HPLC同时测定掌叶半夏中四种含氮化合物的含量

Simultanenous determination of contents of four nitrogenous compounds in Pinellia peda-tisecta by HPLC

目的：用高效液相色谱法同时测定掌叶半夏中葫芦巴碱、胸腺嘧啶、腺嘌呤和6-氧嘌呤4种含氮化合物的含量。方法：采用Diamonsil-C18色谱柱（4.6 mm×150 mm,5μm）;乙腈-纯水-三乙胺梯度洗脱,流速1.0 mL.min-1;柱温25℃;检测波长254 nm。结果：葫芦巴碱在0.21.0μg（r=0.999 6）,胸腺嘧啶在2.613μg（r=0.999 8）,腺嘌呤在0.552.75μg（r=0.999 9）,6-氧嘌呤在0.653.25μg（r=0.999 8）线性关系良好;葫芦巴碱平均回收率100.6%,RSD 2.4%（n=6）;胸腺嘧啶平均回收率99.89%,RSD 0.20%（n=6）;腺嘌呤平均回收率100.53%,RSD 1.1%（n=6）;6-氧嘌呤平均回收率100.0%,RSD1.4%（n=6）。结论：3批样品测定结果表明,该方法简便、准确,可用于掌叶半夏中4种含氮化合物的含量测定。

Objective： To develop a HPLC method for the quantitative determination of four nitrogenous compounds in Pinellia peda-tisecta.Method： The determination was carried out on Diamonsil-C18 column（4.6 mm×250 mm,5μm） eluted with acetonitrile-water-triethylamine in gradient elution acetonitrile-water-triethylamine.The mobile phase was a flow rate of 1.0 mL·min-1 and detection wavelength as 254 nm.Result： The linear ranges of trigonelline,thymine,amidopurine and hypoxanthine were 0.21.0 μg（r=0.999 6）,2.6-13 μg（r=0.999 8）,0.55-2.75 μg（r=0.999 9）,0.65-3.25 μg（r=0.999 8）,respectively.The average recoveries of trigonelline,thymine,amidopurine and hypoxanthine were 100.6%,99.89%,100.5% and 100.0% respectively,and the RSD of the above four compounds were 2.4%,0.20%,1.1%,1.4%,respectively.Conclusion： This method is reliable,accurate and specific.It can be used for quality control of the tube of P.peda-tisecta.