Determination of Alkaloid Content and Limit Check of Aconiti Lateralis Radix Praeparata in Classic Formula Zhenwu Decoction
Objective:To establish a high-performance liquid chromatography(HPLC)method for the simultaneous determination of benzoylmesaconine,benzoylaconine,benzoylhypaconitine,mesaconitine,hypaconitine,and aconitine in Zhenwu Decoction.Methods:HPLC was applied using an XBridge C18 column(250 mm×4.6 mm,5 μm)with gradient elution to measure the content of alkaloid components in Zhenwu Decoction.The method also referred to the 2020 edition of the pharmacopoeia of the people's republic of China for the limit check of diester alkaloids.Results:The mass concentrations of benzoylmesaconine,benzoylaconitine,benzoylhypaconitine,mesaconitine,hypaconitine,and aconitine were in the ranges of 9.314-93.140(r=0.999 6),10.596-105.960(r=0.999 7),6.848-68.480(r=0.999 7),1.515-151.540(r=0.999 7),1.434-143.420(r=0.999 8),and 1.520-152.020 μg·mL-1(r=0.999 7),respectively,showing a good linear relationship with peak areas.The spiked recovery rates ranged from 98.41%-102.17%,with RSDs less than 3.00%.In 15 batches of Zhenwu Decoction,the powder yield of the reference substance was 10.86%-17.73%.The mass fractions of single ester-type alkaloids,namely benzoylmesaconine,benzoylaconitine,and benzoylhypaconitine,were 27.49-109.14,3.48-13.42,and 4.76-17.63 μg·g-1,respectively.The total mass fractions of diester-type alkaloids(mesaconitine,hypaconitine,and aconitine)were 2.55-17.67 μ g·g-1.The average transfer rates of benzoylmesaconine,benzoylaconitine,benzoylhypaconitine,and hypaconitine from decoction pieces to the reference substance were 19.93%,22.34%,17.95%,and 26.39%,respectively.Conclusion:The established HPLC method is simple,reliable,and accurate,and can be used for the determination of single ester-type and diester-type alkaloids in Zhenwu Decoction,which can provide a reference for quality control of related preparations of Zhenwu Decoction.
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