2D-HPLC测定枳实中柚皮苷、橙皮苷、新橙皮苷及辛弗林的含量

Simultaneous Determination of Contents of Naringin,Hesperidin,Neohesperidin and Synephrine in Immature Bitter Orange by 2D-HPLC

目的建立二维高效液相色谱（2D-HPLC）法同时测定枳实中橙皮苷、柚皮苷、新橙皮苷、辛弗林含量的方法。方法采用2D-HPLC法,分离体系1色谱条件：ODS柱（自制预柱）（4.6×30mm,5μm）、Thermo Hypersil-Keystone C1 8柱（4.6×250mm,5μm）;流动相：0.1%磷酸（A）-乙腈（B）=82：18;流速：1.0m L/min;柱温：25℃,检测波长：283nm。分离体系2色谱条件：Waters SCX柱（4.6×250mm,5μm）,流动相：60mmol/L磷酸二氢钾-乙腈=90：10;流速：1.0m L/min;检测波长：275nm;柱温：25℃。切换时间：1.0min。结果柚皮苷、橙皮苷、新橙皮苷、辛弗林分别在进样量0.371～2.968、0.1505～1.204、0.575～4.60、0.191～1.528μg范围内与色谱峰面积线性关系良好,加样回收率（n=6）分别为99.75%、100.81%、99.83%、100.27%,RSD分别为2.53%、1.56%、2.75%、1.43%,具较好的精密度、重复性和稳定性。结论 2D-HPLC法可用于枳实多成分的质量控制。

Obje ctive To develop an 2D- HPLC method for simultaneous determination of contents of four components in Immature Bitter Orange. Me thod The first separation system： The quantitative analysis was carried out on a ODS（ Pre column）（ 4. 6 × 30 mm,5μm） and a hypersil ODS2（ 250 mm × 4. 6mm,5μm） by simultaneous HPLC,using a mobile phase of 0. 1mol / L phosphorous acid（ A）- Acetonitrile（ B） = 82： 18 with a flow rate of 1 m L / min-1 at 25 ℃. The detection wavelength was set at 283 nm. The second system： The quantitative analysis was carried out on a Waters SCX（ 4. 6 × 250 mm,5μm） by simultaneous HPLC,using a mobile phase of 0. 06 mol / L potassium dihydrogen phosphate（ A）- Acetonitrile（ B） = 90： 10 with a flow rate of 1 m L. min- 1at 25 ℃. The detection wavelength was set at 275 nm. The Switching time was 1 minute. Re sult The linear ranges of Naringin,Hesperidin,Neohesperidin and Synephrine fell within the ranges of 0. 371 ～ 2. 968,0. 1505 ～ 1. 204,0. 575 ～ 4. 60,0. 191 ～ 1. 528μg,respectively. The recoveires（ n = 6） of Naringin, Hesperidin, Neohesperidin and Synephrine were 99. 75%,100. 81%,99. 83%,100. 27% respectively,and the relative standard deviations were 2. 53%,1. 56%,2. 75%,1. 43%. Conclusion The four components determined by this method were representative,which could be used for the simultaneous determination of the four constituents in Immature Bitter Orange.