OBJECTIVE:To establish a method for the determination of the content and molar ratio of Ornithine aspartate for in-jection. METHODS:HPLC was performed on the column of Thermo HYPERSIL Aps-2 amino column with mobile phase of aceto-nitrile-0.05mol/L Potassium dihydrogen phosphate solution,the flow rate was 0.9 ml/min,the column temperature was 30℃,the de-tection wavelength was 200 nm,and the injection volume of 20 μl. The results determined by HPLC and potentiometric titration were compared. RESULTS:The linear range of ornithine aspartate was 0.02-10.01 mg/ml(r=0.999 9);RSDs of precision,stabili-ty and reproducibility tests were lower than 2%;recovery was 99.15%-100.15%(RSD=0.35%,n=9). The content of 3 batches of Ornithine aspartate for injection was 100.04%-100.64% and molar ratio was 0.982-0.989. The content is similar to the results de-termined by potentiometric titration with national standards. CONCLUSIONS:The method is simple,accurate,specific and sensi-tive,and suitable for the determination of the content and molar ratio of Ornithine aspartate for injection.

OBJECTIVE:To establish a method for the content determination of carbohydrates in Osseous and cucumis extract injection. METHODS:Ion chromatography was conducted on the column of Dionex CarboPac PA20 with mobile phase of water-1 mol/L Sodium acetate solution-250 mmol/L Sodium hydroxide solution(gradient elution)at flow rate of 0.45 ml/min,column tem-perature was 35 ℃,and injection volume was 10 μl,detector was electrochemical analysis detector. RESULTS:The linear ranges of galactose,glucose,mannose,sucrose and fructose were 0.5-10.0μg/ml(r>0.998 0);RSDs of precision,stability and reproduc-ibility tests were lower than 2%;recovery was 94.8%-99.1%(RSD=0.52%-0.89%,n=9). CONCLUSIONS:The method is sim-ple,fast and accurate,and can be used for the determination of carbohydrates in Osseous and cucumis extract injection.

BACKGROUND: How to establish a stable in vitro culture system, including location of corneal limbal epithelial stem cells, in vitro sample harvest, in vitro culture, vector selection, as well as identification methods, play a key role in corneal limbal epithelial stem cells culture. OBJECTIVE: To culture the isolated rabbit corneal limbal epithelial stem cells and to identify the biological properties of cultured cells. METHODS: The primary rabbit cornel limbal epithelial stem cells were isolated and cultured with tissue inoculation using human amniotic membrane as vector. The growth features of cells were observed under an inverted microscope. The morphology of cells was observed by hematoxylin-eosin staining and a scanning electron microscope. Furthermore, the monoclonal antibody AE5 and P63 two-step immunohistochemical staining were used to identify limbal epithelial stem cell protein expression. RESULTS AND CONCLUSION: The rabbit corneal limbal epithelial stem cells could be successfully cultured and maintained a relatively high value-added potential in vitro. Rabbit corneal limbal epithelial stem cells cultured on the amniotic membrane pull netted cellular layer. The AE5 monoclonal antibody positive rate of primary cultured cells was about 5% and P63 monoclonal antibody positive up to 90%. AE5-positive rate increased and P63-positive rate decreased with the increase in the number of subculture. The rabbit limbal epithelial stem cells can be successful culture and amplified on human amniotic membrane in vitro by limbal tissue culture method. The cultured cells maintain the characteristics of corneal epithelial cells. The rabbit corneal limbal epithelial stem cells can form grafts on the amniotic membrane.