Quantitative LC-MS/MS analysis of seven ginsenosides and three aconitumalkaloids in Shen-Fu decoction
Chemistry Central Journal
Quantitative LC-MS/MS analysis of seven ginsenosides and three aconitum alkaloids in Shen-Fu decoction
Na Guo
Mingtao Liu
Dawei Yang
Ying Huang
Xiaohong Niu
Ruifan Wu 1
Ying Liu
Guizhi Ma 1
Deqiang Dou 0
0 Department of Chinese Medicine Chemistry, Liaoning University of Traditional Chinese Medicine , Dalian 116600 , China
1 College of Pharmacy, Xinjiang Medical University , Urumqi 830011 , China
Background: Shen-Fu decoction is a traditional Chinese medicine prescription with a 3:2 ratio of Radix Ginseng and Fuzi (Radix Aconiti lateralis praeparata). Ginsenosides and alkaloids are considered to be the main active components of Shen-Fu decoction. However, no analytical methods have been used to quantitatively analyse both components in Shen-Fu decoction simultaneously. Results: We successfully developed a rapid resolution liquid chromatography coupled with tandem mass spectrometry (RRLC-MS/MS) method for the simultaneous analysis of seven ginsenosides and three aconitum alkaloids in Shen-Fu decoction, the decoction of Radix ginseng and Fuzi (Radix Aconiti lateralis praeparata). Chromatogrpahic separation by RPLC was achieved using a reversed-phase column and a water/acetonitrile mobile phase, containing 0.05% formic acid and using a gradient system. The method was optimized to allow for simultaneous analysis of all analytes in 11minutes without the need for baseline resolution of the components. Furthermore, the separation demonstrated good linearity (r > 0.9882), repeatability (RSD < 7.01%), intra- and inter-day precisions (RSD < 5.06%) and high yields of recovery (91.13-111.97%) for ten major constituents, namely ginsenoside-Re, Rg1, Rb1, Rc, Rb2, Rd, Rf, aconitine, hypacoitine and mesaconitine. Conclusions: The developed method could be used as a rapid and reliable approach for assessment of the quantity of the major constituents in Shen-Fu decoction.
Ginsenosides; Aconitum alkaloids; Shen-Fu decoction; RRLC-MS/MS
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Background
Decoction is the traditional prescription of traditional
Chinese medicines (TCMs). Based on TCM theory, one
single herb or several kinds of herbs combined are
boiled in water to make the decoction. First documented
in 1465, Shen-Fu decoction is a TCM prescription with
a 3:2 ratio of Radix Ginseng and Fuzi (Radix Aconiti
lateralis praeparata). Both components have been
commonly used as herbal medicines in China for about
1800 years, predominantly used for folk treatment of
diseases with the sign of Yangqi decline or Yang exhaustion.
Shen-Fu decoction is also used to treat cardiovascular
diseases such as circulatory collapse, shock, thoracic
obstruction and acute thoracic pain. Shen-Fu Injection (SFI
for intravenous medication), is a typical form of Shen-Fu
decoction, that has been used for treatment of many
kinds of diseases because of its cardiovascular protective
effectiveness [1-3]. The main active components found
in Shen-Fu decoction are ginsenosides and alkaloids.
Ginsenosides are generally classified into four groups:
protopanaxadiol, protopanaxatriol, ocotillol and oleanolic
acid type [4-6], Currently, more than 150 ginsenosides
have been isolated and identified in the literature.
Among them, ginsenosides-Rb1, Rb2, Rc, Rd, Rg1, Re and
Rf (Figure 1) are the most important compounds in
chemical analysis of ginsengs. At present, about 224 alkaloids
have been isolated and identified from Aconitum [7,8].
-O-Glc6-1Glc
-O-Glc6-1Arap
-O-Glc6-1Araf
-O-Glc
-O-Glc2-1Rha
-O-Glc
-O-Glc2-1Glc
Protopanaxadiol-type
Ginsenoside-Rb1
Ginsenoside-Rb2
Ginsenoside-Rc
Ginsenoside-Rd
Protopanaxadiol-type
Ginsenoside-Re
Ginsenoside-Rg1
Ginsenoside-Rf
-O-Glc2-1Glc
-O-Glc2-1Glc
-O-Glc2-1Glc
-O-Glc2-1Glc
Figure 1 Chemical structures of ginsenosides and Aconitum alkaloids analyzed in Shen-Fu decoction.
These have been classified into four major groups, nonester
alkaloids (NEAs), monoester diterpene alkaloids (MDAs),
diester diterpene alkaloids (DDAs) and lipoalkaloids.
Aconitum alkaloids are mainly constituted of three DDAs,
diester-diterpence called aconitine (AC), measaconitine
(MA) and hypaconitine (HA) (Figure 1). They are known
for their high toxicity and pharmacological activity, as well
as being the target markers of Fuzi. In general, the
curative effect of traditional Chinese medicine is an integrative
result of a number of ginsenosides and alkaloids. In order
to minimize the variability of active ingredients in the
decoction and ensure repeatable and reproducible
therapeutic effects, it is very important to establish quality
control methodology for the decoction. To this end,
analysis of ginsenosides and Aconitum alkaloids is required
to assess the quality of Shen-Fu decoction.
Previous methods that have been used to analyze
ginsenosides and alkaloids include HPLC-DAD (ELSD),
CE, GC-MS and LC-MS [9-19] and alkaloids [20-30]. In
comparison with traditional HPLC, RRLC provides a
higher peak capacity, greater resolution, increased
sensitivity and higher speed o (...truncated)