RP-HPLC同时测定暗紫贝母中10个核苷及碱基类成分的含量

目的:建立RP-HPLC-DAD同时测定暗紫贝母药材中5个核苷类活性成分(尿苷、鸟苷、胸苷、腺苷、肌苷)及5个游离的碱基(腺嘌呤、胸腺嘧啶、鸟嘌呤、尿嘧啶、胞嘧啶)含量的方法。方法:采用Agilent Zorbax Bonus-RP C18色谱柱(4.6 mm×250 mm,5μm);流动相为甲醇和水,梯度洗脱,流速1 mL·min-1;检测波长260 nm;柱温30℃。结果:胞嘧啶、尿苷、鸟苷、胸苷、腺苷、腺嘌呤、肌苷、胸腺嘧啶、鸟嘌呤、尿嘧啶的质量浓度分别在1.78～56.80 mg·L-1、1.84～59.00 mg·L-1、1.58～50.60 mg·L-1、1.73～55.30 mg·L-1、1.78～57.10 mg·L-1、1.63～52.10 mg·L-1、1.79～57.20 mg·L-1、1.80～57.70 mg·L-1、1.63～52.30 mg·L-1、1.63～52.30 mg·L-1范围内与峰面积呈良好线性关系,平均回收率为97.7%～103.2%,RSD≤2.2%。结论:建立了RP-HPLC-DAD同时测定暗紫贝母药材中10个核苷及碱基类成分含量的方法,该方法简单,重复性好,准确可靠;暗紫贝母药材中鸟苷、尿苷、胸苷、腺苷含量较高;青海久治、大通产暗紫贝母药材中核苷含量均高于四川松潘;青海2个产地比较,大通产暗紫贝母药材中核苷含量又明显高于久治。

RP-HPLC simultaneous determination of ten nucleosides and nucleobases in Fritillaria unibracteata

Objective:To develope an HPLC coupled with a photodiode array detector(HPLC-DAD) method for simultaneous determination of 10 nucleosides and nucleobases in Fritillaria unibracteata.Method:The analyses were performed on an Agilent Zorbax-Bonus-RP C18 column(4.6 mm×250 mm,5 μm) eluted with water and methanol in gradient mode.The flow rate was 1.0 mL·min-1;detection wavelength was 260 nm;and column temperature was set at 30 ℃.Result:The investigated compounds including cytidine,uridine,guanosine,thymidine,adenosine,adenine,thymine,guanine,inosine,uracilwere shown good linearity(r≥0.9991) over the tested ranges.The average recoveries were within 97.7%-103.2% with RSD≤2.2%.Conclusion:The developed HPLC method is simple,sensitive and accurate with good repeatability.High content of Fritillaria unibracteata is guanosine,uridine,thymidine,adenosine,and Qinghai Fritillaria unibracteata nucleotide levels were higher than Sichuan Songpan,but compared the two origins of Qinghai,Qinghai Datong nucleoside content was significantly higher than Qinghai Jiuzhi.