Identification of Metabolites of 6'-Hydroxy-3,4,5,2',4'-pentamethoxychalcone in Rats by a Combination of Ultra-High-Performance Liquid Chromatography with Linear Ion Trap-Orbitrap Mass Spectrometry Based on Multiple Data Processing Techniques - PubMed (original) (raw)

Identification of Metabolites of 6'-Hydroxy-3,4,5,2',4'-pentamethoxychalcone in Rats by a Combination of Ultra-High-Performance Liquid Chromatography with Linear Ion Trap-Orbitrap Mass Spectrometry Based on Multiple Data Processing Techniques

Siyi Liu et al. Molecules. 2016.

Abstract

In this study, an efficient strategy was established using ultra-high-performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry (UHPLC-LTQ-Orbitrap MS) to profile the in vivo metabolic fate of 6'-hydroxy-3,4,5,2',4'-pentamethoxychalcone (PTC) in rat urine and feces. The UHPLC-LTQ-Orbitrap method combines the high trapping capacity and MS(n) scanning function of the linear ion trap along with accurate mass measurements within 5 ppm and a resolving power of up to 30,000 over a wider dynamic range compared to many other mass spectrometers. In order to reduce the potential interferences of endogenous substances, the post-acquisition processing method including high-resolution extracted ion chromatogram (HREIC) and multiple mass defect filters (MMDF) were developed for metabolite detection. As a result, a total of 60 and 35 metabolites were detected in the urine and feces, respectively. The corresponding in vivo reactions such as methylation, hydroxylation, hydrogenation, decarbonylation, demethylation, dehydration, methylation, demethoxylation, sulfate conjugation, glucuronide conjugation, and their composite reactions were all detected in this study. The result on PTC metabolites significantly expanded the understanding of its pharmacological effects, and could be targets for future studies on the important chemical constituents from herbal medicines.

Keywords: 6′-hydroxy-3,4,5,2′,4′-pentamethoxychalcone (PTC); UHPLC-LTQ-Orbitrap MS; metabolite identification; multiple data processing method.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1

Figure 1

The proposed major metabolic pathway of PTC in the rat urine and feces.

Figure 2

Figure 2

ESI-MSn spectra of four reference standards and M16: (A) MS2 spectrum of PTC; (B) MS2 spectrum of 6′-hydroxy-3,4,5,2′,3′,4′-hexamethoxychalcone; (C) MS2 spectrum of 4,6′-dihydroxy-3,2′,4′-trimythoxychalcone; (D) MS3 spectrum of 4,6′-dihydroxy-3,2′,4′-trimethoxychalcone (precursor-ion was m/z 149); (E) MS2 spectrum of velutin; (F) MS3 spectrum of velutin (precursor-ion was m/z 298); (G) MS2 spectrum of M16.

Figure 3

Figure 3

Chemical structures of the reference standards.

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