• 제목/요약/키워드: LC-ESI/MS/MS

검색결과 213건 처리시간 0.024초

양파(Allium cepa L.) 유전자원의 Flavonol 배당체 조성 및 함량 (The Compositions and Contents of Flavonol Glycosides in Onion (Allium cepa L.) Germplasm)

  • 이정로;이민기;김헌웅;이성현;이영민;장환희;황경아;곽재균;고호철;김정봉;조진웅
    • 한국작물학회지
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    • 제59권3호
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    • pp.385-389
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    • 2014
  • 국내외 75점의 흰색 및 자색 양파 유전자원으로부터 총 5종의 flavonol 배당체(Q34'diG, Q3G, Q4'G, I4'G, Q)를 분리 동정하였다. 양파 내 존재하는 flavonol 배당체는 aglycone인 quercetin 및 isorhamnetin에 하나 또는 두 개의 glucose가 결합된 형태로 검출되었다. 총 flavonol 함량 범위는 흰색 양파가 0.18-6.47 mg/g, 자색 양파가 2.39-6.47 mg/g 이었으며, 자색 양파가 평균 4.41 mg/g 로 흰색 양파(3.23 mg/g)보다 약 1.4배 높았다. 양파의 주요 성분으로는 Q34'diG 및 Q4'G 총함량 비율이 흰색 및 자색 양파에서 모두 약 85% 이상을 보였다.

꼬리진달래 신초의 폴리페놀 프로파일링과 추출용매에 따른 유용성분 함량 및 항산화 활성 비교 (Polyphenol Profiling and Comparison of Extraction Solvents on Useful Component Content and Antioxidant Activity of Rhododendron micranthum Shoot)

  • 강정목;박용우;정은숙;최대호
    • 한국자원식물학회지
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    • 제37권2호
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    • pp.110-119
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    • 2024
  • 본 연구는 꼬리진달래의 자원식물 개발을 위한 정보를 제공하고자 수행되었다. 꼬리진달래 신초 추출물의 폴리페놀 프로파일링을 통해 유용성분을 선발하고, 추출용매에 따른 유용성분 함량과 항산화 활성을 비교하였다. 꼬리진달래 신초 추출물에서 37개의 폴리페놀 화합물을 확인하였으며, chlorogenic acid, astragalin, myricetin, afzelin의 추출용매별 함량을 비교한 결과, chlorogenic acid의 함량은 에탄올 추출물에서 6.57±0.12 mg/g이었고, astragalin의 함량은 에탄올 추출물에서 2.29±0.02 mg/g, myricetin의 함량은 메탄올 추출물에서 4.77±0.06 mg/g, afzelin의 함량은 메탄올 추출물에서 0.10±0.01 mg/g이었다. 꼬리진달래 에탄올 추출물의 총 폴리페놀 함량은 80.01±2.36 mg/g, 총 플라보노이드 함량은 78.08±3.44 mg/g이었다. 또한 꼬리진달래 메탄올 추출물의 DPPH radical 소거활성 IC50값은 943.57±10.68 mg/L였으며, ABTS radical 소거활성 IC50은 641.60±7.58 mg/L였다. 꼬리진달래는 유기용매 추출 시 유용성분과 항산화 활성이 높았고, 동일 속의 종들과 비교하여 높은 함량의 chlorogenic acid, astragalin, myricetin, afzelin을 함유하고 있어 향후 건강기능성 소재로의 개발이 가능할 것으로 판단된다.

Chemotaxonomy of Trichoderma spp. Using Mass Spectrometry-Based Metabolite Profiling

