HPLC法比较延胡索炮制前后7个生物碱成分的含量

目的: 采用反相高效液相色谱法分别测定中药延胡索生品与醋制品中四氢非洲防己碱、原阿片碱、延胡索乙素、黄连碱、巴马汀、小檗碱、去氢延胡索甲素7个生物碱的含量,并对其进行比较。方法: 采用Benetnach C₁₈色谱柱(4.6 mm×250 mm,5 μm),以乙腈-0.1%磷酸水溶液(含0.08%三乙胺)为流动相,梯度洗脱,流速1.0 mL·min^-1,柱温35℃,检测波长280 nm,洗脱时间为70 min。结果: 四氢非洲防己碱、原阿片碱、延胡索乙素、黄连碱、巴马汀、小檗碱、去氢延胡索甲素7个生物碱线性范围分别为1.12~56.0、1.27~63.5、3.37~168.5、1.02~51.0、0.89~44.5、0.88~44.0、3.22~161.0 μg·mL^-1(r≥0.999 3);平均回收率(n=6)为98.3%~103.6%,RSD≤2.8%。炮制前后7个生物碱含量的质量分数测定结果分别为163.90~376.28、140.24~207.29、973.61~1 158.33、180.49~433.49、68.01~196.76、18.43~46.10、550.57~1 489.84 μg·g^-1。结论: 醋制品中四氢非洲防己碱比生品含量低,原阿片碱、延胡索乙素、巴马汀、小檗碱、去氢延胡索甲素含量均有不同程度的升高,黄连碱含量变化不定。本实验所建立方法经方法学验证,可为延胡索醋制品的质量控制提供科学依据。

Objective: To establish an HPLC method for simultaneous determination of seven alkaloids in crude and processed in traditional Chinese medicine Corydalis Rhizoma,and compare the change of these seven alkaloids. Methods: The methods were performed on an RP-HPLC Benetnach C₁₈ column(4.6 mm×250 mm,5 μm)with the mobile phase consisting of acetonitrile-0.1% phosphoric acidwater solution(containing 0.08% triethylamine).Gradient elution was performed at the flow rate of 1.0 mL·min^-1,the column temperature was 35℃,the detection wavelength was at 280 nm,and the eluting period were 70 min. Results: The investigated seven alkaloids including tetrahydrocolumbamine,protopine,tetrahydropalmatine,coptisine,palmatine,berberine,and dehydrocorydaline showed good linearity(r≥0.999 3)in the range of 1.12-56.0,1.27-63.5,3.37-168.5,1.02-51.0,0.89-44.5,0.88-44.0 and 3.22-161.0 μg·mL^-1,respectively;the average recoveries(n=6)were within 98.3%-103.6% with RSD≤2.8%.The contents of seven alkaloids in crude and processed Corydalis Rhizoma were in the range of 163.90-376.28,140.24-207.29,973.61-1 158.33,180.49-433.49,68.01-196.76,18.43-46.10,550.57-1 489.84 μg·g^-1. Conclusion: Compared to the crude Corydalis Rhizoma,the tetrahydrocolumbamine in the processed nethod was decreased,while the protopine,tetrahydropalmatine,palmatine,berberine,and dehydrocorydaline were increased to a certain degree,and the coptisine was unsteady.This established methods provide an effective means for quality control of vinegar Corydalis Rhizoma.