Abstract: This paper deals with the HPLC determination of four constituents (uridine Ⅰ; uracil Ⅱ; adenosine Ⅲ, adenine Ⅳ) in the mycelia of Ganoderma capense by submerged cultivation. The sample was extracted with 50% aqueous ethyl alcohol by ultrasonic method, defatted with ether and then analysed by HPLC method.Inject.2μl of the sample solution onto an ODS column (25 cm × 4.0 mm,i d) Run the chromatogram with Michaelis phosphate buffer solution(pH 6.98)—tetrahydrofuran(100:1) as mobile phase using UV 260 nm detector. The four constituents were fairly well separated, p-Toluenesulfonic acid was chosen as the internal standard. Peak area was used for quantitative determination. Analytical results obtained by both HPLC and TLC-UV method are in good agreement.