• Title/Summary/Keyword: ultra performance liquid chromatography (UPLC)

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Anthocyanin Composition and Characteristics of 'Gaeryangmeoru', 'Kyoho', and 'Hongisul' Grape Varieties ('개량머루', '거봉' 및 '홍이슬' 품종의 anthocyanin 조성과 특징)

  • Kwon, YongHee;Park, Hee-Seung
    • Horticultural Science & Technology
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    • v.33 no.4
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    • pp.470-478
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    • 2015
  • This study was investigated the anthocyanin composition of 'Gaeryangmeoru', 'Kyoho', and 'Hongisul' grapes cultivated in Korea using ultra-performance liquid chromatography (UPLC) coupled to a mass spectrometer (MS) equipped with an ESI (electrospray ionization) source. 'Gaeryangmeoru' is a dark-blue grape used for winemaking that can reach its coloring in unfavorable weather. The 'Kyoho' and 'Hongisul' varieties are hybrid grapes that feature black and pink skin, respectively. The anthocyanins extracted from the peels of grapes were analyzed using UPLC-ESI-MS/MS. Twenty-five anthocyanins were identified in the 'Gaeryangmeoru' and 'Kyoho' varieties, and 21 were identified in the 'Hongisul' variety. Eight, 14 and five predominant anthocyanins were identified in 'Gaeryangmeoru', 'Kyoho' and 'Hongisul' grape respectively. In all three varieties, mono-glucosides were 2.3-5.9 times more abundant than di-glucoside. Malvidin was the predominant anthocyanidin in 'Gaeryangmeoru' (44.1%) and 'Kyoho' (56.5%), but cyanidin (96.8%) was in 'Hongisul'. The acylated anthocyanins in 'Gaeryangmeoru' (2.0%) were rare, whereas acylated anthocyanins with p-coumaric acid were predominant in 'Kyoho' (40.9%) and 'Hongisul' (70.7%). In particular, cyanidin feruloyl glucoside was found only in the 'Hongisul' cultivar and considered to be useful as a criterion for identification of the variety. As a result, the grape varieties were demonstrated to have variety-specific anthocyanin characteristics, enabling classification based on anthocyanin composition in terms of anthocyanidins, glucosylation and acylation. The taxonomical application of anthocyanin composition confirmed the possibility that 'Gaeryangmeoru' originated from Vitis amurensis or its hybrids, and the 'Hongisul' grape was distinguished from other grapes by cyanidin feruloyl glucoside.

Classification and Comparative Analysis of the Contents of Acorus species and Anemone altaica by UPLC-PDA Analysis (UPLC-PDA를 이용한 창포류의 분류 및 함량 분석)

  • Jo, Ji Eun;Lee, A Yeong;Kim, Hyo Seon;Moon, Byeong Cheol;Ji, Yunui;Chun, Jin Mi;Kim, Ho Kyoung
    • Korean Journal of Food Science and Technology
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    • v.45 no.3
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    • pp.279-284
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    • 2013
  • A quantitative method using ultra performance liquid chromatography with a photodiode array detector (UPLCPDA) was established for the analysis of 2 major plant metabolites: ${\beta}$-asarone and ${\alpha}$-asarone from Acorus gramineus, A. tatarinowii, A. calamus and Anemone altaica, and their contents are compared with other herbs of Acorus species. The method was validated according to the International Conference on harmonization (ICH) guideline for validation of analytical procedures with respect to precision, accuracy, and linearity. The average content of ${\beta}$-asarone in Acorus gramineus was significantly higher than that in others, with the second highest concentration observed in A. tatarinowii, and only a trace amounts found in A. calamus and Anemone altaica. In contrast, the average content of ${\alpha}$-asarone in A. calamus was the highest, followed by that in Acorus gramineus and A. tatarinowii. principle component analysis (PCA) confirmed that ${\beta}$-asarone and ${\alpha}$-asarone content differed among the species. These results suggest that this UPLC-PDA method can be considered as good quality control criteria for Acorus gramineus.

