혜,양승환,이영섭,김금숙,백남인,이문순,이대영 한국응용생명화학회 2019 Journal of Applied Biological Chemistry (J. Appl. Vol.62 No.4

The roots of Scrophularia buergeriana were extracted with 80% aqueous Methanol and the concentrates were partitioned into EtOAc, n-BuOH, and H2O fractions. The repeated silica gel or octadecyl SiO2column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of phenylethanoid glycosides and iridoid glycosides. The chemical structures of these compounds were determined as harpagoside (1), angoroside C (2), aucubin (3) and acetoside (4) based on spectroscopic analyses including nuclear magnetic resonance and MS. A simple and efficient HPLC with UV detection method for the simultaneous determination of the four compounds (1-4) has been developed and applied to their content determination in the S. buergeriana. The roots were extracted by 80% methanol, and the contents of 1, 2, 3, and 4 were determined to 11.5, 7.6, 41.2, and 4.8 mg/g, respectively. Additionally, angoroside C (2) and acetoside (4) exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line. 현삼(Scrophularia buergeriana) 뿌리를 80% Methanol수용액으로 추출한 뒤, 감압 농축한 추출물을 EtOAc, n-BuOH과 H2O 층으로 계통 분획을 실시하였다. n-BuOH분획에 대하여 silica gel, octadecyl SiO2 column chromatograph 및 중압분취(MPLC) 장비를 반복 실시하여 4종의 phenylethanoid glycoside 및iridoid glycoside계의 화합물을 분리하였다. NMR 및 Mass데이터를 해석하여, harpagoside (1), angoroside C (2), aucubin (3) 및 acetoside (4)로 구조 동정하였다. 분리한 4종의 화합물에 대하여 HPLC 분석법을 이용하여 정량분석한 결과, 11.5 mg/g (1), 7.6 mg/g (2), 41.2 mg/g (3), 및 4.8 mg/g (4) 이현삼 뿌리에 함유된 것을 확인하였다. 현삼으로부터 분리된 화합물 중 angoroside C 및 acetoside는 에탄올에 의해 저해된 세포 성장률을 검증한 결과, 간암세포종인 HepG2세포에 대해서간세포를 보호하는 효과가 있음을 확인하였다.

황해쑥(Artemisia agryi)으로부터 flavonoid 화합물들의 분리 동정과 세포 내 GSH 회복능을 통한 항산화 활성 평가

나현선,윤다혜,전형주,이대영,김형근 한국응용생명화학회 2022 Journal of Applied Biological Chemistry (J. Appl. Vol.65 No.4

The arial parts of Artemisia argyi were extracted with methanol : water (70:30), and the concentrates was partitioned into EtOAc (ethyl acetate), n-BuOH (normal butanol), and H2O (water) fractions. The repeated silica gel and ODS (octadecyl silica gel) column chromatographies for EtOAc and n-BuOH fractions led to isolation of four flavonoids without any ambiguity based on intensive interpretation of several spectroscopic data including nuclear magnetic resonance, and mass spectrometry. The chemical structure of the isolated compounds revealed to (2S)-naringenin (1), 3-methylkaempferol (2), 3,3′-dimethylquercetin (3), and 3,3′,4′-trimethylquercetin (4). These four compounds were first isolated from A. argyi through this study. In this study, four compounds isolated from A. argyi showed an increase in glutathione mean and a decrease in glutathione heterogeneity so that the compounds uniformly raised the intracellular glutathione (GSH) level. Based on these results, it is considered that it can be used as a functional pharmacological material.

