The objective of this study is to develop and validate a method of reversed phase high performance liquid chromatography (RP-HPLC) assay with ultraviolet (UV) detector after solid phase extraction (SPE) for simultaneous measure tenofovir and emtricitabine in Chinese population. Detection of analytes was performed with a Phecda C-18 (250 x 4.6 mm, 5 mu m), reversed-phase analytical column and a security guard column C-18 (4 x 3.0 mm, 5 mu m). An obviously extrusive separation was successively followed with the SPE method using BOND ELUT-C-18 Varian columns and a 30 min gradient elution consisting of potassium phosphate monobasic buffer with 0.08% of triethanolamine (pH 3.52) and methanol with UV detector at 270 nm. The results of this study showed that the method used in this study was simple, accurate and sensitive with a wide linear range from 10 to 5000 ng/ml for both tenofovir and emtricitabine when 300 mu l aliquots was analyzed. The intra- and inter-day precision and accuracy for both analytes were lower than +/- 20% at the limit of quantification (LOQ) and +/- 15% at the other quality control (QC) levels. The absolute recoveries for tenofovir and emtricitabine were 80.6 and 86.5%, respectively. This method is suitable for routinely monitor plasma concentrations of tenofovir and emtricitabine in Chinese population.