• Title/Summary/Keyword: UHPLC

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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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Geographical Classification of Angelica gigas using UHPLC-DAD Combined Multivariate Analyses (UHPLC-DAD 및 다변량분석법을 이용한 참당귀의 산지감별법 연구)

  • Kim, Jung-Ryul;Lee, Dong Young;Sung, Sang Hyun;Kim, Jinwoong
    • Korean Journal of Pharmacognosy
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    • v.44 no.4
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    • pp.332-335
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    • 2013
  • Geographical classification of A. gigas was performed in the present study using UHPLC-DAD combined with multivariate data analysis techniques. Six active constituents were isolated from A. gigas; nodakenin, marmesin, decursinol, demethylsuberosin, decursin and decursinol angelate. One hundred sixty eight A. gigas samples were simultaneously determined using UHPLC-DAD. A principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) was used to classify the samples according to geographical origins (Korea and China). The origins of A. gigas from Korea and China were correctly classified by 81.6% and 93.8% using PLS-DA Y prediction. This result demonstrates the potential use of UHPLC-DAD combined with multivariate analysis techniques as an accurate and rapid method to classify A. gigas according to their geographical origin.

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-ESI-qTOF-MS Analysis of Cyclopeptide Alkaloids in the Seeds of Ziziphus jujuba var. spinosa

  • Kang, Kyo Bin;Jang, Dae Sik;Kim, Jinwoong;Sung, Sang Hyun
    • Mass Spectrometry Letters
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    • v.7 no.2
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    • pp.45-49
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    • 2016
  • An UHPLC-ESI-qTOF-MS analytical method was developed for cyclopeptide alkaloids in the seeds of Ziziphus jujuba var. spinosa (Semen Ziziphi Spinosae), which is a commonly used herb in Chinese and Korean traditional medicines. Considering the basicity of cyclopeptide alkaloids, a specific separation method was developed for an UHPLC system. The compounds were detected by DAD and ESI-qTOF-MS, and their fragmentation patterns were also acquired by MSE technologies. Peak-picking of major compounds was performed with nine previously isolated compounds and two reference standard compounds. Tandem MS fragmentation behaviors of type-Ia and -Ib cyclopeptide alkaloids were investigated with the acquired data to develop a strategy for dereplication of other cyclopeptide alkaloid compounds in Z. jujuba var. spinosa. Two more cyclopeptide alkaloids were tentatively identified with UHPLC-ESI-qTOF-MS using this method.

Development of Quality Control Method for a Novel Herbal Medicine, HPL-1 using UHPLC (UHPLC를 이용한 새로운 한약제제 HPL-1의 품질관리법 개발)

  • Kim, Se-Gun;Lamichhane, Ramakanta;Lee, Kyung-Hee;Jung, Hyun-Ju
    • The Korea Journal of Herbology
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    • v.30 no.3
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    • pp.19-24
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    • 2015
  • Objectives : HPL-1, a novel herbal medicine which is composed of five herbs such as Kalopanacis Cortex, Chaenomelis Fructus, Raphani Semen, Atractylodis Rhizoma and Pulvis Aconiti Tuberis Purificatum, was developed for treatment of osteoarthritis. This study is aimed to develop analytical method for consistent quality control of HPL-1 and validate chromatographic method. Methods : Chromatographic analysis was performed using ultra-high performance liquid chromatography - diode array detector (UHPLC-DAD) equipped with RP-amide column, column oven, and auto sampler. Marker compounds [protocatechuic acid, chlorogenic acid, liriodendrin, 3,5-dicaffeoylquinic acid, ${\beta}$-D-(3-O-sinapoyl)-fructofuranosyl-$\alpha$-D-(6-O-sinapoyl)glucopyranoside and benzoylmesaconine] were separated by step gradient elution of acetonitrile and 0.1% phosphoric acid/water. The method validation was evaluated by quantitative validation parameters of linearity, accuracy, precision, limit of detection (LOD) and limit of quantification (LOQ) according to KFDA guideline.Results : An optimized method for six marker compounds in HPL-1 was established by UHPLC-DAD. The correlation coefficient (R2) with each calibration curve was greater than 0.99. The LOD and LOQ were within the range of 0.008-0.090 and $0.023-0.274{\mu}g/mL$, respectively. The relative standard deviation (RSD) of intra- and inter-day variability were less than 4.0%. The result of recovery test was range from 93.3-106.3% with RSD < 4.0%.Conclusions : These results suggest that the quantitative UHPLC method is precise, accurate, effective for quality evaluation of HPL-1. The method may also contribute to improve quality of crude drug preparations used for treatment of various diseases.

Metabolomics Investigation of Cutaneous T Cell Lymphoma Based on UHPLC-QTOF/MS

  • Zhou, Qing-Yuan;Wang, Yue-Lin;Li, Xia;Shen, Xiao-Yan;Li, Ke-Jia;Zheng, Jie;Yu, Yun-Qiu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5417-5421
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    • 2014
  • Objectives: The identification of cutaneous T cell lymphoma (CTCL) biomarkers may serve as a predictor of disease progression and treatment response. The aim of this study was to map potential biomarkers in CTCL plasma. Design and Methods: Plasma metabolic perturbations between CTCL cases and healthy individuals were investigated using metabolomics and ultrahigh performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Results: Principal component analysis (PCA) of the spectra showed clear metabolic changes between the two groups. Thirty six potential biomarkers associated with CTCL were found. Conclusions: Based on PCA, several biomarkers were determined and further identified by LC/MS/MS analysis. All of these could be potential early markers of CTCL. In addition, we established that heparin as a nticoagulant has better pre-treatment results than EDTA with the UHPLC-QTOF/MS appraoch.

