超高效液相色谱法测定风湿骨痛丸中5种成分的含量

目的： 采用超高效液相色谱法（UPLC）测定风湿骨痛丸中乌头碱、次乌头碱、新乌头碱、麻黄碱、伪麻黄碱5种成分的含量。方法： （1）乌头碱、次乌头碱、新乌头碱检测方法：色谱柱为ACQUITY UPLC HSS T3柱（100 mm×2.1 mm，1.8 μm），ACQUITY UPLC HSS T3预柱（5 mm×2.1 mm，1.8 μm），流动相为甲醇-0.3 mol·L^-1三乙胺（65∶35，V/V），流速0.3 mL·min^-1，柱温30℃，检测波长235 nm，进样量10 μL；（2）麻黄碱、伪麻黄碱检测方法：色谱柱为ACQUITY UPLC BEH C₁₈柱（100 mm×2.1 mm，1.7 μm），流动相为乙腈（A）-0.1%磷酸溶液（B），采用梯度洗脱，流速0.3 mL·min^-1，柱温30℃，检测波长210 nm，进样量10 μL。结果： 乌头碱、次乌头碱、新乌头碱、麻黄碱、伪麻黄碱分离度良好；线性范围依次为4.00~80.04 μg·mL^-1（r=0.999 4），4.00~80.02 μg·mL^-1（r=0.999 3），4.03~80.57 μg·mL^-1（r=0.999 6），6.40~640.25 μg·mL^-1（r=0.999 6），6.41~641.34 μg·mL^-1（r=0.999 7）；检出限依次为2.03，1.98，1.93，3.67，3.73 μg·mL^-1；定量限依次为3.98，3.87，4.03，6.37，6.41 μg·mL^-1；平均回收率依次为99.06%，98.87%，99.95%，99.92%，100.54%，RSD依次为0.95%，1.55%，1.58%，1.18%，0.91%；风湿骨痛丸中乌头碱、次乌头碱、新乌头碱、麻黄碱、伪麻黄碱的平均含量依次为0.001%，0.004%，0.004%，0.041%，0.027%。结论： 本研究UPLC方法准确，重复性良好，可测定风湿骨痛丸中乌头碱、次乌头碱、新乌头碱、麻黄碱、伪麻黄碱的含量，为风湿骨痛丸的质量控制提供参考。

Determination of the contents of five components in Fengshi Gutong Pills by ultra performance liquid chromatography

OBJECTIVE To determine the contents of aconitine, hypaconitine, mesaconitine, ephedrine and pseudoephedrine in Fengshi Gutong Pills by UPLC.METHODS The detection methods of aconitine, hypaconitine and mesaconitine were as follows:chromatographic column:ACQUITY UPLC HSS T3 column(100 mm×2.1 mm, 1.8 μm), ACQUITY UPLC HSS T3 precolumn(5 mm×2.1 mm, 1.8 μm), mobile phase:methanol-0.3 mol·L^-1 triethylamine=65:35, flow rate 0.3 mL·min^-1, column temperature 30℃, detection wavelength 235 nm, injection volume 10 μL;the detection methods of ephedrine and pseudoephedrine were as follows:chromatographic column:ACQUITY UPLC BEH C₁₈ column(100 mm×2.1 mm, 1.7 μm), mobile phase:acetonitrile(mobile phase A), 0.1% phosphoric acid solution(mobile phase B), gradient elution, flow rate 0.3 ml·min^-1, column temperature 30℃, detection wavelength 210 nm, injection volume 10 μL.RESULTS The separation of aconitine, hypaconitine, mesaconitine, ephedrine and pseudoephedrine was good;the linear ranges were as follows:4.00-80.04 μg·mL^-1(r=0.999 4), 4.00-80.02 μg·mL^-1(r=0.999 3), 4.03-80.57 μg·mL^-1(r=0.999 6), 6.40-640.25 μg·mL^-1(r=0.999 6), 6.41-641.34 μg·mL^-1(r=0.999 7);the detection limits were 2.03, 1.98, 1.93, 3.67, 3.73 μg·mL^-1, and the quantitation limits were 3.98, 3.87, 4.03, 6.37, 6.41 μg·mL^-1;the average recoveries were 99.06%, 98.87%, 99.95%, 99.92% and 100.54%, respectively, and RSD were 0.95%, 1.55%, 1.58%, 1.18% and 0.91%, respectively;the contents of aconitine, hypaconitine, mesaconitine, ephedrine and pseudoephedrine in Fengshi Gutong Pills were 0.001%, 0.004%, 0.004%, 0.041% and 0.027%, respectively.CONCLUSIONS UPLC in this study is accurate and reproducible, which can be used to determine the contents of aconitine, hypaconitine, mesaconitine, ephedrine and pseudoephedrine in Fengshi Gutong Pills, and provide reference for the quality control of Fengshi Gutong Pills.