During my mouse harvests, I keep serum with intra-cardiac puncture. Then I send some serum samples to pathology for ALT, AST, Trig, and cholesterol results. Besides they also send me a Haem score.
If the IP during the harvest is not very good, my Haem score can be really high and I feel like it also affects on ALT levels. Do you have any idea about this? High score of Haem can give me false ALT results? Then what is the minimum Haem score for mice?
As I remember, ALT is specific only for hepatocytes therefore hemolysis cannot affect it. In contrast it can change AST because this enzyme is also found in erythrocytes but I don't think that this change will be drastic. If you are evaluating liver damage, usually both enzymes are measured simultaneously and ALT can be used as a control.
Yes, if the are grossly haemolysed (not sure what your scoring system is) but mild haemolysis seems OK. See http://www.biochemia-medica.com/content/effects-hemolysis-interference-routine-biochemistry-parameters
hemolysis should be avoided, but its influence is not very significant for ALT (unlike AST)
overflow error for hemogobine > 60 mg / dL (Roche) or > 110 mg / dL (G. Siest G and J. Henny)
Dear Faro,
I perform ALT tests frequently in the lab, and I can assure you that hemolysis interfere with ALT results.
Most methods for ALT dosage now use a kinetic reaction, specially in big labs. In this way, very hemolytic or icteric samples will damper the absorbance loss during ALT assay, giving you a false negative result (you see less ALT than really is).
However, In regular colorimetric assays you may see the opposite, more ALT than there really is in your sample.
The point is, you need to avoid hemolytic and icteric samples, and always dilute your samples if its possible.
Good luck
Dear researchers, thank you very much for your replies!!
I wanna explain what I did yesterday:
Normally I was diluting my serum samples 1/2 with 0.9% NaCl solution and yea, sometimes Haem score was really high as well as ALT level. When I make 1 in 2 dilution, I multiply my ALT levels with 2.
Yesterday in stead 1/2, I diluted my samples 1/5. So Haem score came way smaller. And this time I multiplied my ALT levels with 5. However, it looks like ALT levels do not change with high or low Haem score.
Don't know what to think now.
Dear Colleagues,
I am about to carry out AST and ALT test on my serum samples. I am very grateful for all your contributions.However, I want to know if ddH2O can be used as the diluent instead of NaCl solution as explained by Fahrettin?
Cheers!
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