• Title/Summary/Keyword: UHPLC

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UHPLC/TOFHRMS analysis and anti-inflammatory effect of leaf extracts from Zizyphus jujuba in LPS-stimulated RAW264.7 cells

  • Hyun Ji Eo;Sun-Young Lee;Gwang Hun Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.27-33
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    • 2023
  • Zizyphus jujube is a plant in the buckthorn family (Rhamnaceae) that has been the subject of research into antibacterial, antifungal and anti-inflammatory properties of its fruit and seed. However, few studies have investigated its leaves. In this study, the anti-inflammatory activity of ZJL (an extract of Z. jujube leaf) was evaluated to verify its potential as an anti-inflammatory agent and SARS-CoV-2 medicine, using nitric oxide (NO) assay, RT-PCR, SDSPAGE, Western blotting, and UHPLC/TOFHRMS analysis. We found that ZJL suppresed pro-inflammatory mediators such as NO, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor α (TNF-α) in lipopolysaccharide (LPS)-induced RAW264.7 cells. ZJL acted by inhibiting NF-KB and MAPK signaling pathway activity. We also confirmed that ZJL contains a phenol compound and flavonoids with anti-inflammatory activity such as trehalose, maleate, epigallocatechin, hyperoside, catechin, 3-O-coumaroylquinic acid, rhoifolin, gossypin, kaempferol 3-neohesperidoside, rutin, myricitrin, guaiaverin, quercitrin, quercetin, ursolic acid, and pheophorbide a. These findings suggest that ZJL may have great potential for the development of anti-inflammatory drugs and vaccines via inhibition of NF-κB and MAPK signaling in LPS-induced RAW264.7 cells.

Method Development for Determination of Chlorogenic Acid and Arbutin Contents in Fruits by UHPLC-MS/MS (UHPLC-MS/MS를 이용한 과일류 중 클로로젠산 및 알부틴 동시분석법 개발)

  • Choi, Young-Ju;Jeon, Jong-Sup;Kim, Woon-Ho;Jung, You-Jung;Ryu, Ji-Eun;Choi, Jong-Chul;Chae, Kyung-Suk;Lee, Jin-Hee;Do, Young-Sook;Park, Young-Bae;Yoon, Mi-Hye
    • Journal of Food Hygiene and Safety
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    • v.34 no.5
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    • pp.413-420
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    • 2019
  • In this study, a sample preparation method and a simultaneous determination method by ultra-high performance liquid chromatography coupled with tandem mass spectrometry for 9 isomers of chlorogenic acid and arbutin in fruits were developed. The samples were extracted using 90% methanol (pH 3.0), with the solutions being shaken and then sonicated for 10 min each. After centrifugation at 4,000 rpm for 10 min, the extraction was concentrated under a vacuum at $40^{\circ}C$ using a vacuum evaporator. The residue was dissolved in 5 mL of 5% methanol and filtered through a $0.45{\mu}m$ membrane before UHPLC-MS/MS analysis. The separations were performed on a C18 column with gradient elution of water (containing 0.1% formic acid) and methanol (containing 0.1% formic acid). The specificity, linearity, limit of detection, limit of quantification, accuracy, and precision of the proposed methods were also evaluated.

Induction of Salicylic Acid Production in Pepper by Yeast Cell Wall Extract (효모세포벽추출물에 의한 고추내 salicylic acid 생성유도)

  • Kang, Dae Sun;Cho, Soo Muk;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.299-302
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    • 2012
  • Yeast cell wall extract (YCWE) was treated on leaves and roots of pepper seedlings at the dosage of 4 mg/mL and salicylic acid (SA) production in pepper was detected by ultra high performance liquid chromatography (UHPLC). The SA production in pepper stem was induced by YCWE. SA was produced at the highest level of 20.29 ${\mu}g/g$ after 48 hrs of foliar spray with YCWE, which is 3.7 times higher than that of root perfusion with YCWM. SA production was gradually reduced after 72 hrs of YCWE treatment.

Establishment of analytical methods for allergenic compounds in mouthwashes and sanitary napkins by ultra-high-performance liquid chromatography with tandem mass spectrometry

  • Hee-Jung Sim;Hee-Jin Jeong;Yeong-In Lee;Yu-Jin Cho;Seung-Hoon Baek;Jong-Hwan Kim
    • Analytical Science and Technology
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    • v.36 no.2
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    • pp.89-98
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    • 2023
  • Analytical methods for detecting atranol, chloroatranol, evernic acid, (+)-usnic acid, and atranorin in sanitary napkins and mouthwashes were developed using ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). UHPLC-MS/MS conditions were optimized for rapid, sensitive, and simultaneous analysis of the five allergenic compounds. The methods were validated by assessing their specificity, matrix effects, limit of detection (LOD), limit of quantification (LOQ), linearity, accuracy, and precision. Good linearity was achieved with a determination coefficient of ≥0.99. The LOD and LOQ were 2.1-9.8 and 6.4-29.6 ng/g for sanitary napkins and 0.29-0.48 and 0.87-1.45 ng/mL for mouthwashes, respectively. The accuracy and precision were within an acceptable range according to the criteria reported in the European SANTE/11813/2017 guidelines (70-120 % recovery, <20 % relative standard deviation). Therefore, these methods can be used to analyze atranol, chloroatranol, evernic acid, (+)-usnic acid, and atranorin in sanitary napkins and mouthwashes.

