Aconiti Radix [Chuanwu (CW)] is widely used to treat chronic and intractable diseases due to its remarkable curative effect. CW has been combined with honey for thousands of years to reduce toxicity and enhance efficacy. This study first determined the compatibility mechanism of CW with honey using a comparative pharmacokinetic concept. We developed and validated a simple, sensitive, specific, and accurate UHPLC-MS/MS method to simultaneously determine five Aconitum alkaloids in rat plasma after the oral administration of CW decoction and CW-honey concentrated solution. Pharmacokinetic parameters were significantly different between the two groups (P < 0.01 and P < 0.05). Compared with the CW group, C(max) and AUC(0 --> t) decreased in the CW-honey group for three diester-diterpenoid alkaloids (hypaconitine, mesaconitine, and aconitine); T(max) and T(1/2) were prolonged. However, C(max) and AUC(0 --> t) increased in the CW-honey group for two monoester-diterpenoid alkaloids (benzoylaconine and benzoylmesaconine); T(max) was shortened, and T(1/2) was prolonged. These findings suggest that honey affected the pharmacokinetic behaviors of five Aconitum alkaloids. We speculate that the detoxification and synergism of honey might result from reducing the toxicity of diester-diterpenoid alkaloids and promoting the biological activity of monoester-diterpenoid alkaloids in vivo. This study provides a theoretical basis for the clinical use of CW combined with honey. CI - (c) 2022 John Wiley & Sons Ltd.