Objective This study delves into the pharmacokinetic intricacies of ANM33-loaded liposomal microbubbles within the physiology of SD rats. Methods ANM33 takes the center stage in our investigation. We employ UV spectrophotometry to scrutinize the drug's blood concentrations at discrete intervals post intravenous administration in rats. Subsequently, we meticulously calculate the pertinent pharmacokinetic parameters. Results Our diligent efforts yield a well-fitted UV spectral analysis technique apt for discerning ANM33. The critical pharmacokinetic parameters reveal a t1/2 of (4.70 ± 0.96) h, CL of (0.11 ± 0.02) L/h, AUC(0-6) of (37.11 ± 2.14) μg/L*h, AUC(0-∞)) of (56.01 ± 4.25) μg/L*h, and a Cmax of (7.92 ± 1.95) μg/L. Conclusion The ultraviolet spectrophotometry method offers a straightforward, precise, and highly stable analytical approach, making it a suitable choice for conducting pharmacokinetic investigations of the formulation. In vivo pharmacokinetic studies of dual-targeted liposomal microbubbles carrying anti-miR-33 in rats demonstrate a fundamental alignment with a two-compartment model. These studies indicate a rapid time to reach peak concentration, high peak concentrations, a swift onset of action, and a high level of biocompatibility.