The optimized chromatographic conditions were completed on an Atlantis T3-C18 analytical column (3.0 µm, 2.1 × 100 mm; Waters Co, Milford, CT, USA). The column was equilibrated and eluted under gradient phases containing 0.0075% formic acid in water (phase A, pH4) and in acetonitrile (phase B), and the flow rate was 0.3 mL/min. The mobile phases were degassed automatically using the online degasser system. The gradient variation started with 0% phase B. Within 0.5 min, the phase B escalated to 10% and maintained until 2 min and then sharply rose to 90% at 3 min and maintained until 5 min. The total run time was 5 min. The column temperature was maintained at 35°C. The injection volume was 5 µL with a 3-second needle wash using 5% methanol aqueous solution.
Atlantis t3 c18 analytical column
Manufactured by Waters Corporation
Sourced in United States
The Atlantis T3-C18 analytical column is a reversed-phase high-performance liquid chromatography (HPLC) column. It is designed for the separation and analysis of a wide range of polar and non-polar compounds. The column features a porous silica particle structure with a chemically bonded C18 stationary phase, providing effective separation performance.
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3 protocols using atlantis t3 c18 analytical column
1
Optimized Atlantis T3-C18 Analytical Chromatography
2
Quantifying Trace Elements via RP-ICP-MS
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RP-ICP-MS measurements were performed on an Agilent 1260 Infinity Bio-inert HPLC system (Agilent Technologies, Waldbronn, Germany) hyphenated to an Agilent 8800 ICP-MS/MS instrument (Agilent Technologies, Tokyo, Japan). Chromatographic separations were performed using an Atlantis T3 C18 analytical column (150 mm L × 2.1 mm i.d., 3 μm dp, 100 Å pore size) equipped with an Atlantis T3 guard column (20 mm L × 2.1 mm i.d., 3 μm dp) from Waters (Milford, USA). Chromatographic conditions are summarized in Table S2.† The ICP-MS/MS was equipped with a torch with 1.5 mm inner diameter, platinum cones and the brass-based lens system in order to withstand the high organic solvent load. Oxygen was introduced as the option gas and admixed with the aerosol before entering the ICP-MS to avoid carbon deposition on the cones. Data processing was performed with the Agilent MassHunter software package (Workstation Software, Version C.01.03, 2016). The ICP-MS operation parameters and chromatographic conditions used are summarized in Tables S1 and S2,† respectively. Samples were measured in triplicates and an aqueous solution of complex 3 (50 μM) was measured after every third sample as quality control.
3
Analytical HPLC for Compound Separation
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The chromatographic separation was achieved on an Atlantis T3-C18 analytical column (3.0 μm, 2.1 × 100 mm, Waters, Milford, MA, USA). The column was equilibrated and eluted with a mixed mobile phase consisting of acetonitrile and water containing 0.1% formic acid plus 10 mM ammonium acetate at a flow rate of 0.25 mL/min. The mobile phase was degassed automatically using the online degasser system. Mobile phase A was water containing 0.1% formic acid plus 10 mM ammonium acetate. Mobile phase B was acetonitrile. The gradient variation started with 100% A and maintained for 1 min, then switched to 100% mobile phase B at 1.01 min and lasted until 9 min, and finally switched back to 100% mobile phase A at 9.01 and lasted until 10 min, after which the system was returned to the initial condition. Under these conditions, the analytes coeluted with the IS within 9 min. The column temperature was maintained at 35°C. The injection volume was 10 μL, and the analysis time was 10.0 min.
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