• Title/Summary/Keyword: ultra performance liquid chromatography

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Antimicrobial Effects of Sophorae Radix Extracts against Oral Microorganisms (구강미생물에 대한 고삼의 항균효과)

  • Park, Sook-Jahr;Kim, Sang-Chan;Lee, Jong-Rok
    • The Korea Journal of Herbology
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    • v.25 no.2
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    • pp.81-88
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    • 2010
  • Objectives : Sophorae Radix (SR), the dried root of Sophorae Flavescens Aiton, has been used to treat atherosclerosis, arrhythma and skin diseases including scabies and eczema. The present study was examined to evaluate antimicrobial activities of SR extracts against oral microorganism. Methods : Antimicrobial properties of SR extracts were determined by agar diffusion method and minimum inhibitory concentration (MIC) against Streptococcus mutans, Streptococcus sobrinus and Actinomyces viscosus. Analysis of kurarinone from SR extracts was conducted using UPLC (Ultra Performance Liquid Chromatography). Results : The ethanolic extracts of SR showed stronger antimicrobial effect than methanolic extracts, while the aqueous extracts of SR had no activity. In addition, the higher content of kurarinone was found in ethanolic extracts than methanolic extracts. The purified kurarinone from ethanolic extracts showed potent antimicrobial activity with the MIC value of $3.9{\sim}7.8{\mu}g/m{\ell}$. Conclusion : An ethanolic extract of SR showed antimicrobial properties against several oral microorganisms, and kuranrinone contributed to antimicrobial action of SR. Thus, ethanolic extracts of SR or purified kurarinone should be beneficial for the preparation of the useful agent for treating oral disease including anticaries.

Comparison of 2-D RP-RP MS/MS with 1-D RP MS/MS for Proteomic Analysis (단백체 분석을 위한 일차원 및 이차원 역상크로마토그래피의 비교)

  • Moon, Pyong-Gon;Cho, Young-Eun;Baek, Moon-Chang
    • YAKHAK HOEJI
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    • v.54 no.5
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    • pp.377-386
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    • 2010
  • Single-dimensional (1-D) and two-dimensional (2-D) LC methods were utilized to separate peptides from various sources followed by MS/MS analysis. Two-dimensional ultra-high performance liquid chromatography is a useful tool for proteome analysis, providing a greater peak capacity than 1-D LC. The most popular 2-D LC approach used today for proteomic research combines strong cation exchange and reversed-phase LC. We have evaluated an alternative mode for 2-D LC of peptides using 2-D RP-RP nano UPLC Q-TOF Mass Spectrometry, employing reversed-phase columns in both separation dimensions. As control experiments, we identified 129 proteins in 1-D LC and 322 proteins in 2-D LC from E. coli extract peptides. Furthermore, we applied this method to rat primary hepatocyte and a total of 170 proteins were identified from 1-D LC, and 527 proteins were identified from all 2-D LC system. The in-depth protein profiling established by this 2-D LC MS/MS from rat primary hepatocyte could be a very useful reference for future applications in regards to drug induced liver toxicity.

Constructing Proteome Reference Map of the Porcine Jejunal Cell Line (IPEC-J2) by Label-Free Mass Spectrometry

  • Kim, Sang Hoon;Pajarillo, Edward Alain B.;Balolong, Marilen P.;Lee, Ji Yoon;Kang, Dae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1124-1131
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    • 2016
  • In this study, the global proteome of the IPEC-J2 cell line was evaluated using ultra-high performance liquid chromatography coupled to a quadrupole Q Exactive Orbitrap mass spectrometer. Proteins were isolated from highly confluent IPEC-J2 cells in biological replicates and analyzed by label-free mass spectrometry prior to matching against a porcine genomic dataset. The results identified 1,517 proteins, accounting for 7.35% of all genes in the porcine genome. The highly abundant proteins detected, such as actin, annexin A2, and AHNAK nucleoprotein, are involved in structural integrity, signaling mechanisms, and cellular homeostasis. The high abundance of heat shock proteins indicated their significance in cellular defenses, barrier function, and gut homeostasis. Pathway analysis and annotation using the Kyoto Encyclopedia of Genes and Genomes database resulted in a putative protein network map of the regulation of immunological responses and structural integrity in the cell line. The comprehensive proteome analysis of IPEC-J2 cells provides fundamental insights into overall protein expression and pathway dynamics that might be useful in cell adhesion studies and immunological applications.

