HPLC法同时测定人工蛹虫草不同部位中5种核苷类成分的含量

摘要

目的:建立同时测定人工蛹虫草不同部位中5种核苷类成分含量的方法.方法:采用高效液相色谱法.色谱柱为Inert-Sustain C18,流动相为甲醇-0.02 mol/L磷酸二氢钾溶液(梯度洗脱),流速为0.6 mL/min,检测波长为254 nm,柱温为30℃.结果:尿苷、肌苷、鸟苷、腺苷和虫草素检测进样量线性范围分别为0.568~3.408μg(r=0.9999)、0.284~1.704μg(r=0.9999)、0.264~1.584μg(r=0.9999)、0.232~1.392μg(r=0.9999)、1.672~10.032μg(r=0.9998);精密度、稳定性、重复性试验的RSD<3.0%;加样回收率分别为98.2%~103.9%(RSD=1.97%,n=9)、96.2%~101.6%(RSD=1.76%,n=9)、96.7%~102.0%(RSD=1.94%,n=9)、95.1%~99.4%(RSD=1.43%,n=9)和95.6%~101.3%(RSD=1.82%,n=9).结论:该方法操作简便,精密度、稳定性、重复性好,可用于人工蛹虫草中5种核苷类成分含量的同时测定,蛹虫草不同部位中所含核苷类成分含量不同.%OBJECTIVE:To establish a method for simultaneous determination of 5 nucleoside from different parts of cultured Cordyceps militaris. METHODS:HPLC method was adopted. The determination was performed on InertSustain C18 column with mobile phase consisting of methanol-0.02 mol/L monobasic potassium phosphate solution (gradient elution) at the flow rate of 0.6 mL/min. The detection wavelength was set at 254 nm,the column temperature was 30 ℃. RESULTS:The linear ranges of uridine, inosine,guanosine,adenosine and cordycepin were 0.568-3.408 μg(r=0.9999),0.284-1.704 μg(r=0.9999),0.264-1.584 μg(r=0.9999),0.232-1.392 μg(r=0.9999)and 1.672-10.032 μg(r=0.9998),respectively. RSDs of precision,stability and repeatabili-ty tests were all lower than 3.0%. The recoveries were 98.2%-103.9%(RSD=1.97%,n=9),96.2%-101.6%(RSD=1.76%,n=9),96.7%-102.0%(RSD=1.94%,n=9),95.1%-99.4%(RSD=1.43%,n=9) and 95.6%-101.3%(RSD=1.82%,n=9). CON-CLUSIONS:The method is simple,precise,stable and repeatable,and can be used for simultaneous determination of 5 nucleoside from cultured C. militaris,the content of nucleosides are different in different parts of C. militaris.