Abstract: Objective Tostudy ocular pharmacokinetic of Ray cartilage glycosaminoglycans eye drops (RCG) and determine the RCG concentration by UV-visible spectroscopy in rabbit eyes cornea, iris, vitreous and aqueous humor. Methods Twenty rabbits were rand omly dividedinto10 groups. The cornea, iris, vitreous and aqueous humor was collectedat 10 min, 20 min, 30 min, 60 min, 90 min, 120 min, 180 min, 240 min, 300 min, 360 min for quantification analysis of drugs after application of RCG eye drops in the rabbit eyes. The concentration of RCG was detectedusing UV-visible spectrophotometry and the parameters were calculated. Results The linear regression equation of Glucuronide in cornea, iris, vitreous and aqueous humor were Y= 0.029 1 x + 0.349 3, r² = 0.999 5; Y= 0.036x + 0.223 71, r² = 0.999 2; Y= 0.033 x + 0.284 4, r² = 0.999 1; Y= 0.039 9 x+ 0.255 1, r² =0.999 1 (x: glucuronide concentrations, Y: light absorption value), respectively. The glucuronide hadgoodlinearity with the 1.5-30 μg/ml, 0.5-20 μg/ml, 1.0-5.0 μg/ml, 0.5- 1.5 μg/ml concentration range in cornea, iris, vitreous and aqueous humor, respectively. Peak concentration of the cornea, iris, vitreous and aqueous humor were 29.03±0.38 μg/L, 16.83±0.12 μg/L, 9.62±0.41 μg/L, 1.51±0.43 μg/L, respectively with the peak time of 9.81±0.51 min, 58.53±0.54 min, 60.37±0.19 min, 61.78±0.52 min, respectively. Conclusion RCG has the maximumdrug concentration at about 10 min after administration in the rabbit cornea, and it reaches the maximumconcentration at about 60 min after administration in the aqueous humor.