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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,1.7μm);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(0～12.40min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0.110～0.990μg(r=0.9997),0.012～0.108μg(r=0.9991),0.030～0.270μg(r=0.9993),0.011～0.099μg(r=0.9991),0.011～0.099μg(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 method for simultaneous determination of 5 major active constituents of Rubescens Alabara, Rubescens B, Rosmarinic acid, oleanolic acid and ursolic acid in Rubescens by using ultra performance liquid chromatography. METHODS: The mobile phase consisted of 0.1% formic acid in methanol (A) - 0.1% formic acid in water (B) and gradient elution (0) using a WATERS UPLC system with ACQUITY BEH Shield RP18 (2.1 mm × 100 mm, ~ 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) at a flow rate of 0.2 mL · min-1; detection wavelength was 250 nm (0 ~ 12.40 min) ) And 210 nm (oleanolic acid and ursolic acid measured at 12.41-22.00 min); column temperature 23 ℃. Results: The method can complete the chromatographic analysis of five components within 22 min, and the chromatographic peaks of each component have good resolution. Oridonin, Rubescensine, rosmarinic acid, oleanolic acid (R = 0.9997), 0.030 ~ 0.270μg (r = 0.9993), 0.011 ~ 0.099μg (r = 0.9991), 0.011 ~ 0.099μg ~ 0.099μg (r = 0.9996). The average recoveries (n = 9) were 100.9%, 95.4%, 102.1%, 97.1% and 99.0%, respectively. Conclusion: The established ultra-high performance liquid chromatography is simple, rapid, accurate and reliable, reproducible, and can be used for the quality control and evaluation of Rubescens medicinal herbs.