Optimization of Analytical Methods for Ochratoxin A and Zearalenone by UHPLC in Rice Straw Silage and Winter Forage Crops (UHPLC를 이용한 볏짚 사일리지와 동계사료작물의 오크라톡신과 제랄레논 분석법 최적화)

  • Ham, Hyeonheui;Mun, Hye Yeon;Lee, Kyung Ah;Lee, Soohyung;Hong, Sung Kee;Lee, Theresa;Ryu, Jae-Gee
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.4
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    • pp.333-339
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    • 2016
  • The objective of this study was to optimize analytical methods for ochratoxin A (OTA) and zearalenone (ZEA) in rice straw silage and winter forage crops using ultra-high performance liquid chromatography (UHPLC). Samples free of mycotoxins were spiked with $50{\mu}g/kg$, $250{\mu}g/kg$, or $500{\mu}g/kg$ of OTA and $300{\mu}g/kg$, $1500{\mu}g/kg$, or $3000{\mu}g/kg$ of ZEA. OTA and ZEA were extracted by acetonitrile and cleaned-up using an immunoaffinity column. They were then subjected to analysis with UHPLC equipped with a fluorescence detector. The correlation coefficients of calibration curves showed high linearity ($R^2{\geq_-}0.9999$ for OTA and $R^2{\geq_-}0.9995$ for ZEA). The limit of detection and quantification were $0.1{\mu}g/kg$ and $0.3{\mu}g/kg$, respectively, for OTA and $5{\mu}g/kg$ and $16.7{\mu}g/kg$, respectively, for ZEA. The recovery and relative standard deviation (RSD) of OTA were as follows: rice straw = 84.23~95.33%, 2.59~4.77%; Italian ryegrass = 79.02~95%, 0.86~5.83%; barley = 74.93~97%, 0.85~9.19%; rye = 77.99~96.67%, 0.33~6.26%. The recovery and RSD of ZEA were: rice straw = 109.6~114.22%, 0.67~7.15%; Italian ryegrass = 98.01~109.44%, 1.65~4.81%; barley = 98~113.53%, 0.25~5.85%; rye = 90.44~108.56%, 2.5~4.66%. They both satisfied the standards of European Commission criteria (EC 401-2006) for quantitative analysis. These results showed that the optimized methods could be used for mycotoxin analysis of forages.

The rapid determination of PAHs in foods using ultra high performance liquid chromatography (UHPLC를 이용한 식품 중의 PAHs 신속분석법)

  • Ha, Jae-Ho;Shin, Dong-Bin;Hwang, Jin-Bong;Seo, Hye-Young;Ito, Masahito;Nakagawa, Hiroaki
    • Analytical Science and Technology
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    • v.24 no.4
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    • pp.266-274
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    • 2011
  • Using a Hitachi LaChrom Ultra 2000U, a reverse phase ultra high performance liquid chromatography (u-HPLC) method was developed for the rapid quantification of 14 PAHs in foods. The proposed method for PAH analysis is based on solid phase extraction (SPE) cartridges; the determination was carried out by u-HPLC with fluorimetric detection. The method was very sensitive; PAH concentration levels were in a low ${\mu}g$/kg range and could be detected and quantified. Six samples of food were analyzed. Among PAHs, PHE was found in most of samples, the concentration ranging from 2.5 to 19.9 ${\mu}g$/kg. The contents of benzo[c]fluorine (BCL), pyrene (PYR), benzo[a]anthracene (BaA), chrysene (CHR), benzo[b]fluoranthene (BbF), benzo[k]fluoranthene (BkF) were low at the '${\mu}g$/kg' level or were less than LOD.

Study on biosynthesis of ginsenosides in the leaf of Panax ginseng by seasonal flux analysis

  • Kim, Dongmin;Han, Jaehong
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.315-322
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    • 2019
  • Seasonal ginsenoside flux in the leaves of 5-year-old Panax ginseng was analyzed from the field-grown ginseng, for the first time, to study possible biosynthesis and translocation of ginsenosides. The concentrations of nine major ginsenosides, Rg1, Re, Rh1, Rg2, R-Rh1, Rb1, Rc, Rb2, and Rd, were determined by UHPLC during the growth in between April and November. It was confirmed total ginsenoside content in the dried ginseng leaves was much higher than the roots by several folds whereas the composition of ginsenosides was different from the roots. The ginsenoside flux was affected by ginseng growth. It quickly increased to 10.99±0.15 (dry wt%) in April and dropped to 6.41±0.14% in May. Then, it slowly increased to 9.71±0.14% in August and maintained until October. Ginsenoside Re was most abundant in the leaf of P. ginseng, followed by Rd and Rg1. Ginsenosides Rf and Ro were not detected from the leaf. When compared to the previously reported root data, ginsenosides in the leaf appeared to be translocated to the root, especially in the early vegetative stage even though the metabolite translocated cannot be specified. The flux of ginsenoside R-Rh1 was similar to the other (20S)-PPT ginsenosides. When the compositional changes of each ginsenoside in the leaf was analyzed, complementary relationship was observed from ginsenoside Rg1 and Re, as well as from ginsenoside Rd and Rb1+Rc. Accordingly, ginsenoside Re in the leaf was proposed to be synthesized from ginsenoside Rg1. Similarly, ginsenosides Rb1 and Rc were proposed to be synthesized from Rd.