Discrimination and Hordein Polypeptide Patterns of Malting Barley Varieties Using UPLC (UPLC 분석을 이용한 맥주보리 품종의 호데인 단백질 분석 및 품종 판별)

  • Yoon, Young-Mi;Kim, Yang-Kil;Kang, Chon-Sik;Park, Jin-Cheon;Park, Tae-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.4
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    • pp.326-338
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    • 2021
  • Ultra-performance liquid chromatography (UPLC) was used to assess the hordein protein fraction of malting barley. C-hordeins (barley prolamins) were extracted with 70% ethanol (EtOH) and 55% isopropyl alcohol (IPA, 2-propanol), and B-hordeins were extracted with the same alcohols in 1.0% dithiothreitol (DTT). High molecular weight (HMW) prolamins (D-hordeins) were extracted with 50% IPA with 1M Tris-HCl (pH 8.0). The same protein patterns were observed in both the experimental extraction solutions (EtOH and IPA). However, the patterns of hordein, extracted with EtOH and IPA containing 1.0% DTT, differed slightly. C- and B-hordeins extracted from those solutions were analyzed. Twenty-six malting barley varieties developed in Korea were analyzed using UPLC. The varieties were divided into seven groups according to hordein patterns of retention time 16 min to 18 min, and 20 varieties showed unique patterns.

Anti-Inflammatory and Enzyme Inhibitory Activities of Polyphenols from Peanut (Arachis hypogaea L.) Hull

  • Mihyang Kim;Yeo Ul Cho;Narae Han;Jin Young Lee;Yu-Young Lee;Moon Seok Kang;Hyun-Joo Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.312-312
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    • 2022
  • Peanut hull as by-product has been discarded during peanut processing. However, peanut hull contains plenty of polyphenols that shows various physiological activities. The objectives of this study were to investigate anti-inflammatory and enzyme inhibitory activities of polyphenols from 'Sinpalkwang' peanut (Arachis hypogaea L.) hull. Compounds were isolated from methanol extracts of peanut hull by preparative-high performance liquid chromatography after identifying and quantifying polyphenols using Ultra performance liquid chromatography (UPLC) and UPLC-Quadrupole time-of-flight-mass spectrometry profiling. The structures of compounds were elucidated by one-dimensional [1H, 13C] nuclear magnetic resonance (NMR) and two-dimensional NMR (correlated spectroscopy, heteronuclear single quantum coherence and heteronuclear multiple bond correlation). Three compounds were identified as 5,7-dihydroxy-4H-chromen-4-one (peak 2), luteolin (peak 4) and eriodictyol (peak 5). Significant differences in inflammatory mediator such as nitric oxide (NO), interleukin-6 (IL-6) and interleukin-1β (IL-lβ) in lipopolysaccharide stimulated Raw 264.7 macrophages and in enzyme (xanthine oxidase [XO] and α-glucosidase [AG]) inhibitory activities were observed between three compounds (p < 0.05). Peak 5 treated Raw 264.7 macrophages showed lower content of NO (16.4 uM), IL-6 (7.0 ng/mL), and IL-1β (60.6 pg/mL) than peak 2 (NO: 28.3 uM, IL-6: 11.3 ng/mL, IL-1β: 66.9 pg/mL) and peak 4 (NO: 24.7 uM, IL-6: 9.3 ng/mL, IL-1β: 62.6 pg/mL). Peak 5 showed higher XO inhibitory activity (84.7%) and higher AG inhibitory activity (52.4%) than peak 2 (XO inhibitory activity: 45.4%, AG inhibitory activity: 21.6%) and peak 4 (XO inhibitory activity: 37.9%, AG inhibitory activity: 37.5%) at concentration of 0.5mg/mL. This study suggests that peanut hull could be a potential source of anti-inflammatory and physiological materials while creating new use of discarded peanut hull as by-products concomitantly.

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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.

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.

A Study of Cutting Methods by Comparing the Contents of Cinnamic acid and Cinnamaldehyde in Different Parts of Cinnamomi Ramulus (Cinnamic acid, Cinnamaldehyde의 부위별 정량을 통한 계지(桂枝)의 음편(飮片) 규격 연구)

