Zanamivir is in a class of medications called neuraminidase inhibitors. It works by stopping the growth and spread of the flu virus in your body. Zanamivir helps shorten the time you have flu symptoms such as nasal congestion, sore throat, cough, muscle aches, tiredness, weakness, headache, fever, and chills. To establish a simple, sensitive and completely validated HPLC-MS-MS approach for the analysis of Zanamivir in Human plasma samples. The method was developed after protein precipitation by the Mobile phase was a mixture of MeOH: Ammonium Formate 30:70 v/v. was selected and directed at an isocratic flow stream of 1.0 ml/min with 10μl of sample injection volume. The retention times of Zanamivir and Remdesivir were 2.600 min and 3.956 min, correspondingly, with 6 min run time. The approach developed showed fine calibration curve in the quantity range of 4-80 ng/ml with correlation coefficient (r2) of ≥ 0.9994 and The intra-day and inter-day accuracy was ranged from 98.50% to 102.88%. The intra-day and inter-day precision was ranged from 0.81 to 1.98 % according to FDA guidelines.