I Tried HPLC of the sample with the following set of parameters set:

Instrument: LC AD 20

Flow rate: 1ml/min

Temp: 30 degree

Sample Injection: 30 microliter

Protein analyte conc: 5mcg/ml, 10mcg/ml, 20mcg/ml, 30mcg/ml.

Time for running: 15min

Pressure: 4293 Psi

Solvent used: 30% Acetonytrile(ACN:D.H2O)

Prior to the use I washed the column with solvent for 1 hour and found that the peaks were not clear and same upon repetetion. Can anyone suggest how to perform HPLC for a short aminoacid sequence(11 aminoacids) structure.

Thankyou

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