Analyses were performed using the analytical reverse phase column C18 CLC-ODS (M) 25 cm × 4.6 mm (Shimadzu) with a particle diameter of 5 μm.
A HP5-MS capillary column (30 m × 0.25 mm × 0.25 μm) was used as the analytical column.
A Cadenza CD-C18 (150 × 2.0-mm inner diameter; 3.0-μm particle size; Imtakt Corp ,Kyoto, Japan) was used as the analytical column.
In the first stage of the work, the composite column was designed as an alternative to the existing steel column using the analytical method.
Comparing the force developed in each PT bar at the peak strength of the columns using the analytical approach, T Analysis, and experimental results, T EXP, revealed that using the analytical approach, the PT force in the bars at the peak strength was within ±12% of the average of the test results.
The predicted ductility versus the ductility of the columns obtained using the analytical method is presented in Fig. 18a.
A Zorbax guard fittings kit packed with a replaceable Eclipse Plus C18 Guard column (12.5 × 4.6 mm i.d.; 5 μm) was used to protect the analytical column.
A Zorbax guard fittings kit packed with replaceable Eclipse Plus C18 Guard column (12.5 × 4.6 mm i.d.; 5 μm) was used to protect the analytical column.
The LC column was a Luna C-18 column (25 cm × 4.6 mm; particle size, 5 μm) (Phenomenex, Torrance, CA, USA) with a small pre-column (2.5 cm × 3 mm) containing the same filling used to protect the analytical column.
The solvent system used includes 0.1% aqueous formic acid as solvent A and 100% acetonitrile, 0.1% formic acid as solvent B. The peptides were loaded on the trap column using 97% solvent A, followed by resolution on the analytical column using a linear gradient of 5-30% solvent B for 70 min at a constant flow rate of 0.35 μL/min.