공인중개사의 권리금계약 중개에 관한 연구 : 행정사법 위반 문제를 중심으로

나현선,김학환 한국부동산법학회 2024 不動産法學 Vol.28 No.1

공인중개사의 권리금거래에 관한 업무는 오랜 관행임에도 불구하고 공인중개사법에 제도화되지 못함으로 인하여 행정사법위반 문제가 제기되고 있다. 실무상으로 인터넷상에서 광고되고 있는 전국 공인중개사의 상가임대차 중개대상물에는 권리금이 포함되어 있으며, 이러한 권리금 중개가 공인중개사의 상관행이라고 하는 점은 수십년간의 기존 여러 연구를 통해서도 언급되고 있다. 이는 판례와 국토교통부의 공문에도 나타나고 있다. 일본에서는 택지건물거래업자의 권리금 중개가 업무로서 인정되고 있으며, 그에 관한 보수 규정을 두고 있다. 공인중개사의 권리금계약 중개는 임대차계약의 임대료 조율의 성격이 있고 이는 임대차계약의 일부를중개하는 것이 된다. 이처럼 임대차계약이나 임차권양도계약은 공인중개사법상 중개대상물이지만, 이에부수하는 권리금계약이 임차권양도계약의 대가적 성격이거나 실질상 임대료의 성격을 갖고 있음에도 공인중개사법상 중개대상물로 규정되지 않은 점은 공인중개사법령상의 미비점이라고 할 것이다. 권리금 계약을 중개하는 행위는 행정사의 업무영역이 아니다. 더욱이 권리금 거래업무의 핵심은 권리금 산정과 조정인 중개행위이며, 권리금 계약서 작성은 계약당사자의 중요한 합의를 서면에 옮기는 단순한 요식행위에 불과하다. 가사 권리금 거래계약서 작성이 행정사의 업무라고 하더라도 공인중개사의 권리금 거래의 중개는 행정사법에서 예외를 인정하는 “다른 법률에 따라 허용되는 경우”에 해당한다. 권리금이 공인중개사의 중개대상물, 중개업무 영역이라는 점은 사실인 관습이기에 이는 공인중개사의전속적 업무라고 볼 수 있다. 따라서 행정사는 권리금에 대하여 중개업무를 할 수 없으며, 단지 권리금 거래당사자가 직접 거래하여 합의한 내용에 대하여 거래계약서의 대필을 할 수 있을뿐이라고 해석하여야 한다. 이러한 권리금을 둘러싼 전문자격사간의 업무영역 충돌을 막기 위한 개선방안으로 일본 택지건물거래업자의 권리금 중개보수 법규정을 참고하여, 공인중개사법령 상 ‘권리금에 대한 중개대상물 신설’과 중개보수 신설 법령개정안을 제안하였고, ‘중개대상물 확인설명서 상 권리금 규정 신설’을 제시하였다. 권리금에 대해서 더 이상 전문자격사 간의 업무영역에 대한 충돌을 막고, 보다 전문지식이 있는 거래의 전문가인공인중개사에 의해 안전한 권리금 거래가 행하여질 수 있도록 하려면 국토부 등의 적극적이고 신속한 입법적 해결을 기할 필요가 있다. Licensed real estate agents have been involved in premium transactions for a long time. However, since it is not institutionalized in the Licensed Real Estate Agents Act, there are raising concerns about violations of the Licensed Administrative Agent Act. In practice, the commercial lease brokerage objects of licensed real estate agents advertised nationwide on the Internet include a premium. Many previous studies published in past decades show that this premium brokerage is a business practice of licensed real estate agents. Judicial precedents and official documents by the Ministry of Land, Infrastructure, and Transport also demonstrate it. In Japan, the brokerage of premiums is recognized as the work of real estate brokers, and compensation regulations are in place. The brokerage of premium contracts by a licensed real estate agent has the nature of adjusting the rent of a lease contract, which is brokering a part of the lease contract. As such, a lease contract or a lease transfer agreement is a brokerage object under the Licensed Real Estate Agents Act. However, although a premium contract is the price of a lease transfer agreement or has the nature of rent in substance, the Licensed Real Estate Agents Act does not stipulate it as a brokerage object, which is a deficiency in the Licensed Real Estate Agents Act. The act of brokering premium contracts is not within the scope of a licensed administrative agent’s business. Moreover, the core of the premium transaction business is the estimation of the premium and the brokerage of the mediator, and the preparation of the premium contract is a mere formality of putting the important agreement of the contracting parties in writing. Even if the preparation of a premium contract is the work of a licensed administrative agent, the brokerage of a premium transaction by a licensed real estate agent falls within “cases permitted under other laws” that allow exceptions in the Licensed Administrative Agent Act. It is a factual convention that premiums are the brokerage object and brokerage business area of licensed real estate agents. Therefore, it can be considered the exclusive business of licensed real estate agents. Therefore, it should be interpreted that the licensed administrative agent cannot perform brokerage services for premiums but can only write the transaction contract for the contents agreed upon by the parties to the premium transaction directly on behalf of these parties. Therefore, this study proposed “amendments to the Licensed Real Estate Agents Act to create a new brokerage object of for premium and to create a new brokerage fee” by referring to the Premium Brokerage Fee Law of the Japanese Real Estate Brokers Act and suggested to “establish a Premium Regulation in the Confirmation Document for Brokerage Objects” as an improvement measure to prevent conflicts between professional licensed agents over premiums. There is a need for an aggressive and swift legislative resolution by the Ministry of Land, Infrastructure and Transport and other ministries to prevent further conflicts over the scope of practice between professional licensed agents over premiums and ensure safe premium transactions by a licensed real estate agent, a transaction expert with more professional knowledge.

