Agilent 1200 series hplc chip system

Tandem MS data of peptides and glycopeptides were obtained by injecting 2 μL of sample into an Agilent 1200 series HPLC-Chip system coupled to an Agilent 6520 Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA). The microfluidic chip consisted of C18 (300 Å, 5 μm) enrichment (4 mm, 40 nL) and separation (43 mm × 75 μm) columns with a nanoelectrospray tip. LC separation was performed using a 60-min binary gradient at a flow rate of 0.3 μL/min. Solvent A consisted of 3% acetonitrile and 0.1% formic acid in nanopure water (v/v); solvent B consisted of 90% acetonitrile and 0.1% formic acid in nanopure water (v/v). The mass spectrometer was operated in the positive mode. Collision energies (V_collision) were calculated on the basis of m/z values using equation (1) for peptides and equation (2) for glycopeptides.

The glycan mapping was performed on an Agilent 1200 series HPLC-Chip system coupled to an Agilent 6520 Q-TOF (Agilent Technologies, Santa Clara, CA). Agilent Zorbax C18 stationary phase (300 Å, 5 μm) and porous graphitized carbon (250 Å, 5 μm) stationary phase were used to separate the trypsin digest and pronase digest, respectively. The microfluidic C18 chip consists of two columns: one for enrichment (4 mm, 40 nL) and one for separation (150 mm × 75 μm). The microfluidic PGC chip also consists of one enrichment column (4 mm, 40 nL) and one separation column (43 mm × 75 μm).
The quantitative analyses were performed on an Agilent 1290 infinity LC system coupled to an Agilent 6490 triple quadrupole (QqQ) mass spectrometer (Agilent Technologies, Santa Clara, CA). An Agilent Eclipse plus C18 (RRHD 1.8 μm, 2.1 × 100 mm) coupled with an Agilent Eclipse plus C18 pre-column (RRHD 1.8 μm, 2.1 × 5 mm) was used for UPLC separation.