• 생약학회지
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• 제47권1호
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• pp.92-101
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• 2016

In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was established for simultaneous determination of the 25 marker components, including chlorogenic acid, gallic acid, oxypaeoniflorin, homogentisic acid, methyl gallate, caffeic acid, 3,4-dihydroxybenzaldehyde, paeoniflorin, albiflorin, liquiritin, nodakenin, ferulic acid, ginsenoside Rg1, liquiritigenin, coumarin, cinnamic acid, benzoylpaeoniflorin, ginsenoside Rb1, cinnamaldehyde, paeonol, glycyrrhizin, 6-gingerol, evodiamine, rutecarpine, and spicatoside A in traditional Korean formula, Onkyung-tang. All analytes were separated on a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. The MS analysis was carried out using a Waters ACQUITY TQD LC-MS/MS coupled with an electrospray ionization (ESI) source in the positive and negative modes. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}L$, respectively. The correlation coefficient of all analytes in the test ranges was greater than 0.98. The limits of detection and quantification values of the 25 marker compounds were in the ranges 0.03-19.43 and 0.09-58.29 ng/mL, respectively. As a result, methyl gallate, 3,4-dihydroxybenzaldehyde, evodiamine, and rutecarpine were not detected in this sample and the concentrations of the 21 compounds except for the above 4 compounds were $33.09-3,496.32{\mu}g/g$ in Onkyung-tang decoction. Among these compounds, paeonol was detected at the highest amount as a $3,496.32{\mu}g/g$.

• Journal of Ginseng Research
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• 제40권4호
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• pp.382-394
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• 2016

Background: Ginsenosides are the characteristic and principal components which manifest a variety of the biological and pharmacological activities of the roots and rhizomes of Panax ginseng (GRR). This study was carried out to qualitatively and quantitatively determine the ginsenosides in the cultivated and forest GRR. Methods: A rapid and sensitive ultra-high-performance liquid chromatography coupled with diode-array detector and quadrupole/time of flight tandem mass spectrometry (UPLC-DAD-QTOF-MS/MS) was applied to the qualitative analysis of ginsenosides and a 4000 QTRAP triple quadrupole tandem mass spectrometer (HPLC-ESI-MS) was applied to quantitative analysis of 19 ginsenosides. Results: In the qualitative analysis, all ingredients were separated in 10 min. A total of 131 ginsenosides were detected in cultivated and forest GRR. The method for the quantitative determination was validated for linearity, precision, and limits of detection and quantification. 19 representative ginsenosides were quantitated. The total content of all 19 ginsenosides in the forest GRR were much higher than those in the cultivated GRR, and were increased with the growing ages. Conclusion: This newly developed analysis method could be applied to the quality assessment of GRR as well as the distinction between cultivated and forest GRR.

• Molecules and Cells
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• 제27권5호
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• pp.601-608
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• 2009

The 2-deoxystreptamine and paromamine are two key intermediates in kanamycin biosynthesis. In the present study, pSK-2 and pSK-7 recombinant plasmids were constructed with two combinations of genes: kanABK, and kanABKF and kacA respectively from kanamycin producer Streptomyces kanamyceticus ATCC12853. These plasmids were heterologously expressed into Streptomyces lividans TK24 independently and generated two recombinant strains named S. lividans SK-2/SL and S. lividans SK-7/SL, respectively. ESI/ MS and ESI-LC/MS analysis of the metabolite from S. lividans SK-2/SL showed that the compound had a molecular mass of 163 $[M+H]^+$, which corresponds to that of 2-deoxystreptamine. ESI/MS and MS/MS analysis of metabolites from S. lividans SK-7/SL demonstrated the production of paromamine with a molecular mass of $324[M+H]^+$. In this study, we report the production of paromamine in a heterologous host for the first time. This study will evoke to explore complete biosynthetic pathways of kanamycin and related aminoglycoside antibiotics.