Analytical Method Validation to Distinguish Angelicae dahuricae Radix

Hyeon Seon Na(나현선),Boram Choi(최보람),Da Hye Yoon(윤다혜),Dae Young Lee(이대영),Geum Soog Kim(김금숙),Kyung Sook Han(한경숙) 한국약용작물학회 2021 한국약용작물학회 학술대회논문집 Vol.2021 No.1

도라지 (Platycodon grandiflorum (Jacq.) A. DC.) 부정아 형성을 통한 식물체 재분화

김주영,나현선,최필선,Kim, Ju Young,Na, Hyun Sun,Choi, Pil Son 한국식물생명공학회 2017 식물생명공학회지 Vol.44 No.3

도라지(Platycodon grandiflorum (Jacq. A. DC.)에서 신초발생에 대한 최적의 호르몬 조건을 조사하기 위하여 잎 및 배축절편을 1.0 mg/L 2,4-D와 0.1, 0.5, 1.0, 2.0 mg/L BA 또는 kinetin과 각각 조합 첨가한 MS 기본배지에 6주 동안 명 배양 하였다. 배양절편에 따라 신초발생율은 잎 절편에서 56.38%로 배축절편(28.20%)보다 높았다. Kinetin과 BA를 2,4-D와 조합첨가 하였을 때 kinetin의 경우 잎에서 70.38%로 BA (42.38%)보다 더 효과적이었으며, 반면 BA의 경우 배축에서는 35.56%로 kinetin (20.83%)보다 더 높은 빈도를 보였다. 가장 높은 신초형성 빈도(94.20%)는 1.0 mg/L 2,4-D와 1.0 mg/L kinetin이 조합첨가된 MS 배지에서 잎 절편을 배양하였을 때 얻을 수 있었다. 대부분의 신초는 1/2 MS기본배지에서 뿌리를 유도한 후 토양에서 완전한 식물체를 얻을 수 있었다. 이러한 결과는 최적의 기내배양 조건을 통해 도라지의 대량생산 가능성을 보여주었다. To investigate optimal conditions for plant regeneration in Platycodon grandiflorum (Jacq. A. DC.).Both leaf and hypocotyl explants were cultured on Murashige& Skoog's (MS) medium supplemented with combinations of 0.1, 0.5, 1.0, or 2.0 mg/L cytokinins (BA and kinetin) and 1.0 mg/L 2,4-D for 6 weeks, respectively. According to the type of explant, the total shoot organogenesis (56.38%) in leaf explants was higher than in hypocotyls (28.20%). In comparison with kinetin and BA for the plant regeneration, the frequency (70.38%) of leaf explants was higher in combination with kinetin and 2,4-D than of BA with 2,4-D (42.38%), whereas the frequency (35.56%) of hypocotyls explants was higher in BA combination than kinetin combination (20.83%). Thehighest frequency (94.20%) was observed from the cultures of leaf explants on the MS medium supplemented with 1.0 mg/L kinetin and 1.0 mg/L 2,4-D. Upon transfer onto 1/2 MS basal medium containing 3% sucrose, shoots developed into plantlets with roots, and were well grown in soil in the greenhouse. These results lead us to speculate that the optimization of culture conditions was responsible for the mass propagation from in vitro cultures of Platycodon grandiflorum (Jacq. A. DC.).

