• Title/Summary/Keyword: HPLC fingerprinting

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Investigating chemical features of Panax notoginseng based on integrating HPLC fingerprinting and determination of multiconstituents by single reference standard

  • Yang, Zhenzhong;Zhu, Jieqiang;Zhang, Han;Fan, Xiaohui
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.334-342
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    • 2018
  • Background: Panax notoginseng is a highly valued medicine and functional food, whose quality is considered to be influenced by the size, botanical parts, and growth environments. Methods: In this study, a HPLC method integrating fingerprinting and determination of multiconstituents by single reference standard was established and adopted to investigate the chemical profiles and active constituent contents of 215 notoginseng samples with different sizes, from different botanical parts and geographical regions. Results: Chemical differences among main root, branch root, and rotten root were not distinct, while rhizome and fibrous root could be discriminated from other parts. The notoginseng samples from Wenshan Autonomous Prefecture and cities nearby were similar, whereas samples from cities far away were not. The contents of major active constituents in main root did not correlate with the market price. Conclusion: This study provided comprehensive chemical evidence for the rational usage of different parts, sizes, and growth regions of notoginseng in practice.

A study on the optimal HPLC condition for peptides complex analysis using mass spectrometry (질량분석기를 단백질 분석에 적용하기 위한 고성능액체크로마토그래피 최적조건 연구)

  • Kwon, Sung Won;Park, Chul Hong
    • Analytical Science and Technology
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    • v.16 no.1
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    • pp.78-81
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    • 2003
  • Peptides separation in high performance liquid chromatography (HPLC) is very important for the analysis of total proteins using mass spectrometry rather than two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). In this study, we investigated the optimal HPLC condition of peptides for the use of mass fingerprinting. As a result of pursuing a combination of solvent additives for HPLC, water and acetonitile containing both 0.1% trifluoroacetic acid and 0.1% acetic acid respectively showed the most efficient resolution and sensitivity.

Anti-inflammatory Activities of Lupane-triterpenoids In Vitro and Their Phytochemical Fingerprinting from Leaves of Acanthopanax gracilistylus

  • Li, Xiao Jun;Dai, Ling;Li, Zhi;Zhang, Xiao Dan;Liu, Xiang Qian;Zou, Qin Peng;Xie, Xia
    • Natural Product Sciences
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    • v.21 no.2
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    • pp.104-110
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    • 2015
  • The activities on the inhibition of NO on LPS-induced RAW 264.7 macrophages were investigated in this work. A simple and sensitive method has been developed and validated for fingerprinting analysis of leaves of Acanthopanax gracilistylus W.W. Smith (AGS). The cytotoxicity and inhibition of NO on LPS-induced RAW 264.7 cells of the extract and triterpenoids were determined. Optimal conditions of HPLC analysis were established as follows. The separation was performed with an ODS-C18 column at $30^{\circ}C$, the detected wavelength was 210 nm, the flow rate was 1 mL/min, and the mobile phase consisted of acetonitrile (0.05% phosphoric acid)-0.05% phosphoric acid solution with gradient elution. Our results showed that impressic acid and acankoreaogenin was more effective on the inhibition of NO than the methanol extract and other compounds. There were seventeen peaks coexisted with similarities above 0.95 and nine lupane-triterpenoids including acankoreaogenin and impressic acid detected and identified. The result of anti-inflammatory activities provides a potential explanation for the use of AGS leaves as a herbal medicine in the treatment of inflammatory diseases. Our results also show that acankoreanogenin and impressic acid may be potentially useful in developing new anti-inflammatory agents. In addition, the fingerprint chromatography clearly illustrated and confirmed the material basis for the anti-inflammatory activities of this plant.

Simultaneous Quantification Analysis of Multi-components on Erycibae Caulis by HPLC (HPLC를 이용한 정공등의 다성분 동시함량분석)

  • Jeon, Hye Jin;Liu, Ting;Whang, Wan Kyunn
    • YAKHAK HOEJI
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    • v.57 no.4
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    • pp.272-281
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    • 2013
  • In this study, we developed and validated the HPLC method using the isolated components from Erycibae caulis. Their structures were elucidated by spectroscopic methods including UV, $^1H$-NMR, $^{13}C$-NMR, FAB-Mass and ESI-Mass as Compound 1 (crypto-chlorogenic acid), Compound 2 (scopolin), Compound 3 (neochlorogenic acid) and Compound 4 (3,4-di-O-caffeoylquinic acid). Major three compounds and scopoletin were decided as representative components of Erycibae caulis. We established HPLC analytical method by using the representative components and 20 commercial samples which were collected considering to various cultivated area. The HPLC fingerprinting was successfully achieved with an AKZO NOBEL Kromasil 100-5C18 column. The mobile phase consisted of 0.5% acetic acid in water (A) and methanol (B) using gradient method of 85(A) to 50(A) for 35min. The fingerprints of chromatograms were recorded at an optimized wavelength of 330 nm. This developed analytical method was validated with specificity, selectivity, accuracy and precision. And it is suggested that scopolin, scopoletin, neochlorogenic acid, 3,4-di-O-caffeoylquinic acid were more than 0.162%, 0.133%, 0.057%, 0.044%, respectively. In addition, principal component analysis (PCA) was performed on the analytical data of 20 different Erycibae caulis samples in order to classify samples collected from different regions. We hope that this assay can be readily utilized as quality control method for Erycibae caulis.

