•A curcuminoid profile is established by two complementary LC–MS/MS platforms.•Ninety-six curcuminoids were fully characterized using the profile.•The curcuminoid profile is successfully applied to the quality evaluation of turmeric.Turmeric and curcuminoids are used as natural food coloring and functional food additives in various parts of the world. In this study, ninety-six curcuminoids were fully characterized using a targeted curcuminoid profile, which established by integrated use of two complementary LC–MS/MS platforms (liquid chromatography–quadrupole time of flight mass spectrometry (LC–QTOF-MS/MS) and liquid chromatography–quadrupole linear ion trap mass spectrometry (LC–QTRAP-MS/MS)). The curcuminoid profile was represented in the form of a multiple reaction monitoring (MRM) mode based on LC–QTRAP-MS/MS analysis. It facilitated the qualitative and relative quantitative analysis of curcuminoids in a single injection. Meanwhile, the profile was successfully applied to the quality evaluation of raw materials of turmeric from different regions in China and Myanmar. The structural identification procedures of curcuminoids and the integrated strategy provide a suitable method to analyze targeted plant metabolites which occur in a high number but sharing either structural similarities or similar functional groups.

•This work is the most comprehensive characterization of triterpenes in Alismatis rhizome (AR) to date.•Eighty triterpenes were identified in AR by QTOF-MS/MS.•A characteristic chemical profile (CCP) of triterpenes was established by QTOF and QTRAP-MS/MS.•Seven more triterpenes were discovered using MRM-based CCP in the processed AR.It was reported that triterpenes compounds in Alismatis rhizoma (AR) contributed to the lipid lowering effect on high fat diet (HFD)-induced hyperlipidemia. To date only 24 triterpenes (including the isomers) were characterized by LC-QTOF-MS/MS due to the lack of strategies for systematic discovery, classification and identification of triterpenes in AR. In this study, an integrated strategy combining various QTOF-MS/MS and QTRAP-MS/MS scan functions was developed for systematic identification and specific characterization of triterpenes in AR and processed AR. First, MS/MS fragmentation behaviors of different types of triterpenes were investigated and their diagnostic product ions were systematically summarized for discovery and classification of triterpenes. Second, diagnostic product ions were used to filter the data acquired by UHPLC-QTOF MS/MS for efficient identification of targeted triterpenes. Third, MRM-based characteristic chemical profile (CCP) of triterpenes was established using 30 MRM transitions by UHPLC-QTRAP-MS/MS. Fourth, MRM-based CCP was applied for comparative analyses of triterpenes in AR from different regions and from two other processed AR (salt processed AR and bran processed AR). Consequently, a total of 80 triterpenes including 14 novel compounds were identified in the AR, and 7 more triterpenes compounds were discovered using MRM-based CCP in the processed AR. This work is the most comprehensive characterization of triterpenes compounds in AR to date. The established MRM-based CCP of triterpenes compounds can be instructive for qualitative analyses and relative quantitative analyses of triterpenes in AR and its related medicinal products for potential applications including quality control and classification of different AR materials.

•Quantification of LC-B and its metabolite, LC-A, in rat urine and feces samples.•A SPE procedure was optimized to clean-up urinary and fecal samples.•Samples acidified with formic acid allow a high extraction recovery.•CHAPS was used to overcome nonspecific adsorption in urine.Capilliposide B, a novel oleanane triterpenoid saponin isolated from Lysimachia capillipes Hemsl, showed significant anti-tumor activities in recent studies. To characterize the excretion of Capilliposide B, a reliable liquid chromatography-tandem mass spectrometry (LC–MS/MS) method was developed and validated for simultaneous determination of Capilliposide B and its active metabolite, Capilliposide A in rat urine and feces. Sample preparation using a solid-phase extraction procedure was optimized by acidification of samples at various degrees, providing extensive sample clean-up with a high extraction recovery. In addition, rat urinary samples were pretreated with CHAPS, an anti-adsorptive agent, for overcoming nonspecific analytes adsorption during sample storage and process. The method validation was conducted over the curve range of 10.0–5000 ng/ml for both analytes. The intra- and inter-day precision and accuracy of the QC samples showed ≤11.0% RSD and −10.4–12.8% relative error. The method was successfully applied to an excretion study of Capilliposide B following intravenous administration.

•Oxidative and reactive metabolites were simultaneously characterized.•On-line UV detection was employed for semi-quantitation of major metabolites.•Multiple data mining methods were used for efficient discovery of metabolites.The use of liquid chromatography (LC) coupled with triple quadrupole linear ion trap (Qtrap) mass spectrometry (MS) for both quantitative and qualitative analysis in drug metabolism and pharmacokinetic studies is of great interest. Here, a new Qtrap-based analytical methodology for simultaneous detection, structural characterization and semi-quantitation of in vitro oxidative metabolites and glutathione trapped reactive metabolites was reported. In the current study, combined multiple ion monitoring and multiple reaction monitoring were served as surveying scans to trigger product ion spectral acquisition of oxidative metabolites and glutathione adduct, respectively. Then, detection of metabolites and recovery of their MS/MS spectra were accomplished using multiple data mining approaches. Additionally, on-line ultraviolet (UV) detection was employed to determine relative concentrations of major metabolites. Analyses of metabolites of clozapine and nomifensine in rat liver microsomes not only revealed multiple oxidative metabolites and glutathione adducts, but also identified their major oxidative metabolism and bioactivation pathways. The results demonstrated that the LC/UV/MS method enabled Qtrap to perform the comprehensive profiling of oxidative metabolites and glutathione adducts in vitro.

