以离子液体作流动相添加剂的高效液相色谱法测定复方苦参注射液中4种生物碱

将离子液体1-丁基-3-甲基咪唑四氟硼酸盐(［BMIM］BF4)作为流动相添加剂建立了同时测定复方苦参注射液中4种主要生物碱的HPLC分析方法。以Agilent TC-C18柱(250 mm×4.6 mm, 5 μm)为分离柱,甲醇-0.1%磷酸水溶液(含2.2×10-4mol/L ［BMIM］BF4)(5:95, v/v)为流动相,流速1.0 mL/min,柱温30 ℃,进样量20 μL,在205 nm下检测。结果表明,苦参碱、槐定碱、氧化槐果碱以及氧化苦参碱的质量浓度分别在25.8～155.0 mg/L, 40.0～240.0 mg/L, 21.7～130.0 mg/L和37.5～225.0 mg/L范围内线性关系良好,相关系数均大于0.9990,平均回收率(n=9)在96.2%和98.9%之间。离子液体的加入能明显改善C18柱分离生物碱的色谱峰形并提高分离度。本法简便、快速、重复性好,可用于复方苦参注射液中生物碱的分离与测定。

Determination of four alkaloids in Compound Kushen Injection by high performance liquid chromatography with ionic liquid as mobile phase additive

In order to improve the resolution and peak shape, 1-butyl-3-methylimidazolium tetrafluoroborate (［BMIM］BF4) was used as mobile phase additive to develop an HPLC method for the determination of four main alkaloids in Compound Kushen Injection. An Agilent TC-C18 column was used with the mobile phase of methanol-0.1% phosphoric acid solution (containing 2.2×10-4mol/L ［BMIM］BF4) (5:95, v/v), flow rate of 1.0 mL/min, column temperature of 30 ℃, and injection volume of 20 μL, detected at 205 nm. The calibration curves of matrine, sophoridine, oxymatrine and oxysophocarpine were in good linearity over the ranges of 25.8-155.0 mg/L, 40.0-240.0 mg/L, 21.7-130.0 mg/L and 37.5-225.0 mg/L, respectively, with correlation coefficients above 0.9990. The mean recoveries (n=9) were between 96.2% and 98.9%. Ionic liquid, ［BMIM］BF4, can obviously improve the resolution and peak shape. The method is simple, rapid, and reliable, and can be used for the determination of alkaloids in Sophora flavescens Ait in Compound Kushen Injection.