HPLC一测多评法同步测定玳玳果黄酮降脂提取物4个效应组分含量

Simultaneous Determination of Four Effect Components of Flavonoids Lipid-Lowering Extract from Daidai Fruit by QAMS

目的建立HPLC一测多评法(QAMS)同步测定玳玳果黄酮降脂提取物4个效应组分柚皮苷、新橙皮苷、橙皮内酯水合物、枳属苷含量。方法采用高效液相色谱法,选择主成分柚皮苷为内参物,建立HPLC一测多评法,并与HPLC外标法同时测定的4个组分含量结果比较。色谱柱Lichrocart C18柱(250 mm×4.6 mm,5μm),乙腈(A)-0.2%冰醋酸水溶液(B)为流动相,梯度洗脱;流速1.0 m L/min,检测波长284 nm;柱温30℃;进样量10μL。结果玳玳果黄酮降脂提取物柚皮苷、新橙皮苷、橙皮内酯水合物、枳属苷的质量浓度分别在31.377~159.188μg/mL(r=0.999 5)、63.632~208.123μg/m L(r=0.999 8)、3.899~12.241μg/m L(r=0.999 6)、2.696~17.366μg/mL(r=0.999 4)范围内呈良好的线性关系;平均回收率(n=6)分别为97.99%、101.50%、98.21%、95.56%,RSD分别为2.07%、2.80%、2.92%、2.32%。结论以柚皮苷为内参物,HPLC一测多评法同步测定玳玳果黄酮降脂提取物4个效应组分含量结果准确,可用于玳玳果黄酮降脂提取物多指标质量监控。

Objective: To establish a method for simultaneous determination of four effect components including naringin, neohesperidin, hesperidin hydrate, and poncirin in the lipid-lowering extract of flavonoids from the Daidai fruit by QAMS. Methods:HPLC was applied to establish a multiple-evaluation method for the simultaneous determination of naringin, neohesperidin, hesperidin hydrate, and poncirin with naringin representative component as internal reference material. The results were compared with those of the four components determined by HPLC external standard method. Using Lichrocart C18 columns(250 mm×4.6 mm, 5 μm), the mobile phase was consisted of acetonitrile(A)-0.2% and glacial acetic acid(B) with gradient elution, flow rate at 1.0 m L/min, detection wavelength at 284 nm, column temperature at 30 ℃ and 10 μL injection volume. Results: There were good linearities for naringin, neohesperidin, hesperidin hydrate, and poncirin in the concentration ranges of 31.377-159.188 μg/m L(r=0.999 5), 63.632-208.123 μg/m L(r=0.999 8), 3.899-12.241 μg/m L(r=0.999 6), and 2.696-17.366 μg/m L(r=0.999 4), respectively. The average recoveries(n=6) were 97.99%, 101.50%, 98.21%, 95.56%, with RSDs of 2.07%, 2.80%, 2.92%, 2.32%, respectively. Conclusion:Using naringin as the internal reference, the content of four effect components were accurately determined by the simultaneous determination method which provide a scientific experimental basis for the establishment of a multi-index quality monitoring method for flavonoid lipid-lowering extracts of Daidai fruit.