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Purpose The aim of this study was to examine the utility of fluoroscopic bony landmarks in predicting the location of the left adrenal vein during adrenal vein sampling (AVS). Methods Eighty-six AVS procedures were performed in 81 patients between August 2013 and March 2020. A selectivity index was calculated for each case by dividing the measured left adrenal vein cortisol level by the peripheral vein cortisol level. Successful “target” left adrenal vein catheterization was confirmed in cases with a selectivity index of three or greater. Intraprocedural AVS fluoroscopic images were selected that demonstrated catheter position in the left adrenal vein. Lateral distance from the catheter tip in the left adrenal vein to the lateral margin of the left pedicle at the associated vertebral body level was measured. Results Mean patient age was 56.4 years (range: 19–80 years) and 48 (59.3%) patients were male. Target sampling in the left adrenal vein was confirmed in 82 (95.3%) cases. In 78 (95.1%) targeted cases, the catheter terminated less than 25 mm from the left lateral pedicle at a mean distance of 11.2 mm. The catheter was most frequently placed at the T12 and L1 vertebral body levels. Four (4.7%) cases demonstrated nontarget catheter positioning, two (50.0%) of these cases were within 25 mm. Conclusion The position of the left adrenal vein is generally located in a predictable position relative to bony landmarks. By utilizing these landmarks, positioning of the sampling catheter during AVS can be more reliable with the potential to avoid repeat procedures and delays in patient care. [ABSTRACT FROM AUTHOR]