反相离子对高效液相色谱法测定疏风活络丸中4种生物碱的含量

Ion pair RP-HPLC determination of 4 alkaloids in Shufeng Huoluo pills

目的:建立疏风活络丸中伪麻黄碱、麻黄碱、马钱子碱和士的宁的含量测定方法。方法:采用5%氢氧化钠溶液浸润样品后以三氯甲烷于70℃水浴中回流提取,以反相离子对高效液相色谱法测定,采用C18柱(4.6mm×250mm,5μm)分离,柱温30℃;流动相:A为0.01mol.L-1庚烷磺酸钠与0.01mol.L-1磷酸二氢钾等量混合溶液(用磷酸调pH2.8),B为乙腈,梯度洗脱(0min时,A为81%,B为19%;30min时,A为73%,B为27%);流速1.0mL.min-1,检测波长为206nm。结果:4种生物碱成分色谱行为良好,平均回收率(n=6)分别为伪麻黄碱97.36%(RSD=2.1%),麻黄碱97.22%(RSD=2.5%),马钱子碱98.65%(RSD=1.3%),士的宁97.69%(RSD=1.6%)。结论:所建立的方法简单易行,专属性强,准确性好,可用于疏风活络丸中活性兼毒性生物碱类成分的质量控制。

Objective：To develop a method for the determination of ephedrine, pseudoephedrine, brucine and strychnine in Shufeng Huoluo pills, a traditional Chinese patent medicine. Method：The samples were infiltrated with 5% NaOH solution following a reflux extraction with chloroform at 70 ℃, a ion pair RP - HPLC method was employed to determinate the target components in a step gradient elution, a column （4.6 mm × 250 mm, 5 μm） of Cl8 with a temperature at 30 ℃ was used to separate the alkaloids, the mixed solution（ pH was adjusted to 2. 8 with phosphate acid）composing of 0. 01 mol · L^-1 sodium- heptanesulfonate solution and equal quantity of 0.01 mol · L^-1 potassium dihydrogen phosphate solution was chosen as mobile phase A and acetonitrile as B to elute in a gradient program of A - B （ 81 ： 19 ） at 0 min and A - B （73： 27 ） at 30 min, the flow rate was 1.0 mL · min^-1, the detection wavelength was at 206 nm. Results： 4 alkaloids were separated perfectly, the average recoveries （ n = 6 ） were 97.36% （RSD = 2. 1% ）for pseudoephedrine,97.22% （RSD = 2. 5% ）for ephedrine,98.65% （RSD = 1.3% ）for brucine,and 97.69% （ RSD = 1.6% ） for strychnine. Conclusions： The developed method, which is simple and practical, demonstrates a good accuracy and specificity, it can be adopted to monitor the active and toxic alkaloids in Shufeng Huoluo pills.