I had someone recently ask me about validated methods to measure HbA1c in rats, and because I once had the same question and could not find any such methods at the time, I followed the suggestion of a coworker and used the HbA1c. Now meters, which to my mind worked quite well, but the question sparked my curiosity to know what others have done. HbA1c in ZDF rats correlated well with increased 3 month average BG and went from undetectable (13% at 17 weeks of age, in diabetic animals.
Hi Pavan, Have you used HPLC to determine HbA1c in rats? How much blood would one need? I always send human samples out to a lab, so I do not have experience doing my own measurements via HPLC. If you have a protocol to share using HPLC to measure HbA1c in rats, I'd love to see it and to know what validation you have done. Thank you for your input.
Nikola: I would imagine that you would want to run a standard, but that is the tricky thing. In humans there are several Hb variants that can cause erroneously higher or lower HbA1c levels. I have no idea if this is the case in rats as well. I hope that my rats, being an inbred strain at least have the same Hb variant (if this is a consideration at all in rodents), such that the method I used was reasonable for my purposes. Any suggestions, Pavan?
Oh well thats intresting...I guess that inbred rats have same Hb variant...not sure. I was just curious when I asked how is done...tnx for answers any way
I m sorry for replying late as I was out of station for last 10 daysYes we do need a standard before every run as far as quantity of blood is concerned it is 20 ul whole blood required. Even the same can be done using photometric assays the results are comparable but the only drawback is that it does not give the graphic representation I am using both the methods. Since your querry is about HbA1C levels in rodents the standard needs to be used before every batch is run. Photometric method is fast and relaible but gold standard is chromatography
@Pavan Malhotra Sir,
Can u please share good paper and/or protocol for the same....
LPLC (Low pressure Liquid Ion Chromatography) is the varient of HPLC, its accurate and precise, less expensive, need just 5 ul sample whole blood
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