Abstract: A method for determination of pro-xylane in anti-wrinkle cosmetics by high performance liquid chromatography with water condensation particle laser counting detector was proposed. 0.25 g of anti-wrinkle cosmetics was taken and placed in a 50 mL-centrifuge tube. 3 mL of methanol was added, and the mixture was vortexed for 1 min. The solution was made its volume up to 25 mL with water, and ultrasound for 15 min. The mixed solution was shaken well and passed through a 0.45 μm filter membrane. The filtrate was analyzed by the instrument. CAPCELL PAK ADME-HR S5 chromatographic column was used as the stationary phase and the mixed solution composed of 0.1% (volume fraction) formic acid solution and acetonitrile at different volume ratios was used as the mobile phase for gradient elution. The solution was determined by water condensation particle laser counting detector. As shown by the results, linear relationship between the corresponding the sum of peak areas of diastereomers and mass concentration of pro-xylane was kept in the range of 10-1 000 mg·L^-1, with detection limits (3S/N) of 0.013% (mass fraction). RSDs (n=6) of the determined values of test of sample for precision was less than 2.0%. Test for recovery was made according to the standard addition method, giving results in the range of 99.5%-100%. This method was used to determine the pro-xylane in actual anti-wrinkle cosmetics, with detection amounts in the range of 2.33%-9.89% (mass fraction).