Exionlc high flow lc

Analysis was carried out in the Proteomics and Metabolomics Unit, Children’s Cancer Hospital (57357), Basic Research Department, Cairo, Egypt. Adopting the methods previously reported by Negm et al. [21 (link)]. In-line filter disks (0.5 µm × 3.0 mm, Phenomenex^®, Torrance, CA, USA) and X select HSS T3 (2.5 µm, 2.1 mm × 150 mm, Waters^®, 40 °C, Milford, MA, USA) were used as a pre-column and analytical column, respectively. The mobile phases consisted of buffer A (5 mM ammonium formate buffer pH 3 containing 1% methanol), buffer B (5 mM ammonium formate buffer pH 8 containing 1% methanol), and buffer C (100 % acetonitrile). The flow rate was adjusted at 0.3 mL/min. The liquid chromatography (ExionLC -High flow LC, Sciex^®) was programmed to use a mobile phase composition of buffer A and C in positive mode, and buffer B and C in negative mode. The mobile phase composition started with 90 (A or B): 10 (C) for the first 20 min, which was inversed from 21–25 min, and finally returning to the starting values for the last 3 min until the end of the protocol at 28 min. In addition, the instrument (ExionLC -High flow LC-, Sciex®, Framingham, MA, USA) was coupled with Triple TOF 5600+ (Sciex^®) for IDA acquisition and Analyst TF 1.7.1 (Sciex^®) for LC-Triple TOF control.

Analysis was conducted in a Proteomics and Metabolomics Unit, Children’s Cancer Hospital (57357), Basic Research Department, Cairo, Egypt. In-line filter disks (0.5 µm × 3.0 mm, Phenomenex^®, Torrance, CA, USA) and X select HSS T3 (2.5 µm, 2.1 × 150 mm, Waters^®, Milford, MA, USA, 40 °C) were used as a pre-column and analytical column, respectively. The mobile phases consisted of buffer A (5 mM ammonium formate buffer pH 3 containing 1% methanol), buffer B (5 mM ammonium formate buffer pH 8 containing 1% methanol), and buffer C (100% acetonitrile). The flow rate was adjusted at 0.3 mL/min. The liquid chromatography (ExionLC -High flow LC-, Sciex^®, Framingham, MA, USA) was programmed to use a mobile phase composition of buffer A and C, in positive mode, while it was programmed to use a mobile phase composition of buffer B and C in negative mode. The mobile phase composition started with 90 (A or B): 10 (C) for the first 20 min, which was inversed from 21 to 25 min, and finally returned back for the last 3 min until the end of the protocol at the 28 min. In addition, the instrument was coupled with Triple TOF 5600+ (Sciex^®) for IDA acquisition and Analyst TF 1.7.1 (Sciex^®) for LC-Triple TOF control.