OBJECTIVE:To establish a method for the simultaneous determination of uracil,hypoxanthine,xanthine,uridine and adenosine in Eisenia foetida. METHODS:HPLC was performed on the column of Venusil XBP C18 with mobile phase of 50%methanol-0.01 mol/L potassium dihydrogen phosphate solution(gradient elution)at a flow rate of 1.0 ml/min. The detection wave-length was 254 nm with a column temperature at 30 ℃,and the injection volume was 10 μl. RESULTS:The linear range was 0.05-1.60 μg for uracil(r=0.9991),0.05-1.60 μg for hypoxanthine(r=0.9993),0.002-0.010 μg for xanthine(r=0.9991), 0.0075-0.24 μg for uridine(r=0.9999)and 0.0250-0.80 μg for adenosine(r=0.9990);RSDs of precision,stability and accuracy tests were lower than 3%;recoveries were 98.30%-100.76%(RSD=0.8%,n=9),98.68%-100.96%(RSD=0.8%,n=9), 95.16%-97.67%(RSD=0.9%,n=9),96.15%-99.57%(RSD=1.5%,n=9)and 96.39%-101.93%(RSD=1.8%,n=9),respective-ly. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determina-tion of uracil,hypoxanthine,xanthine,uridine and adenosine in E. foetida.

OBJECTIVETo develop an assay methodology for determination of lumbrukinase potency in Rongshuan capsules.

METHODThe agarose-fibrin plate assay methodology for determination of Lumbrukinase potency in Rongshuan capsules was studied including durability, specificity, linearity range, product's handling method, accuracy , repetitiveness, solution stability, recovery and statistical method. The method of parallel line assay based on quantitative responses in statistical methods for biological assays was used in the statistics of potency assay.

RESULTThe durability and specificity of assay accord with the requirement; The linearity range was 12.5 to approximately 400 U, the RSD of accuracy tests was 3.2%, the RSD of repetitiveness tests was 8.3%, the solution is stable under 4 degrees C for 72 hours, the recovery rate was 97.0% and the RSD of recovery assays was 16.5%.

CONCLUSIONThe agar-fibrin plate assay is rapidly, feasible, simple, convenient and accurate way for determining the Lumbrukinase potency. The method of parallel line assay based on quantitative responses in statistical methods for biological assays can control the error of determination.

Animals ; Biological Assay ; methods ; Capsules ; analysis ; Endopeptidases ; analysis ; Pharmaceutical Preparations ; analysis