Abstract Background Calculated LDL cholesterol (LDL-C) is a standard component of the lipid panel and an important risk factor in managing atherosclerotic cardiovascular disease (ASCVD). Due to the assumptions in the derivation of the Friedwald formula, fasting (≥8 hours) and triglyceride levels ≤400mg/dL have long been recommended. More recent data have shown that in most cases fasting has minimal effect on results, leading to the endorsement of non-fasting lipid panels for routine evaluation of ASCVD risk in clinical practice guidelines. Furthermore, newer equations (e.g. Sampson-NIH) allow for more accurate estimation of LDL-C even in the presence of hypertriglyceridemia. Mayo Clinic has implemented the Sampson-NIH LDL-C calculation and removed fasting/non-fasting designation from lipid panel orders. We hypothesized that the change to fasting designation would allow more afternoon draws for routine lipid panels and that increased non-fasting collections may increase triglyceride levels, but that the Sampson-NIH equation would prevent bias in LDL-C estimation. In order to test these hypotheses, we analyzed patient data from 1 year prior and 1 year post-implementation. Methods Results of lipid panels performed for one year preceding (n=334,719) and one year after (n=381,804) removal of the fasting/non-fasting designation and implementation of Sampson-NIH equation were collected. Median, 10th, and 90th percentile of draw time, triglyceride, and LDL-C calculated by either Sampson-NIH or Friedwald formulas were compared by quantile regression. Results Draw times shifted post-implementation at the 10th percentile (7:06 vs. 7:10, p<0.0001), median (8:49 vs. 9:08, p<0.0001) and 90th (12:02 vs. 13:46, p<0.0001). No difference in median triglyceride measurement was observed between pre-and post-implementation (110mg/dL), but slight differences were observed in the 10th and 90th percentile values (60 vs 59mg/dL and 224 vs 229mg/dL, p<0.0001). No difference was observed in 10th, 50th, or 90th percentile LDL values when calculated by Sampson-NIH equation (56, 99, 153mg/dL); however, if the Friedwald formula is used, a slight shift is observed in the 10th and 50th percentile LDL values (54 vs. 53, 97 vs. 96; p<0.0001) but not the 90th (150mg/dL). Conclusions Changing to the Sampson/NIH formula for LDL-C and removing fasting vs. non-fasting designation from routine lipid panels had minimal effect on reported triglyceride and LDL-C values, while allowing increased flexibility in draw time and reporting of LDL-C on patients with triglycerides ≥400mg/dL. Patients with TG ≥400mg/dL represented 1.7% of the dataset overall, including 5,387 patients before the change in which LDL-C could not be reported and 6,697 patients in the year post-implementation for whom LDL-C was reported based on the Sampson-NIH formula. Using an equation that is robust in patients with elevated triglyceride levels and removing fasting requirements from routine lipid panels allows greater flexibility and convenience for patients while allowing for more even distribution of phlebotomy workload and reporting of LDL-C results for patients with hypertriglyceridemia without evidence of clinically significant bias in reported results.