Assignment:
Conduct HPLC analysis of myoglobin and hemoglobin using a C18 and C4 column. Both proteins were in a buffer solution only. On the C4 column the order of the peaks was a reverse of the C18 column.
The chromatographic output showed two peaks. Looking at the UV absorbance, I can attribute one of the peaks to the heme group (400 nm) and the other protein (280 nm), but I have no clue why this would have happened and can find nothing in the literature to further verify that this happens regularly or why it happens.
Provide complete and step by step solution for the question and show calculations and use formulas.