• 원예과학기술지
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• 제33권4호
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• pp.470-478
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• 2015

본 연구는 국내에서 재배중인 '개량머루', '거봉', '홍이슬' 품종의 anthocyanin 조성을 UPLC-ESI-MS/MS를 이용하여 조사하였다. '개량머루'는 착색에 불량하게 작용하는 기상 조건에서도 착색이 우수한 흑포도 품종이고, '거봉'과 '홍이슬'은 교잡종 포도로 각각 검정색과 분홍색 과피를 갖고 있다. 과피에서 추출된 anthocyanin의 조성은 UPLC-ESI-MS/MS를 이용하여 분석하였다. 각 품종에서 검출된 anthocyanin은 '개량머루'와 '거봉'에서는 각각 25가지, '홍이슬'에서는 21가지였고, 각 품종에서 검출량이 많았던 anthocyanin은 8(개량머루), 15(거봉), 5(홍이슬)가지였다. 세 품종 모두 mono-glucoside가 di-glucoside보다 많았으며, '개량머루'와 '거봉'은 malvidin이 많았던 반면, '홍이슬'은 cyanidin이 가장 많았다. '개량머루'는 acylated anthocyanin(2.0%)은 거의 존재하지 않았고 '거봉'과 '홍이슬'에서는 p-coumaric acid의 acylation이 가장 많았다. Cyanidin feruloyl glucoside가 '홍이슬'에서만 검출되어 품종 지표로 활용이 가능하였다. 결론적으로 anthocyanin의 조성을 anthocyanidin, 결합된 당의 개수, acylation 및 특정 anthocyanin을 기준으로 분류하여 품종의 특징이 확인되었다. 이러한 품종별 anthocyanin 특성을 이용하여 '개량머루'가 Vitis amurensis 혹은 그 교잡종의 가능성과 '홍이슬'에서 품종 구별의 기준으로 활용 가능성이 확인되었다.

• Bulletin of the Korean Chemical Society
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• 제30권2호
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• pp.397-401
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• 2009

The complexes of $C_2-\;and\;C_6$-dihydroceramides with transition metal ions have been investigated by using Electrospray ionization-tandem mass spectrometry (ESI-MS/MS). The formation and fragmentation pathways of several doubly charged cluster ions as well as singly charged cluster ions of $C_2-\;and\;C_6$-dihydroceramides with transition metal ions have studied by ESI-MS/MS in the positive mode. Under ESI conditions, dihydroceramides form singly and doubly charged complexes with transition metal ions $(Mn^{2+},\;Fe^{2+},\;Co^{2+},\;Ni^{2+},\;and\;Zn^{2+}\;except\;Cu^{2+})$ with the compositions of $[DHCer+M+2H^2O-H]^+,\;[2DHCer+M+2H2O-H]^+,\;[3DHCer+M+2H2O-H]^+,\;[2DHCer+M]^{2+},\;[3DHCer+M]^{2+},\;[4DHCer+M]^{2+},\;[5DHCer+M]^{2+},\;and\;[6DHCer+M]^{2+}\;(DHCer\;=\;C_2-\;or\;C_6$-dihydroceramide, M = transition metal ion). The different complexation behavior of copper is responsible for relatively lower affinity of dihydroceramides to copper compared to those of other transition metals. It is also found that in the mass spectrum of the dihydroceramide complexes with copper(II), [2DHCer+Cu-H]$^+$ was observed with considerable intensity as well as [2DHCer+Cu+2$H_2O-H]^+$ due to its different geometry from those of other metals.

• 대한화장품학회지
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• 제34권3호
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• pp.157-165
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• 2008

