UPLC_法同时测定冬凌草中5种有效成分的含量

目的: 建立超高效液相色谱法同时测定冬凌草药材中冬凌草甲素、冬凌草乙素、迷迭香酸、齐墩果酸和熊果酸5个主要有效成分的含量。 方法: 采用WATERS UPLC色谱系统,ACQUITY BEH Shield RP₁₈ 色谱柱(2.1 mm×100 mm,1.7 μm);流动相为0.1%甲酸甲醇溶液(A)-0.1%甲酸水溶液 (B),梯度洗脱(0~9 min,70%A→57%A;9~10 min,57%A→48%A;10~11 min,48%A→48%A;11~14 min,48%A→14%A;14~22 min,14%A→14%A),流速0.2 mL·min^-1 ;检测波长为250 nm (0~12.40 min测定冬凌草甲素、冬凌草乙素和迷迭香酸)和210 nm(12.41~22.00 min测定齐墩果酸和熊果酸);柱温23 ℃。 结果: 本方法可在22 min内完成5个成分的色谱分析,且各成分色谱峰之间具有良好的分离度;冬凌草甲素、冬凌草乙素、迷迭香酸、齐墩果酸和熊果酸的线性范围分别为0.110~0.990 μg(r=0.9997),0.012~0.108 μg(r=0.9991),0.030~0.270 μg(r=0.9993),0.011~0.099 μg(r=0.9991),0.011~0.099 μg(r=0.9996);平均加样回收率(n=9)分别为100.9%,95.4%,102.1%,97.1%,99.0%。 结论: 所建立的超高效液相色谱法简便快速,结果准确可靠,重复性好,可用于冬凌草药材的质量控制与评价。

Objective: To establish a UPLC method for simultaneous determination of five effective constituents(oridonin,ponicidin,rosmarinic acid,oleanolic acid and ursolic acid) in different parts of Rabdosia rubescens from various sources. Methods: Five effective constituents were simultaneously determined by UPLC with ACQUITY BEH Shield RP₁₈ column(2.1 mm 100 mm,1.7 μm) by gradient elution(0-9 min,70%A→57%A;9-10 min,57%A→48%A;10-11 min,48%A→48%A;11-14 min,48%A→14%A;14-22 min,14%A→14%A) using 0.1% methanoic acid in methanol(A) and 0.1% methanoic acid(B) as the mobile phase at the flow rate of 0.2 mL·min^-1;The detection wavelength was 250 nm(0-12.40 min) for oridonin,ponicidin,rosmarinic acid,and 210 nm(12.41-22 min) for oleanolic acid and ursolic acid with column temperature at 23 ℃. Results: Five effective constituents were separated clearly and respectively in 22 min,the linear ranges of oridonin,ponicidin,rosmarinic acid,oleanolic acid and ursolic acid was 0.110-0.990 μg(r=0.9997),0.012-0.108 μg(r=0.9991),0.030-0.270 μg(r=0.9993),0.011-0.099 μg(r=0.9991),0.011-0.099 μg(r=0.9996),respectively;The average recoveries(n=9) were 100.9%,95.4%,102.1%,97.1%,99.0%,respectively. Conclusion: The method is simple,accurate and can be used for quality control and evaluation of Rabdosia rubescens.