Abstract: OBJECTIVE To establish a UHPLC-MS/MS method for simultaneous determination of the concentrations of sodium valproate, carbamazepine, lamotrigine and levetiracetam in human plasma and carry out clinical practice. METHODS After methanol precipitation of plasma samples, the Waters Acquity UPLC BEH C₁₈ column(100 mm×2.1 mm, 1.7 μm) was used, the mobile phase was 0.1% formic acid water acetonitrile with gradient elution, and 1 μL was injected. Select the positive and negative ion simultaneous monitoring mode for analysis, the MRM channels of carbamazepine, levetiracetam, lamotrigine, sodium valproate, and tinidazole(internal standard) were m/z 236.9→193.9, m/z 171.0→154.2, m/z 256.0→145.0, m/z 143.1→143.1, m/z 248.1→120.9, respectively. Declustering voltage and collision voltage were 70 V/21 V, 90 V/10 V, 90 V/23 V, -50 V/-10 V, 90 V/42 V, respectively. RESULTS In plasma samples, the metabolites of sodium valproate(linear range 20-150 µg·mL^-1), carbamazepine(linear range 2-15 µg·mL^-1), lamotrigine(linear range 0.5-15 µg·mL^-1), levetiracetam(linear range 2-60 µg·mL^-1) showed a good linear relationship under this method, r²>0.997, the precision and accuracy of each analyte in the plasma sample tested met the requirements(RSD<6.7%, −12.9%<RE<10.5%). A total of 77 patients with clinical epilepsy patients from January 2021 to June 2021 were collected for drug concentration detection and analysis. The concentration of sodium valproate, carbamazepine, lamotrigine and levetiracetam was treated. The treatment window pass rates were 69.57%, 70.00%, 87.50% and 36.11% respectively. CONCLUSION This method is suitable for clinical analysis of large sample sizes, and provides a convenient, reliable and scientific detection method for the detection of therapeutic drug concentration in the clinical medical technology departments.