Analysis of Exenatide Variants

Columns: 2 of PolySULFOETHYL A 300Å, 3µm, 100~4.6mm connected in series
Sample: Exenatide (refer to the above sequence)
Eluent A: 0.1M KH2PO4, pH 3.3 (adjusted after filtration and before addition of ACN) containing 43.6% ACN
Eluent B: Same thing but with 0.23M NaClO4 (adjust pH to 3.3 after dissolving the NaClO4 and KH2PO4 but before the addition of the ACN)
Gradient: 0% B in 0-5 minutes
0-60% B in 5-45 minutes
60-100% B in 45-50 minutes
100% B in 50-52 minutes
100-0% B in 52-54 minutes
Flow rate: 1.2ml / min
Temp.: 40ºC
Detector: UV214nm