Beh hilic column

Manufactured by Waters Corporation

Sourced in United States

The BEH HILIC column is a hydrophilic interaction liquid chromatography (HILIC) column manufactured by Waters Corporation. It is designed for the separation and analysis of polar and hydrophilic compounds. The BEH HILIC column utilizes a proprietary stationary phase to provide efficient and reproducible separation of a wide range of analytes.

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Lab products found in correlation

Acquity uplc system, by Waters Corporation (2 mentions) Waters acquity triple quadrupole mass spectrometer, by Waters Corporation (1 mentions) Ms grade methanol, by Merck Group (1 mentions) C18 column, by Agilent Technologies (1 mentions) Acetonitrile, by Merck Group (1 mentions) Formic acid, by Merck Group (1 mentions) Xevo tq xs mass spectrometer, by Waters Corporation (1 mentions) 4000 qtrap mass spectrometer, by AB Sciex (1 mentions) Zorbax eclipse xdb c18 column, by Agilent Technologies (1 mentions) Triple quadruple tsq quantum mass spectrometer, by Thermo Fisher Scientific (1 mentions) Acquity uplc 1 class, by Waters Corporation (1 mentions) Qtrap 6500, by AB Sciex (1 mentions) Multiquant software, by AB Sciex (1 mentions)

7 protocols using beh hilic column

Optimization of UHPLC-ESI-MS/MS for Metabolite Analysis

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TMA was analyzed separately from the other compounds, but with the same UHPLC-ESI-MS/MS method. Briefly, compound separation was achieved on a Waters Acquity UPLC system (Milford, MA, USA) with an ACQUITY BEH HILIC column (1.7 µm, 2.1 × 100 mm) coupled to an ACQUITY BEH HILIC pre-column (1.7 µm, 2.1 × 5 mm) (Waters). Mobile phases consisted of 15 mM ammonium formate in water (pH 3.5) (A) and acetonitrile (B). The gradient was set to isocratic at 80% B for 3 min, with a flow rate of 0.65 mL/min. Column temperature was set at 30 °C, and autosampler at 10 °C. Quantification was achieved by coupling the above system with a Waters Acquity triple quadrupole mass spectrometer. Source and capillary temperatures were set at 150 and 400 °C, respectively. Capillary voltage was set at 0.60 kV, and desolvation and cone gas flow (both N₂) were set at 800 and 20 L/h, respectively. Electrospray ionization (ESI) was operated in the positive mode, and data were acquired using the multiple reaction monitoring (MRM) mode. Multi-reaction monitoring (MRM) fragmentation conditions of choline, L-carnitine, betaine, γ-butyrobetaine, TMAO and TMA, as well as IS compounds can be found in Table S1.

Iglesias-Carres L., Essenmacher L.A., Racine K.C, & Neilson A.P. (2021). Development of a High-Throughput Method to Study the Inhibitory Effect of Phytochemicals on Trimethylamine Formation. Nutrients, 13(5), 1466.

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UHPLC-QTOF-MS/MS Biofilm Analysis

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Using agilent UHPLC/6545 QTOF-DAD-MS/MS spectrometer to measure electron spray ionization (ESI) spectra. Column chromatography (CC) used C₁₈ columns (2.1 mm × 50 mm, 1.8 μm, Agilent Technologies) and BEH HILIC column (2.1 × 50 mm, 1.7 μm, Waters). MS-grade methanol, acetonitrile, and formic acid were purchased from Merck (USA). Bacteria Biofilms were analyzed under an inverted fluorescence microscopy (LeiCa, DM 2000) and stained by thiazoyl blue tetrazolium bromide (MTT). Ampicillin sodium, cefoxitin, fluconazole, vancomycin, berberine chloride, 5-(4, 6-dichlorotriazinyl) aminofluorescein (5-DTAF), crystal violet, and MTT were got from Macklin Reagents (Shanghai, China). The standard compounds (purity ≥98 %) are purchased from Mai De Sheng (Cheng Du, China). Fresh pork purchased in Haoxiansheng Supermarket in KunMing.

Wang Z.J., Huang H., Zhu Y.Y., Zhou Z.S., Liu T., He X.C., Zhang T.L, & Luo X.D. (2023). Antimicrobial ingredients of Zanthoxylum motuoense and potential in fresh pork meat preservation. Heliyon, 10(1), e22963.

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Quantification of Tranexamic Acid in Plasma

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Plasma concentrations of TXA were determined by ultrahigh-performance liquid chromatography-tandem mass spectrometry. 4-Aminocyclohexanecarboxylic acid was used as the internal standard^{41 (link),42} with modifications. Specifically, plasma samples were prepared for analysis using protein precipitation with acetonitrile. Sample extracts were analyzed using normal phase chromatography with a Waters BEH HILIC column (2.1 × 100 mm, 1.7 μm, Waters Corp.) followed by detection with a Waters Xevo TQ-XS mass spectrometer. The mobile phase was A (10 mmol/L ammonium formate in water/isopropanol/formic acid 50/50/0.1):B (10 mmol/L ammonium format in acetonitrile/water/formic acid 90/10/0.1) = 40:60 with 0.25 mL/min flow rate. The mass spectrometer used an electrospray ionization source and a positive ion multiple reaction monitoring mode. For quantification of TXA, mass-to-charge ratios were set to 158.2 > 95.2 for TXA and 144.2 > 109.1 for the internal standard, respectively. Measurements for samples containing over 25 μg/mL TXA were obtained after diluting the samples into control plasma. The lower limit for TXA quantification was 0.04 μg/mL. Intra-assay (within-day) precision (% coefficient of variation [% CV]) and accuracy (% bias) were 1.3% to 8.7% and 0.9% to 7.6%, respectively. Inter-assay (between-day) % CV and % bias were 0.7% to 6.7% and 1.2% to 15.2%, respectively.

