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目的:建立同时测定赤子爱胜蚓药材中尿嘧啶、次黄嘌呤、黄嘌呤、尿苷、腺苷含量的方法。方法:采用高效液相色谱法。色谱柱为Venusil XBP C_(18),流动相为50%甲醇-0.01 mol/L磷酸二氢钾溶液(梯度洗脱),流速为1.0 ml/min,检测波长为254 nm,柱温为30℃,进样量为10μl。结果:尿嘧啶、次黄嘌呤、黄嘌呤、尿苷、腺苷检测进样量线性范围分别为0.05~1.60μg(r=0.999 1)、0.05~1.60μg(r=0.999 3)、0.002~0.010μg(r=0.999 1)、0.007 5~0.24μg(r=0.999 9)、0.025 0~0.80μg(r=0.999 0);精密度、稳定性、重复性试验的RSD<3%;加样回收率分别为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)、96.39%~101.93%(RSD=1.8%,n=9)。结论:该方法操作简便、稳定、重复性好,可用于赤子爱胜蚓药材中尿嘧啶、次黄嘌呤、黄嘌呤、尿苷、腺苷含量的同时测定。
Objective: To establish a method for the simultaneous determination of uracil, hypoxanthine, xanthine, uridine and adenosine in Eisenia foetida. Methods: Using high performance liquid chromatography. The chromatographic column was Venusil XBP C 18 with a mobile phase of 50% methanol-0.01 mol / L potassium dihydrogen phosphate (gradient elution) at a flow rate of 1.0 ml / min with a detection wavelength of 254 nm and a column temperature of 30 ° C , Injection volume of 10μl. Results: The linear ranges of uracil, hypoxanthine, xanthine, uridine and adenosine were 0.05-1.60μg (r = 0.999 1), 0.05-1.60μg (r = 0.999 3), 0.002-0.010 (r = 0.999 1), 0.007 5 ~ 0.24μg (r = 0.999 9) and 0.025 0 ~ 0.80μg (r = 0.999 0). The RSD of precision, stability and reproducibility was less than 3% Rates were 98.30% to 100.76% (RSD = 0.8%, n = 9), 98.68% to 100.96% (RSD = 0.8%, n = 9), 95.16% to 97.67% , 96.15% ~ 99.57% (RSD = 1.5%, n = 9), 96.39% ~ 101.93% (RSD = 1.8%, n = 9). Conclusion: The method is simple, stable and reproducible. It can be used for the simultaneous determination of uracil, hypoxanthine, xanthine, uridine and adenosine in Eisenia foetida.