HPLC测定半枝莲中野黄芩苷、黄芩素、木犀草素和芹菜素的含量

Determination of scutellarin,baicalein,luteolin and apigenin in Herba Scutellariae barbatae by HPLC

建立半枝莲药材中野黄芩苷、黄芩素、木犀草素和芹菜素含量同时测定的HPLC方法。色谱条件为DiamonsilC18柱(250 mm×4.6 mm,5μm)色谱柱分离,检测波长为335 nm,流动相为乙腈(A)-1%冰醋酸水溶液(B)进行梯度洗脱,流速为1.0 m L/min,柱温为30℃。野黄芩苷、黄芩素、木犀草素和芹菜素浓度分别在2.0~200、0.5~50、0.5~50、0.5~50μg/m L范围内与峰面积积分值线性关系良好。在精密度实验、稳定性实验、重复性实验中,野黄芩苷、黄芩素、木犀草素和芹菜素峰面积的相对标准偏差(RSD)<2.0%;野黄芩苷、黄芩素、木犀草素和芹菜素的加样回收率范围及平均值和RSD分别为97.73%~101.7%、99.72%、1.36%(n=9),97.78%~102.8%、99.53%、1.99%(n=9),97.40%~102.4%、99.39%、1.84%(n=9)和97.80%~102.0%、99.24%、1.49%(n=9)。所测12份不同产地半枝莲药材中,野黄芩苷、黄芩素、木犀草素和芹菜素的含量范围分别为2.41~7.08、0.26~0.56、0.31~0.90、0.28~0.95 mg/g。本方法具有良好的专属性和重现性,加样回收率实验符合要求,能准确、稳定地对半枝莲药材中野黄芩苷、黄芩素、木犀草素和芹菜素含量进行同时定量测定。

To establish HPLC methods for simultaneous determination of scutellarin, baicalein,luteolin and apigenin in Herba Scutellariae barbatae.The chromatographic conditions were as follows- A Diamonsil- C is （ 250 mm × 4.6 mm, 5 μm） was choosed as the chromatographic column, the mobile phase was the mixture of acetonitril （A） and 1% glacial acetic acid in water （B） with gradient elution, the detection wavelength at 335 nm,the flow velocity of 1.0 mL/min, the column temperature at 30 ℃.The linear relationship between content and peak area was obtained during the range of 2.0-200 μg/mL for scntellarin ,0.5-50 μg/mL for baicalein ,0.5-50 μg/mL for luteolin and 0.5-50 μg/mL for apigenin.RSD＇ s founds of the peak area of scutellarin, baicalein, luteolin and apigenin for the precision test, the stability test and the repeatability test were less than 2.0%.The recoveries, average recovery and RSDs of scutellarin, baicalein, luteolin and apigenin obtained were 97.73% -101.7% ,99.72%, 1.36% （ n =9） ,97.78% - 102.8% , 99.53% , 1.99% （ n = 9 ） , 97.40% ~ 102.4%, 99.39% , 1.84% （ n = 9 ） and 97.80% - 102.0% , 99.24% , 1.49% （ n= 9） .By using the established HPLC methods, 12 samples of Herba ScuteUariae barbate in different districts were analyzed.The resuhs showed that the contents of scutellarin,baicalein,luteolin and apigenin were 2.41-7.08,0.26-0.56, 0.31-0.90 mg/g and 0.28-0.95 mg/g, respectively. The method could be applied to sinmltaneous quantitative assay of scutellarin, baicalein, luteolin and apigenin in Herba Scutellariae barbatae with good specificity, reproducibility and stability. The sampling recovery test is in compliance with the requirement.