  • Kim, Young-Sik;Lee, Guem-San;Kim, Jung-Hoon;Choi, Go-Ya;Jeong, Seung-Il;Cho, Su-In;Ju, Young-Sung;Kim, Hong-Jun
    • The Korea Journal of Herbology
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    • v.26 no.2
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    • pp.11-15
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    • 2011
  • Objectives : In the present study, the contents of cinnamic acid and cinnamaldehyde in three different parts of Cinnamomi Ramulus (CR) (the whole body, the bark part, and the wood part) was evaluated using UPLC (ultra performance liquid chromatography) in order to investigate a suitable cutting method. Methods : Analysis was performed on SMART LC with UV detector. Reference compounds were separated on Inertsil ODS-4 column ($2.1mm{\times}50mm$, $3{\mu}m$, GL Science, Japan) using isolation elution with water and acetonitrile each containing acetic acid at a flow rate of $500{\mu}L/min$. Additionally, samples of CR were purchased from pharmacy of medicinal herb. Results : The correlation coefficients of the cinnamic acid and cinnamaldehyde levels showed good linearity ($r^2{\geq}0.9999$) over the linear ranges. Furthermore, the bark part exhibited higher concentration levels of reference compounds than the wood part in all samples. In addition the bark exfoliation rates in oblique and perpendicular-long cut samples of CR were lower than the perpendicular-short cut samples. Conclusions : These results suggested that the optimal cutting method would be able to reduce the bark exfoliation. Therefore, the oblique or perpendicular-long cutting method is considered to be a better cutting type than the perpendicular-short cutting method.

Quantitative Determination and Antioxidant Effects of Cheonwangbosimdan (LC-MS/MS를 이용한 천왕보심단(天王補心丹)의 함량분석 및 항산화 효능 연구)

  • Seo, Chang-Seob;Kim, Ohn Soon;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.45 no.4
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    • pp.300-314
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    • 2014
  • An ultra-performance liquid chromatography-electrospray ionization-mass spectrometer (UPLC-ESI-MS) method was established for the simultaneous quantification of eighteen marker compounds in traditional Korean formula, Cheonwangbosimdan (CWBSD). In addition, we evaluated the antioxidant effects of CWBSD. Eighteen marker components were separated on a UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) and kept at $45^{\circ}C$ by gradient elution with 0.1% (v/v) formic acid in water and acetonitrile as mobile phase. The flow rate was 0.3 mL/min and the injection volume was $2.0{\mu}L$. The antioxidant activities of CWBSD were assessed by measuring free radical scavenging activities on 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The calibration curves of all analytes showed good linearity (correlation coefficient ${\geq}0.9937$) within the test ranges. The limits of detection and quantification for the 18 marker compounds were 0.01-4.71 ng/mL and 0.03-14.13 ng/mL, respectively. The contents of the 18 compounds in CWBSD extract ranged from none to $1701.00{\mu}g/g$. The CWBSD showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction ($RC_{50}$) against ABTS and DPPH radicals were $149.42{\mu}g/mL$ and $339.24{\mu}g/mL$.

Quantification Analysis and Antioxidant Activity of Leejung-tang (LC-MS/MS를 이용한 이중탕(理中湯)의 정량분석 및 항산화 활성)

  • Seo, Chang-Seob;Kim, Ohn Soon;Kim, Yeji;Shin, Hyeun-Kyoo
    • Herbal Formula Science
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    • v.21 no.1
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    • pp.177-185
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    • 2013
  • Objectives : Leejung-tang (Lizhong-tang) has been used for treatment of gastrointestinal disorders in Korea. In this study, we performed quantification analysis of five marker components, liquiritin, ginsenoside Rb1, ginsenoside Rg1, glycyrrhizin, and 6-gingerol in Leejung-tang using a ultra performance liquid chromatography- electrospray ionization-mass spectrometer (UPLC-ESI-MS). In addition, we evaluated antioxidant activity of Leejung- tang. Methods : The column for separation of five constituents used a UPLC BEH C18 ($100{\times}2.1mm$, $1.7{\mu}m$) maintained at $45^{\circ}C$. The mobile phase consisted of two solvent systems, 0.1% (v/v) formic acid in H2O (A) and CH3CN (B) by gradient flow. The flow rate was 0.3 mL/min with detection at mass spectrometer. The antioxidative activities conduct an experiment on 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of Leejung-tang. Results : Calibration curves of five marker compounds were acquired with r2 values > 0.99. The amount of the five compounds in Leejung-tang were 0.07 - 0.84 mg/g. The concentration required for 50% reduction (RC50) against ABTS radical was 119.02 ug/mL. In addition, the scavenging against DPPH radical of Leejung-tang was 11.4%, 14.5%, 19.8%, 29.6%, and 49.2% at 25 ug/mL, $50{\mu}g/mL$, $100{\mu}g/mL$, $200{\mu}g/mL$, and $400{\mu}g/mL$, respectively. Conclusions : The established LC-MS/MS method will be helpful to improve quality control of Leejung-tang. In addition, Leejung-tang is a potential antioxidant therapeutic agent.