Anti-oxidant Activities and Identification of Standard Compounds from Cibotii Rhizoma (구척의 항산화 활성 및 지표성분 동정)

  • Kim, So-Hwa;Kim, Eun-Young;Whang, Wan-Kyunn
    • YAKHAK HOEJI
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    • v.58 no.5
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    • pp.314-321
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    • 2014
  • Cibotii Rhizoma, the dried rhizome of Cibotium barometz J. Smith (C. barometz), has long been used to treat bone or nervous system disorders. In this regard, we isolated three main phenolic compounds, onitin-4-O-${\beta}$-D-glucopyranoside (1), irisdichototins E & F epimeric mixture (2), and protocatechuic acid (3) from C. barometz methanol extract. In addition, we screened their antioxidative activities by DPPH, ABTS radical, and superoxide scavenging assays. Among these three compounds, irisdichototins E & F and protocatechuic acid showed strong antioxidant activities. Also, the antioxidant activities of the C. barometz extracts were proportional to the contents of irisdichototins E & F and protocatechuic acid, thus these two phenolic compounds could be main active compounds of C. barometz. In addition, onitin-4-O-${\beta}$-D-glucopyranoside is considered as a marker compound of C. barometz because this compound is specifically contained in C. barometz which belongs to Pteridophyta order. A rapid analysis method for the simultaneous determination of phenolic compounds was also developed by UPLC (Ultra Performance Liquid Chromatography). Using the developed method, the two active compounds (irisdichototins E & F and protocatechuic acid) and a marker compound (onitin-4-O-${\beta}$-D-glucopyranoside) were successfully quantified in 14 commercial samples that were collected from different regions.

Simultaneous Determination of Anthraquinone, Flavonoids, and Phenolic Antidiabetic Compounds from Cassia auriculata Seeds by Validated UHPLC Based MS/MS Method

  • Girme, Aboli;Saste, Ganesh;Chinchansure, Ashish;Joshi, Swati;Kunkulol, Rahul;Hingorani, Lal;Patwardhan, Bhushan
    • Mass Spectrometry Letters
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    • v.11 no.4
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    • pp.82-89
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    • 2020
  • A systematic isolation and characterization study for Cassia auriculata (CA) seeds resulted in identifying antidiabetic compounds 1,3,8-trihydroxyanthraquinone and quercetin, quercetin-3-O-rutinoside, gallic acid, caffeic acid, ferulic acid, and ellagic acid. The ultra-high-performance liquid chromatography based triple quadrupole mass spectrometry methodology was developed and validated for simultaneous identification and confirmation of these compounds from CA seeds. Multiple reaction monitoring (MRM) based quantification method was developed with MRM optimizer software for MS1 and MS2 mass analysis. The method was optimized on precursor ions and product ions with the ion ratio of each compound. The calibration curves of seven bioactive analytes showed excellent linearity (r2 ≥ 0.99). The quantitation results found precise (RSD, < 10 %) with good recoveries (84.58 to 101.42%). The matrix effect and extraction recoveries were found within the range (91.66 to 102.11%) for the CA seeds. This is the first MS/MS-based methodology applied to quantifying seven antidiabetic compounds in CA seeds and its extract for quality control purposes.

