HPLC

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NNGG
NNGG's picture
HPLC

Hello ,

I am using the column 2.6 micron column , mobile phase is water: methanol.
trying to analyze vitamin. But with each chromatogram by area fluctuates .

Please suggest.
Flow rate 0.5ml /min, shimadzu, injection volume 40microliters, detector 550nm.temperature 50degree celsius.

Thank you

Dr. Analytical
Dr. Analytical's picture
Which vitamin are you trying

Which vitamin are you trying to analyze?  Vitamin A?

What are the column dimensions - diameter X length.  What is the column packed with?  How much methanol in water?  What is the retention time of the vitamin?

A 40 uL injection is very large.  Try a smaller volume.