Rapid identification of absorbed components and metabolites of Gandou decoction in rat plasma and liver by UPLC-Q-TOF-MSE
Graphical abstract
Introduction
Wilson’s disease (WD), also known as hepatolenticular degeneration, is a genetic disorder of copper metabolism due to ATP7B gene mutation. It is an autosomal recessive illness characterized by excessive accumulation of copper in the liver, brain and other tissues, leading to various neurologic, psychiatric and hepatologic symptoms [1], [2]. Typical Western medicines, such as D-penicillamine, dimercaptosuccinic acid, sodium dimercaptosuccinate and tetrathiomolybdate, can be used for treating WD, but their toxic side effects seriously affect the patient's condition and prognosis [3], [4], [5]. In China, Gandou decoction (GDD), a traditional Chinese medicine prescription (TCMP), has been widely used to treat WD for decades. Due to its remarkable clinical effect, GDD has been recommended as the basic prescription recorded in “The Diagnosis and Treatment Guidelines of WD” in China [6], [7].
GDD is composed of six medicinal materials, including Rheum palmatum L. (Da-Huang), Coptis chinensis Franch. (Huang-Lian), Curcuma longa L. (Jiang-Huang), Lysimachia christinae Hance (Jin-Qian-Cao), Alisma orientale (Sam.) Juzep. (Ze-Xie) and Panax notoginseng (Burk.) F. H. Chen (San-Qi). Modern pharmacological researches have demonstrated that GDD appears to be effective, safe, mild adverse reactions and well-tolerated [8]. Our previous studies found that GDD could reduce the hepatic copper accumulation and improve liver pathological characteristics by restoring the impaired lipid metabolism, amino metabolism and glucose metabolism [9]. For the constituent research in our lab, 96 compounds have been isolated from GDD as well as its relative individual herbs [10]. However, the absorbed components and metabolites of GDD in vivo still remain unclear. Therefore, there is an urgent need for systematic research on the metabolic profiling of GDD.
It is an arduous task to identify the prototypes and metabolites of herbal medicines and TCMP in biologic samples due to their very low levels of concentration in vivo, diverse structure types, unpredictable metabolites and the interference from endogenous substances, resulting in incomprehensive and unsystematic constituent characterization [11], [12]. In recent years, ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MSE) has been used as an effective analytical tool for chemical characterization and metabolite identification because of its high resolution, sensitivity and accuracy. TOF-MSE technology uses two interleaved scan functions with different collision energies, allowing for the simultaneous acquisition of precursor ions with low collision energy and fragment ions with high collision energy [13]. In the present study, an efficient approach was applied to rapidly characterize the metabolic profiling of GDD in plasma and liver between normal and copper-laden rats based on UPLC-Q-TOF-MSE. It is expected that the current work could provide a detailed basis for revealing the potential effective materials in vivo, and contribute to the further pharmacology and curative mechanism research on GDD.
Section snippets
Chemicals and reagents
Rheum palmatum L., Coptis chinensis Franch., Curcuma longa L., Lysimachia christinae Hance, Alisma orientale (Sam.) Juzep. and Panax notoginseng (Burk.) F. H. Chen were purchased from Beijing Tongrentang Pharmacy (Hefei, China). The medicinal plants were authenticated by Doctor Rongchun Han (College of Pharmacy, Anhui University of Chinese Medicine, Hefei, China). All voucher specimens were deposited at the authors' laboratory. Eleven compounds were used as reference standards (purity > 98%),
Determination of serum and hepatic copper levels
The abnormal elevation of serum and hepatic copper levels can be used as an indicator of copper-laden rat model [[15], [18]]. As shown in Table 1, serum copper levels showed nonsignificant changes in 30-day copper-laden model group compared to those in control group, but copper levels in 60-day and 90-day model groups were significantly different from those in control group (P < 0.05). Hepatic copper accumulation increased considerably during the experiment, in which copper levels in 30-day,
Conclusion
In this study, we successfully established a mature copper-laden rat model by intragastric administration of CuSO4. A rapid and high-throughput UPLC-Q-TOF-MSE was applied for systematic research on the metabolic profiling of GDD in vivo between normal and copper-laden rats. As a result, a total of 89 GDD-related xenobiotics, including 83 (36 prototypes and 47 metabolites) in plasma and 52 (21 prototypes and 31 metabolites) in liver, were detected in normal dosed rats; a total of 77 GDD-related
CRediT authorship contribution statement
Li Xu: Data curation, Validation, Writing - review & editing. Yi Liu: Methodology, Data curation, Writing - original draft. Hongfei Wu: Resources, Supervision, Visualization, Investigation. An Zhou: Conceptualization, Supervision, Project administration, Funding acquisition.
Acknowledgments
This research was supported by the National Natural Science Foundation of China (No: 81872976, 81873038, 81303245), Key Project of the National Science Fund of Anhui Province (No: KJ2016A398), Key Project Foundation of Oversea Visiting and Research for the Excellent Young and Middle-aged Faculties in Universities of Anhui Province (No: gxgwfx2018042), and Primary Research & Development Plan of Anhui Province (No: 201904a07020051).
