Objective To determine the parathyroid hormone (PTH) content in ultrasound-guided fine-needle aspiration sample of normal parathyroid based on ultrasound characteristics previously screened out, so as to verify the reliability of the ultrasound characteristics of normal parathyroid. Methods Between Oct. 2012 and Jan. 2013, 10 healthy volunteers were enrolled in the present study after obtaining written informed consent and approval by the ethics committee of our institution. Ultrasound-guided percutaneous aspiration from both the ultrasonic parathyroid area and well-recognized neck lymph node was conducted by using 22G fine needles. The aspirates were taken for cytology purpose and the washing of the left-over in the canula was taken for PTH evaluation by electrochemiluminescence. Simultaneously samples of venous blood were obtained also for PTH evaluation as controls. Results The aspiration procedures were successfully for all the 10 subjects, and cytological examination all found cells, with no intraoperative or and postoperative complications. PTH level in the washing of left-over extracted from the parathyroid aspirates averaged (287.12±35.41) pg/mL and ranged 232.53-357.38 pg/mL, and that from the lymph node averaged (16.03±8.47) pg/mL and ranged 8.67-34.28 pg/mL; while the serum PTH level averaged (37.44±7.26) pg/mL and ranged 32.64-47.85 pg/mL. Three of the ten aspirates of parathyroid did not have enough cells to make predication, and the other seven aspirates had enough cells, with four likely from parathyroid and three from indefinite origins. All the ten aspirates from the lymph node areas were confirmed to have definite lymphocytes. Conclusion PTH evaluation results by fine-needle washing after ultrasound-guided aspiration are highly consistent with the ultrasonic parathyroids and lymph nodes, which preliminarily proves the reliability of the previously identified ultrasound characteristics of normal parathyroids.