Analysis of bee venom residues in milks of dairy cattle using UHPLC with newly developed pre-processing method (봉독 분석을 위한 전처리 방법 개발 및 이를 이용한 젖소 원유 중의 봉독 잔류물질 조사)

  • Han, Sang-Mi;Hong, In-Pyo;Woo, Soon-Ok;Kim, Se-Gun;Jang, Hye-Ri
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.25-30
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    • 2015
  • Bee venom has been used as to prevent and treat bovine mastitis as natural antimicrobial compounds in some dairy cattle farms in Korea. It is needed to determine the residual of bee venom in milks of dairy cattle treated with bee venom. Since bee venom is not approved as a raw material for animal drugs, the preprocessing method to detect bee venom residual in milk and the tolerance limit for its residue has not been established yet in Korea. Therefore, the purpose of this study was to develop pre-processing method not affecting major component of bee venom for detection of its residue in milks using ultra-high performance liauid chromatography (UHPLC). In addition, bee venom residue was also analyzed in milk samples of dairy cattle treated for mastitis with bee venom using UHPLC with the developed pre-processing method in this study. As a result, melittin, histamin and phospolipase A2, the major components of bee venom, were all detected by UHPLC with the pre-processing method developed in this study. The results of this study suggest that the pre-processing method developed in this study can be useful to detect bee venom residue in dairy cattle milk. We also found that no bee venom residues were detected in milk samples collected from dairy cattle treated with bee venom after 1 and 3 days, respectively.

Multi-Residue Analysis of Fipronil and Its Metabolites in Eggs by SinChERS-Based UHPLC-MS/MS

  • Han, Keguang;Hua, Jin;Zhang, Qi;Gao, Yuanhui;Liu, Xiaolin;Cao, Jing;Huo, Nairui
    • Food Science of Animal Resources
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    • v.41 no.1
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    • pp.59-70
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    • 2021
  • A method for simultaneous detection of fipronil (F) and its metabolites fipronil desulfinyl (FD), fipronil sulfide (FS), fipronil sulfone (FSO) in chicken eggs was applied and validated. It includes single-step, cheap, effective, rugged, safe-based method (SinChERS) for sample preparation and ultra high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS) for chemical analysis. Results suggested that formic acid enhanced the recovery of 4 target residues and 1% supplementation to acetonitrile gained higher recoveries than that of 5%. SinChERS integrated extraction and clean-up steps into one, with shorter time (1.5 h) to operate and higher recoveries (97%-100%) than HLB, Envi-Carb-NH2 and quik-easy-cheap-effective-rugged-safe method (QuEChERS), and it consumed the smallest volume of extracting solvent (10 mL) as QuEChERS. Quantitative analyses using external standard method suggested the linear ranges of 4 target compounds were 1-20 ㎍/L with R2 >0.9947. The limit of detection (S/N>3) and quantification (S/N>10) were 0.3 ㎍/kg and 1 ㎍/kg. Recoveries ranged from 89.0% to 104.4%, and the relative standard deviations (n=6) at 1, 10, and 20 ㎍/kg were lower than 6.03%. Thirty batches of domestic eggs (500 g each) were detected by the established SinChERS-based UHPLC-MS/MS and no target residues were detected in all samples. The method developed in this study is a rapid, sensitive, accurate and economic way for multi-residue analysis of fipronil and its metabolites in eggs.

Cultivation, UHPLC Pattern Analysis, and Inhibitory Effect on Respiratory Inflammation of Lysimachia mauritiana Lam. (갯까치수염(Lysimachia mauritiana Lam.)의 재배 및 UHPLC 패턴 분석, 호흡기염증 억제 효과)

  • Dong-Seon Kim;Heung Joo Yuk;JungMi Kim;Chaeseok Koh;Younjung Jang;Yoon-Young Sung
    • The Korea Journal of Herbology
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    • v.39 no.3
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    • pp.77-84
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    • 2024
  • Objectives : Lysimachia mauritiana Lam. is known as a medicinal plant native to Korea that has antioxidant, anticancer, antibacterial, and antiviral activities. However, until now, research on the cultivation technology of L. mauritiana is insufficient, and there are no research data on the systematic cultivation method and mass production of L. mauritiana. Therefore, this study aims to establish a cultivation system of L. mauritiana. Methods : The cultivation environment of open land and facilities according to the growth of L. mauritiana was compared and tested. In addition, the equivalence of the origin collection extract and the cultivation extract was evaluated through Ultra high performance liquid chromatography (UHPLC) patterns analysis according to cultivation and comparison of the effect of inhibiting respiratory inflammation using BEAS-2B human bronchial epithelial cells. Results : The cultivation technology system was established through cultivation research of L. mauritiana raw materials. In addition, as a result of comparing and evaluating the equivalence of cultivated plants and L. mauritiana raw materials for suppressing respiratory inflammation, the same results were confirmed, and the equivalence was confirmed as a result of analyzing the UHPLC pattern with L. mauritiana raw materials. Conclusions : This study suggests that extract from cultivation research of L. mauritiana plants, which are native to Korea, can be used as a health functional food or medicine to improve respiratory health.