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目的:采用HPLC双波长法测定补骨脂-肉豆蔻药对不同比例配伍前后水提液中有效成分的含量变化,优选出最佳配伍比例。方法:采用Agilent Eclipse Plus C18(4.6 mm×250 mm,5μm),以乙腈(A)-水(B)为流动相,梯度洗脱;双波长检测(λ_1=254 nm,λ_2=360 nm);流速1.0mL·min~(-1);柱温30℃;进样10μL。结果:补骨脂素、异补骨脂素、补骨脂乙素、去氢二异丁香酚、补骨脂酚的进样量与峰面积分别在0.080~0.798μg(r=0.999 8)、0.063~0.626μg(r=0.999 8)、0.014~0.054μg(r=0.999 8)、0.012~0.050μg(r=0.999 5)、0.239~1.431μg(r=0.999 8)内呈良好的线性关系。加样回收率96.95%~102.29%,优选出最佳配伍比例为补骨脂、肉豆蔻2∶1。结论:所建立的HPLC双波长法操作简便,结果准确,重复性好,回收率高,优选出发挥最佳作用的配伍比例为2∶1,为其质量评价提供研究基础。
OBJECTIVE: To determine the optimal composition of psoralea-nutmeg medicine by HPLC with dual-wavelength method for the content changes of active ingredients in aqueous extracts before and after compatibility. Methods: The gradient elution was carried out with Agilent Eclipse Plus C18 (4.6 mm × 250 mm, 5 μm) with gradient elution with acetonitrile (A) - water (B); dual wavelength detection (λ_1 = 254 nm, λ_2 = 360 nm) The flow rate was 1.0mL · min ~ (-1); the column temperature was 30 ℃; 10μL was injected. Results: The injection volume and peak area of psoralen, isopsoralen, psoralen, dehydrogenated diisonol and bakuchiol were 0.080 ~ 0.798μg (r = 0.999 8), 0.063 ~ 0.626μg (r = 0.999 8), 0.014 ~ 0.054μg (r = 0.999 8), 0.012 ~ 0.050μg (r = 0.999 5) and 0.239 ~ 1.431μg (r = 0.999 8). The recoveries of samples ranged from 96.95% to 102.29%. The optimal ratio of pectin and nutmeg was 2: 1. Conclusion: The established HPLC dual wavelength method is simple, accurate, reproducible and has high recovery rate. The best ratio of 2:1 is the best, which provides the basis for its quality evaluation.