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

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 C₁₈（250 mm×4.6 mm，5 μm）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、15.00~150.04 μg·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 C₁₈ 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.