Zorbax rapid resolution 300sb c18

An Agilent 1290 UPLC system (Agilent, Santa Clara, CA) was utilized for intact adducted protein analysis. The analytical column utilized was a Zorbax rapid resolution 300SB-C₁₈ (part number 863974-302, Agilent) 3 × 150 mm-i.d. (3.5 μm particle size). The aqueous mobile phase (A) was 0.1% acetic acid/water, and the organic phase (B) was 0.1% acetic acid/acetonitrile (ACN). After injection of 2 μL sample onto the column the sample was eluted at 400 μL/min from the column using a solvent gradient that initially consisted of 99% A and 1% B for 1 min, and then a 5.5% increase in B for the next 13.5 min to a final concentration of 75% B. The column eluent was introduced into an Agilent 6530 QTOF (Agilent) mass spectrometer with an electrospray ionization interface. The instrument, operated in full scan mode, was utilized for intact adducted protein analysis. The instrument was operated in the positive ion mode with a survey scan range from 800 to 3000 Da. Instrument parameters were as follows: gas temperature 350 °C, gas flow 10 L/min, nebulizer 60 psi, fragmentation voltage was 250 V and the capillary voltage was 3500 V. Instrument control and data processing were performed with the Mass Hunter software version for B.02.01 data acquisition and B.05.00 for qualitative analysis.