基于BP-ANN优化提取HPLC特征图谱测定3种黄精辐照前后黄酮含量变化

徐胜; 屈曾义; 毕健丽; 李婷婷; 甘小凤; 韦国良; 杜纪富; 何敏; 范宝磊

doi:10.13386/j.issn1002-0306.2020050171

基于BP-ANN优化提取HPLC特征图谱测定3种黄精辐照前后黄酮含量变化

Optimization of Extraction by BP-ANN and Determination of Flavonoids in Three Varieties of Polygonati Rhizoma Before and After Irradiation by HPLC Chromatographic Fingerprint

摘要

摘要: 目的：设计反向传播人工神经网络（BP-ANN）优化鸡头黄精、竹节黄精和马鞭黄精中7种黄酮的得率，明确各黄精电子束辐照前后黄酮的含量变化，建立指纹图谱。方法：检测方法使用高效液相色谱法（HPLC），采用Inertsil ODS-3，C₁₈色谱柱（250 mm×4.6 mm，5 μm），流动相0.02%磷酸溶液（A）-乙腈（B），梯度洗脱，柱温43℃，流速1.2 mL/min，检测波长270 nm。并建立在不同辐照剂量下的黄精黄酮指纹图谱，比较不同辐照剂量下黄精黄酮的含量变化。结果：建立的HPLC法能准确测定黄精中芦丁、杨梅素、甘草素、槲皮素、芹菜素、山奈酚、异鼠李素的含量。黄精中的7种化学成分能完整出峰，分离度均>1.5，各成分在相应的浓度范围内与峰面积有良好的线性关系（R²均>0.999），加样回收率97.95%~103.71%。BP人工神经网络技术优化提取结果显示，在固液比为1：50（g：mL），提取时间为2 h，提取温度为90℃，乙醇浓度为70%的提取条件下，能得到黄精中黄酮的最大得率0.0622%。样品测定结果显示，只有鸡头黄精中含所有的7种黄酮，其余两种黄精中的黄酮成分各不相同。黄精中黄酮的含量随着辐照剂量的增大而减小。结论：使用BP人工神经网络拟合黄精中黄酮的得率精密度高，拟合效果好。高效液相色谱法检测黄精中的7种黄酮类成分灵敏度高、专属性强，可用于黄精中黄酮的含量测定。

Abstract: Objective:Back propagation artificial neural network(BP-ANN)was designed to optimize the yield rate of seven flavonoids from Polygonatum sibiricum,Polygonatum odoratum and Polygonatum filipes,to determine the content changes of flavonoids before and after electron beam irradiation of different varieties of Polygonati Rhizoma,and to establish chromatographic fingerprint. Methods:The determination was performed by high performance liquid chromatography(HPLC),Inertsil ODS-3,C₁₈ column(250 mm×4.6 mm,5 μm)was used,the mobile phase consisted of 0.02% phosphoric acid(A)-acetonitrile(B),gradient elution at a flow rate of 1.2 mL/min,and the column temperature was 43 ℃,the detection wavelength was set at 270 nm. The fingerprint of Polygonati Rhizoma flavonoids under different irradiation doses was established,and the content changes of Polygonati Rhizoma flavonoids under different irradiation doses were compared. Results:The established HPLC method could accurately determine the contents of rutin,myricetin,liquiritigenin,quercetin,apigenin,kaempferol and isorhamnetin in Polygonati Rhizoma. All the 7 chemical constituents in the Polygonati Rhizoma could produce complete peaks,with the separation degree was more than 1.5,and the components had a good linear relationship within the corresponding concentration range(R²>0.999),and the recovery rate was 97.95%~103.71%. The optimized extraction results by BP-ANN technology showed that under the conditions of solid-liquid ratio of 1:50,extraction time of 2 h,extraction temperature of 90 ℃,and ethanol concentration of 70%,the maximum extraction efficiency(0.0622%)of flavonoids from Polygonati Rhizoma could be obtained. The results of the sample determination showed that only the Polygonatum sibiricum contained all 7 kinds of flavonoids,while the flavonoids in the other two kinds of Polygonati Rhizoma were not the same. The content of flavonoids decreased with the increase of irradiation dose. Conclusion:BP-ANN has high precision and good fitting effect to fit yield rate of flavonoids in Polygonati Rhizoma. The HPLC for the determination of seven flavonoids in Polygonati Rhizoma has high sensitivity and specificity,and can be used for the determination of flavonoids in Polygonati Rhizoma.