• Title/Summary/Keyword: high performance liquid chromatography

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Development of high-performance liquid chromatography methods for the anticoccidials: toltrazuril and diclazuril (고속액체크로마토그래피를 이용한 항원충제 분석법 개발: 톨트라주릴 및 디클라주릴)

  • Jeong, Kyung Hun;Jeong, Miyoung;Park, Hae-Chul;Hossain, Md Akil;Kim, Daegyun;Lee, Kwang-Jick;Kang, Jeong Woo
    • Korean Journal of Veterinary Research
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    • v.57 no.4
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    • pp.223-226
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    • 2017
  • This study was undertaken to develop new analytical methods for assessment of anticoccidials. High-performance liquid chromatography (HPLC) was found to be a fast, reliable, and practical method. The anticoccidials used in this experiment were toltrazuril and diclazuril, and the analysis factors were specificity, linearity, accuracy, repeatability, and intermediate precision. The linearity of each anticoccidial was better than 0.99, and the accuracies were 99.5% and 99.1% with relative SD of 0.5 and 0.4, respectively. To assess whether the developed HPLC method could be effectively applied, toltrazuril and diclazuril post-market veterinary products (five products) that are currently sold were tested. The results revealed no non-compliant items and the method was applied successfully. Therefore, the newly developed HPLC method for anticoccidial assessment described in this study may be useful as a reference method in the Korean Standards of Veterinary Pharmaceuticals for the analysis of toltrazuril and diclazuril.

Mass Balance on the Pulping Extracts of Maple Hardwood using High Performance Liquid Chromatography (HPLC 분석기를 이용한 펄프용 단풍나무의 펄핑 추출액에 관한 물질수지)

  • Um, Byung-Hwan
    • Journal of the Korean Wood Science and Technology
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    • v.36 no.4
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    • pp.102-108
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    • 2008
  • At the University of Maine, a hemicellulose pre-extraction technology is now being investigated to improve pulp yields, reduce the organic and inorganic load for liquor recovery, and create a feed stream for the generation of new biomaterials. It is important to understand the composition of unextracted wood, extracted wood, and pulping extracts in the design of an economically viable pilot-scale ethanol plant. For analysis of wood pulp composition, the total analytical mass balance closure was 100.6, 100.3, and 81.6% for unextracted chips, extracted chips, and pulping extracts from HPLC-H column analysis. Meanwhile, the total analytical mass balance from the HPLC-P column was 97.8, 86.3, and 80.7%, respectively. This slight variability between H- and P-column results for analytical mass balance may be within the experimental error of the measurement. The data generated by this analysis are important to further design work in commercializing this process.

Determination of Ginseng Saponins by Reversed-Phase High Performance Liquid Chromatography (역상 고속 액체크로마토그래피를 이용한 인삼 사포닌의 분석)

  • Jeong, Seung-Il;Kim, Choen-Suk;Lee, No-Woon;Choi, Kang-Ju;Lee, Yong-Gu;Kim, Il-Kwang
    • Analytical Science and Technology
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    • v.11 no.6
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    • pp.436-439
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    • 1998
  • Ginseng saponins were analysed using reversed-phase high performance liquid chromatography with several columns. The optimum conditions were as following : reverse phase column; Novapak $C_{18}$ ODS column ($3.9mm{\times}150mm$, $5{\mu}m$), acetonitrile/water binary mobile phase gradient controller system, solvent flow rate; 1.5 mL/min, and UV (203 nm) detector. The complete separation of ginsenoside $Rb_1$, $Rb_2$, Rc, Rd, Re, Rf and $Rg_1$ was achieved within 50 min. The regression coefficients of the calibration curves for seven ginsenosides were 0.98~0.99.

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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.

