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Acquity uplc csh c18 vanguard precolumn

Manufactured by Waters Corporation

The Acquity UPLC CSH C18 VanGuard precolumn is a component designed for use in ultra-performance liquid chromatography (UPLC) systems. It is intended to protect the primary analytical column from particulates and other contaminants, helping to extend the column's lifespan and maintain system performance.

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4 protocols using acquity uplc csh c18 vanguard precolumn

1

UPLC-QToF MS Lipid Analysis Protocol

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The lipid analysis was conducted by UPLC-QToF MS (Agilent 6530 QToF MS coupled with an Agilent 1290 UPLC system; Agilent, Los Angeles, CA, USA). Dried samples were reconstituted with 100 μL of a methanol/toluene mixture (9:1 v/v) and then injected into the UPLC system using an Acquity UPLC SCH C18 column (2.1 × 100 mm, 1.7 μm) combined with an Acquity UPLC CSH C18 VanGuard precolumn (2.1 × 5 mm, 1.7 μm) (Waters, Milford, MA) for separation. The gradient conditions and solvents (A: 60:40 (v/v) acetonitrile:water and B: 90:10 (v/v) isopropanol:acetonitrile with 10 mM ammonium formate and 0.1% formic acid in both mobile phases) were adopted and modified from previous studies [27 (link)]. The information was acquired in positive ion mode. The MS condition was established following a previous study using auto MS/MS data-dependent acquisition mode and scan mode with proper modifications [28 (link)].
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2

LC-MS Analysis of Metabolite Profiles

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For LC-MS analysis, dried samples were re-dissolved in 300 μl acetonitrile:water (80:20; v/v). All measurements were carried out on a Q Exactive HF mass spectrometer coupled with a Vanquish LC system (Thermo Scientific). A sample volume of 5 μl were separated on a Waters Acquity UPLC CSH C18 column (100 × 2.1 mm; 1.7 μm) coupled to an Acquity UPLC CSH C18 VanGuard precolumn (5 × 2.1 mm; 1.7 μm). The column was maintained at 65 °C at a flow rate of 0.6 ml min− 1. The mobile phases consisted of A: acetonitrile:water (60:40, v/v) with ammonium formate (10 mM) and formic acid (0.1%) and B: 2-propanol:acetonitrile (90:10, v/v) with ammonium formate (10 mM) and formic acid (0.1%). The 15 min separation was conducted under the following gradient: 0 min 15% B; 0–2 min 30% B; 2–2.5 min 48% B;2.5–11 min 82% B; 11–11.5 min 99% B; 11.5–12 min 99% B; 12–12.1 min 15% B; 12.1–15 min 15% B. Orbitrap MS instrument was operated in electrospray ionization (ESI) in positive mode with the following parameters: mass range 60–900 m/z; spray voltage 3.6 kV, sheath gas (nitrogen) flow rate 60 units; auxiliary gas (nitrogen) flow rate 25 units, capillary temperature 320 °C, full scan MS1 mass resolving power 120,000, data-dependent MSMS (dd-MSMS) 4 scans per cycle, dd-MSMS mass resolving power 30,000. Thermo Xcalibur 4.0.27.19 was used for data acquisition and analysis [73 (link)].
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3

Untargeted Lipidomics Analysis Protocol

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The non-polar extracts were reconstituted in 100 μl methanol:toluene (9:1). Untargeted lipidomics analysis was adapted from reference (66 (link)). Briefly, lipid separation was accomplished by chromatography on an ACQUITY UPLC CSH C18 column (1.7 μm, 100 × 2.1 mm) equipped with an ACQUITY UPLC CSH C18 VanGuard pre-column (1.7 μm, 5 × 2.1 mm) (Waters) using the same LC-HRMS instrument as for untargeted metabolomics analysis. The column temperature was set at 65°C and the mobile phase flow rate at 0.6 ml/min. Mobile phases (A) 60:40 (vol/vol) acetonitrile:water with ammonium formate (10 mM) and formic acid (0.1%) and (B) 90:10 (vol/vol) isopropanol:acetonitrile with ammonium formate (10 mM) and formic acid (0.1%) were mixed according to the following gradient program: 0 min, 15% (B); 0–2 min, 30% (B); 2–2.5 min, 48% (B); 2.5–11 min, 82% (B); 11–11.5 min, 99% (B); 11.5–12 min, 99% (B); 12–12.1 min, 15% (B); and 12.1–15 min, 15% (B). The sample temperature was maintained at 4°C. Peak picking, lipid MS2 annotation, and data normalization by the LOESS algorithm were performed following the protocol for lipid analysis using MS-DIAL (version 4.48) (67 (link), 68 (link)).
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4

Lipidomics Analysis by UPLC-QTOF-MS

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The lipidomics analyses were performed using an ACQUITY UPLC system coupled to a XEVO-G2 Q-TOF (quadrupole time of flight) mass spectrometer (Waters Corporation, Milford, MA, USA) with electrospray ionization (ESI) source and an autosampler maintained at 4 °C.
Separation of lipids was performed on a Acquity UPLC CSH C18 column (100 × 2.1 mm, 1.7 μm, Waters) coupled to an Acquity UPLC CSH C18 VanGuard pre-column (5 × 2.1mm, 1.7 μm, Waters), operating at 55 °C. Injection volume of 4 µL sample.
The Acquity UPLC operated at a flow rate of 0.350 µL/min and the mobile phase consisted of acetonitrile—water (60:40; v/v) with 10 mM ammonium formate (solution A) and isopropanol-acetonitrile (90:10; v/v) with 10 mM ammonium formate (solution B). Initial conditions started with 40% B, and immediately a linear gradient from 40 to 100% B in 0 to 13 min, the eluent composition returned to the initial conditions in 0.1 min, and the column was equilibrated at the initial conditions for 3 min before the next injection, leading to a total run time of 16 min for each sample. For both positive and negative ionization modes, the same chromatographic conditions were used. Data collection and system control were performed using Waters MassLynx software (version 4.1, Waters Corporation, Milford, MA, USA).
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