Parathyroid hormone (PTH) concentrations are routinely measured in the diagnosis and management of bone and kidney diseases, but reference ranges can be overestimated if determined in otherwise healthy individuals for whom vitamin D deficiency was not evaluated. We establish PTH reference ranges in apparently healthy, normocalcemic, normophosphatemic individuals categorized by 25-hydroxyvitamin D (25(OH)D) status using the Elecsys® PTH (cobas e 601) and Elecsys® Vitamin D total II electrochemiluminescence immunoassays (cobas e 411). This prospective, non-interventional study measured PTH in serum from 653 apparently healthy adults [56.7% female; 68.2% white/Caucasian; 28.6% African American; median age 44 years (range 21-83)] from three diverse geographic sites across the USA during summer and winter months. Subjects were classified by concomitant vitamin D sufficiency (≥ 30 ng/mL), insufficiency (> 20 to < 30 ng/mL) or deficiency (≤ 20 ng/mL). In vitamin D sufficiency, median PTH was 31.9 pg/mL [range (2.5th-97.5th percentile) 17.9-58.6] compared with 35.5 pg/mL (17.0-60.4) for insufficiency, and 39.8 pg/mL (19.5-86.4) for deficiency. A significant inverse relationship was found between PTH and 25(OH)D (P < 0.001). After accounting for vitamin D, potential effects of race or season as covariates were relatively small or absent. Upper reference limits (URL) for PTH in vitamin D sufficiency/insufficiency were similar and lower than current values. Clinically important PTH elevations were observed in vitamin D deficiency, where revised reference ranges with a higher URL may be appropriate. These data may help to distinguish vitamin D-related PTH elevations from other causes [e.g., primary (normocalcemic) or secondary hyperparathyroidism].