First measure the hematocrit, then measure glucose in plasma and in whole blood (hemolysed blood). Then you can calculate glucose in RBC. You shuld be able to derive the corresponding formula.

Sighart Golf

I agree with Walter Golf, but I want to add that hemolysis of whole blood will be achieved by freezing of blood 12 hours at -20 ° C or so to take the blood in the syringe and blood repeatedly pushed through the needle of small diameter. You must be sure that all erythrocytes were destroyed. Also, you have to aware that the freezing destroys other blood cells (leukocytes and platelets) too, and they also contain intracellular glucose.

Whole blood usually contains ca. 20% lower glucose concentration than plasma. This is mainly due to the volume from proteins and lipids contained in the whole blood sample, which is added to the cuvette. In forensic blood samples, for example for ethanol determination, a factor of 1,2 is usually used to correct for this analyte-free volume.

Since leucocyte number and thrombocyte volume and number are much lower than from red blood cells, their contribution to whole-blood glucose concentration is usually neglected.

However, if one has an experimental model of erythrocytes, in which all other variables (cell numer of leucocytes and thrombocytes and plasma proteins) remain unchanged, changes in erythrocyte glucose concentration will be correctly reflected by changes in whole blood glucose.

Sighart Walter Golf

Dear Walter Golf,

I want to thank you for the answer was because it helped me.

Jasna

you can centrifuge the blood sample collected on anticoagulant, wash cells with saline to remove all traces of plasma, measure the hematocrit of the suspension and measure the glucose after fully hemolyzed these cells as described by Jasna Lenicek Krieza. You shall know and the glucose concentration of blood cells in your sample after correcting the concentration measured by hematocrit of your suspension (which should be high). You must ensure that the technique of determination of glucose is not influenced by the presence hemoglobin and work in the presence of inhibitor of glycolysis. It is rather difficult

When considering the method from Nicole Gillain-Martin, you have to take into account, that there exists a considerable efflux of glucose from eryrhocytes, which might be as high as 1000 mmol glucose/liter cells and min together with Oxygen. while washing the redblood cells as described, As long as you are not sure, that this efflux decreases intracellular glucose in the red blood cells, you cannot use this method.

Sighart Golf

Dear Advisers (All above),

Actually, We thought to measure intracellular glucose in RBCs to understand that the transport of glucose through RBCs (either increased or decreased) in between non-diabetic and diabetic subjects after food intake. Now, I can understand that after glucose is transported into the erythrocyte, it is rapidly phosphorylated as glucose-6-phosphate. Even if the transport of glucose increased, the intracellular concentration of free glucose does not increase as glucose is taken up by the cell. Hence, what i have to do?. Suppose if i measure the glucose-6-phosphate within the RBCs of the those two conditions and can i claim that transport of glucose in increased through RBCs while intracelluar glucose-6-phosphate is increased.

For Sighart Golf: in fact, I have not thought about the efflux of glucose

but then how to determine RBC glucose?

centrifuge the blood, remove plasma, measure the hematocrit of the RBC suspension, lyse the RBC, mesure glucose on lysed RBC and correction of the value from hematocrit and glucose value in plasma (provided that one uses a technique which is not influenced by hemoglobin)

For Nicole Gillain-Martin!

How about that. Take EDTA-Blood, measure HK in whole blood as well as Glucose, the prepare Plasma and measure again Glucose and calculate Glucose in RBC as suggested. Here, you have an exact volume of RBC, sincde RBC is calculated from number and volume of RBC. Measuring of HK fom centrifuged RBC contains still a variable rest of Plasma depending on centrifugal force..

Sighart Golf