peak separation for chilli

Jul 25, 2017
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good day, i have problem in separating the peaks from 2 minutes to 6 minutes. There are at least 9 peaks that are combined with each other. The condition of hplc are as follows :

Isocratic.

Mobile phase : (A) Acetonitrile - 41% (b) 0.5% formic acid (59%)

Flow : 1 min/ml ; Injection : 20 microlit; Temperature : 30 degrees

Retention time : 40 mins

Solvent extracts : methanol

Column : C18 (250mm x 4.0mm I.D., 5 micronmeter diameter)

I have run the standard and it came out nicely.

Please help...