Determination of polyphenolic compounds in grape seed extracts using reverse-phase high performance liquid chromatography

  • Wang, Chong-Zhi;Osinski, Joachim;Shao, Zuo-Hui;Basila, Daniel;Kim, Stephen;Yuan, Chun-Su
    • Advances in Traditional Medicine
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    • v.4 no.4
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    • pp.267-273
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    • 2004
  • Oxidative stress is associated with many kinds of chronic diseases. Antioxidants such as polyphenols are compounds that protect cells against the damaging effects of reactive oxygen species. Grape seeds are considered good resources of polyphenols, and grape seed extracts have a very strong antioxidant effect. In the present study, we established a simple gradient reverse-phase high performance liquid chromatography method to determine polyphenol content from three different grape seed resources. An ODS (2), $150\;{\times}\;3.2\;mm$ column has been employed, and six polyphenols have been determined: gallic acid, protochatechuic acid, (+)-catechin, (-)-epicatechin, procyanidin B2, and epicatechin gallate. Catechin and epicatechin were the main polyphenol compounds in all three extracts. The amount of procyanidin B2 was higher in Extract 1 (from a company of China), while Extract 2 (extracted in our lab) and Extract 3 (from a company of USA) contained higher proportions of epicatechin gallate. For the total polyphenol content, Extract 1 was much higher than that of Extract 2 and 3. The results suggest that the dietary dose of grape seed extracts from different resources should be adjusted according to polyphenol content.

Preparative separation of minor saponins from Panax notoginseng leaves using biotransformation, macroporous resins, and preparative high-performance liquid chromatography

  • Liu, Fang;Ma, Ni;Xia, Fang-Bo;Li, Peng;He, Chengwei;Wu, Zhenqiang;Wan, Jian-Bo
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.105-115
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    • 2019
  • Background: Ginsenosides with less sugar moieties may exhibit the better adsorptive capacity and more pharmacological activities. Methods: An efficient method for the separation of four minor saponins, including gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, and notoginsenoside Fd, from Panax notoginseng leaves (PNL) was established using biotransformation, macroporous resins, and subsequent preparative high-performance liquid chromatography. Results: The dried PNL powder was immersed in the distilled water at $50^{\circ}C$ for 30 min for converting the major saponins, ginsenosides Rb1, Rc, Rb2, and Rb3, to minor saponins, gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, and notoginsenoside Fd, respectively, by the enzymes present in PNL. The adsorption characteristics of these minor saponins on five types of macroporous resins, D-101, DA-201, DM-301, X-5, and S-8, were evaluated and compared. Among them, D-101 was selected due to the best adsorption and desorption properties. Under the optimized conditions, the fraction containing the four target saponins was separated by D-101 resin. Subsequently, the target minor saponins were individually separated and purified by preparative high-performance liquid chromatography with a reversed-phase column. Conclusion: Our study provides a simple and efficient method for the preparation of these four minor saponins from PNL, which will be potential for industrial applications.

Multiclass Method for the Determination of Anthelmintic and Antiprotozoal Drugs in Livestock Products by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry

  • Hyunjin Park;Eunjung Kim;Tae Ho Lee;Sihyun Park;Jang-Duck Choi;Guiim Moon
    • Food Science of Animal Resources
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    • v.43 no.5
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    • pp.914-937
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    • 2023
  • The objective of this study was to establish a multi-residue quantitative method for the analysis of anthelmintic and antiprotozoal drugs in various livestock products (beef, pork, and chicken) using ultra-high-performance liquid chromatography-tandem mass spectrometry. Each compound performed validation at three different levels i.e., 0.5, 1, and 2× the maximum residue limit according to the CODEX guidelines (CAC/GL 71-2009). This study was conducted according to the modified quick, easy, cheap, effective, rugged, and safe procedure. The matrix-matched calibrations gave correlation coefficients >0.98, and the obtained recoveries were in the range of 60.2%-119.9%, with coefficients of variation ≤32.0%. Furthermore, the detection and quantification limits of the method were in the ranges of 0.03-3.2 and 0.1-9.7 ㎍/kg, respectively. Moreover, a survey of residual anthelmintic and antiprotozoal drugs was also carried out in 30 samples of beef, pork, and chicken collected in Korea. Toltrazuril sulfone was detected in all three samples. Thus, our results indicated that the developed method is suitable for determining the anthelmintic and antiprotozoal drug contents in livestock products.

