线叶菊有效部位HPLC特征图谱研究及5种成分含量测定

Study on HPLC specific chromatograms of effective parts and determination of five components of Filifolium sibiricum

目的建立10个不同批次线叶菊有效部位醋酸乙酯层的HPLC特征图谱,并测定5种成分的含有量。方法线叶菊有效部位的分析采用色谱柱Thermo Acclaim ^(TM)120 C_(18)(250 mm×4.6 mm,5μm),流动相:乙腈(B)-PBS(A)系统(0.1 mol/L磷酸二氢钠和2%冰醋酸1∶1混合),梯度洗脱,检测波长360 nm,体积流量0.8 mL/min,柱温35℃。结果建立了线叶菊有效部位HPLC特征图谱,确立了10个共有峰。测定的5种成分异荭草素、异牡荆苷、异槲皮苷、木犀草苷、异鼠李素-3-O-β-D-葡萄糖分别在0.018~0.108、0.066 8~0.400 8、0.088~0.528、0.118 4~0.710 4、0.017 6~0.105 6μg范围内线性关系良好,平均加样回收率分别为98.67%、97.93%、98.95%、99.81%、97.33%,RSD分别为1.10%、0.93%、1.10%、0.62%、1.48%。10批药材中8个批次样品的相似度在0.9以上,其中2批样品的相似度为0.688、0.695,含量偏低,差异性较大。各采收期样品中5种成分含有量差异明显,开花率高的药材黄酮类成分的含量高,提示线叶菊药材黄酮类成分的高低与采收期开花率有关。结论该方法建立了线叶菊有效部位的HPLC特征图谱,确定了共有特征峰,可用于线叶菊药材的质量控制。

Objective To establish the HPLC specific chromatograms of the ethyl acetate layer in ten batches of effective parts of Filifolium sibiricum and to determine the contents of five components. Methods The analysis of effective parts of F. sibiricum was performed on a Thermo AcclaimTM120 C18 column（250 mm × 4.6 mm, 5 μm） with acetonitrile（B）-PBS（A）（0.1 mol/L sodium dihydrogen phosphate-2% glacial acetic acid, 1∶1） as the mobile phase in a gradient elution mode, the detection wavelength was set at 360 nm, the flow rate was 0.8 mL/min, and the column temperature was 35 ℃. Results The specific chromatograms of F. sibiricum effective parts were established and ten common peaks were designated. Among them, five components including isorientin, isovitexin, isoquercitrin, luteoloside and isorhamnetin-3-O-β-D-glucose all showed good linear relationship within the ranges of 0.018—0.108, 0.066 8—0.400 8, 0.088—0.528, 0.118 4—0.710 4 and 0.017 6—0.105 6 μg, respectively. The average recovery was 98.67%, 97.93%, 98.95%, 99.81%, and 97.33% with the RSD value at 1.10%, 0.93%, 1.10%, 0.62%, and 1.48%, respectively. Moreover, the similarity of the eight batches of samples was above 0.9 in the ten batches of medicinal herbs, the similarity of the two batches of which was 0.688 and 0.695, indicating that its content was lower and the difference was greater. In addition, there were significant differences in the content of five components in each harvest time. The content of flavonoids in medicinal herbs was higher with high flower percentage. It was suggested that the content of flavonoids in F. sibiricum was related to the flower percentage of harvest period. Conclusion The HPLC specific chromatograms of the F. sibiricum effective parts were established and the common characteristic peaks were determined, which could be used for quality control of the F. sibiricum.