Metabolic fate analysis of Huang–Lian–Jie–Du Decoction in rat urine and feces by LC–IT-MS combining with LC–FT-ICR-MS: a feasible strategy for the metabolism study of Chinese medical formula

Abstract

1. Huang-Lian-Jie-Du Decoction (HLJDD) is widely used for the treatment of hypertension, diabetes, inflammation and neural system diseases in clinic. In the present study, the comprehensive metabolic profile of HLJDD was demonstrated reliably and rapidly followed by the metabolic pathway analysis of six typical pure compounds (four alkaloids, one flavonoid and one iridoid) in HLJDD using LC-IT-MS combined with high resolution LC-FT-ICR-MS. 2. Totally, 85 compounds, including 32 prototype components and 53 biotransformed metabolites were detected and characterized in the urine and feces after oral administration of HLJDD and six pure compounds to rats, respectively. Among them, 17 prototypes were identified definitely with standard references. 3. Hydroxylation, demethylation and glucuronidation reactions of alkaloids, as well as glucuronidation and sulfonation reactions of iridoids and flavonoids, were observed as the major metabolic pathways of HLJDD. Flavonoids, iridoids and their metabolites were mainly excreted from urine. However, amount of alkaloids were detected in feces. 4. In general, the distinctive metabolic process of three kinds of representative components in HLJDD was clarified. The in vivo metabolic network of HLJDD was demonstrated. Meanwhile, the investigation of representative pure compounds in metabolic study provided a valuable strategy to elucidate the full-scale metabolic fate of HLJDD. This might be helpful to understand the in vivo mechanism of Traditional Chinese medicine (TCM).

Metabolic profile of Rhizoma coptidis in human plasma determined using ultra‐high‐performance liquid chromatography coupled with high‐resolution mass spectrometry

“…Their MS 3 fragment at m/z 307.0855 attributed to [M + − C 6 H 8 O 6 − ▪CH 3 ] + was observed as well. The full MS and MS n fragments of M1 , M2 and M7 (Figure and Table ) were the same as those previously reported for the metabolites of Rhizoma coptidis . Thus, by comparing with the literature, M1 was determined to be dehydrocheilanthifoline 2‐ O ‐glucuronide or groenlandicine 3‐ O ‐glucuronide, while M2 and M7 were deduced to be thalifendine 10‐ O ‐glucuronide and berberrubine‐9‐ O ‐glucuronide, respectively.…”

“…The product ion at m/z 338.1401 was observed in their MS/MS spectra corresponding to a loss of 176 Da, which indicated that a GluA unit was attached to the aglycone moiety (Figure S4, supporting information). Their MS 3 fragments at m/z 323.1167 and 294.1138 were the same as those reported in the literature, and they were assigned as jatrorrhizine‐3‐ O ‐glucuronide and columbamine‐2‐ O ‐glucuronide, respectively. However, the accurate site of glucuronidation of M3 and M8 could not be confirmed.…”

“…However, the structures of the compounds in TCMs are so complex and many metabolites contain similar moieties, that it would be difficult to unambiguously confirm all the constituents solely by mass spectrometry. 49 Thus, the metabolite identification in the present study is preliminary and speculative. In addition, to obtain a more scientific and comprehensive metabolite profile of Rhizoma coptidis in vivo, our study collected human plasma at 14 different time points (0.08, 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 7, 9 and 12 h) following administration.…”

“…Based on the metabolites detected in human plasma, a possible metabolic pathway of Rhizoma coptidis in vivo was proposed (Figure ). However, the structures of the compounds in TCMs are so complex and many metabolites contain similar moieties, that it would be difficult to unambiguously confirm all the constituents solely by mass spectrometry . Thus, the metabolite identification in the present study is preliminary and speculative.…”

“…, MS 2 spectra and B, MS 3 spectra of M1, M2 and M7 [Color figure can be viewed at wileyonlinelibrary.com] groenlandicine 3-O-glucuronide,36,49 while M2 and M7 were deduced to be thalifendine 10-O-glucuronide and berberrubine-9-Oglucuronide,53,54 respectively.M3 and M8 were isomers that eluted at t R = 10.3 min and 11.4 min, respectively (Figure S3, supporting information). They showed the same [M] + ion at m/z 514.1708 (C26 H 28 O 10 N, calculated 514.1708; error, 0.13 ppm) in positive ion mode.…”