Rapid and Simultaneous Determination of Ginsenosides Rb1, Rb2, Rc and Re in Korean Red Ginseng Extract by HPLC using Mass/Mass Spectrometry and UV Detection

  • Kwon, Young-Min;Lee, Sung-Dong;Kang, Hyun-Sook;Cho, Mu-Gung;Hong, Soon-Sun;Park, Chae-Kyu;Lee, Jong-Tae;Jeon, Byeong-Seon;Ko, Sung-Ryong;Shon, Hyun-Joo;Choi, Dal-Woong
    • Journal of Ginseng Research
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    • v.32 no.4
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    • pp.390-396
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    • 2008
  • For evaluating the quality of ginseng, simple and fast analysis methods are needed to determine the ginsenoside content of the ginseng products. The aim of this study was therefore to optimize conditions for fast analysis of the ginsenosides, the active ingredients in extracts of Korean red ginseng. When tandem HPLC mass spectrometry (HPLC-MS/MS) was used, four forms of ginsenoside, Rb1, Rb2, Rc, and Re, were readily separated in seven minutes using a gradient mobile phase (acetonitrile and water containing acetic acid). This is the shortest separation time reported among the studies of major ginsenoside analysis. When gradient HPLC with UV detection was used, the detection limit was high, but separation of these four ginsenosides required 25 minutes using acetonitrile and water containing formic acid as a mobile phase. HPLC-MS/MS was able to separate ginsenoside Rg1 easily regardless of the mobile phase condition, but the HPLC-UV could not separate Rg1 because acetonitrile concentration in the mobile phase had to be maintained below 20%. Ginsenoside peaks were clearer and had more sensitive detection limits when Korean red ginseng extract was analyzed by the HPLC-MS/MS, but the UV detection was useful for chromatographic fingerprinting of all four major ginsenosides of the extract: Rb1, Rb2, Rc, and Re. Extracts were found to contain 2.17 mg, 1.51 mg, 1.29 mg, and 0.46 mg of ginsenoside Rb1, Rb2, Rc, Re, respectively, per gram weight. The ratios of each ginsenoside in the extracts were 1.0 : 0.7 : 0.6 : 0.2, respectively. Taken together, the results indicate that HPLC-MS/MS spectrometry could be the most useful method for rapid analysis of even small amounts of major ginsenosides, while HPLC with UV detection could also be used for rapid analysis of major ginsenosides and for quality control of ginseng products.

Application of Metabolomics to Quality Control of Natural Product Derived Medicines

  • Lee, Kyung-Min;Jeon, Jun-Yeong;Lee, Byeong-Ju;Lee, Hwanhui;Choi, Hyung-Kyoon
    • Biomolecules & Therapeutics
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    • v.25 no.6
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    • pp.559-568
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    • 2017
  • Metabolomics has been used as a powerful tool for the analysis and quality assessment of the natural product (NP)-derived medicines. It is increasingly being used in the quality control and standardization of NP-derived medicines because they are composed of hundreds of natural compounds. The most common techniques that are used in metabolomics consist of NMR, GC-MS, and LC-MS in combination with multivariate statistical analyses including principal components analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). Currently, the quality control of the NP-derived medicines is usually conducted using HPLC and is specified by one or two indicators. To create a superior quality control framework and avoid adulterated drugs, it is necessary to be able to determine and establish standards based on multiple ingredients using metabolic profiling and fingerprinting. Therefore, the application of various analytical tools in the quality control of NP-derived medicines forms the major part of this review. $Veregen^{(R)}$ (Medigene AG, Planegg/Martinsried, Germany), which is the first botanical prescription drug approved by US Food and Drug Administration, is reviewed as an example that will hopefully provide future directions and perspectives on metabolomics technologies available for the quality control of NP-derived medicines.

