HPLC法同时测定连钱草中3个黄酮类成分和4个酚酸类成分的含量

Simultaneous determination of three flavonoids and four phenolic acids in Glechomae Herba by HPLC

目的:建立同时测定连钱草中3个黄酮类成分(木犀草素-7-O-β-D-葡萄糖醛酸(1→2)-O-β-D-葡萄糖醛酸苷、芹菜素-7-O-β-D-葡萄糖醛酸(1→2)-O-β-D-葡萄糖醛酸苷、木犀草素)及4个酚酸类成分(绿原酸、隐绿原酸、咖啡酸、迷迭香酸)含量的高效液相色谱法。方法:采用Waters SunFire C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱,流速1.0 mL·min^(-1),检测波长330 nm,柱温30℃。结果:7个成分在各自范围内线性关系良好(r≥0.9996);精密度(RSD≤0.87%)、稳定性(RSD≤1.1%)良好;平均加样回收率(n=6)为99.1%~101.1%,RSD为0.59%~1.7%。18批样品中绿原酸、隐绿原酸、咖啡酸、木犀草素-7-O-β-D-葡萄糖醛酸(1→2)-O-β-D-葡萄糖醛酸苷、芹菜素-7-O-β-D-葡萄糖醛酸(1→2)-O-β-D-葡萄糖醛酸苷、迷迭香酸和木犀草素含量范围分别为0.046~0.602、0.071~0.817、0.102~1.159、0.728~4.817、0.592~4.081、0.850~13.060和0.019~0.198 mg·g^(-1)。结论:该方法简便易行,重复性好,可用于连钱草药材的质量控制。

Objective:To develop an HPLC method for simultaneous determination of three flavonoids(luteolin7-O-β-glucuronyl-(1→2)-O-β-glucuronide,apigenin 7-O-β-glucuronyl-(1→2)-O-β-glucuronide and luteolin)and four phenolic acids(chlorogenic acid,cryptochlorogenic acid,caffeic acid and rosmarinic acid)in Glechomae Herba.Methods:The Waters SunFire C_(18)column(250 mm×4.6 mm,5μm)was used,the mobile phase was acetonitrile(A)-0.1% phosphoric acid(B)with gradient elution,at the flow rate of 1.0 mL·min^(-1),the detection wavelength was 330 nm,and the column temperature was 30℃.Results:Seven constituents showed good linear relationships within their own ranges(r≥0.9996),and the precision(RSD≤0.87%)and stability(RSD≤1.1%)were good.The average recoveries(n=6)were 99.1%-101.1% with the RSDs of 0.59%-1.7%.The contents of chlorogenic acid,cryptochlorogenic acid,caffeic acid,luteolin 7-O-β-glucuronyl-(1→2)-O-β-glucuronide,apigenin 7-O-β-glucuronyl-(1→2)-O-β-glucuronide,rosmarinic acid and luteolin in eighteen batches of samples were 0.046-0.602 mg·g^(-1),0.071-0.817 mg·g^(-1),0.102-1.159 mg·g^(-1),0.728-4.817 mg·g^(-1),0.592-4.081 mg·g^(-1),0.850-13.060 mg·g^(-1)and 0.019-0.198 mg·g^(-1),respectively.Conclusion:This method is specific,accurate and reproducible,and can be used to control the quality of the Glechomae Herba.