Glycogen storage disease type II, also known as Pompe disease, is a disorder of glycogen metabolism resulting from defect in activity of the lysosomal acid hydrolase a-1,4 glycosidase in all tissues of affected individuals. The enzyme deficiency results in intralysosomal accumulation of glycogen in numerous tissues. The disorder is transmitted as an autosomal recessive trait. The enzyme deficiency results in two major presentations; disorder of infancy and disorder of adult-onset. The former is a disorder of infancy, dominated by massive cardiomegaly, macroglossia, progressive muscle weakness and marked hypotonia and the death occurs before the child reaches the second year of live. The second is an adult-onset disease, dominated by progressive proximal muscle weakness including major impairment of respiratory function. Death usually results from respiratory failure. The objective of this work is the measure the enzymatic activity of acid a 1- 4 glycosidase in dried blood spots samples on filter paper, in order to determine the reference curve of the population. In order to reach this objective, samples of health individuals from both genders were used. The methodology of this work involves the measurement of the acid a-1,4 glycosidase enzyme (AaG), inhibited acid a-1,4 glycosidase enzyme (AaGIA), neutral a-1,4 glycosidase enzyme (NaG), as well as blanks for each person. From these results it was calculated the relation between NaG and AaGIA and the percentage of inhibition (PI) of acid a 1- 4 glycosidase. Until now, we have evaluated 50 people. The mean result of NaG/AaGIA (N/A) was 11.99 SD 3.53 and mean PI 71.58% SD 6.37. The maximum and the minimum values were 20.09 and 5.64 for N/A and 86.62% and 59.47% for PI. The establishment of a standard curve for a -glycosidase activity for health subjects will make future diagnostics easier, faster and more accurate.