OBJECTIVE： To study the pharmacokinetics and brain targeting of Toutongning nasal spray in rats in vivo. METHODS： 84 SD rats were divided into nasal administration group and vein administration group， 42 in each group， with dose of 1.2 mL/kg. 5 mL sample blood was taken in abdominal aorta after 5， 10， 15， 30， 60， 90， 120 min of administration， and brain tissue was taken （6 rats in each time point）. HPLC-MS was adopted to determine the concentration of prim-o-glucosylcimifugin and 5-O-methylvisammioside in plasma and brain tissue of rats in each group， and DAS 2.0 software was used to calculate the pharmacokinetic parameters and brain targeting indexes. RESULTS： The cmax  of prim-o-glucosylcimifugin and 5-O-methylvisammioside in plasma of rats in nasal administration group were （0.202 4±0.015 8）， （0.373 8±0.085 7） μg/mL； tmax  were （10.000 0±0.000 0） min； and AUC0-∞ were （16.542 9±2.110 3）， （27.452 7±5.572 1） μg·h/mL， respectively. The cmax of prim-o-glucosylcimifugin and 5-O-methylvisammioside in brain tissue of rats were （0.180 2±0.038 4）， （0.320 4±0.027 7） μg/g； tmax  were （10.000 0±0.000 0） min； and AUC0-∞ were （17.105 3±2.432 9）， （24.541 6±3.753 4） μg·h/g， respectively. The cmax  of prim-o-glucosylcimifugin and 5-O-methylvisammioside in plasma of rats in vein administration group were （0.300 2±0.016 1）， （0.526 7±0.044 1）    μg/mL； tmax  were （10.000 0±0.000 0） min； and AUC0-∞ were （28.010 5±4.112 8）， （60.294 1±11.290 2） μg·h/mL， respectively. The cmax  of prim-o-glucosylcimifugin and 5-O-methylvisammioside in brain tissue of rats were （0.149 8±0.031 5）， （0.199 8±0.040 1） μg/g； tmax were （15.000 0±0.000 0） min； and AUC0-∞ were （22.643 4±2.883 1）， （36.721 8±14.885 6） μg·h/g， respectively. The brain targeting indexes of prim-o-glucosylcimifugin and 5-O-methylvisammioside were 2.387 0 and 2.176 1， respectively. CONCLUSIONS： After nasal administration of Toutongning nasal spray， parts of drugs can directly transport to the brian by nasal absorption. It is scientific and reasonable to make nasal spray.