Abstract Disclosure: K. Kucharczyk: None. A. Drynda: None. J. Podlewski: None. M. Trofimiuk-Muldner: None. A. Hubalewska-Dydejczyk: None. Introduction. Primary hyperparathyroidism (PHPT) is a prevalent endocrinopathy typically identified through biochemical testing. PHPT is characterized by hypercalcemia accompanied by increased or inappropriately normal plasma parathyroid hormone (PTH) concentrations. Preoperative imaging is conducted when surgery is indicated to pinpoint parathyroid adenomas. [99mTc]Tc-Sestamibi scintigraphy is highly specific and sensitive in diagnosing PHPT, with a higher adenoma detection rate demonstrated for patients with larger lesions or higher blood calcium concentrations. The study aimed to investigate the relationship between the results of biochemical tests and clinical manifestations (PHPT “phenotype”) and the rate of positive results of parathyroid [99mTc]Tc-Sestamibi scintigraphy. Materials and methods. This was a retrospective study of patients suspected of PHPT who underwent parathyroid [99mTc]Tc-Sestamibi SPECT/CT scintigraphy between 2010 and 2022. Patients’ cluster analysis was conducted based on PTH, serum calcium (Ca), and phosphate (Pi) concentrations to find distinct patterns in these parameters. Furthermore, we compared the distributions of additional patients’ attributes, including gender, lesion size, serum vitamin D, ALP concentrations, urinary calcium, urolithiasis prevalence, as well as parathyroid scintigraphy results. Results. 513 patients were divided into 5 clusters using different algorithms. Finally, clusters from K-means methods were selected based on statistical metrics and expert interpretation. Those were organized in ascending order of positive scintigraphy results, with Cluster 5 reaching a 79.5% positivity rate and median values of PTH - 315.5 pmol/L, Ca - 2.96 mmol/L, and Pi - 0.7 mmol/L. As the proportion of positive imaging outcomes declined, median PTH and calcium concentrations decreased while phosphate concentrations increased. In Cluster 1, with 30% of positive imaging results, medians were as follows: PTH - 79.06 pmol/L, Ca - 2.55 mmol/L and Pi - 1.1 mmol/L. A comparative analysis of various clinical and biochemical parameters revealed statistically significant differences between clusters. Patients from Clusters with a higher frequency of positive scintigraphy were significantly more likely to be males, vitamin D deficient, with higher ALP concentrations, diagnosed with urolithiasis, and harbouring larger lesions. Conclusions. PHPT “phenotypes” correlated with disease severity are evidently linked to parathyroid scintigraphy detection rate. Therefore, it is crucial to know that false-negative imaging is more likely in PHPT patients with minor deviations in laboratory tests. Judicious assessment of biochemical and clinical features may limit the number of redundant parathyroid scintigraphies and, in consequence, reduce healthcare costs and unnecessary radiation exposure. Presentation: 6/2/2024