藏药麻花秦艽花和线叶龙胆花中10种核苷类成分的含量分析

Determination of 10 Nucleosides in Tibetan Medicine of Flower of Gentiana Straminea and Flower of Gentiana Farreri

关键词:  麻花秦艽花  线叶龙胆花  核苷和碱基  含量测定  高效液相色谱法

Key words:  flower of Gentiana straminea  flower of Gentiana farreri  nucleosides and bases  content determination  HPLC

摘要:

目的 建立藏药麻花秦艽花和线叶龙胆花中同时测定胞嘧啶、胞苷、尿嘧啶、腺嘌呤、鸟嘌呤、尿苷、胸腺嘧啶、鸟苷、胸苷和腺苷10种核苷类成分含量的HPLC-DAD方法，并对产自青海的6批次麻花秦艽花和6批次线叶龙胆花的这10种核苷成分进行含量测定和比较分析。方法 Bonus RP色谱柱（250 mm×4.6 mm，5 μm），甲醇-0.04%乙酸水溶液梯度洗脱，流速1.0 mL·min^-1，柱温35℃，检测波长260 nm，进样量20 μL。结果 10种核苷类成分线性关系、精密度、稳定性、重复性均符合含量测定方法学的要求，加样回收率为95.1%~98.5%。不同产地线叶龙胆花中10种核苷类成分总含量849.38~1 014.37μg·g^-1，麻花秦艽花为256.27~822.52μg·g^-1。腺苷、鸟苷、尿苷和胞苷在麻花秦艽花和线叶龙胆花中的含量均较高，是其主要核苷成分，而尿嘧啶、鸟嘌呤和胸苷在2种样品中含量均较低。结论 该法简便实用、重复性好，适用于藏药麻花秦艽花和线叶龙胆花中10种核苷类成分的测定。青海不同产地2种藏药中10种核苷类成分的分析数据可为其质量控制和品质评价提供参考。

Abstract:

OBJECTIVE To establish an HPLC-DAD method for simultaneously determination of cytosine, cytidine, uracil, adenine, guanine, uridine, thymine, guanosine, thymidine and adenosine in Tibetan medicine of flower of Gentiana stramiinea and flower of Gentiana farreri. Determination and comparison the ten nucleoside compounds of the two Tibetan medicine samples which come from six different origins of Qinghai province. METHODS A Bonus RP column(250 mm×4.6 mm, 5 μm) was used, methanol and 0.04% acetic acid solution was used as the mobile phase, the flow rate was 1.0 mL·min^-1, the column temperature was 35℃ and the detector wavelength was set at 260 nm with injection volumn of 20 mL. RESULTS Ten nucleoside compounds had good linear relationship, precision, stability, and repeatability according to the requirements of the quantitative analysis methodology. The recoveries of ten nucleoside compounds were 95.1%-98.5%. Furthermore, the total contents of the ten nucleoside compounds in different origins were 849.38-1 014.37 μg·g^-1 and 256.27-822.52 μg·g^-1 for the flower of Gentiana farreri and Gentiana straminea respectively. Adenosine, guanosine, uridine and cytidine were the main nucleoside compounds in the two Tibetan medicine samples which were higher than others in content, while uracil, guanine and thymidine were lower. CONCLUSION The developed HPLC-DAD method is simple, available, and has good repeatability, which is suitable for the determination of above ten nucleoside compounds of flower of Gentiana straminea and Gentiana farreri. Moreover, the quantitative data of ten nucleoside compounds in the two Tibetan medicine samples origin from Qinghai province can provide helpful information for the quality control and evaluation.