Enzymatic Synthesis of Resveratrol α-Glucoside by Amylosucrase of Deinococcus geothermalis

  • Moon, Keumok;Lee, Seola;Park, Hyunsu;Cha, Jaeho
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1692-1700
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    • 2021
  • Glycosylation of resveratrol was carried out by using the amylosucrase of Deinococcus geothermalis, and the glycosylated products were tested for their solubility, chemical stability, and biological activities. We synthesized and identified these two major glycosylated products as resveratrol-4'-O-α-glucoside and resveratrol-3-O-α-glucoside by nuclear magnetic resonance analysis with a ratio of 5:1. The water solubilities of the two resveratrol-α-glucoside isomers (α-piceid isomers) were approximately 3.6 and 13.5 times higher than that of β-piceid and resveratrol, respectively, and they were also highly stable in buffered solutions. The antioxidant activity of the α-piceid isomers, examined by radical scavenging capability, showed it to be initially lower than that of resveratrol, but as time passed, the α-piceid isomers' activity reached a level similar to that of resveratrol. The α-piceid isomers also showed better inhibitory activity against tyrosinase and melanin synthesis in B16F10 melanoma cells than β-piceid. The cellular uptake of the α-piceid isomers, which was assessed by ultra-performance liquid chromatography (UPLC) analysis of the cell-free extracts of B16F10 melanoma cells, demonstrated that the glycosylated form of resveratrol was gradually converted to resveratrol inside the cells. These results indicate that the enzymatic glycosylation of resveratrol could be a useful method for enhancing the bioavailability of resveratrol.

Anti-bacterial properties and safety evaluation of disinfectant using Dendropanax morbifera (Hwangchil) extract for passenger cabin in the subway (지하철 객실 적용을 위한 황칠 추출물 소독제의 항균특성 및 안전성 평가)

  • Bui, Vu Khac Hoang;Park, Jae-Seok;Lee, Young-Chul
    • Particle and aerosol research
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    • v.18 no.2
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    • pp.37-50
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    • 2022
  • Due to the syndrome coronavirus 2 (SARS-CoV-2) pandemic, the subway passenger cabin should be continuously sterilized. However, a disinfectant such as chlorine is toxic and can lead to different issues to human health. In this paper, we introduced a novel disinfectant based on natural product (Dendropanax morbifera extract). Via ultra-high performance liquid chromatography - mass spectrometer (UHPLC-MS), different compounds from Dendropanax morbifera extract showed antivirus potentials. Antimicrobial experiments confirmed that the air-disinfectant containing Dendropanax morbifera can eliminate harmful microorganisms including Gram (-), Gram (+), and yeast within 5 mins. The as-prepared air-disinfectant also showed high antivirus activity against H1N1, HRV, and EV71. Deodorization test also indicates that the as-prepared air-disinfectant can lower the harmful gas such as ammonia and trimethylamine in the atmosphere. To evaluate the potential of air-disinfectant containing Dendropanax morbifera in practical applications, different safety tests including acute oral toxicity, acute skin irritation, and eye irritation were conducted. Results showed that the as-prepared disinfectant did not negatively affect tested animals during these safety investigations.

Genetic and biochemical evidence for redundant pathways leading to mycosporine-like amino acid biosynthesis in the cyanobacterium Sphaerospermopsis torques-reginae ITEP-024

  • Geraldes, Vanessa;de Medeiros, Livia Soman;Lima, Stella T.;Alvarenga, Danillo Oliveira;Gacesa, Ranko;Long, Paul F.;Fiore, Marli Fatima;Pinto, Ernani
    • ALGAE
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    • v.35 no.2
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    • pp.177-187
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    • 2020
  • Cyanobacteria have been widely reported to produce a variety of UV-absorbing mycosporine-like amino acids (MAAs). Herein, we reported production of the unusual MAA, mycosporine-glycine-alanine (MGA) in the cyanobacterium Sphaerospermopsis torques-reginae ITEP-024 using a newly developed UHPLC-DAD-MS/HRMS (ultra-high performance liquid chromatography-diode array detection-high resolution tandem mass spectrometry) method. MGA had previously been first identified in a red-algae, but S. torques-reginae strain ITEP-024 is the first cyanobacteria to be reported as an MGA producer. Herein, the chemical structure of MGA is fully elucidated from one-dimensional / two-dimensional nuclear magnetic resonance and HRMS data analyses. MAAs are unusually produced constitutively in S. torques-reginae ITEP-024, and this production was further enhanced following UV-irradiance. It has been proposed that MAA biosynthesis proceeds in cyanobacteria from the pentose phosphate pathway intermediate sedoheptulose 7-phosphate. Annotation of a gene cluster encoded in the genome sequence of S. torques-reginae ITEP-024 supports these gene products could catalyse the biosynthesis of MAAs. However, addition of glyphosate to cultures of S. torques-reginae ITEP-024 abolished constitutive and ultra-violet radiation induced production of MGA, shinorine and porphyra-334. This finding supports involvement of the shikimic acid pathway in the biosynthesis of MAAs by this species.

