Exionlc series uhplc system

The UHPLC separation was carried out using a SCIEX ExionLC series UHPLC System. The mobile phase A consisted of 40% water, 60% acetonitrile, and 10 mmol/L ammonium formate. The mobile phase B consisted of 10% acetonitrile and 90% isopropanol, and 10mmol/L ammonium formate. The column temperature was maintained at 45°C, and the auto-sampler temperature was set at 6°C, with an injection volume of 2 μL. The AB Sciex QTrap 6500+ mass spectrometer was used for assay development, with typical ion source parameters including IonSpray Voltage at 5,500/−4,500 V, curtain Gas at 40 psi, temperature at 350°C, ion source gas 1 at 50 psi, ion source gas 2 at 50 psi, and DP at ±80V. The measurement of the specific compounds was carried out using Biobud-v2.1.4.1. The exact quantity of each lipid was determined by analyzing the peak area and the actual concentration of the internal standard from the same lipid class.

A SCIEX ExionLC series UHPLC System was used to perform ultrahigh-performance liquid chromatography (UHPLC) separation. Mobile phase A consisted of water/acetonitrile (40%/60%) with 10 mmol/L ammonium acetate. Mobile phase B consisted of acetonitrile/isopropanol (10%/90%) with 10 mmol/L ammonium acetate. A gradient elution procedure was used (0.0–16.0 min, 80–2% mobile phase A; 16.01–18.0 min, 80% mobile phase A). The autosampler temperature was set as 6°C, and the injection volume was set as 2 μl. The column temperature was 40°C. The flow rate was set as 0.3 ml/min.
Assay development was performed with an AB Sciex QTrap 6500+ mass spectrometer. The ion spray voltage was +5,500/−4,500 V, ion source gas 1 was 50 psi, ion source gas 2 was 50 psi, the DP was ±80 V, the curtain gas was 40 psi, and the temperature was 350°C.