Agilent 6230 tof system

All the MS experiments were conducted on an Agilent 6230 TOF system
(Agilent Technologies, Wilmington, DE, U.S.A.), equipped with a DUAL AJS ESI
source operating in positive or negative ion mode. MS spectra were acquired from
m/z 100 to 500 at a scan rate of 1 spectrum per second. The electrospray
ionization (ESI) source parameters were used as follows: gas temp, 325
°C; gas flow, 11 L/min; nebulizer, 35 psi; Vcap, 3500 V; nozzle,1000 V;
fragmentor, 175 V.

High-performance liquid chromatography was performed using an Agilent 1290 Infinity II LC system (Agilent Technologies, Wilmington, DE, USA) equipped with a Diode-Array Detection (DAD), binary pump, multicolumn thermostat, and autosampler. The mobile phases used for the separation were MS-grade water with 0.1% formic acid (solvent A) and MS-grade acetonitrile with 0.1% formic acid (solvent B). Gradient elution was performed at a flow rate of 0.4 mL/min, and the mobile phase was 15% solvent B for 3 min, then from 15% to 45% solvent B for 12 min, 45% solvent B to 90% solvent B for 0.5 min and maintained at 90% solvent B for 3 min, followed by 3.5 min for column re-equilibration. Separations were performed at a column temperature of 60 °C with a total run time of 22 min. A varying volume of sample was injected onto the column for each experimental run.
The HPLC-TOF-MS experiments were conducted on an Agilent 6230 TOF system (Agilent Technologies, Wilmington, DE, USA), equipped with a DUAL Jet Stream electrospray ionization (AJS ESI) source operating in positive ion mode. MS spectra were acquired from m/z 150 to 1700 at the scan rate of 1 spectrum per second. The ESI source parameters were used as follows: Gas Temp: 325 °C; Gas Flow: 11 L/min; Nebulizer: 35 psi; Vcap: 3500 V; Nozzle V: 1000; Fragmentor: 175V.

Liquid chromatography was performed on an Agilent 1290 Infinity II LC system (Agilent Technologies, Wilmington, DE, U.S.A.) equipped with a diode array detector (DAD), binary pump, multicolumn thermostat, and autosampler. The mobile phases used for the separation were MS-grade water with 0.1% formic acid (solvent A) and MS-grade acetonitrile with 0.1% formic acid (solvent B). Gradient elution of proteins from the analytical column was performed using a gradient starting at 15% B at a flow rate of 0.6 mL/min. The mobile phase was then 15–95% B for 3.5 min, and maintained at 95% B for 0.5 min, followed by 15% B for 0.5 min and re-equilibration of column with 15% B for 1.5 min. Separations were performed at a column temperature of 50 °C with a total run time of 6 min. A varying volume of sample was injected onto the column for each experimental run.
The MS experiment was conducted on an Agilent 6230 TOF system (Agilent Technologies, Wilmington, DE, U.S.A.), equipped with a DUAL AJS ESI source operating in positive ion mode. MS spectra were acquired from m/z 600 to 5000 at a scan rate of 1 spectrum per second with Profile format. The electrospray ionization (ESI) source parameters were used as follows: gas temperature, 290 °C; gas flow, 13 L/min; nebulizer, 25 psi; sheath gas temperature: 375 °C; Vcap, 4500 V; nozzle, 1000 V; fragmentor, 175 V.