5ppb of phenobarbital have been spiked into blank blood. This have been replicated by 10 times. However, ELISA results show that all 10 of the replicates contains LESS than 5ppb (average of 1.198ppb). Note that the blood samples have been centrifuged before being placed in ELISA. Is it possible that some of the phenobarbital binded with the proteins present in blood?
50ppb or phenobarbital have been spiked into blank urine and replicated by 10 times. Results show that all 10 of the replicates contains MORE than 50ppb (average of 61.18ppb).
*From your experience, should the quantitative result of ELISA be taken into account although ELISA was meant to be a screening procedure which provides QUALITATIVE results?
Enzyme-linked Immunosorbent Assay (ELISA) : BIO-RAD PhD FlexTek2 System.
ELISA kit: Barbiturates
It is not clear that estimation of phenobarbital is done in whole blood or serum or plasma .
After spiking phenobarbital or any other compound generally get bind to serum or plasma proteins. To get released bound phenobarbital do the estimation at low pH (4 to 5). Urine pH is low that is why you have got equal or more amount of spiked one when estimated
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