Background: We assessed cumulative antiretroviral exposure—using tenofovir diphosphate (TFV‐DP) in dried blood spots (DBS)—in persons with HIV (PWH) receiving tenofovir disoproxil fumarate (TDF)‐based antiretroviral therapy (ART) as single‐tablet regimens (STR) or multiple‐tablet regimens (MTR). Methods: Blood for DBS was prospectively collected in PWH on TDF during 1144 person visits (n = 523). Linear mixed‐effects models, adjusted for baseline characteristics, were used to compare TFV‐DP in STR versus MTR. Models adjusted for ART regimen using either anchor drug class, pharmacokinetic booster status (unboosted [u/] or boosted [b/]), or a combined STR/MTR and booster categorical variable. Results: In the anchor class‐adjusted model, STR had 19% (95% confidence interval [CI]: 3%–37%; p = 0.02) higher TFV‐DP concentrations than MTR. However, in the booster‐adjusted model, STR was not significantly higher than MTR (estimate 5%, 95% CI: −9% to 21%; p = 0.48), although PWH on b/ART had 35% (95% CI: 16%–58%; p = 0.0001) higher TFV‐DP than u/ART. In the STR/MTR‐boosted variable model, when compared to u/MTR, b/STR, b/MTR, and u/STR had 25% (95% CI: 7%–47%; p = 0.005), 37% (95% CI: 17%–59%; p < 0.0001), and 7% (95% CI: −7% to 24%; p = 0.34) higher TFV‐DP, respectively. Compared with b/MTR, b/STR had 9% (95% CI: −31% to 10%; p = 0.37) lower TFV‐DP. In a sensitivity analysis of PWH with HIV viral load <20 copies/ml at all visits, b/STR and b/MTR had 34% (95% CI: 16%–55%; p < 0.0001) and 12% (95% CI: −2% to 27%; p = 0.09) higher TFV‐DP, respectively, compared with u/MTR, while u/STR had 4% (95% CI: −15% to 8%; p = 0.50) lower TFV‐DP. Compared with b/MTR, b/STR had 17% (95% CI: 2%–30%; p = 0.03) higher TFV‐DP. Conclusions: Persons with HIV on b/TDF‐based ART had higher TFV‐DP than u/ART, regardless of STR or MTR use. No significant differences in TFV‐DP between regimens of the same boosting status (i.e., b/STR vs. b/MTR; u/STR vs. u/MTR) were observed in the full cohort. Future research should examine the clinical utility of these findings in patient‐tailored ART selection. [ABSTRACT FROM AUTHOR]