UPLC波长切换法同时测定桑叶中6种指标成分的含量

Simultaneous Determination of Six Index Components in Mori Folium by UPLC Wavelength Switching Method

目的采用超高效液相色谱(UPLC)波长切换法同时测定桑叶中新绿原酸、隐绿原酸、绿原酸、芦丁、异槲皮苷、紫云英苷的含量。方法以Waters BEH shield RP18为色谱柱(100 mm×2.1 mm,1.7μm),以乙腈(A)-0.1%磷酸(B)为流动相,梯度洗脱;柱温40℃,全波长扫描(190~400 nm),波长切换(0~3 min,325 nm,检测新绿原酸、隐绿原酸、绿原酸;3~8 min,358 nm,检测芦丁、异槲皮苷、紫云英苷)。结果桑叶中新绿原酸、隐绿原酸、绿原酸、芦丁、异槲皮苷、紫云英苷6种活性成分在8 min内分离效果理想,质量浓度分别在17.24~33.91μg/ml(r=0.999),23.01~45.68μg/ml(r=0.999),50.93~121.93μg/ml(r=0.999),11.91~54.11μg/ml(r=0.999),13.99~60.00μg/ml(r=0.999),4.83~24.67μg/ml(r=0.999)范围内与峰面积呈良好的线性关系;平均加样回收率(n=6)为99.28%~103.75%,RSD≤3.3%,供试品溶液在室温条件下24 h内稳定,RSD<2.4%。11个不同产地桑叶中新绿原酸、隐绿原酸、绿原酸、芦丁、异槲皮苷、紫云英苷6个成分的含量分别为0.86~1.7,1.15~2.28,2.55~6.10,0.60~2.71,0.70~3.00,6.1~16.97 mg/g。结论该方法操作简单,结果准确,专属性强,可为桑叶药材的质量控制标准提高提供参考。

Objective To develop a method for simultaneous determination of nochlorogenic acid,cryptochlorogenic acid,chlorogenic acid,rutin,isoquercetin and astragalin in Mori folium by UPLC with wavelength switching.Methods The chromatographic column was Waters BEH shield RP18(100 mm×2.1 mm,1.7μm).Gradient elution was performed,using acetonitrile(A)-0.1%phosphoric acid(B)as mobile phase.The flow rate was 0.3 ml/min,the column temperature was 40℃.The detection wavelength was set at 325 nm for neochlorogenic acid,cryptochlorogenic acid and chlorogenic acid in the first 3 min,and 358 nm for rutin,isoquercetin and astragalin during 3-8 min after full wavelength scanning(190-400 nm).Results The good saperation of six components was achieved within 8 min.Good linearity was obtained for neochlorogenic acid,cryptochlorogenic acid,chlorogenic acid,rutin,isoquercetin and astragalin in the ranges of 17.24-33.91μg/ml(r=0.999),23.01-45.68μg/ml(r=0.999),50.93-121.93μg/ml(r=0.999),11.91-54.11μg/ml(r=0.999),13.99-60.00μg/ml(r=0.999)and 4.83-24.67μg/ml(r=0.999),respectively.The average recoveries(n=6)were 99.28%-103.75%,with RSDs not more than 3.3%.The six analytes were stable at room temperature within 24 h,with RSDs less than 2.4%.The contents of six index components in Mori folium from 11 different origins were in the ranges of 0.86-1.7 mg/g for nochlorogenic acid,1.15-2.28 mg/g for cryptochlorogenic acid,2.55-6.10 mg/g for chlorogenic acid,0.60-2.71 mg/g for rutin,0.70-3.00 mg/g for isoquercetin and 6.1-16.97 mg/g for astragalin.Conclusion The proposed method is simple,accurate and repeatable,which can provide reference for the improvement of the quality control standard of Mori folium.