Stanton D. answered • 02/23/22
Tutor to Pique Your Sciences Interest
Hi Amritpal K.,
I hope you realize that these two questions are unrelated. If you know differently, I would be extremely interested to know how that works.
re/ (1), how many different times did you run HPLC on your sample? Your question seems to imply that you did injections several times, rather than that several different peaks eluted of different hemoglobins at different times in the same chromatogram?
You, and other species as well, may have more than one specific primary sequence for hemoglobin molecules, which may vary with developmental age. But it gets much more complicated, since 4 hemoglobin chains customarily aggregate (this is quarternary structure). So even between only 2 different primary chain types, you could have 5 assortments, assuming no special steric stacking occurs. Also, any of those chains could be oxygenated, or not. (since oxygenation is allosteric, you could probably throw it one way or the other by ambient O2 saturation level). You have some serious sorting, if you leave those chains aggregated!
For (2), "treatment method" refers to soaking up the oil into the fine structure of the diatom shell. It's a structural thing, not a chemical attraction, I think. (As far as I know, most of the diatoms are SiO2 based). Look up some photomicrographs of diatoms and describe what you see.
--Cheers, --Mr. d.
Stanton D.
On rereading "how are", the different hemoglobin chains have different physical properties, with slightly differing attractions to the stationary phase in the HPLC column. So they tend to spend differing amounts of time attached to the stationary phase (as opposed to moving in the bulk mobile phase, where they all move at the same speed), and therefore elute at different times.02/23/22