UPLC-MS/MS测定复方甘草片中4种生物碱含量

目的: 采用超高效液相色谱-质谱联用方法同时测定复方甘草片中吗啡、可待因、罂粟碱、可卡因等4种生物碱的含量。方法: 采用Waters BEH C₁₈(50 mm×2.1 mm,1.7 μm)色谱柱,以含0.1%甲酸的0.02 mol·L^-1乙酸铵溶液(A)-甲醇(B)为流动相,梯度洗脱(0~1.5 min,92%A;1.5~2.0 min,92%A→60%A;2.0~4.5 min,60%A;4.5~4.6 min,60%A→92%A;4.6~5.0 min,92%A),流速0.2 mL·min^-1,柱温35 ℃;质谱采用电喷雾离子源,多反应监测模式(正离子)。结果: 吗啡、可待因、罂粟碱、可卡因质量浓度分别在2.84~56.7、1.04~20.85、0.53~10.58和0.16~3.13 μg·L^-1的范围内线性关系良好,r值在0.998 4~0.999 9之间,平均回收率在96.0%~102.8%之间,分析方法的定量下限分别为0.5、0.5、0.05、0.01 μg·L^-1,检测限为0.2、0.2、0.02、0.005 μg·L^-1;3批复方甘草片中吗啡、可待因的含量分别为0.39~0.40 mg·片^-1和0.18~0.19 mg·片^-1,没有检出罂粟碱和可卡因。结论: 该方法前处理简单,分析速度快,灵敏度高,是复方甘草片质量控制的一种可行方法。

Objective: To establish a method for the quantitative determination of 4 alkaloids,including morphine,codeine,papaverine and cocaine,in compound liquorice tablets by UPLC-MS/MS. Methods: Chromatography was carried on a Waters BEH C₁₈ column (50 mm×2.1 mm,1.7 μm) with the mobile phase consisting of 0.02 mol·L^-1 ammonium acetate water solution (containing 0.1% formic acid) (A) and methanol (B) in a programme of gradient elution (0-1.5 min,92%A;1.5-2.0 min,92%A→60%A;2.0-4.5 min,60%A;4.5-4.6 min,60%A→92%A;4.6-5.0 min,92%A). The flow rate was 0.2 mL·min^-1 with a column temperature at 35 ℃. Determination was performed by multiple reaction monitoring (MRM) mode using electrospray ionization in positive ion mode. Results: The linear ranges of morphine, codeine,papaverine and cocaine were 2.84-56.7,1.04-20.85,0.53-10.58 and 0.16-3.13 μg·L^-1(r=0.998 4-0.999 9); the average recovery were 96.0%-102.8%; the limits of quantitation and detection were 0.5,0.5,0.05,0.01 μg·L^-1 and 0.2,0.2,0.02,0.005 μg·L^-1,respectively. The content of morphine and codeine in 3 batches of compound liquorice tablets were 0.39-0.40 mg and 0.18-0.19 mg per tablet,respectively. Pavaverine or cocaine was not detected. Conclusion: The pretreatment of the method is simple,rapid,sensitive,and can be an appropriate technique for quality control of compound liquorice tablets.