• Title/Summary/Keyword: Ultra-high performance liquid chromatography

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Avantor® ACE® UltraCore HPLC and UHPLC Columns (Avantor® ACE® UltraCore HPLC/UHPLC 칼럼 가이드)

  • Peter Bridge;Ian Phillips;Gemma Lo;Cassandra Rusher
    • FOCUS: LIFE SCIENCE
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    • no.1
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    • pp.4.1-4.15
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    • 2024
  • The Avantor® ACE® UltraCore series encompasses High Performance Liquid Chromatography (HPLC) and Ultra High Performance Liquid Chromatography (UHPLC) columns designed to deliver high throughput and high-efficiency ultra-fast separations. Utilizing ultra-inert solid-core silica particles with monodisperse particle distribution, these columns combine the high efficiency of UHPLC with the operability of HPLC instrumentation, yielding lower backpressure and high-resolution separations suitable for a broad spectrum of analytes. The Avantor® ACE® UltraCore range includes three primary product types: • UltraCore BIO: Designed for large biomolecules (≥5 kDa), these columns offer exceptional performance in separating biologically derived compounds. • UltraCore: Ideal for standard small organic molecules, providing rapid separations for both synthetic and natural mixtures. • UltraCore Super: Equipped with encapsulated bonding technology for small organic molecules in extreme pH conditions, optimal for high pH buffer requirements. The Avantor® ACE® UltraCore columns present a versatile and high-efficiency solution for chromatographic separation needs, accommodating a wide range of molecular sizes and providing enhanced resolution and reduced analysis time. Their adaptability to both HPLC and UHPLC systems, combined with the advantages of solid-core technology, makes them an invaluable tool in analytical and preparative chromatography.

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UHPLC System Shutdown and Reactivation Advice (UHPLC 시스템 종료 및 재가동 시 가이드)

  • Mark Fever;Gemma Lo
    • FOCUS: LIFE SCIENCE
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    • no.1
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    • pp.8.1-8.3
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    • 2024
  • Ultra-high performance liquid chromatography (UHPLC) systems are integral to modern analytical laboratories, necessitating careful maintenance and operation protocols to ensure optimal performance. This document provides comprehensive guidelines for the proper shutdown and reactivation of UHPLC systems to prevent damage and maintain operational efficiency. • Shutdown: Remove the column and replace it with a union to avoid blockages. Flush the system with a compatible solvent mix, clean mobile phase reservoirs to prevent microbial growth, flush the pump with storage solvent, and clean the autosampler, including the needle and injection port. • Reactivation: Inspect the system for wear or damage, gradually reintroduce mobile phases starting with a weak solvent, reinstall the column securely, and perform system checks on baseline stability, pressure consistency, and detector performance. By adhering to these guidelines, laboratories can ensure the longevity and reliability of their UHPLC systems, maintaining high analytical performance and minimizing downtime. These procedures help prevent common issues such as blockages, contamination, and component wear, thereby supporting efficient and accurate analytical operations.

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Improved Calibration for the Analysis of Emerging Contaminants in Wastewater Using Ultra High Performance Liquid Chromatography and Time-of-Flight Mass Spectrometry

  • Pellinen, Jukka;Lepisto, Riikka-Juulia;Savolainen, Santeri
    • Mass Spectrometry Letters
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    • v.9 no.3
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    • pp.77-80
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    • 2018
  • The focus of this paper is to present techniques to overcome certain difficulties in quantitative analysis with a time-of-flight mass spectrometer (TOF-MS). The method is based on conventional solid-phase extraction, followed by reversed-phase ultra high performance liquid chromatography of the extract, and mass spectrometric analysis. The target compounds included atenolol, atrazine, caffeine, carbamazepine, diclofenac, estrone, ibuprofen, naproxen, simazine, sucralose, sulfamethoxazole, and triclosan. The matrix effects caused by high concentrations of organic compounds in wastewater are especially significant in electrospray ionization mass spectroscopy. Internal-standard calibration with isotopically labeled standards corrects the results for many matrix effects, but some peculiarities were observed. The problems encountered in quantitation of carbamazepine and triclosan, due to nonlinear calibration were solved by changing the internal standard and using a narrower mass window. With simazine, the use of a quadratic calibration curve was the best solution.

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.

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.

Phytochemical analysis of Panax species: a review

  • Yang, Yuangui;Ju, Zhengcai;Yang, Yingbo;Zhang, Yanhai;Yang, Li;Wang, Zhengtao
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.1-21
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    • 2021
  • Panax species have gained numerous attentions because of their various biological effects on cardiovascular, kidney, reproductive diseases known for a long time. Recently, advanced analytical methods including thin layer chromatography, high-performance thin layer chromatography, gas chromatography, high-performance liquid chromatography, ultra-high performance liquid chromatography with tandem ultraviolet, diode array detector, evaporative light scattering detector, and mass detector, two-dimensional high-performance liquid chromatography, high speed counter-current chromatography, high speed centrifugal partition chromatography, micellar electrokinetic chromatography, high-performance anion-exchange chromatography, ambient ionization mass spectrometry, molecularly imprinted polymer, enzyme immunoassay, 1H-NMR, and infrared spectroscopy have been used to identify and evaluate chemical constituents in Panax species. Moreover, Soxhlet extraction, heat reflux extraction, ultrasonic extraction, solid phase extraction, microwave-assisted extraction, pressurized liquid extraction, enzyme-assisted extraction, acceleration solvent extraction, matrix solid phase dispersion extraction, and pulsed electric field are discussed. In this review, a total of 219 articles published from 1980 to 2018 are investigated. Panax species including P. notoginseng, P. quinquefolius, sand P. ginseng in the raw and processed forms from different parts, geographical origins, and growing times are studied. Furthermore, the potential biomarkers are screened through the previous articles. It is expected that the review can provide a fundamental for further studies.