References (27)
- et al.
Wilson's disease
Lancet
(2007) - et al.
Wilson's disease and other neurological copper disorders
Lancet Neurol.
(2015) - et al.
Novel perspectives on Wilson disease treatment
Handb. Clin. Neurol.
(2017) - et al.
Clinical efficacy and safety of Chinese herbal medicine for Wilson's disease: a systematic review of 9 randomized controlled trials
Complement. Ther. Med.
(2012) - et al.
Metabolic profiling of copper-laden hepatolenticular degeneration model rats and the interventional effects of Gandou decoction using UPLC-Q-TOF/MS
J. Pharm. Biomed. Anal.
(2019) - et al.
UPLC-Q-TOF-MSE analysis of the constituents of Ding-Zhi-Xiao-Wan, a traditional Chinese antidepressant, in normal and depressive rats
J. Chromatogr. B
(2016) - et al.
Relationship of antioxidant and oxidative stress markers in different organs following copper toxicity in a rat model
Toxicol. Appl. Pharmacol.
(2016) - et al.
Identification and analysis of components in Shen-Fu-Shu granule extract and in rat plasma after oral administration by UPLC-ESI/Q-TOF-MS
J. Pharm. Biomed. Anal.
(2019) Wilson disease - liver pathology
Handb. Clin. Neurol.
(2017)- et al.
Metabolites profile of Gualou Xiebai Baijiu decoction (a classical traditional Chinese medicine prescription) in rats by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry
J. Chromatogr. B
(2018)
An untargeted metabolomics-driven approach based on LC-TOF/MS and LC-MS/MS for the screening of xenobiotics and metabolites of Zhi-Zi-Da-Huang decoction in rat plasma
J. Pharm. Biomed. Anal.
Rapid characterization of chemical constituents and rats metabolites of the traditional Chinese patent medicine Gegen-Qinlian-Wan by UHPLC/DAD/qTOF-MS
J. Pharm. Biomed. Anal.
Use of ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry system as valuable tool for an untargeted metabolomic profiling of Rumex tunetanus flowers and stems and contribution to the antioxidant activity
J. Pharm. Biomed. Anal.
Cited by (17)
Identification and characterization of quinoline alkaloids from the root bark of Dictamnus dasycarpus and their metabolites in rat plasma, urine and feces by UPLC/Qtrap-MS and UPLC/Q-TOF-MS
2021, Journal of Pharmaceutical and Biomedical AnalysisCitation Excerpt :During the past decades, UPLC coupled with MS/MS, such as quadrupole-time of flight MS (Q-TOF-MS) and hybrid quadrupole-liner ion trap MS (Qtrap-MS), has been developed as a frequently-used method for the identification of herb components and metabolites due to its superior separation efficiency, detection sensitivity, and structural characterization potency [11–13]. Q-TOF-MS has been shown to be intrinsically capable of comprehensively acquiring accurate mass spectral data based on MS1 full scan and MSE-based data-independent acquisition (DIA), indicating promising potential for the global chemical profiling of complex system and minor components such as plasma and urine [14,15]. Although mass defect filter (MDF) had been developed to simplify the data processing [16], it is still a complicated task to exactly assign the fragment ions to their precursor for the co-eluting components.
Liver and urine metabolomics reveal the protective effect of Gandou decoction in copper-laden Hepatolenticular degeneration model rats
2021, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life SciencesCitation Excerpt :First, 96 chemical constituents including flavonoids, anthraquinones, and triterpenoid saponins, were identified from GDD using UPLC-Q-TOF-MSE and UNIFI informatics platform [13]. Subsequently, we identified major bioactive components and metabolites of GDD in the liver, plasma, and urine between normal and copper-laden rats (an animal model of HLD) [14,15]. Our research further indicated that GDD reduces copper accumulation and alleviated liver damage in copper-laden rats by regulating the serum metabolic profile and copper levels [15].
Deciphering the metabolic profile and pharmacological mechanisms of Achyranthes bidentata blume saponins using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry coupled with network pharmacology-based investigation
2021, Journal of EthnopharmacologyCitation Excerpt :These characteristics of UPLC-QTOF/MSE make it uniquely suited for structural characterization (Fu et al., 2019), metabolomics analysis (Wu et al., 2018b) and metabolites profiling (Gu et al., 2019). Furthermore, a number of powerful post-acquisition data processing platforms (such as UNIFI and MetaboLynx) have been successfully developed for prediction of biological transformation and mining of exogenous metabolites in complex biological samples (Ameline et al., 2019; Fu et al., 2019; Xu et al., 2020). In a previous study by our group, fingerprint analysis of AB (Li et al., 2018) and chemical and in vitro intestinal metabolic profiles of ABS (Fu et al., 2019) were evaluated.
Determination of complexation ability of rhubarb with copper ions by ultra-high performance liquid chromatography
2023, Chinese Journal of Chromatography (Se Pu)