이전 연구에서 저자들은 나문재 추출물의 항산화 작용과 추출물 함유 크림의 유화 안정성에 대한 결과를 보고한 바 있다[1,2]. 본 연구에서는 thin layer chromatography(TLC), high performance liquid chromatography (HPLC)와 liquid chromatography/Electrospray Tandem mass spectrometry (LC/ESI/MS/MS), $^1H$-NMR을 이용하여 나문재 추출물에 대한 성분 분석을 수행하였다. 나문재 추출물 중 ethyl acetate분획의 TLC는 5개의 띠($SA1{\sim}SA5$)로 분리되었다. Ethyl acetate 분획의 당 제거반응 후 얻어진 aglycone 분획에 대한 HPLC 크로마토그램은 2개의 피이크(SAA 2 및 SAA 1)를 나타냈고, 각각 그 용리 순서는 quercetin, kaempferol이었으며 조성비는 quercetin 16.88%, kaempferol 83.12%로 kaempferol의 함량이 큰 것으로 나타났다. 또한 LC/ESI-MS/MS를 통해서 SA 2는 kaempferol-3-O-gluco-side로 SA 3는 quercetin-e-O-glucoside, SA 4는 kaempferol-3-O-rutinoside, SA 5는 quercetin-3-O-rutinoside로 확인되었다. LC/ESI-MS/MS의 스펙트럼에서 SAA 1은 탈양성자화된 aglycone 분획에 상용하는 분자이온 $[M-H]^$-(m/z285) 피이크를 나타냈으며, $^1$v 분석을 실시한 결과 [${\delta}$ 6.19 (1H, d, J=1.8Hz, H-6), ${\delta}$ 6.44 (1H, d, J=1.8Hz, H-8), ${\delta}$ 6.92 (2H, d, J=9.0Hz, H-3',5'), ${\delta}$ 8.04 (2H, d, J=9.0Hz, H-2',6')]에서 피이크들이 나타났다. 따라서 SAA 1은 kaempferol임이 확인되었다. SAA 2는 aglycone 분획에 상응하는 분자이온 $[M-H]^-$ (m/z 301)을 생성하였고, $^1H$-NMR 스펙트럼은 [${\delta}$ 6.20 (1H, d, J=2.0Hz, H-6), ${\delta}$ 6.42 (1H, d, J=2.0Hz, H-8), ${\delta}$ 6.90 (1H, d, J=8.6Hz, H-5'), ${\delta}$ 7.55 (1H, dd, J=8.6, 2.2Hz, H-6'), ${\delta}$ 7.69 (1H, d, J=2.2Hz, H-2')]에서 피이크들을 나타냈고, 따라서 SAA 2는 quercetin으로 확인되었다. 결론적으로, 이미 보고된 나문재 추출물의 항산화 작용 그리고 안정성 실험과 더불어 나문재 추출물의 성분분석은 새로운 기능성 화장품원료로서 응용이 가능함을 시사한다.

• 한국식품과학회지
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• 제39권4호
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• pp.372-379
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• 2007

부정유해물질 총 13종에 대한 신속하고 고감도의 LC-ESI-MS-MS 동시분석 방법을 개발하였으며 바데나필을 포함한 11종은 ESI positive 모드에서 타다라필을 포함한 2종은 ESI negative 모드에서 검출하는 방법으로서 시료 전처리는 간단히 메탄올 추출법을 사용하였다. 기능성표방식품 중 부정유해물질의 확인은 한번 시료를 주입함으로써 15분 이내에 13종의 분석이 가능하고 크로마토그램의 분리는 아세토니트릴과 10mM ammonium formate가 들어있는 탈이온수를 이용한(pH 7.0) 기울기 용매 조건으로 수행하였다. 확립한 13종에 대한 부정유해물질 분석방법은 검출 한계(LOD)는 0.1-5 ng/mL이고, 정량한계(LOQ)는 0.1-10 ng/mL으로서 평균 상관계수$(r^2)$는 0.9853로서 ppb 수준에서 정량성을 가지며 회수율은 87.5-98.5%, 변동계수는 15% 이하임을 확인할 수 있었다. 또한 확립한 시험방법 LC/MS/MS를 이용하여 147건의 기능성표방식품 중의 부정유해물질의 검증을 실시한 결과, 유해물질의 검출이 나타타지 않음을 알 수 있었다. 기능성표방식품 중의 실데나필과 그 유사물질을 포함한 13종의 부정유해물질에 대한 스크리닝 방법으로 MRM 모드를 이용한 LC-ESI-MS-MS 방법을 개발하였으며, 이는 유해물질에 대한 고성능액체크로마토그래피/자외선흡광광도법의 선택성등의 제한성을 극복한 부정유해 물질의 스크리닝에 신속하고 미량까지 검출 가능한 가치 있는 방법임을 확인할 수 있었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2010년도 정기총회 및 춘계학술발표회
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• pp.18-18
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• 2010

