高效液相色谱-荧光检测法同时测定多烯酸乙酯软胶囊中4种生育酚的含量

目的: 建立多烯酸乙酯软胶囊中4种生育酚的HPLC含量测定方法,同时对不同厂家产品中4种生育酚(α、β、γ、δ-生育酚)的含量进行比较。方法: 采用色谱柱为MERCK Lichro CART 250-4硅胶柱(250 mm×4.6 mm,5 μm),流动相为正己烷-异丙醚(80:20),流速1.0 mL·min^-1,柱温25 ℃,荧光检测器激发波长298 nm,发射波长325 nm。结果: 4种生育酚的质量浓度均在40~1 000 ng·mL^-1范围内线性关系良好(r> 0.999);检出限分别为7.0、6.0、6.2、6.2 ng·mL^-1;加样平均回收率为90.1%~99.7%。国内不同厂家的α-生育酚的含量范围为1.92~32.95 μg·g^-1,γ-生育酚范围为1.23~21.22 μg·g^-1,δ-生育酚范围为0.83~13.05 μg·g^-1,β-生育酚均未检出,国外同类品种中α-生育酚的含量约为1 800 μg·g^-1。结论: 本文建立的生育酚的高效液相色谱-荧光检测法具有较高的灵敏度、精密度和准确性,可作为有效控制ω-3多不饱和脂肪酸乙酯类药物的产品质量的检测方法。

Objective: To establish an HPLC method for determining four kinds of tocopherol in ethyl polyenoate soft capsules, and to compare the contents of them from different pharmaceutical factories.Methods: The analysis was carried out on a MERCK Lichro CART 250-4 silica column (250 mm×4.6 mm, 5 μm) with hexane-isopropyl ether (80:20) as the mobile phase at a flow rate of 1.0 mL·min^-1.The column temperature was set at 25 ℃.The fluorescence excitation and emission wavelengths were λ_ex=298 nm and λ_em=325 nm.Results: The calibration showed good linearity for four kinds of tocopherol in the range of 40-1 000 ng·mL^-1 (r> 0.999).The minimum detection limits were 7.0, 6.0, 6.2 and 6.2 ng·mL^-1 and the average recovery was in the range of 90.1%-99.7%.The content ranges were 1.92-32.95 μg·g^-1 for α-tocopherol, 1.23-21.22 μg·g^-1 for γ-tocopherol, and 0.83-13.05 μg·g^-1 for δ-tocopherol;but β-tocopherol was not detected.The content of tocopherol in EPA ethyl ester was about 1 800 μg·g^-1.Conclusion: The established method of HPLC with fluorescence detection is accurate, sensitive, and reliable for the determination of tocopherol in ethyl polyenoate soft capsules, which could be used for the research of quality control for ω-3 polyunsaturated fatty acid ethyl ester drug in future.