*2.4. Diastase Analysis*

The diastatic activity was determined by the Phadebas method in which the α-amylase activity is expressed as the diastase number, and is reported in Schade units [40]. One Schade unit corresponds to the enzyme activity contained in 1 g of honey, which can hydrolyze 0.01 g of starch in 1 hour at 40 ◦C. The procedure was as follows: first, 1 g of the analyzed honey was weighed, transferred to a 100 mL volumetric flask, and filled up to its volume with 0.1 M acetate buffer at pH = 5.2. Then, 5 milliliters of the sample was transferred to the test tube, placed in a water bath at 40 ◦C, and after 15 min a Phadebas tablet was added to the solution. The solution was mixed and heated again in a water bath for 30 min. After this time, 1 mL of 0.5 M sodium hydroxide solution was added to complete the enzymatic reaction. Next, the solution was filtered through a filter paper (ϕ = 70 mm) and the absorbance at 620 nm was measured using a Specord 200 spectrophotometer (Analytic Jena, Jena, Germany) as it is in proportion to the enzyme activity in the analyzed honey sample. The measurements were collected in triplicate and the final result is their average.
