微乳液相色谱法测定夏天无中生物碱的含量

Simultaneous Determination of Five Alkaloids in Corydalis Decumbentis Rhizoma by Microemulsion Liquid Chromatography

目的：建立微乳液相色谱同时测定夏天无中药根碱、巴马汀、小檗碱、延胡索乙素、原阿片碱含量的方法.方法：以微乳为流动相,通过对影响分离度和保留时间的因素进行考察,优化微乳组成体系,得到最佳的微乳体系组成为2％ SDS-0.5％正辛醇-4.0％正丁醇-磷酸氢二钠（磷酸调pH至4.0）.选择Sinochrom ODS-BP色谱柱（4.6 mm×250 mm,5μm）,柱温25℃,流速0.8 mL· min^-1,检测波长280 nm.结果：在优化条件下,5种生物碱类成分在20 min内达到基线分离,分离度及峰形良好.其中,药根碱、巴马汀、小檗碱、延胡索乙素和原阿片碱分别在0.800 9～ 16.01,0.800 9～ 16.01,0.200 2 ～ 4.004,8.013 ～160.3,12.09 ～240.7 mg·L^-1线性关系良好,相关系数均＞0.999 5,检出限（S/N =3）分别为0.201,0.254,0.053,1.27,1.05 mg·L^-1,平均加样回收率为97.34％ ～99.51％.结论：所建立的微乳液相色谱法简单、快速、准确,可用于夏天无中5种生物碱类成分的含量测定,为中药的质量控制提供了新选择.

Objective： To develop the accurate and rapid method for determination of jatrorrhizine, palmatine, berberine, tetrahydropulmatine and protopine in Corydalis Decumbentis Rhizoma by microemulsion liquid chromatography （MELC）. Method： The mobile phase was microemulsion that was optimized by inspecting the influence factors of separation and retention time. The optimized microemulsion was 2% sodium dodecyl sulfate- 0.5% noctyl alcohol-4% nbutanol-phosphate buffer （pH was adjusted by phosphoric acid to 4.0）. Sinochrom ODS-BP Chromatography columns （4.6 mm ×250 mm, 5 μm） was selected, with the column temperature of 25℃. The contents of five alkaloids in Corydalis Decumbentis Rhizoma were determined simultaneously at a flow rate of 0. 8 mL·min^-1, and the detection wavelength was 280 nm. Result： Under the optimized conditions, baseline separation of five alkaloids was achieved in 20 min, and the resolution is well. The calibration curves of jatrorrhizine, palmatine, berberine, tetrahydropulmatine and protopine were linear in the ranges of 0. 800 9-16.01, 0. 800 9-16.01, 0. 200 2-4. 004, 8. 013-160.3 and 12.09-240.7 mg · L^-1 , respectively, with correlation coefficients of 0. 999 5. Their limits of detection （ LOD, S/N = 3 ） were 0. 201 , 0. 254, 0. 053, 1.27 and 1.05 mg·L^-1 , respectively. The spiked recoveries of three compounds ranged from 97.34% to 99.51%. Conclusion：The method is simple, rapid and accurate, and can be used for the quality control of five alkaloids in Corydalis Decumbentis Rhizoma.