HPLC-ELSD法测定附子水溶性生物碱提取物和蒸附片中5种生物碱的含量

Determination of five representative components in the extract of water-soluble alkaloids from Aconiti Lateralis Radix Praeparata and Zhengfu tablets by HPLC-ELSD method

目的:建立同时测定附子水溶性生物碱提取物和蒸附片中新乌头原碱、次乌头原碱、附子灵、尼奥灵、宋果灵含量的方法。方法:采用HPLC-ELSD法,Extend C_(18)(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈(A)-0.1%三乙胺水溶液(B),梯度洗脱(0~8 min,10%A→20%A;8~15 min,20%A→25%A;15~25 min,25%A→25%A;25~30 min,25%A→28%A;30~31 min,28%A→60%A;31~42 min,60%A→65%A),流速1.0 mL·min^(-1),柱温35℃,蒸发光散射检测器(蒸发温度50℃,载气流速1.5 L·min^(-1),增益为1)。结果:在此色谱条件下,5种生物碱分离度良好,新乌头原碱、次乌头原碱、附子灵、尼奥灵、宋果灵分别质量浓度在50.01~500.08、30.00~300.02、30.01~300.13、50.01~500.07、15.00~150.04μg·mL^(-1)范围内线性关系良好(r≥0.999 1);平均加样回收率(n=6)分别为98.2%(RSD=1.9%),98.3%(RSD=1.6%),98.5%(RSD=1.3%),97.2%(RSD=1.3%),97.7%(RSD=1.7%)。结论:该方法简便、准确,可用于附子水溶性生物碱提取物与蒸附片的质量控制与评价。

Objective：To establish a quantitative method for simultaneous determination of mesaconine,hypaconine,fuziline,neoline,and songorine in the extract of water-soluble alkaloids from Aconiti Lateralis Radix Praeparata and Zhengfu tablets.Methods：HPLC-ELSD was adopted in the assay.The analysis was performed on an Extend C（18） column（250 mm × 4.6 mm,5 μm）.The mobile phase was acetonitrile（A）-0.5%（v/v）triethylamine solution（B）with gradient elution（0-8 min,10%A → 20%A;8-15 min,20%A → 25%A;15-25 min,25%A → 25%A;25-30 min,25%A → 28%A;30-31 min,28%A → 60%A;31-42 min,60%A → 65%A）.The flow rate was 1.0 mL·min^-1 and the column temperature was 35 ℃.The evaporative light scattering detector（ELSD）was selected as the detector（the temperature of evaporation of 50 ℃,the flow rate of carrier gas of 1.5 L·min^-1,and the gain was 1）.Results：Five alkaloids（mesaconine,hypaconine,fuziline,neoline,songorine）were completely separated under the above condition.Satisfactory linearities of mesaconine,hypaconine,fuziline,neoline,songorine were in the ranges of 50.01-500.08,30.00-300.02,30.01-300.13, 50.01-500.07,and 15.00-150.04 μg·mL^-1,respectively（r ≥0.999 1）,and the average recoveries（n=6）were 98.2%（RSD=1.9%）,98.3%（RSD=1.6%）,98.5%（RSD=1.3%）,97.2%（RSD=1.3%）,97.7%（RSD=1.7%）,respectively.Conclusion：The method is simple and accurate,and provides a reliable approach to the quality control and evaluation of water-soluble alkaloids extract from Aconiti Lateralis Radix Praeparata Zheng fu tablets.