The chromatographic separation was accomplished with an ACQUITY UPLC BEH Amide column, 2.1 × 100 mm, 1.7 μm (Waters, Ireland, Dublin), equipped with an ACQUITY UPLC BEH Amide VanGuard Pre-column, 1.7 µm, 2.1 mm × 5 mm (Waters, Ireland, Dublin). The data were acquired by implementing the Dionex UltiMate 3000 RSLC UHPLC system (Thermo Fischer Scientific, Dreieich, Germany) coupled to a Maxis Impact QTOF mass spectrometer (Bruker Daltonics, Bremen, Germany) through an electrospray ionization source (ESI) capable of both positive and negative ionization. The column temperature was maintained at 30 °C. The gradient elution program is the same in both ionization modes. The conditions of liquid chromatography and the settings of ESI–QTOF instrumentation are described in Table 4 .
Acquity uplc beh amide vanguard pre column
Manufactured by Waters Corporation
Sourced in Ireland
The ACQUITY UPLC BEH Amide VanGuard Pre-column is a laboratory equipment product designed to be used in conjunction with Waters Corporation's ACQUITY UPLC system. It serves as a pre-column, providing sample separation and protection for the primary analytical column.
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2 protocols using acquity uplc beh amide vanguard pre column
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Optimized UPLC-QTOF-MS Analytical Protocol
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Quantification of Sialic Acid Metabolites
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Ammonium acetate (≥98% purity) and acarbose (≥95% purity) were purchased from Sigma-Aldrich. Ammonium hydroxide solution p.a. was obtained from Honeywell, UPLC water (ULC-MS grade) from Biosolve and acetonitrile (hypergrade) from Merck. N-acetyl-D-[1,2,3-13 C 3 ] neuraminic acid ( 13 C 3 -NANA) and hexaacetyl-chitohexaose were purchased from Omicron Biochemicals Inc. and Neogen, respectively. The ACQUITY UPLC BEH Amide Column, 130 Å, 1.7 μm, 2.1 mm  50 mm and ACQUITY UPLC BEH Amide VanGuard Pre-column, 130 Å, 1.7 μm, 2.1 mm  5 mm were purchased from Waters.
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