高效液相色谱与QDA质谱联用同时测定附子理中丸中6个单酯及双酯型生物碱含量

目的：建立同时测定附子理中丸中6个单酯及双酯型生物碱含量的方法。方法：应用高效液相色谱（HPLC）与QDA质谱联用同时测定附子理中丸中苯甲酰新乌头原碱、苯甲酰次乌头原碱、苯甲酰乌头原碱、新乌头碱、次乌头碱和乌头碱6个成分的含量。使用Phenomenex Luna C₁₈色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.2%乙酸水（用三乙胺调pH 6.2）为流动相，梯度洗脱，流速1.0 mL·min^-1，分流比为1∶4，使用ESI⁺离子源，选择性离子扫描（SIM）模式下对苯甲酰新乌头原碱（m/z 590.0）、苯甲酰乌头原碱（m/z 604.0）、苯甲酰次乌头原碱（m/z 574.0）、新乌头碱（m/z 632.0）、次乌头碱（m/z 616.0）和乌头碱（m/z 646.0）进行测定。结果：苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、次乌头碱、新乌头碱和乌头碱6个成分的质量浓度分别在0.000 2~51.75 μg·mL^-1（r=0.999 2）、0.000 2~50.95 μg·mL^-1（r=0.999 5）、0.000 2~50.25 μg·mL^-1（r=0.999 8）、0.000 2~49.80 μg·mL^-1（r=0.999 9）、0.000 2~50.60 μg·mL^-1（r=0.999 5）和0.000 2~52.00 μg·mL^-1（r=0.999 7）范围内线性关系良好；平均回收率（n=6）分别为101.8%、101.3%、98.2%、104.8%、99.9%和89.9%，RSD分别为6.4%、2.6%、2.7%、6.1%、3.8%和5.7%。3批附子理中丸中6个生物碱成分即苯甲酰新乌头原碱、苯甲酰次乌头原碱、苯甲酰乌头原碱、新乌头碱、次乌头碱和乌头碱的含量范围分别为7.095~7.366、1.007~1.214、0.594~0.742、1.261~1.438、1.492~1.578和0.135~0.163 μg·g^-1。结论：该方法可测定附子理中丸中6个生物碱成分含量，为附子理中丸的质量标准提高打下了基础。

Objective:To establish the quantitative method of six diterpenoid alkaloids in Fuzilizhong pills. Methods:The contents of benzoylmesaconitine,benzoylaconitine,benzoylhypaconitine,mesaconitine,aconitine,and hypaconitine in Fuzilizhong pills were determined by HPLC coupled with QDA mass detector.The chromatographic column was Phenomenex Luna C₁₈(4.6 mm×250 mm,5 μm).The mobile phase was the mixture of acetonitrile-0.2% acetic acid(adjust pH to 6.2 with triethylamine)in gradient mode,the flow rate was 1.0 mL·min^-1,and the split ratio was 1:4.The ESI⁺ source of the mass spectrometer was used in selected ion monitoring(SIM)mode for the detection of benzoylmesaconitine(m/z 590.0),benzoylaconitine(m/z 604.0),benzoylhypaconitine(m/z 574.0),mesaconitine(m/z 632.0),aconitine(m/z 646.0)and hypaconitine(m/z 616.0).Results:The linear ranges of benzoylmesaconitine,benzoylaconitine,benzoylhypaconitine,hypaconitine,mesaconitine,and aconitine were 0.000 2-51.75 μg·mL^-1(r=0.999 2),0.000 2-50.95 μg·mL^-1(r=0.999 5), 0.000 2-50.25 μg·mL^-1(r=0.999 8),0.000 2-49.80 μg·mL^-1(r=0.999 9),0.000 2-50.60 μg·mL^-1(r=0.999 5)and 0.000 2-52.00 μg·mL^-1(r=0.999 7),respectively.The average recovery rates(n=6)of the above compounds were 101.8%,101.3%,98.2%,104.8%,99.9% and 89.9% with RSD of 6.4%,2.6%,2.7%,6.1%,3.8% and 5.7%,respectively.The contents of benzoylmesaconitine,benzoylhypaconitine,benzoylaconitine,mesaconitine,aconitine and hypaconitine were 7.095-7.366,1.007-1.214,0.594-0.742,1.261-1.438,1.492-1.578 and 0.135-0.163 μg·g^-1,respectively.Conclusion:The method can be used to determine the content of six diterpenoid alkaloids in Fuzilizhong pills.The present study provides a scientific basis for the quality control of Fuzilizhong pills.