HPLC法测定蒙药嘎日迪-13味丸中6种乌头类生物碱的含量

Simultaneous determination of the 6 kinds of aconitum alkaloid contents in Mongolian medicine Garidi- 13 pill by HPLC method

目的建立测定蒙药嘎日迪-13味丸中6种生物碱的含量的高效液相色谱法。方法 Diamonsi I C18色谱柱,以乙腈-四氢呋喃（25∶15）为流动相A,以0.1 mol·L^-1醋酸铵溶液为流动相B,梯度洗脱,流速为0.8 m L·min-1,柱温为30℃,检测波长为235 nm,检测专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性。结果乌头碱、次乌头碱、新乌头碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、苯甲酰新乌头原碱分别在3.48×10^-3-0.17（r=0.999 5）,3.24×10^-3-0.16（r=0.999 6）,6.96×10^-3-0.35（r=0.999 5）,2.06×10^-3-0.10（r=0.999 9）,4.04×10^-3-0.20（r=0.999 9）,6.66×10^-3-0.33（r=0.999 9）mg·m L^-1呈良好的线性关系,RSD分别为3.56%,1.87%,4.86%,2.74%,3.21%,4.59%,平均回收率分别为101.11%,98.07%,97.81%,99.24%,104.55%,104.84%。结论本方法简便、准确、重复性好,可用于嘎日迪-13味丸中单酯型生物碱和双酯型生物碱的含量测定。

Objective To establish a method for simultaneous determination of 6 alkaloid in Mongolia medicine Garidi- 13 pill. Methods Diamonsi I C18 column was used. The acetonitrile- tetrahydrofuran（ 25 ∶15） and 0. 1 mol·L^-1of ammonium acetate solution were used as mobile phase by gradient elution. The volume flow rate was 0. 8 m L · min- 1,and column temperature was 30 ℃,wavelength was 235 nm. The specificity,lower limit of quantification and standard curve,precision and recovery rate and stability as well as the matrix effect were investigated.Results The aconitine,hypaconitine,mesaconitine,benzoylaconitine,benzoylhypacoitine and benzoylmesaconine showed a good linear relationship in the 3. 48 × 10^-3- 0. 17（ r = 0. 999 5）,3. 24 × 10^-3- 0. 16（ r =0. 999 6）,6. 96 × 10^-3- 0. 35（ r = 0. 999 5）,2. 06 × 10^-3- 0. 10（ r =0. 999 9）,2. 02 × 10^-3- 0. 20（ r = 0. 999 9） and 6. 66 × 10^-3- 0. 33（ r = 0. 999 9） mg·m L^-1. The RSD was 3. 56%,1. 87%,4. 86%,2. 74% and 3. 21%,4. 59%,respectively,and the average recovery rate was 101. 11%, 98. 07%, 97. 81%,9 9. 24%, 104. 55% and104. 84%. Conclusion The method is simple,accurate and reproducible,which can simultaneously determinate the mono ester alkaloid and double ester alkaloid in the Mongolia medicine Garidi- 13 pills.