Levetiracetam is one of the new generation anti-epileptic agents (also known as anticonvulsants or antiseizure drugs). Following its approval for marketing in 2000, levetiracetam has been widely used in the treatment of epilepsy due to its broad spectrum effects. One of the advantages of this antiseizure drug is its rapid and complete absorption after oral administration. It has also minimal drug-drug and food interactions, and shows more than 95% bioavailability. The determination of levetiracetam in various samples is carried out using several analytical methods including HPLC-based methods. HPLC-based methods are used for different pharmaceutical analyses and play an important role in drug monitoring during patient follow-ups. This review provides a summary of the HPLC-based methods used in the determination and quantification of levetiracetam in biological fluids and pharmaceutical preparations.