Introduction: One of the main ingredients of Anar-5 tablets is Piper longium L. Piperine alkaloids are the main active ingredients of the Piper longum and have anti-inflammatory, antioxidant and gastric protection properties.In the framework of the standardization study of Anar-5 tablets, a method was developed to determine the content of piperine in highly perpormance liquid chromatography, and then it was sought to include it in the method of analysis of Anar-5 drugs. Goal: Quantitative determination of piperine in Anar-5 tablets and validate the method Material and Methods: The research was conducted in the Chemistry and Chemical Technology Laboratory of the Research Center of the Institute of Traditional Medicine and Technology. And Anar-5 tablets (serial number 04012020) that are produced for experimental were used in the research. The standard substance, piperine alkaloids, was purchased from Green Chemistry.Purification of HPLC (organic solvent methanol, 99.9%, distilled water) was used. The EX 1600 HP/ PUMP high-performance liquid chromatography instrument (column Arcus EP+-C18, 5µm, 4.6x250 mm) and the organic solvent filter 0.45 μm syringe filter were used. The methodology related to this research was discussed and approved at the online meeting of the Ethics Committee of the Academy of Sciences on January 26, 2021. SPSS 16 software was used to statistically program the survey results. Results : According to the above method, the retention time of the standard piperine is 10.38± 0.02 minutes, and the retention time of the piperine in Anar-5 tablets is 10.42±0.033 minutes. Relative velocity deviation RSD 1.077%, accuracy 0.65446±0.0068mg, stability 0.61298±0.013mg, capture time 10.42±0.033 minutes, relative standard deviation RSD≤2%, specificity 10.35 minutes, The equation of a line constructed with a standard curve is y=43360x-33587 and the correlation coefficient R2=0.9989. The piperine content of Anar-5 tablets was determined to be 0.61298±0.013 mg. The LOD and LOQ for piperine were in 2.268 μg/ml and 6.873 μg/ml, respectively. Conclusion The content of piperine in Anar-5 tablets can be determined by the HPLC method, and the appropriate conditions for this method have been established. The HPLC method is unique, accurate, linear, and stable, and meets ICH Q2 (R1) guideline criteria.