  • Kang, Dae-Jung;Kim, Ji-Young;Choi, Jung-Nam;Liu, Kwang-Hyeon;Lee, Choong-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제21권1호
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    • pp.5-13
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    • 2011
  • In this study, seven Trichoderma species (33 strains) were classified using secondary metabolite profile-based chemotaxonomy. Secondary metabolites were analyzed by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS) and multivariate statistical methods. T. longibrachiatum and T. virens were independently clustered based on both internal transcribed spacer (ITS) sequence and secondary metabolite analyses. T. harzianum formed three subclusters in the ITS-based phylogenetic tree and two subclusters in the metabolitebased dendrogram. In contrast, T. koningii and T. atroviride strains were mixed in one cluster in the phylogenetic tree, whereas T. koningii was grouped in a different subcluster from T. atroviride and T. hamatum in the chemotaxonomic tree. Partial least-squares discriminant analysis (PLS-DA) was applied to determine which metabolites were responsible for the clustering patterns observed for the different Trichoderma strains. The metabolites were hetelidic acid, sorbicillinol, trichodermanone C, giocladic acid, bisorbicillinol, and three unidentified compounds in the comparison of T. virens and T. longibrachiatum; harzianic acid, demethylharzianic acid, homoharzianic acid, and three unidentified compounds in T. harzianum I and II; and koninginin B, E, and D, and six unidentified compounds in T. koningii and T. atroviride. The results of this study demonstrate that secondary metabolite profiling-based chemotaxonomy has distinct advantages relative to ITS-based classification, since it identified new Trichoderma clusters that were not found using the latter approach.

Development and Validation of Primary Method for the Determination of Glucose in Human Serum by Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry and Comparison with Field Methods

  • Lee, Hwa Shim;Lee, Jong Man;Park, Sang Ryoul;Lee, Je Hoon;Kim, Yong Goo
    • Bulletin of the Korean Chemical Society
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    • 제34권6호
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    • pp.1698-1702
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    • 2013
  • Glucose is a common medical analyte measuring in human serum or blood samples. The development of a primary method is necessary for the establishment of traceability in measurements. We have developed an isotope dilution liquid chromatography tandem mass spectrometry as a primary method for the measurement of glucose in human serum. Glucose and glucose-$^{13}C_6$ in sample were ionized in ESI negative mode and monitored at mass transfers of m/z 179/89 and 185/92 in MRM, respectively. Glucose was separated on $NH_2P$-50 2D column, and the mobile phase was 20 mM $NH_4OAc$ in 30% acetonitrile/70% water. Verification of this method was performed by the comparison with NIST SRMs. Our results agreed well with the SRM values. We have developed two levels of glucose serum certified reference material using this method and distributed them to the clinical laboratories in Korea as samples for proficiency testings. The expended uncertainty was about 1.2% on 95% confidence level. In proficiency testings, the results obtained from the clinical laboratories showed about 3.6% and 3.9% RSD to the certified values. Primary method can provide the traceability to the field laboratories through proficiency testings or certified reference materials.

적조 살상 해양 미생물 Hahella chejuensis의 유전체 구조 (Lessons from the Sea : Genome Sequence of an Algicidal Marine Bacterium Hahella chehuensis)

  • 정해영;윤성호;이홍금;오태광;김지현
    • 한국미생물·생명공학회지
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    • 제34권1호
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    • pp.1-6
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    • 2006
  • Harmful algal blooms (HABs or red tides), caused by uncontrolled proliferation of marine phytoplankton, impose a severe environmental problem and occasionally threaten even public health. We sequenced the genome of an EPS-producing marine bacterium Hahella chejuensis that produces a red pigment with the lytic activity against red-tide dinoflagellates at parts per billion level. H. chejuensis is the first sequenced species among algicidal bacteria as well as in the order Oceanospirillales. Sequence analysis indicated a distant relationship to the Pseudomonas group. Its 7.2-megabase genome encodes basic metabolic functions and a large number of proteins involved in regulation or transport. One of the prominent features of the H. chejuensis genome is a multitude of genes of functional equivalence or of possible foreign origin. A significant proportion (${\sim}23%$) of the genome appears to be of foreign origin, i.e. genomic islands, which encode genes for biosynthesis of exopolysaccharides, toxins, polyketides or non-ribosomal peptides, iron utilization, motility, type III protein secretion and pigment production. Molecular structure of the algicidal pigment was determined to be prodigiosin by LC-ESI-MS/MS and NMR analyses. The genomics-based research on H. chejuensis opens a new possibility for controlling algal blooms by exploiting biotic interactions in the natural environment and provides a model in marine bioprospecting through genome research.

