Samples were analyzed by LC-MS using positive-ion electrospray ionization and reverse-phase chromatography in full-scan mode with MS/MS product-ion spectra generated for detected metabolite masses. This was performed using the MS2 scan mode of the G6410B mass spectrometer to produce MS/MS product-ion spectra for masses detected at sufficient intensities. Chromatography was performed using a ZORBAX Eclipse XDB-C18 4.6 × 50 mm, 5 µm, column, with gradient elution at 1 mL min−1. Mobile phase was comprised of A = MilliQ Water with 0.1% formic acid, B = acetonitrile. Six metabolites were detected (M1, M2, M3a, M3b, M4, M5) and chromatography resolved.
Eclipse xdb c18 4
Manufactured by Agilent Technologies
The Eclipse XDB-C18 4 is a high-performance liquid chromatography (HPLC) column designed for a wide range of analytical applications. It features a 4.6 mm internal diameter and a 150 mm length, with a 5 μm particle size. The column packing material is a dimethyloctadecylsilyl-bonded amorphous silica, providing a stable and efficient platform for reversed-phase chromatography.
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Lab products found in correlation
2 protocols using eclipse xdb c18 4
1
LC-MS Analysis of Metabolites
2
Quantitative LC-MS/MS Analysis of Compounds
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Agilent 1200 series liquid chromatography system equipped with applied biosystems (API 5500 Q trape and API 4000 Q trape) tandem mass spectrometry with electrospray ionization Interface (ESI) source in the positive mode was used. Separation was performed on a C18 ZORBAX Eclipse XDB-C 18 4.6 × 150 mm shaft, 5 mm particle size. A gradient elution program was used at a flow rate of 0.3 ml/min, with one reservoir containing the mobile phase which was 10 mM of ammonium format solution in methanol: water ________________________________________________ Egypt. J. Chem. 65, No. 7 (2022) 431 (1:9 v/v). Nitrogen was used as nebulizer gas, curtain gas, heating gas and impingement gas according to the manufacturer's settings; the source temperature was 450 °C, and the ion spray potential was 5500 V-. The injection volume was 2 µl. Multiple Reaction Monitoring (MRM) mode was used where one MRM was used for quantification and the other was used for confirmation.
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