Miszta A., Ahmadzia H.K., Luban N.L., Li S., Guo D., Holle L.A., Berger J.S., James A.H., Gobburu J.V., van den Anker J., de Laat B, & Wolberg A.S. (2020). Application of a plasmin generation assay to define pharmacodynamic effects of tranexamic acid in women undergoing cesarean delivery. Journal of thrombosis and haemostasis : JTH, 19(1), 221-232.

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UHPLC-Q-TOF Metabolite Profiling

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The metabolite extracts were run on an Agilent 1290 Ultra High-Performance Liquid Chromatograph (UHPLC)-coupled to an Agilent 6538 Quadrupole-Time of Flight (Q-TOF) mass analyzer (Agilent; Santa Clara, CA, USA). An autosampler was used to inject 5 µL of sample onto a BEH-HILIC column (Waters Inc., Millford, MA, USA) with a pressure limit of 600 bar. The mobile phase consisted of A (HPLC grade water with 0.1% formic acid) and B (Acetonitrile with 0.1% formic acid) set to a flow rate of 0.400 mL/min. Gradient elution proceeded as follows: 100% B (initial), 55% B (15 to 18 min), 30% B (18 to 21 min), 100% B (21 to 25 min). The column temperature was held constant at 40 °C. Mass spectra were acquired in positive ionization mode.

Kempa J., O’Shea-Stone G., Moss C.E., Peters T., Marcotte T.K., Tripet B., Eilers B., Bothner B., Copié V, & Pincus S.H. (2022). Distinct Metabolic States Are Observed in Hypoglycemia Induced in Mice by Ricin Toxin or by Fasting. Toxins, 14(12), 815.

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Quantitative Analysis of Adenosine Metabolites

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Targeted quantitative analyses of adenosine and methylated adenosine were carried out on an Acquity UPLC system (Waters, Milford, MA, United States) coupled with a 4000 QTRAP mass spectrometer (AB SCIEX, Foster City, CA, United States). A Waters BEH HILIC column (2.1 × 100 mm, 1.7 μm) was implemented for chromatographic separation at room temperature. Mass spectrometer was operated in electrospray ionization (ESI) positive ion mode, and data were acquired by using multiple-reaction monitoring (MRM) mode. Data acquisition and processing were controlled by Analyst 1.6.3 software.

Hu Y., Fang Z., Mu J., Huang Y., Zheng S., Yuan Y, & Guo C. (2021). Quantitative Analysis of Methylated Adenosine Modifications Revealed Increased Levels of N6-Methyladenosine (m6A) and N6,2′-O-Dimethyladenosine (m6Am) in Serum From Colorectal Cancer and Gastric Cancer Patients. Frontiers in Cell and Developmental Biology, 9, 694673.

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Quantitative Analysis of Bile Acids and Ketogenic Metabolites

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The quantitative analysis of bile acids (CA, DCA, GCA) was performed on a ZORBAX Eclipse XDB-C18 column (150 mm×2.1 mm, 3.5 μm, Agilent, CA, USA) and detected by a triple quadruple TSQ Quantum mass spectrometer with ESI source (Thermo Fisher, Palo Alto, CA, USA). Blood sample preparation and analytical methods were shown in Supporting Information Methods 1.3.
For ketogenic amino acids (Phe, Lys, Trp) and BHB quantitative analysis, samples were separated on a BEH HILIC column (100 mm×2.1 mm, 1.7 μm, Waters, Ireland) and detected by a triple quadruple TSQ Quantum mass spectrometer with ESI source (Thermo Fisher, Palo Alto, CA, USA). Blood sample preparation and analytical methods were shown in Supporting Information Methods 1.4.
All the quantification methods were developed and validated in accordance with FDA guidance for Bioanalytical Method Validation (2013) (http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM368107.pdf) in terms of accuracy, precision, linearity, matrix effects, extraction recovery and stability.

Gao Y., Li W., Chen J., Wang X., Lv Y., Huang Y., Zhang Z, & Xu F. (2018). Pharmacometabolomic prediction of individual differences of gastrointestinal toxicity complicating myelosuppression in rats induced by irinotecan. Acta Pharmaceutica Sinica. B, 9(1), 157-166.

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Quantification of Fecal Metabolites

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Fecal trimethylamine and related precursors, including betaine, creatinine, L-carnitine, and choline hydroxide, were quantified by LC-MS/MS. In this study, an Acquity UPLC I-Class (Waters, Milford, MA, USA) + QTRAP 6500+ (Sciex, Framingham, MA, USA) was used as the analytical instrument and MultiQuant software (Sciex) was used for automatic identification and integration of each multi-reaction monitoring (MRM) transition (ion pair). Liquid chromatography was performed using a Waters BEH HILIC column (100 mm × 2.1 mm, 1.7 um) with a mobile phase A of ultrapure water + 0.15% FA + 10 mM ammonium formate and a mobile phase B of acetonitrile.

Liu Z., Luo G., Du R., Kan G., Han X., Zhong G., Xing W., Cui Y., Sun W., Li J., Li Y., Zhao D., Yuan X., Jin X., Han Y., Sun H., Ling S, & Li Y. (2023). Simulated spaceflight-induced cardiac remodeling is modulated by gut microbial-derived trimethylamine N-oxide. iScience, 26(12), 108556.

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