Simultaneous qualitative and quantitative analysis of morroniside and hederacoside D in extract mixture of Cornus officinalis and Stauntonia hexaphylla leaves to improve benign prostatic hyperplasia by HPLC-UV

  • Dan, Gao;Cho, Chong Woon;Vinh, Le Ba;Kim, Jin Hyeok;Cho, Kyoung Won;Kim, Young Ho;Kang, Jong Seong
    • 분석과학
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    • 제33권5호
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    • pp.224-231
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    • 2020
  • With the improvement in the standard of living and extension of life expectancy, the incidence of prostate diseases has increased yearly, thus becoming a serious disease affecting the health of men. The extract mixture of Cornus officinalis and Stauntonia hexaphylla leaves is a developed functional food formula to improve prostate health. This study developed a simultaneous analytical method of bioactive compounds for quantifying the mixture of Cornus officinalis and S. hexaphylla leaves using high-pressure liquid chromatography-ultraviolet (HPLC-UV). HPLC analytical condition was performed on a Hector C18 column with a mobile phase of 0.1 % formic acid in water (A) and 0.1 % formic acid in acetonitrile (B) under the following gradient conditions: 0-50 min, 12 %-40 % (B) at a flow rate of 1.0 mL/min. Meanwhile, this method was validated properly and successfully used to quantify the bioactive components of morroniside and hederacoside D in 20 sample batches and assess the quality of different ages and seasons of S. hexaphylla leaves. The result showed that the content of morroniside in the extract mixture of Cornus officinalis and S. hexaphylla leaves ranged from 1.38-1.62 mg/g, and the hederacoside D ranged from 28.42-32.02 mg/g, suggesting that this novel analytical method will be suitable for the quality control of the extract mixture to improve benign prostatic hyperplasia.

Antioxidative and Antiaging Activities and Component Analysis of Lespedeza cuneata G. Don Extracts Fermented with Lactobacillus pentosus

  • Seong, Joon Seob;Xuan, Song Hua;Park, So Hyun;Lee, Keon Soo;Park, Young Min;Park, Soo Nam
    • Journal of Microbiology and Biotechnology
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    • 제27권11호
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    • pp.1961-1970
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    • 2017
  • Lespedeza cuneata G. Don is a traditional herb that has been associated with multiple biological activities. In this study, we investigated the antioxidative/antiaging activities and performed an active component analysis of the non-fermented and fermented (using Lactobacillus pentosus) extracts of Lespedeza cuneata G. Don. The antioxidative activities of the fermented extract were higher than those of non-fermented extracts. The elastase inhibitory activity, inhibitory effects on UV-induced MMP-1 expression, and ability to promote type I procollagen synthesis were investigated in Hs68 human fibroblasts cells. These tests also revealed that the fermented extract had increased antiaging activities compared with the non-fermented extract. A component analysis of the ethyl acetate fractions of non-fermented and fermented extracts was performed using TLC, HPLC, and LC/ESI-MS/MS to observe changes in the components before and after fermentation. Six components that were different before and after fermentation were investigated. It was thought that kaempferol and quercetin were converted from kaempferol glucosides and quercetin glucosides, respectively, via bioconversion with the fermentation strain. These results indicate that the fermented extract of L. cuneata G. Don has potential for use as a natural cosmetic material with antioxidative and antiaging effects.

Search for Novel Stress-responsive Protein Components Using a Yeast Mutant Lacking Two Cytosolic Hsp70 Genes, SSA1 and SSA2

