一测多评法同时测定制川乌中9种生物碱的含量

目的:建立一测多评法同时测定制川乌中9种乌头类生物碱含量.方法:采用高效液相色谱四级杆飞行时间串联质谱(HPLC-QTOF-MS)法.色谱柱为Agilent ZORBAX Extend-C18 RRHT(2.1 mm×50 mm,1.8μm);甲醇为流动相A,以水(含0.1%甲酸和2.5 mmol·L-1醋酸铵溶液)为流动相B,梯度洗脱;流速为0.21 mL·min-1;柱温为30℃.以乌头碱为内参物,测定次乌头碱、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、乌头原碱、次乌头原碱、新乌头原碱对于其的相对校正因子,并对相对校正因子进行重现性考察.利用相对校正因子计算其它8种生物碱成分的含量,同时利用外标法测定这9种成分,比较两种测定方法的差异,验证一测多评法的准确性和可行性.结果:建立QAMS法测定乌头类中药材9种生物碱的含量,其测定值与计算值无显著差异.结论:经方法学验证,本方法可用于制川乌药材中9种生物碱的含量测定.

Simultaneous Determination of Nine Aconitum Alkaloids in Radix Aconiti Praeparata with Quantitative Analysis of Multi-Components by Single Marker

Objective: To develop a method for the quantitative analysis of multi-components by single marker (QAMS) for nine alkaloids in Radix Aconiti Praeparata. Methods: An HPLC-QTOF-MS method with an Agilent ZORBAX Extend-C18 RRHT column (2.1 mm×50 mm, 1.8μm) was applied. The flow rate was 0.21 mL·min-1. The column temperature was 30℃. The mobile phase was methanol (A)-water (contain-ing 0.1% formic acid and 2.5 mmol·L-1 ammonium acetate) (B) in gradient elution. Aconitine was used as internal reference substance, to respectively establish the relative correction factor (RCF) of hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, benzoylmesaconine, aconine, hypaconine and mesaconine, and inspect the reproducibility of RCFs; the contents of the other 8 aconitum alkaloids were calculated ac-cording to the RCFs, respectively. At the same time, an external standard method (ESM) was performed for the content determination of the nine alkaloids. The two methods were compared. The feasibility and accu-racy of the QAMS method were verified. Results: The QAMS method was established for determining alka-loids. There was no significant difference between the QAMS method and the external standard method (ESM). Conclusion: With the validation of methodology, the method established in our study can be used for content determination of aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, ben-zoylmesaconine, aconine, hypaconine and mesaconine.