Rapid separation and identification of 31 major saponins in Shizhu ginseng by ultra-high performance liquid chromatography-electron spray ionization-MS/MS

  • Sun, Ting-Ting;Liang, Xin-Lei;Zhu, He-Yun;Peng, Xu-Ling;Guo, Xing-Jie;Zhao, Long-Shan
    • Journal of Ginseng Research
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    • v.40 no.3
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    • pp.220-228
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    • 2016
  • Background: Among the various ginseng strains, Shizhu ginseng is endemic to China, mainly distributed in Kuandian Manchu Autonomous County (Liaoning Province, China); however, not much is known about the compounds (especially saponins) in Shizhu ginseng. Methods: A rapid, sensitive, and reliable ultra-high performance liquid chromatography coupled with MS/MS (UHPLC-MS/MS) method was developed to separate and identify saponins in Shizhu ginseng. Results: The separation was carried out on a Waters ACQUITY UPLC BEH $C_{18}$ column ($100mm{\times}2.1mm$, $1.7{\mu}m$) with acetonitrile and 0.1% formic acid aqueous solution as the mobile phase under a gradient elution at $40^{\circ}C$. The detection was performed on a Micromass Quattro Micro API mass spectrometer equipped with electrospray ionization source in both positive and negative modes. Under the optimized conditions, a total of 31 saponins were identified or tentatively characterized by comparing retention time and MS data with related literatures and reference substances. Conclusion: The developed UHPLC-MS/MS method was suitable for identifying and characterizing the chemical constituents in Shizhu ginseng, which provided a helpful chemical basis for further research on Shizhu ginseng.

Rapid determination and quantification of hair-growth compounds in adulterated products by ultra HPLC coupled to quadrupole-orbitrap MS

  • Lee, Ji Hyun;Park, Han Na;Kang, Gihaeng;Kim, Nam Sook;Park, Seongsoo;Lee, Jongkook;Kang, Hoil
    • Analytical Science and Technology
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    • v.32 no.2
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    • pp.56-64
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    • 2019
  • Recently, a number of adulterated products, which are advertised as hair-growth enhancer have been emerged among those who suffer hair loss disease. For continuous control of illegal products, in this study, a rapid and sensitive method for simultaneous screening of 12 compounds that enhance hair-growth was established to protect public health by ultrahigh-performance liquid chromatography coupled to quadrupole-orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS). Fragmentation pathways of them were proposed based on $MS^2$ spectral data obtained using the established method. In this analysis, the LODs and LOQs ranged from 0.05 to 50 ng/mL and from 0.17 to 167 ng/mL, respectively. The square of the linear correlation coefficient ($R^2$) was determined as more than 0.995. The intra- and inter-assay accuracies were respective 88-112 % and 88-115 %. Their precision values were measured within 5 % (intra-day) and 10 % (inter-day). Mean recoveries of target compounds in adulterated products ranged from 84 to 115%. The relative standard deviation of stability was less than 12 % at $4^{\circ}C$ for 48 h. The method was employed to screen 14 dietary supplements advertised to be effective for the treatment of hair loss. Some of the products (~21 %) were proven to contain synthetic drugs that promote hair growth such as triaminodil, minoxidil, and finasteride.

HPLC-UVD method validation for quantitative analysis of camelliaside A in hot-water extract of soybean (Glycine max L.) leaves (콩잎 열수추출물의 지표성분인 camelliaside A의 정량분석을 위한 HPLC-UVD 분석법 밸리데이션)

  • Kim, Jeong Ho;Lee, Seung Hwan;Moon, Si Won;Park, Ki Hun
    • Journal of Applied Biological Chemistry
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    • v.65 no.3
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    • pp.195-202
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    • 2022
  • Soybean (Glycine max L.) leaves have been researched as functional food stuff actively, but there is no validation method to control quality of soybean leaves (SL). In this study, we annotated seven kaempferol derivatives to confirm camelliaside A as index metabolite in SL using UHPLC-ESI-TOF-MS. HPLC-UVD validation method of camelliaside A in hot-water extract of SL was established according to validation guideline of functional foods from the Ministry of Food and Safety of Korea. The HPLC-UVD method was validated with reliable parameters for examining specificity, accuracy, precision, limit of detection and quantification and linearity. The established method gave the suitable ranges to qunatitate camelliaside A from the hot-water extract of soybean leaves.