Planta Med 2010; 76 - P16
DOI: 10.1055/s-0030-1251778

Evaluation of Processing Degree of Aconitum carmichaeli Debx. for Simultaneous Determination of Six Main Components with Liquid Chromatography-tandem Mass Spectrometry

XL Cheng 1, XY Xiao 1, ZL Bai 2, YM Zhang 1, F Wei 1, YZ Ren 2, SC Ma 1, RC Lin 1
  • 1Research and Inspection Center of Traditional Chinese Medicine and Ethnomedicine, National Institute for the Control of Pharmaceutical & Biological Products, State Food and Drug Administration, 2 Tiantan Xili, 100050 Beijing, China
  • 2Research and development department, China National Group Corp. of Traditional & Herbal Medicine, No. 45, Shuntong Road, Linhe Industry Development Area, Shunyi District, Beijing, China, 101300, Beijing, P.R.China

Common Monkshood Root (Aconitum carmichaeli Debx.) is widely distributed over China. Its main roots are called chuangwu while its lateral roots are called fuzi in Chinese, which are useful herbal medicines. The raw chuangwu and fuzi cannot be used directly because of their high contents and toxicity. Diester diterpene-type Aconitum alkaloids (e.g.aconitine (AC), hypaconitine (HA), and mesaconitine (MA)) [1] are the main compounds of the raw chuangwu and fuzi. The raw chuangwu and fuzi are clinically used only after cautiously processed so that AC, HA, and MA will be hydrolyzed to BMA, BAC, BHA in order to reduce their toxicity [2]. So the different preparations of crude drugs of Aconitum species should be fully controlled for their quality and safety.

A specific and sensitive liquid chromatography tandem mass spectrometry (LC/MS/MS) method has been developed for simultaneous determination of six Aconitum alkaloids in Common Monkshood Root and their processing products. The separation was applied on an Agilent ZORBAX Extend C18 column (2.1mm×50mm, 1.8µm) and a gradient elution of acetonitrile and 40 mmol/L CH3COONH4 in water (pH 10.5, adjust the pH by NH4 · OH) was used as mobile phase. LC/MS/MS system coupled with an electrospray ionization (ESI) source was performed in multiple reaction monitoring (MRM) mode. The transitions of the Aconitum alkaloids executed as follows: m/z 590.1→540.0 for BMA; m/z 604.2→554.0 for BAC; m/z 574.1→542.3 for BHA; m/z 632.5→572.8 for MA; m/z 646.3→586.7 for AC; m/z 616.2→566.7 for HA. The concentrations ranged from 1 to 200ng/mL for all six Aconitum alkaloids and showed a good linearity with the correlation coefficient (r2) >0.995. The limits of detection (LODs) for these six standards were 1.012, 1.022, 1.04, 2.012, 1.014, 2.144 pg. The validated method was employed to simultaneous determination of six main components to evaluate the processing degree of Aconitum carmichaeli Debx. The ratio of BMA, BAC, BHA and a AC, HA, MA will show the processing degree of Common Monkshood Root.

Fig.1

Name

R1

R2

R3

AC

C2H5

OH

acetyl

MA

CH3

OH

acetyl

HA

CH3

H

acetyl

BAC

C2H5

OH

H

BMA

CH3

OH

H

BHA

CH3

H

H

References: [1] Wong SK, Tsui SK, et al. (2002)J Pharm Biomed Anal 30: 161–170. [2] Bisset NG, (1981)J Ethnopharmacol. 4: 247.