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. 2015 Oct-Dec;11(44):740-4.
doi: 10.4103/0973-1296.165574.

High performance liquid chromatography time of flight electrospray ionization mass spectrometry for quantification of sesquiterpenes in Chrysanthemi indici Flos active extract

Affiliations

High performance liquid chromatography time of flight electrospray ionization mass spectrometry for quantification of sesquiterpenes in Chrysanthemi indici Flos active extract

Ling Fu et al. Pharmacogn Mag. 2015 Oct-Dec.

Abstract

Background: Chrysanthemi indici Flos, a traditional herbal medicine is used to clearing heat-toxicity, removing the liver fire, and improving eyesight. In our preliminary work, an active extract of CTC in C. An indici Flos with anti-hepatitis B virus and liver protective activity was found by HepG2.2.1.5 test and experiment of protein synthesis in mice's injured liver. In this work, we aimed to study the active faction CTC further by qualitative and quantitative analysis method.

Materials and methods: High performance liquid chromatography time of flight electrospray ionization mass spectrometry (HPLC TOF ESI-MS) analysis method of the CTC was established. Cumambrin A and angeloylcumambrin B in CTC were analyzed by high performance liquid chromatography-ultraviolet-evaporative light scattering detector (HPLC-UV-ELSD) analysis methods. A binary gradient elution program was conducted for chromatographic separation with acetonitrile (A) and ultrapure water (B) as follows: 0-10 min, 42-46% A; 10-20 min, 46-55% A; 20-25 min, 55-60% A; and 25-35 min, 60-75% A. The column temperature and UV wavelength were set at 30°C and 205 nm.

Result: Ten constituents including (3R, 5R, 6S, 7S, 10R)-7-(2-hydroxy-2-propyl)-10-methyl-4-methyleneperhy, dronaphthal-ene-3, 5, 6-triol acetone solvate; (+)-edusmance-4, (14)-ene-11, 13-diol; linarin; luteolin; apigenin; tricin; 5, 3',4'- trimethyl-6,7-dimethoxy flavones; cumambrin A; acacetin; and angeloylcumambrin B in CTC were identified by HPLC TOF ESI-MS. The contents of sesquiterpenes in CTC were decreased by storing years.

Conclusions: The results showed that both UV and ELSD methods were feasible, accurate, and the determination results were in good consistency. The contents of two sesquiterpenes decreased with storing years. Two sesquiterpenes could be used as quality control for C. indici flos CTC.

Keywords: Angeloylcumambrin B; Chrysanthemi indici Flos; cumambrin A extract high performance liquid chromatography time of flight electrospray ionization mass spectrometry; high performance liquid chromatography-ultraviolet-evaporative light scattering detector.

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Conflict of interest statement

Conflict of Interest: None declared.

Figures

Figure 1
Figure 1
Bifendate Pills group, active extract in Chrysanthemi indici Flos low dosage group, active extract in Chrysanthemi indici Flos middle dosage group, active extract in Chrysanthemi indici Flos high dosage group. Left: Effects of active extract in Chrysanthemi indici Flos on the index of liver and spleen in mice injured by D-gal. Right: Effects of active extract in Chrysanthemi indici Flos on the total protein, albumin, and alanine transaminase in serum of mice injured by D-gal
Figure 2
Figure 2
The high performance liquid chromatography chromatograms of active extract in Chrysanthemi indici Flos and structures formulas of Peak 1, (3R, 5R, 6S, 7S, 10R)-7-(2-hydroxy-2-propyl)-10-methyl-4-methyleneperhy, dronaphthal-ene-3, 5, 6-triol acetone solvate; Peak 2, (+)-edusmance-4, (14)-ene-11, 13-diol; Peak 3, linarin; Peak 4: Luteolin; Peak 5, apigenin; Peak 6, tricin; Peak 7, 5, 3’, 4’-trimethyl-6,7-dimethoxy flavones; Peak 8, cumambrin A; Peak 9, acacetin; Peak 10, angeloylcumambrin B

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