이야기책 듣기와 이야기 다시 말해보기가 유아의 어휘력 향상에 미치는 영향

마송희,나현선 미래유아교육학회 1999 미래유아교육학회지 Vol.6 No.1

The Purpose of this study is to investigate the effect of listening to the storybook and retelling the story on young children's vocabulary acquisition from context. Fifty five five-year-old children participated in this study. They were assigned to one of three groups, and listened to a story two times. The story was constructed to introduce 21 target words which were unfamiliar to preschoolers. Experimental 1 group retold the storybook after each listening to the story, while experimental 2 group listened to the storybook twice without retelling the story. Control group listened to the storybook which do not have target words. Both Listening to the storybook and retelling the story had an effect on 5 year old children's vocabulary acquisition. Experimental 1 group showed a greater gain in receptive and expressive vocabulary than experimental 2 group. The receptive vocabulary of experimental 2 group improved more than that of control group. There was a significant difference in the acquisition of expressive vocabulary according to young children's entering vocabulary. Children who have a higher level of entering vocabulary gained more in expressive vocabulary. Analyses of vocabulary pattern used in children's 1st and 2nd retelling showed that the number of the target words and synonyms increased, while that of faulty or no reponses decreased.

건강기능식품 원료로서 구릿대의 지표성분 분석법 검증

최보람,윤다혜,나현선,김금숙,한경숙,이수경,이대영 한국응용생명화학회 2022 Journal of Applied Biological Chemistry (J. Appl. Vol.65 No.4

This study was performed to establish an analytical method for the standardization of Angelicae Dahuricae Radix as a functional ingredient. We established six compounds including oxypeucedanin hydrate (1), byakangelcol (2), oxypeucedanin (3), imperatorin (4), phellopterin (5) and isoimperatorin (6) as marker compounds of Angelicae Dahuricae Radix. An analytical method using Ultra Performance Liquid Chromatography (UPLC) was established and validated for marker compounds of Angelicae Dahuricae Radix. The specificity was confirmed by the chromatogram from UPLC and the value of coefficient determination was also higher than 0.999, indicating high linearity. The relative standard deviation (RSD) and recovery of marker compounds were less than 5% and in the range of 90- 110%, respectively, which means that this method has high accuracy and precision. Therefore, this analytical method could be used as basic data for the development of functional ingredients for health functional food of Angelicae Dahuricae Radix

인삼(Panax ginseng C.A. Meyer)로부터 Malonyl ginsenoside의 분리 및 정량분석

신우철,정지윤,나현선,황보전,김형근,윤다혜,최보람,이영섭,김금숙,백남인,이이,이대영 한국응용생명화학회 2019 Journal of Applied Biological Chemistry (J. Appl. Vol.62 No.4

The root of Panax ginseng C.A. Meyer were extracted with 70% aqueous EtOH and the concentrates were partitioned into MeOH and H2O fractions using Diaion HP-20. The repeated SiO2 or octadecyl SiO2 column, and MPLC for the MeOH fraction led to isolation of four malonyl ginsenosides. The chemical structures of these compounds were determined as malonyl ginsenoside Rd (1) malonyl ginsenoside Rc (2) malonyl ginsenoside Rb2 (3) malonyl ginsenoside Rb1 (4) based on spectroscopic analyses including Nuclear magnetic resonance and HR-TOF/MS. The contents of malonyl ginsenoside Rb1 was highist as 5.44 mg/g of five years of ginseng. And malonyl ginsenoside Rd was lowest as 0.11 mg/g of six years of ginseng. Additionally, the malonyl ginsenoside Rd exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line. 고려인삼(Panax ginseng C.A. Meyer)을 70% EtOH 수용액으로 저온 추출한 뒤, 감압 농축한 추출물을SiO2, ODS column chromatograph 및 중압분취(MPLC) 장비를 반복 실시하여 4종의 인삼 사포닌 화합물을 분리 및 정제하였다. NMR 및 고분해능 질량분석 장비를 이용하여 malonyl ginsenoside Rd (1), Rc (2), Rb2 (3), 및 Rb1 (4)로 구조 동정하였다. 분리한 4종의화합물에 대하여 UPLC-MS/MS 질량분석기를 이용하여 수삼의5년 및 6년근 뿌리의 동체를 정량분석 하였으며, malonyl ginsenoside의 총 함량의 합은 각각 6.62 및 2.34 mg/g으로 5년근이 약 2.8배 높은 것을 확인하였다. 인삼으로부터 분리된 화합물 중 malonyl ginsenoside Rd의 경우, 알코올에 의해 저해된 HepG2세포에 대해서 간세포를 보호하는 효과가 있음을 확인하였다.

