混合模式色谱相测定四神丸中的吲哚类生物碱与木脂素类化合物

Simultaneous Determination of Alkaloids and Lignins in Sishen Pills by Mixed-mode Chromatography

目的:建立混合模式固定相的高效液相色谱法测定四神丸中的吲哚类生物碱与木脂素类化合物。方法:Hypersil Duet C_(18)/SAX (250 mm×4. 6 mm,5μm)色谱柱,以乙腈-0. 5%磷酸为流动相,梯度洗脱,流速为1. 0 ml·min^(-1),检测波长为240 nm,柱温为35℃,进样量10μl。结果:吴茱萸碱、吴茱萸次碱、补骨脂素、五味子乙素与异补骨脂素在混合模式色谱中完全分离;线性关系良好;线性范围分别为0. 110～2. 120μg·ml^(-1)(r=0. 999 9),0. 119～2. 387μg·min^(-1)(r=0. 999 8),0. 121～2. 417μg·min^(-1)(r=0. 999 4),0. 119～2. 387μg·ml^(-1)(r=0. 999 2),0. 104～2. 084μg·ml^(-1)(r=0. 998 7),加样回收率分别为97. 1%(RSD=0. 9%),101. 7%(RSD=1. 5%),96. 8%(RSD=1. 1%),94. 9%(RSD=0. 4%),98. 6%(RSD=1. 2%)(n=6)。结论:该方法准确性、耐用性良好,可用于四神丸质量控制。

Objective:To establish an HPLC method for the determination of the alkaloids and lignins in Sishen pills using a mixed-mode column.Methods:The samples were separated on a Hypersil Duet C18/SAX column(250 mm×4.6 mm,5μm)with gradient elution using acetonitile and 0.5%phosphoric aicd as the mobile phase.The flow rate was 1.0 ml·min-1,the column temperature was 35℃,the detection wavelength was 240 nm and the sample size was 10μl..Results:Evodiamine,rutaecarpine,psoralen,schisandrin B and isopsoralen were well separated;the linearity of the 5 components was 0.110-2.120μg·ml^-1(r=0.999 9),0.119-2.387μg·ml^-1(r=0.999 8),0.121-2.417μg·ml^-1(r=0.999 4),0.119-2.387μg·ml^-1(r=0.999 2)and 0.104-2.084μg·ml^-1(r=0.998 7),respectively.The average recovery of the compounds was 97.1%(RSD=0.9%),101.7%(RSD=1.5%),96.8%(RSD=1.1%),94.9%(RSD=0.4%)and 98.6%(RSD=1.2%)(n=6),respectively.Conclusion:The optimized method is simple and specific,which can be used for the quality control of Sishen pills.