The loading capacity and strategy of layered double hydroxides (LDHs) determine their suitability for drug delivery applications. In this study, two sizes of MgAl‐CO3 LDHs are converted into MgAl‐NO3 LDHs and MgAl‐Cl LDHs by an acid‐salt method. The influence of the size and type of interlayer anions of LDHs on their loading capacity are studied and compared. Here, Methylene Orange (MO) served as the anionic structure to determine the loading capacity. Due to the strongest anion‐exchange capacity of NO3− and larger interlayer distance of MgAl‐NO3 LDHs, they have impressive high loading capacities, which make these useful as absorbents or drug carriers. Additionally, the pH‐dependent release behaviors of MO‐loaded MgAl‐NO3 LDHs are investigated, which dictates both the release profile and maximum release. Above all, based on the study, it is found that the release of LDHs with agents loaded under mildly acidic environments is not based on its partial dissolution, which is commonly indicated. This study provides a better understanding of the loading and release process of LDHs, which allows for better incorporation of LDHs in biomedical‐related applications.