Factors contributing to stability and instability in alpha-amylase activity in diluted saliva samples over time.

For the measurement of salivary alpha-amylase (sAA) activity, saliva samples first have to be diluted. There is some evidence for instability, that is, a decline of sAA activity in diluted samples. It is not clear which factors during dilution may contribute to this phenomenon and how quickly this decline of sAA activity occurs. Several experiments were conducted to investigate whether and how the material of the container (polystyrene (PS), polypropylene (PP), glass; experiment 1) and the diluent (saline (NaCl) solution, phosphate buffer saline (PBS), ultra-pure water; experiment 2) may affect sAA stability in diluted samples over a broad time window of up to 5 h. To study the velocity of the phenomenon in a fine-grained temporal resolution, sAA activity during the dilution process was studied (experiment 3). The results suggest that the (in)stability of sAA activity in diluted samples is determined by the interaction of material, diluent, and time. The sAA activity was relatively stable if saliva samples were diluted with a NaCl solution or PBS in glass tubes. However, sAA activity in diluted samples decreased in plastic containers (PS, PP), or if ultra-pure water was used as the diluent. There was a clear time effect on this decline. However, the decline appears to require some time to evolve and may not occur immediately during the dilution process. To conclude, the dilution of saliva samples should preferably be conducted with NaCl solution or PBS in glass containers. If glass containers are not available, PS and PP containers can be used if the dilution is processed quickly (within 25 min) and the measurement is initiated immediately upon dilution.