© 2022, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.Objective: The purpose of this study was to examine the effect of applying two different prophylaxis pastes with various protocols on the degree of bleaching and color stability in the office bleaching process with a bleaching agent containing 40% hydrogen peroxide. Material and method: The 300 teeth of 15 (9 female, 6 male) patients were included in our study, and 5 different study groups were randomly formed. The bleaching process with Ultradent Opalescence Boost (Ultradent, South Jordan, USA) was performed according to the manufacturer’s instructions. Two different prophylaxis pastes, Ultrapro (Ultradent, Cologne, Germany) and Cleanic (Kerr, Rastatt, Germany), were applied with protocols determined according to the groups. The effect on color stability at 6-, 12-, and 24-month follow-up was determined using the CIE L* a* b* color system. The changes in the parameters were analyzed using the one-way ANOVA, chi-square, and Tukey test (p > 0.05). Results: The evaluation rate of the treatments was 100% at the end of 24 months. Control group 6-month ΔE (0–6) and 12-month ΔE (0–12) values were higher than 24-month ΔE (0–12) values. UU + UU, KC + KC, and UU group 6-month ΔE (0–6) values were higher than 12-month ΔE (0–12) values. In addition, 6-month ΔE (0–6) and 12-month ΔE (0–12) values were higher than 24-month ΔE (0–24) values. KC group 6-month ΔE (0–6) and 12-month ΔE (0–12) values were higher than 24-month ΔE (0–24) values. Control, UU + UU, KC + KC, UU, and KC group ΔE (1–6) values at 6 months were lower than 12-month ΔE (1–12) and 24-month ΔE (1–12) values in all groups. In addition, 12-month ΔE (1–12) values were lower than 24-month ΔE (1–24) values. According to the study data, there was no statistically significant difference between the groups according to the prophylaxis paste and prophylaxis procedure (p > 0.05). However, it was observed that whiteness decreased in all groups over time. Conclusions: Discoloration was observed at 6 months, 12 months, and 24 months after bleaching. However, at the end of 24 months, the tooth color was significantly whiter than the initial color. Clinical relevance: Pre-bleaching prophylaxis application and pre-bleaching and post-bleaching prophylaxis application have no effect on color stability.