Identification of Spilled Oils in the Marine Environment by Fluorescence Fingerprints and Library Search System (해양유출유의 형광지문에 의한 식별연구)

  • PARK, YONG-CHUL;KIM, YOUNG-HEE;LEE, CHANG-SUP;LEE, KI-BOCK
    • 한국해양학회지
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    • v.26 no.4
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    • pp.295-303
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    • 1991
  • Multi-spectral analyses of excitation and emission fluorescence was applied to spilled crude oils in characterization of their specific fluorescence patterns which is called oil fingerprints. In the present study, oil fingerprints of 33 crude, 4 fuel and 2 other oils were analyzed to establish data base library search system. Cluster analysis showed that crude oils could be classified into two large groups according to their fluorescence characteristics. In simulated experiments, all the spilled sources was identifiable by the present library search system. In the natural environment this system could identify the exact source of weathered crude oil slicks upton 10 days. The present study shows that the fluorescence fingerprinting method with the library search system is reliable and superior to toutine GC/HPLC analyses in identification of the source of weathered spilled oils in the marine environment.

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HPLC-tandem Mass Spectrometric Analysis of the Marker Compounds in Forsythiae Fructus and Multivariate Analysis

  • Cho, Hwang-Eui;Ahn, Su-Youn;Son, In-Seop;Hwang, Gyung-Hwa;Kim, Sun-Chun;Woo, Mi-Hee;Lee, Seung-Ho;Son, Jong-Keun;Hong, Jin-Tae;Moon, Dong-Cheul
    • Natural Product Sciences
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    • v.17 no.2
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    • pp.147-159
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    • 2011
  • A high-performance liquid chromatography-electrospray ionization-tandem mass spectrometric method was developed to determine simultaneously eight marker constituents of Forsythiae fructus, and subsequently applied it to classify its two botanical origins. The marker compounds of Forsythia suspensa were phillyrin, pinoresinol, phillygenin, lariciresinol and forsythiaside; those of F.viridissima were arctiin, arctigenin and matairesinol. Separation of the eight analytes was achieved on a phenyl-hexyl column (150${\times}$2.0 mm i.d., 3 ${\mu}M$) using gradient elution with the mobile phase: (A) 10% acetonitrile in 0.5% acetic acid, (B) 40% aqueous acetonitrile. A few fragment ions specific to the types of lignans, among the product ions generated by collisonally induced dissociation (CID) of molecular ion clusters, such as [M-H]$^-$ or [M+OAc]$^-$ were used not only for fingerprinting analysis but for the quantification of each epimer by using multiple-reaction monitoring mode. It was shown good linearity ($r^2{\geq}$ 0.9998) over the wide range of all analytes; intra- and inter-day precisions (RSD, %) were within 9.14% and the accuracy ranged from 84.3 to 115.1%. The analytical results of 40 drug samples, combined with multivariate statistical analyses - principal component analysis (PCA) and hierarchical cluster analysis (HCA) - clearly demonstrated the classification of the test samples according to their botanical origins. This method would provide a practical strategy for assessing the authenticity or quality of the herbal drug.

Anti-oxidant and Anti-inflammatory Effects of the Fermented Rhododendron weyrichii Flower Extracts in Shindari, a Traditional Jeju Fermented Drink

  • Lee, Nari;Hyun, Su Bin;Yun, Suk Hyun;Chung, You Chul;Hyun, Chang-Gu
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.471-479
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    • 2020
  • The aim of this study was to investigate the anti-oxidant and anti-inflammatory activities of the Rhododendron weyrichii flower extract fermented using Shindari, a traditional Jeju barley Nuruk-based fermentation. In this study, we examined the antioxidant potential of R. weyrichii flower extracts (RF) and R. weyrichii flower extracts fermented with Nuruk or Shindari (RFFN or RFFS, respectively) using various in vitro antioxidant assays including DPPH and ABTS radical scavenging assays, total phenol content and FRAP assays. We also evaluated the anti-inflammatory activity of the RF and RFFS on murine RAW 264.7 cells. The anti-inflammatory activity was evaluated by treating the RAW 264.7 cells with various concentrations (6.25, 12.5, 25, and 50 ㎍/ml) of RF or RFFS. As a result, we observed that the ABTS radical scavenging activity and total phenol content of RFFS was higher than that of RF and RFFN. Additionally, lipopolysaccharide-induced nitric oxide (NO) production was significantly lower in RFFS-treated cells when compared to the LPS-treated control. In addition, RFFS-treated cells exhibited decreased expression of inducible NO synthase (iNOS) proteins and high-performance liquid chromatography (HPLC) fingerprinting showed that both the quercetin and quercetin glucoside (quercitrin and isoquercitrin) levels were affected by the fermentation process. In conclusion, our data suggests that traditional fermentation could be an important strategy in improving the biological properties of raw materials including their antioxidant and anti-inflammatory activities. Finally, RFFS may be a candidate for developing topical antioxidant and anti-inflammatory agents.