Hello all, I need to separate blood plasma from 400 micro-liter of rat blood for HPLC analysis. I need to get at least 200 micro-liter to extract the drug. I tried centrifugation but I could not separate more than 200 micros. Is there any special methods to separate plasma from small volumes with maximum yield?
I need to quantify the plasma drug concentration after each h for 12 hours.
Dear Robin,
The normal hematocrit level in the rat`s blood is 36-48 % and plasma volume is 64 - 52 %. Thus, 200 - 250 micro-liter is the maximal volume of plasma in your blood sample (400 micro-liter).
Gregory Barshtein and Carles Monterrubio.
If I can separate 200 microliter it is ok for me. But I could only collect something like less than 150 microliter. After centrifugation, I could not completely collect the plasma since the blood is mixing with plasma.
You can provide separation by two steps
First - centrifugation of the blood sample. Second - transferring of upper portion of blood column (plasma with upper layer of RBCs, approximately 0.300 ml) to small eppendorf (0.5 ml) for additional centrifugation.
Good Luck
- Badges
- Science topic
- Similar topics
- Computer Communications (Networks)
- Simulators