Purpose To evaluate the serum level of zonulin, which is an intestinal permeability (IP) biomarker, in primary hyperparathyroidism (PHPT) and to investigate the relationship between zonulin, calcium, and parathormone (PTH) levels. Methods The study included 34 healthy control (HC) and 39 patients with PHPT. Serum calcium, phosphorus, magnesium, creatinine, albumin, and 24 h urine calcium levels were measured in all groups. Serum levels of zonulin were measured quantitatively by enzyme-linked immunosorbent assay (ELISA). Urinary ultrasonography (to assess the presence of nephrolithiasis) and dual energy X-ray absorptiometry (to assess the presence of osteoporosis) were used to evaluate complications related to PTHP. Results Serum zonulin levels were significantly higher in the PHPT group than the HC group (p < 0.001). Zonulin levels were significantly positively correlated with plasma PTH and serum calcium levels (r = 0.600,p < 0.001 andr = 0.610,p < 0.001; respectively). There was no correlation between serum zonulin levels and adenoma volume. Conclusion Serum zonulin level increases in patients with PHPT. Serum zonulin levels show a moderate/strong positive correlation with serum calcium and plasma PTH levels. This suggests that IP increase may play a role in the development of hypercalcemia in patients with PHPT.