  • Matsumoto, Rena;Rakwal, Randeep;Agrawal, Ganesh Kumar;Jung, Young-Ho;Jwa, Nam-Soo;Yonekura, Masami;Iwahashi, Hitoshi;Akama, Kuniko
    • Molecules and Cells
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    • 제21권3호
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    • pp.381-388
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    • 2006
  • Heat shock proteins (Hsp) 70 are a ubiquitous family of molecular chaperones involved in many cellular processes. A yeast strain, ssa1/2, with two functionally redundant cytosolic Hsp70s (SSA1 and SSA2) deleted shows thermotolerance comparable to mildly heatshocked wild type yeast, as well as increased protein synthesis and ubiquitin-proteasome protein degradation. Since mRNA abundance does not always correlate well with protein expression levels it is essential to study proteins directly. We used a gel-based approach to identify stress-responsive proteins in the ssa1/2 mutant and identified 43 differentially expressed spots. These were trypsin-digested and analyzed by nano electrospray ionization liquid chromatography tandem mass spectrometry (nESI-LC-MS/MS). A total of 22 non-redundant proteins were identified, 11 of which were confirmed by N-terminal sequencing. Nine proteins, most of which were up-regulated (2-fold or more) in the ssa1/2 mutant, proved to be stress-inducible proteins such as molecular chaperones and anti-oxidant proteins, or proteins related to carbohydrate metabolism. Interestingly, a translational factor Hyp2p up-regulated in the mutant was also found to be highly phosphorylated. These results indicate that the cytosolic Hsp70s, Ssa1p and Ssa2p, regulate an abundance of proteins mainly involved in stress responses and protein synthesis.

Proteomic Analysis of Protein Expression in Streptococcus pneumoniae in Response to Temperature Shift

  • Lee Myoung-Ro;Bae Song-Mee;Kim Tong-Soo;Lee Kwang-Jun
    • Journal of Microbiology
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    • 제44권4호
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    • pp.375-382
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    • 2006
  • From its initial colonization to causation of disease, Streptococcus pneumoniae has evolved strategies to cope with a number of stressful in vivo environmental conditions. In order to analyze a global view of this organism's response to heat shock, we established a 2-D electrophoresis proteome map of the S. pneumoniae D39 soluble proteins under in vitro culture conditions and performed the comparative proteome analysis to a 37 to $42^{\circ}C$ temperature up-shift in S. pneumoniae. When the temperature of an exponentially growing S. pneumoniae D39 culture was raised to $42^{\circ}C$, the expression level of 25 proteins showed changes when compared to the control. Among these 25 proteins, 12 were identified by MALDI-TOF and LC-coupled ESI MS/MS. The identified proteins were shown to be involved in the general stress response, energy metabolism, nucleotide biosynthesis pathways, and purine metabolism. These results provide clues for understanding the mechanism of adaptation to heat shock by S. pneumoniae and may facilitate the assessment of a possible role for these proteins in the physiology and pathogenesis of this pathogen.

A Simple Carbamidomethylation-Based Isotope Labeling Method for Quantitative Shotgun Proteomics

  • Oh, Donggeun;Lee, Sun Young;Kwon, Meehyang;Kim, Sook-Kyung;Moon, Myeong Hee;Kang, Dukjin
    • Mass Spectrometry Letters
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    • 제5권3호
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    • pp.63-69
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    • 2014
  • In this study, we present a new isotope-coded carbamidomethylation (iCCM)-based quantitative proteomics, as a complementary strategy for conventional isotope labeling strategies, with providing the simplicity, ease of use, and robustness. In iCCM-based quantification, two proteome samples can be separately isotope-labeled by means of covalently reaction of all cysteinyl residues in proteins with iodoacetamide (IAA) and its isotope (IAA-$^{13}C_2$, $D_2$), denoted as CM and iCCM, respectively, leading to a mass shift of all cysteinyl residues to be + 4 Da. To evaluate iCCM-based isotope labeling in proteomic quantification, 6 protein standards (i.e., bovine serum albumin, serotransferrin, lysozyme, beta-lactoglobulin, beta-galactosidase, and alpha-lactalbumin) isotopically labeled with IAA and its isotope, mixed equally, and followed by proteolytic digestion. The resulting CM-/iCCM-labeled peptide mixtures were analyzed using a nLC-ESI-FT orbitrap-MS/MS. From our experimental results, we found that the efficiency of iCCM-based quantification is more superior to that of mTRAQ, as a conventional nonisobaric labeling method, in which both of a number of identified peptides from 6 protein standards and the less quantitative variations in the relative abundance ratios of heavy-/light-labeled corresponding peptide pairs. Finally, we applied the developed iCCM-based quantitative method to lung cancer serum proteome in order to evaluate the potential in biomarker discovery study.