I am developing an HPLC assay for daptomycin using gradient elution. My analytic conditions are as follows: Waters Atlantis T3 C18, 5 microns, 4.6x 150 mm, mobile phase starts with 30% acetonitrile for 4 minutes, gradually increases to 45% ACN in 15 minutes returns to 30% in 2 minutes and then stabilizes for additional 2 minutes. The aqueous counterpart is 40mM of KH2PO4 pH 5.5. lamdamax is 223. flow rate 1 ml/min.
Daptomycin elutes around 14 minutes but has a weird shape. I am skeptical of impurities or enantiomers of daptomycin. Is there a way to attach a snapshot of the chromatogram?
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