摘要:Abstract In this study determination of a compound commonly known as acetaminophen in the leaf extract of Aquilaria malaccencis was studied and discussed. Aquilaria species are well known for its valuable agarwood. Acetaminophen or 4’-hydroxyacetanilide in its synthesis name is the main compound found in synthetic drugs. This synthetic drug is referred in treating fever, migraine and other type of illness. Traditionally, it is known that Aquilaria species able to treat various illnesses, including fever in which the leaves is treated as tea and believed to cure many other illnesses. A simple and validated method was developed for the study of 4’-hydroxyacetanilide by High Performance Liquid (HPLC). This analysis is well conducted with C18 column to separate the compounds. The method is validated with different parameters in terms of linearity, range, accuracy and robustness. The mobile phase used in this study is acetonitrile and water (20:80). The pH was adjusted to 3.5 with the aid of orthophosphoric acid. The flow rate was set to 0.8 ml/min and a spectrophotometric condition at 193 NM. The percentage of recovery from this method is within 98% to 101%, which indicates this method is reliable and can be applied to an analysis of 4’-hydroxyacetanilide from Aquilaria malaccencis. The concentration of 4’-hydroxyacetanilide determined in the leaves extract of Aquilaria malaccencis at 60oC were 0.807μg/ml, 1.398μg/ml and 1.811μg/ml for varied ratio (raw materials: water) 1:80, 1:40 and 1:27 respectively.