Necessity of high-performance liquid chromatography validation in good laboratory practice systems (비임상시험관리기준 시스템에서 고성능액체크로마토그래피 분석법 정확도 검증의 필요성)

  • Su Bin Yang;Changyul Kim
    • Journal of the Korean Applied Science and Technology
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    • v.41 no.2
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    • pp.172-187
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    • 2024
  • In good laboratory practice (GLP), analytical method validation prior to toxicity testing is important in terms of concentration setting and sample preparation. The testing agency performed validation of the two requested test substances using high-performance liquid chromatography (HPLC), and as a result, the analysis satisfied the criteria for specificity, system suitability, linearity, intra-day reproducibility, homogeneity, stability, concentration analysis, and quality control. The method was established and validated. However, the content of standard substances in the test report of the requesting institution was found to be 1.34 and 1.17 times higher than the results of the testing institution, confirming that deriving results through validation of analysis methods suitable for GLP is important in terms of securing reliability and stability.

Evaluation of Anti-Oxidant from Natural Products (천연물로부터 유래한 천연 항산화제 규명)

  • Kwon, Jin-A;Yang, Yoon-Jung;Park, Jong-Hyuk;Kang, Se-Chan
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2011.10a
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    • pp.20-20
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    • 2011
  • In this study, we analyzed 80%MeOH extract of fruits of sorbaria sorbifolia var. stellipila MAX. to measure the total antioxidant capacity by oxygen radical absorbance capacity (ORAC) assay, individual flavonoid content by high-performance liquid chromatography (HPLC). n-Hexane ($1.02{\pm}0.036$), $CH_2Cl_2$ ($0.95{\pm}0.025$), EtOAc ($1.94{\pm}0.065$), n-BuOH ($1.98{\pm}0.054$), D.W. ($1.2{\pm}0.032$) fractions were examined antioxidative activity by ORAC assay. It was revealed that EtOAc($1.94{\pm}0.065$), n-BuOH($1.98{\pm}0.054$) fractions had significant antioxidative activity. The isolation and separation were facilitated using open column chromatography, while separation, purification and identification were accomplished by using high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR).

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Determination of Theophylline and its Metabolites in Human Urine by High-Performance Liquid Chromatography

  • Kim, Kyeong-Ho;Park, Young-Hwan;Park, Hyo-Kyung;Kim, Ho-Soon;Lee, Min-Hwa
    • Archives of Pharmacal Research
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    • v.19 no.5
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    • pp.396-399
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    • 1996
  • High-performance liquid chromatographic method with UV detecction was developed for the determination of theophylline and its metabolites in human urine using ${beta}$-hydroxyethyl theophylline$({beta} -HET)$ as an internal standard. For extraction of urine sample, quality control sample and xanthine-free blank urine were mixed with decylamine (ion-paring reagent) and ${beta}$-HET. After saturation with ammonium sulfate, the mixture was then extracted with organic solvent at pH values of 4.0-4.5. All separations were performed with ion-pair chromatography using decylamine as an ion-pairing reagent and 3mM sodium acetate buffered mobile phase (pH 4.0) containing 1% (v/v) acetonitrile and 0.75 mM decylamine. The detection limits of theophylline, 1, 3-DMU, 1-MU, 3-MX and 1-MX in human urine were 0.17, 0.17, 0.39, 0.19 and 0.19 ${\mu}g$/ml, based on a signal-to-noise ratios of 3.0. The mean intraday coefficients of variation (C.V.s) of each compound on nine replicates were lower than 2.0%, while mean interday C.V.s on three days were lower than 1.6%. All separations were finished within 40miutes.

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