Determination of Aconitum Alkaloids in Dietary Supplements and Raw Botanical Materials by Liquid Chromatography/UV Detection with Confirmation by Liquid Chromatography/Tandem Mass Spectrometry: Collaborative Study

Abstract

An interlaboratory study was conducted to evaluate a method for the determination of 3 Aconitum alkaloids, viz., aconitine, mesaconitine, and hypaconitine, in raw botanical material and dietary supplements. The alkaloids were extracted with diethyl ether in the presence of ammonia. After cleanup by solid-phase extraction to remove matrix interferences, the alkaloids were determined by reversed-phase liquid chromatography (LC)/UV detection at 235 nm with confirmation by LC/tandem mass spectrometry (MS/MS). A total of 14 blind duplicates were successfully analyzed by 12 collaborators. For repeatability, the relative standard deviation (RSDr) values ranged from 1.9 to 16.7, and for reproducibility, the RSDR values ranged from 6.5 to 33. The HorRat values were all <2 with only one exception at 2.3. All collaborating laboratories had calibration curves with correlation coefficients of >0.998. In addition, 6 collaborators performed the confirmation and were able to verify the identities of the alkaloids by using LC/MS/MS.

Quantification and Simplified Detoxification Investigation on Fuzi, Root of Aconitum carmichaelii

“…The standard solutions of aconitine, hypaconitine, and mesaconitine were prepared through procedures for analysis. 18 UPLC operating conditions were as follows: injection volume: 2 µL; TQ detector was set to selected reaction monitoring mode with positive electrospray ionization. The analytical column for DDA analysis was an ACQUITY UPLC BEH C18 column (1.7 µm, 50 mm × 2.1 mm) (Waters).…”

“…The concentration of phase A decreased linearly to 20% within 30 minutes. Aconitine, mesaconitine, and hypaconitine precursor ions ([M + H] + ) and their corresponding fragments were monitored (m/z 646→586, m/z 632→572, and m/z 616→556) 18 ( Figure 3).…”

“…The preparation of extract for investigation of DDA analysis was as follows 18 : Accurately weigh 1.0 g sample and put it into a 50 mL centrifuge tube. Add 1 mL 10% ammonium hydroxide solution.…”