HPLC法同时测定补益中药中尿苷、腺嘌呤、鸟苷和腺苷的含量

Simultaneous HPLC determination of uridine,adenine,guanosine,and adenosine in six tonic traditional Chinese medicines

目的:运用高效液相色谱法分析3个核苷类即尿苷、鸟苷、腺苷以及腺嘌呤在6种常用补益中药人参、黄芪、熟地黄、当归、麦冬和肉苁蓉中的含量。方法:采用Zorbax SB-Aq(250 mm×4.6 mm,5μm);以0.05%甲酸水溶液-乙腈为流动相梯度洗脱,流速0.5 mL.min-1;检测波长260 nm;柱温30℃。结果:尿苷、腺嘌呤、鸟苷和腺苷分别在0.30～77.32μg.mL-1(r=1.000)、0.23～58.29μg.mL-1(r=0.9998)、0.35～90.73μg.mL-1(r=1.000)和0.61～157.07μg.mL-1(r=1.000)范围内线性关系良好,加样回收率(n=3)分别为98.3%(RSD=0.6%)、103.4%(RSD=1.7%)、97.9%(RSD=0.8%)和97.2%(RSD=1.8%);人参、黄芪、熟地黄、当归、麦冬和肉苁蓉均含有核苷类成分,其中人参、黄芪和当归同时含有尿苷、腺嘌呤、鸟苷和腺苷,并且含量较高。结论:核苷类成分可能是补益中药的一类重要活性成分。

Objective： To analyze nucleosides（uridine,guanosine,and adenosine）and nucleobase（adenine） in 6 tonic traditional Chinese medicines（Radix Ginseng,Radix Astragali,Radix Rehmanniae Preparata,Radix Angelicae Sinensis,Radix Ophiopogonis,and Herba Cistanches） using HPLC-DAD.Methods： The HPLC analysis was performed on a Zorbax SB-Aq（250 mm×4.6 mm,5 μm） column using gradient elution with mobile phase of 0.05% formic acid and acetonitrile.The assay was carried out at a flow rate of 0.5 mL·min-1 at 30 ℃ with the diode-array detection at 260 nm.Results：Uridine,adenine,guanosine,and adenosine showed good linearity in the ranges of 0.30-77.32 μg·mL-1（r=1.000）,0.23-58.29 μg·mL-1（r=0.9998）,0.35-90.73 μg·mL-1（r=1.000）,and 0.61-157.07 μg·mL-1（r=1.000）,respectively;The average recoveries（n=3）were 98.3%（RSD=0.6%）,103.4%（RSD=1.7%）,97.9%（RSD=0.8%）,and 97.2%（RSD=1.8%）.These compounds could be found in 6 tonic traditional Chinese medicines,but their contents were high in Radix Ginseng,Radix Astragali and Radix Angelicae Sinensis.Conclusions：The nucleosides may be bioactive compounds in tonic traditional Chinese medicines.