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.

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.

Avantor® ACE® Wide Pore HPLC Columns for the Separation and Purification of Proteins in Biopharmaceuticals (바이오의약품의 단백질 분리 및 정제를 위한 Avantor® ACE® 와이드 포어 HPLC 컬럼 가이드)

  • Matt James;Mark Fever;Tony Edge
    • FOCUS: LIFE SCIENCE
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    • no.1
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    • pp.3.1-3.7
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    • 2024
  • The article discusses the critical role of chromatography in the analysis and purification of proteins in biopharmaceuticals, emphasizing the importance of comprehensive characterization for ensuring their safety and efficacy. It highlights the use of Avantor® ACE® HPLC columns for the separation and purification of proteins, focusing on the analysis of intact proteins using reversed-phase liquid chromatography (RPLC) with fully porous particles. This article also details the application of different mobile phase additives, such as TFA and formic acid, and emphasizes the advantages of using type B ultra-pure silica-based columns for efficiency and peak shape in biomolecule analysis. Additionally, it addresses the challenges of analyzing intact proteins due to slow molecular diffusion and introduces the concept of solid-core (or superficially porous) particles, emphasizing their benefits over traditional porous particles for the analysis of therapeutic proteins. Furthermore, it discusses the development of Avantor® ACE® UltraCore BIO columns, specifically designed for the high-efficiency separation of large biomolecules, such as proteins, and demonstrates their effectiveness in achieving high-resolution separations, even for higher molecular weight proteins like monoclonal antibodies (mAbs). In addition, it underscores the complexity of analyzing and characterizing intact protein biopharmaceuticals, requiring a range of analytical techniques and the use of wide-pore stationary phases, operated at elevated temperatures and with relatively shallow gradients. It highlights the comprehensive range of options offered by Avantor® ACE® wide pore columns, including both fully porous and solid-core particles, bonded with a variety of complementary stationary phase chemistries to optimize selectivity during method development. The use of ultrapure and highly inert base silica is emphasized for enabling the use of lower concentrations of mobile phase modifiers without compromising analyte peak shape, particularly beneficial for LC-MS applications. Then the article concludes by emphasizing the significance of reversed-phase liquid chromatography and its compatibility with mass spectrometry as a valuable tool for the separation and analysis of intact proteins and their closely related variants in biopharmaceuticals.

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Analysis of oligosaccharides from Panax ginseng by using solid-phase permethylation method combined with ultra-high-performance liquid chromatography-Q-Orbitrap/mass spectrometry

  • Li, Lele;Ma, Li;Guo, Yunlong;Liu, Wenlong;Wang, Yang;Liu, Shuying
    • Journal of Ginseng Research
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    • v.44 no.6
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    • pp.775-783
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    • 2020
  • Background: The reports about valuable oligosaccharides in ginseng are quite limited. There is an urgent need to develop a practical procedure to detect and analyze ginseng oligosaccharides. Methods: The oligosaccharide extracts from ginseng were permethylated by solid-phase methylation method and then were analyzed by ultra-high-performance liquid chromatography-Q-Orbitrap/MS. The sequence, linkage, and configuration information of oligosaccharides were determined by using accurate m/z value and tandem mass information. Several standard references were used to further confirm the identification. The oligosaccharide composition in white ginseng and red ginseng was compared using a multivariate statistical analysis method. Results: The nonreducing oligosaccharide erlose among 12 oligosaccharides identified was reported for the first time in ginseng. In the comparison of the oligosaccharide extracts from white ginseng and red ginseng, a clear separation was observed in the partial least squares-discriminate analysis score plot, indicating the sugar differences in these two kinds of ginseng samples. The glycans with variable importance in the projection value large than 1.0 were considered to contribute most to the classification. The contents of oligosaccharides in red ginseng were lower than those in white ginseng, and the contents of maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose, maltoheptaose, maltooctaose, maltononaose, sucrose, and erlose decreased significantly (p < 0.05) in red ginseng. Conclusion: A solid-phase methylation method combined with liquid chromatography-tandem mass spectrometry was successfully applied to analyze the oligosaccharides in ginseng extracts, which provides the possibility for holistic evaluation of ginseng oligosaccharides. The comparison of oligosaccharide composition of white ginseng and red ginseng could help understand the differences in pharmacological activities between these two kinds of ginseng samples from the perspective of glycans.