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目的:建立高效液相色谱测定黑草药材中木犀草素和芹菜素的含量的方法。方法:样品用甲醇回流提取90 min,用Agilent Extend-C18(4.6 mm×250 mm,5μm)色谱柱分离,以乙腈-0.7%冰醋酸(30∶70)为流动相,流速1.0 mL.min-1,检测波长350 nm;柱温35℃,进样量10μL,以外标法定量。结果:木犀草素在1.126～112.6 mg.L-1呈良好的线性关系(r=0.999 7),平均回收率101.3%,RSD 1.2%(n=6);芹菜素在0.886～88.6 mg.L-1呈良好的线性关系(r=0.999 5),平均回收率99.4%,RSD 2.2%(n=6)。结论:该法操作简便,结果准确可靠,重复性及稳定性良好,可作为控制黑草药材质量的方法。
Objective: To establish a HPLC method for the determination of luteolin and apigenin in black herbs. Methods: The sample was extracted with methanol for 90 min and separated on an Agilent Extend-C18 (4.6 mm × 250 mm, 5 μm) column with acetonitrile-0.7% glacial acetic acid (30:70) as mobile phase at a flow rate of 1.0 mL · min- 1, detection wavelength 350 nm; column temperature 35 ℃, injection volume 10μL, external standard method. RESULTS: Luteolin had a good linearity (r = 0.999 7) between 1.126 and 112.6 mg.L-1, with an average recovery of 101.3% and a RSD of 1.2% (n = 6). Apigenin was in the range of 0.886 to 88.6 mg.L -1 (r = 0.999 5). The average recovery was 99.4% and RSD was 2.2% (n = 6). Conclusion: The method is simple, accurate, reliable, reproducible and stable, which can be used as a method to control the quality of black herbs.