Glucosinolates(GSLs) are major secondary products($\beta$-thioglucoside N-hydroxysulfates) containing sulfates that are found mainly in Cruciferae family such as cabbage, broccoli, radish, turnip, kale, mustard and rapeseed as the important crops in agronomic and economic aspects. Especially, isothiocyanates(ITCs) have been shown to inhibit carcinogenesis with regulation cancer cell development followed by regulating target enzymes, inducing cell cycle arrest and apoptosis against various human cancer cell lines. Total desulfo(DS)-GSLs of nine type in 210 Korean leaf mustard accessions were isolated and confirmed using electrospray ionization mass spectrometric(ESI-MS/MS) method and DS-GSLs powerful library maked from several reference data. The isolated individual DS-GSLs were identified by removed $\beta$-D-glucopyranose residue ($C_6H_{11}O_5$, MW 163) from MS/MS data. Among them sinigrin(41.7%), glucoiberverin(21.7%) and gluconasturtiin(12.6%) were major components, Especially, despite gluconapin was minor component, accession K046197-1 and K046197-2 showed higher content of 4.11 and 3.31 mg/g(DW), respectively. The total GSLs contents in 210 accessions were ranged from 5.3 to 23.2 mg/g(DW) with a mean value of 13.0 mg/g(DW). As a result of principal component analysis(PCA), the individual GSLs loading plots were composed of three groups and components belonged to each group showed correlationship in quantitative pattern.

• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.207-217
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• 2007

The Saccharomyces0 cerevisiae KNU5377 strain, which was isolated from spoilage in nature, has the ability to convert biomass to alcohol at high temperatures and it can resist against various stresses [18, 19]. In order to understand the defense mechanisms of the KNU5377 strain under menadione (MD) as oxidative stress, we used several techniques for study: peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) followed by two-dimensional (2D) gel electrophoresis, liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), and surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) technology. Among the 35 proteins identified by MALDI-TOF MS, 19 proteins including Sod1p, Sod2p, Tsa1p, and Ahp1p were induced under stress condition, while 16 proteins were augmented under normal condition. In particular, five proteins, Sod1p, Sod2p, Ahp1p, Rib3p, Yaf9p, and Mnt1p, were induced in only stressed cells. By LC-ESI-MS/MS analysis, 37 proteins were identified in normal cells and 49 proteins were confirmed in the stressed cells. Among the identified proteins, 32 proteins were found in both cells. Five proteins including Yel047cp and Met6p were only upregulated in the normal cells, whereas 17 proteins including Abp1P and Sam1p were elevated in the stressed cells. It was interesting that highly hypothetical proteins such as Ynl281wp, Ygr279cp, Ypl273wp, Ykl133cp, and Ykr074wp were only expressed in the stressed cells. SELDI-TOF analysis using the SAX2 and WCX2 chips showed that highly multiple-specific protein patterns were reproducibly detected in ranges from 2.9 to 27.0 kDa both under normal and stress conditions. Therefore, induction of antioxidant proteins, hypothetical proteins, and low molecular weight proteins were revealed by different proteomic techniques. These results suggest that comparative analyses using proteomics might contribute to elucidate the defense mechanisms of KNU5377 under MD stress.

• Bulletin of the Korean Chemical Society
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• 제30권9호
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• pp.1996-2000
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• 2009

Alkali metal salts were introduced to enhance the ionization efficiency of glucose and maltooligoses in electrospray ionization-mass spectrometry (ESI-MS). A mixture of the same moles of glucose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose, and maltoheptaose was used. Salts of lithium, sodium, potassium, and cesium were employed as the cationizing agent. The ionization efficiency varied with the alkali metal cation types as well as the analyte sizes. Ion abundance distribution of the [M+$cation]^+$ ions of the carbohydrates varied with the fragmentor voltage. The maximum ion abundance at low fragmentor voltage was observed at maltose, while the maximum ion abundance at high fragmentor voltage shifted to maltotriose or maltotetraose for Na, K, and Cs. Variation of the ionization efficiency was explained with the hydrated cation size and the binding energy of the analyte and alkali metal cation.