Quantification of Arsenic Species in Some Seafood by HPLC-AFS (HPLC-AFS를 이용한 해산물 중 비소 화학종 분리정량)

  • Jeong, Seung-Woo;Lee, Chae-Hyeok;Lee, Jong-Wha;Jang, Bong-Ki
    • Journal of Environmental Health Sciences
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    • v.47 no.5
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    • pp.496-503
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    • 2021
  • Background: Considering the expenses of and difficulties in arsenic speciation by high performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS), alternative measurement methods should be useful, especially for large-scale research and projects. Objectives: A measurement method was developed for arsenic speciation using HPLC-atomic fluorescence spectrometry (HPLC-AFS) as an alternative to HPLC-ICP-MS. Methods: Total arsenic and toxic arsenic species in some seafoods were determined by atomic absorption spectrometry coupled with hydride vapor generation (AAS-HVG) and HPLC-AFS, respectively. Recovery rate of arsenic species in seafood was evaluated by ultra sonication, microwave and enzyme (pepsin) for the optimal extraction method. Results: Limits of detection of HPLC-AFS for As3+, dimethylarsinate (DMA), monomethylarsonate (MMA) and As5+ were 0.39, 0.53, 0.60 and 0.64 ㎍/L, respectively. The average accuracy ranged from 97.5 to 108.7%, and the coefficient of variation was in the range of 1.2~16.7%. As3+, DMA, MMA and As5+ were detected in kelp, the sum of toxic arsenic in kelp was 40.4 mg/kg. As3+, DMA, MMA and As5+ were not detected in shrimp and squid, but total arsenic (iAS and oAS) content in shrimp and squid analyzed by AAS-HVG were 18.1 and 24.7 mg/kg, respectively. Conclusions: HPLC-AFS was recommendable for the quantitative analysis method of arsenic species. As toxic arsenic species are detected in seaweeds, further researches are needed for the contribution degree of seafood in arsenic exposure.

Phytochemicals and antioxidant activity in the kenaf plant (Hibiscus cannabinus L.)

  • Ryu, Jaihyunk;Kwon, Soon-Jae;Ahn, Joon-Woo;Jo, Yeong Deuk;Kim, Sang Hoon;Jeong, Sang Wook;Lee, Min Kyu;Kim, Jin-Baek;Kang, Si-Yong
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.191-202
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    • 2017
  • Chemical compounds from four different tissues of the kenaf plant (Hibiscus cannabinus), a valuable medicinal crop originating from Africa, were examined to determine its potential for use as a new drug material. Leaves, bark, flowers, and seeds were harvested to identify phytochemical compounds and measure antioxidant activities. Gas chromatography mass spectrometry analyses identified 22 different phytocompounds in hexane extracts of the different parts of the kenaf plant. The most abundant volatile compounds were E-phytol (32.4%), linolenic acid (47.3%), trisiloxane-1,1,1,5,5,5-hexamethyl-3,3-bis[(trimethylsilyl)oxy] (16.4%), and linoleic acid (46.4%) in leaves, bark, flowers, and seeds, respectively. Ultra-high performance liquid chromatography identified the major compounds in the different parts of the kenaf plant as kaemperitrin, caffeic acid, myricetin glycoside, and p-hydroxybenzoic acid in leaves, bark, flowers, and seeds, respectively. Water extracts of flowers, leaves, and seeds exhibited the greatest DPPH radical scavenging activity and SOD activity. Our analyses suggest that water is the optimal solvent, as it extracted the greatest quantity of functional compounds with the highest levels of antioxidant activity. These results provide valuable information for the development of environmentally friendly natural products for the pharmaceutical industry.