HPLC lab separation of peptides

HPLC-UV-Vis detector.
at RP-HPLC mode
eluent: 20mM TFA in water mixed with methanol 30:70, filtered and degassed before used.
Column: Kromasil 100-3.5 C18, dimension length 150mm x inner diameter 4.5,  equilibration in 15 min, flow rate 1mL/min,
set base line at 280nm or 214 nm
observe the system pressure, avoid excedding 20MPa
Note: system pressure, injection volume, column type and dimension,3500psi varied around
requirement for capacity factor optimally between 2--5.
HILIC mode
Eluent: 20mM ammonium formate and 10mM formic acide in water mixed 30:70 acetonitrile.
Column: HILIC dimension: 100x4.6, 200A, 5um.
Back pressure 19-20 bar.
pH = 4.5
flowrate = 0.5 mL/min
pressure = 19-20 Bar
the signal 
50ppm toluen is injected to determine t0 values.
50ppm sample was prepared for analysis