Lipemia interferes with the measurement of analytes by spectrophotometry, the most common is lipemia postprandial. This study aimed to compare two methods of removing lipemia from the biochemical profiles of dogs. Sixteen dogs were used, and blood samples were collected at 2, 4, 6, and 12 hours after feeding (Experiment 1), and lipemic samples from 32 dogs with various clinical conditions (Experiment 2). In Experiment 1, each sample was processed without treatment (control), with treatments by a lipemia removal reagent (LRR) and high-speed centrifugation (HSC). In Experiment 2, control and LRR were evaluated. In Experiment 1, all lipid profile results in the control group remained within the reference range. In Experiment 2, hypertriglyceridemia accompanied by elevated very low-density lipoproteins (VLDL) levels was observed. Both methods effectively eliminated lipemia, with a greater reduction in triglyceride levels compared to cholesterol. In the LRR, the largest reduction in triglycerides occurred in the 12-hour samples (−76.24%), whereas in the HSC was observed in 4-hour (−33.5%). LRR treatment negatively interfered with gamma-glutamyl transferase (GGT) and creatinine measurements, for which HSC was the better option. Both techniques effectively prevented the negative effect of lipemia on AST measurement at 4 hours. For alkaline phosphatase - ALP (Experiment 2), the LRR values were higher than the control's, indicating a negative interference. We conclude that lipemic samples must be treated with lipemia removal techniques, using the most appropriate method based on the specific requirements of each analysis.