도라지 (Platycodon grandiflorum (Jacq.) A. DC.) 부정아 형성을 통한 식물체 재분화

김주영,최필선,나현선 한국식물생명공학회 2017 JOURNAL OF PLANT BIOTECHNOLOGY Vol.44 No.3

To investigate optimal conditions for plant regeneration in Platycodon grandiflorum (Jacq. A. DC.).Both leaf and hypocotyl explants were cultured on Murashige& Skoog’s (MS) medium supplemented with combinations of 0.1, 0.5, 1.0, or 2.0 mg/L cytokinins (BA and kinetin) and 1.0 mg/L 2,4-D for 6 weeks, respectively. According to the type of explant, the total shoot organogenesis (56.38%) in leaf explants was higher than in hypocotyls (28.20%). In comparison with kinetin and BA for the plant regeneration, the frequency (70.38%) of leaf explants was higher in combination with kinetin and 2,4-D than of BA with 2,4-D (42.38%), whereas the frequency (35.56%) of hypocotyls explants was higher in BA combination than kinetin combination (20.83%). Thehighest frequency (94.20%) was observed from the cultures of leaf explants on the MS medium supplemented with 1.0 mg/L kinetin and 1.0 mg/L 2,4-D. Upon transfer onto 1 /2 MS basal medium containing 3% sucrose, shoots developed into plantlets with roots, and were well grown in soil in the greenhouse. These results lead us to speculate that the optimization of culture conditions was responsible for the mass propagation from in vitro cultures of Platycodon grandiflorum (Jacq. A. DC.).

조팝나무로부터 분리된 화합물의 동정 및 정량분석

오선민,최보람,나현선,이재원,이영섭,김금숙,이승은,이대영 한국약용작물학회 2018 한국약용작물학술대회 발표집 Vol.2018 No.05

Background : Spiraea prunifolia var. simpliciflora (Rosaceae) called “Brial wreath” is a deciduous latifoliate shrub that is widely distributed throughout in Northeast Asia. Phytochemical and biological investigation of S. prunifolia have led to the discovery of biologically active compounds. Pharmacological studies revealed that the extract of the root of S. prunifolia possess antioxidant, antipyretic and anti-inflammatory activities. Some chemical constituents such as sterols, phenolics, terpene and fatty acid, as well as ethanolic extracts from the roots of S. prunifolia, have previously been reported to modulate the deleterious effects of diabetes, to prevent high-fat diet-induced obesity, and to prevent cisplatin-induced nephrotoxicity. Our continuing research was carried out to search for other phytochemical constituents from the leavess of S. prunifolia. The chemical structures of compounds were determined by NMR and FAB/MS spectroscopic data interpretation. Methods and Results : Multiple-preparative liquid chromatography (MPLC) purifications were carried out on YMC LC-forte/R instrument (YMC Kyoto, Japan) equipped with YMC-Pack ODS-A columns (ODS gel : 5 ㎛, 10 ㎜ × 250 ㎜). High-performance liquid chromatography (HPLC) was performed on Agilent Technologies instrument (Aglient Tec., Santa clara, CA, USA) equipped with YMC–Pack Pro C18 columns (ODS gel : 5 ㎛, 4.6 ㎜ × 250 ㎜). Next, quantitative analysis was carried out on UPLC-QqQ/MS 3200 Q-TRAP instrument (AB SCIEX Toronto, Canada) using a ACQUITY UPLC (waters corp.) with an ACQUITY BEH C18 column (2.1 ㎜ × 100 ㎜, 1.7 ㎛). The metabolite samples was applied to preparative reversed-phase HPLC and UPLC using gradient method, solvent A [water + 0.1% formic acid (v/v)] and solvent B [acetonitrile + 0.1% formic acid (v/v)]. Conclusion : In this study, we isolated the major metabolites from the stem of Spiraea prunifolia var. simpliciflora by using MPLC and HPLC. UPLC-QqQ/MS was also used to quantify target compounds. Finally, we established methodology and performed the quantitative analysis on target compounds from the stem of Spiraea prunifolia var. simpliciflora.