•An UHPLC-QTOF-MS/MS method for screening bioactive glycoside metabolites group in rat intestinal microflora.•Bioactivity of glycoside metabolites was evaluated by cell-based assay.•Deglycosylation and esterolysis were proposed as the major metabolic pathways of LC-C in rat intestinal microflora.•M4, an esterolysis product of LC-C, was obtained and evaluated for its bioactivity in vitro model.Many plant-derived glycosides are used as medications. It is common that these glycosides show poor intestinal absorption but their metabolites generated by intestinal microflora demonstrate strong bioactivity. Hence, it is crucial to develop a method for the identification and characterization of the metabolites, and consequently reveal the pathway in which the glycosides are processed in gut. In this study, cell-based assays in combination with ultra-high performance liquid chromatography-quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS/MS) were developed for rapid discovery and evaluation of the metabolites of a glycoside compound, capilliposide C (LC-C). 92.7% of LC-C was biotransformed by rat intestinal microflora after 36-h incubation at 37 °C. Human cancer cell lines HepG2, PC-3 and A549 was treated with metabolites pool, respectively, which was followed by cell viability assays and characterization of metabolites using UHPLC-QTOF-MS/MS. As a result, significant cytotoxicity was observed for the metabolites pool, from which six metabolites were identified. Based on the metabolites identified, deglycosylation and esterolysis were proposed as the major metabolic pathways of LC-C in rat intestinal microflora. In addition, M4, an esterolysis product of LC-C, was obtained and evaluated for its bioactivity in vitro. As a result, M4 exhibited a reduction in cell viability in HepG2 with an IC50 value of 17.46 ± 1.55 μg/mL.

•An analytical method that quantifies 25 bile acids in biological samples was developed.•Bile acid profiles were systematically compared between db/db and wt mice.•Plasma bile acid profile can best differentiate the two groups.•Hepatic gene Cyp7b1 was downregulated and Hsd3b7 upregulated in db/db mice.This study aimed to obtain information on bile acid profiles in diabetic (db/db) mice and their wild type (wt) littermates for the understanding of pathogenesis and discovery of potential biomarkers of type 2 diabetes. Analytical methods based on protein precipitation or solid-phase extraction together with liquid chromatography-tandem mass spectrometry were developed for the determination of 25 bile acids in plasma, urine and feces samples collected from db/db and wt mice. GLP-1 concentration and hepatic genes related to bile acid synthesis were also investigated. The results showed that the concentrations of individual bile acids varied notably both interindividually and temporally. However, plasma, urine and feces samples displayed discriminating bile acid profiles between the db/db and wt groups, with the plasma profile showing the best differentiation capacity. In plasma and urine, the concentration variation of taurine-conjugated bile acids was more correlated with that of other taurine-conjugated bile acids, and vice versa for the unconjugated bile acids. Transcription of hepatic gene Cyp7b1 was downregulated, and Hsd3b7 upregulated in db/db mice. In conclusion, the bile acid profile, particularly that in plasma, can distinguish the two animal groups and is a promising biomarker for type 2 diabetes.

•Diagnostic and characteristic product ions of shikonins and shikonofurans were determined.•Various mobile phase compositions and UHPLC elution programs were evaluated.•A data mining strategy was developed in complex samples.•Comparative analyses in three Zicao species samples were conducted.Shikonin, shikonofuran and their derivatives are the main bioactive components of Zicao, a traditional Chinese herbal medicine prepared with the dried roots of Lithospermum erythrorhizon, Arnebia euchroma or Arnebia guttata. However, approaches on the systematic discovery and identification of shikonins and shikonofurans, especially unknown ones, are still not available. To address this issue, the gas-phase CID-fragmentation routes for the shikonins and shikonofurans were established by using ESI-QTOF-MS in the negative ion mode and low-energy collision induced dissociation tandem mass spectrometry (CID-MS/MS) in this study using seventeen standards. As a result, diagnostic product ions for rapid discovery and classification of shikonins and shikonofurans were determined. In addition, various mobile phase compositions and UHPLC elution programs were evaluated to achieve optimal separation efficiency and detection response of these types of analytes. Based on these findings, an integral approach using UHPLC-ESI-QTOF-MS and CID-MS/MS analyses together with a novel two steps data mining strategy was developed for systematic analysis of shikonins and shikonofurans in complex samples. Consequently, 58 compounds including 32 novel ones were efficiently discovered and identified from the crude extract of Zicao. Moreover, comparative analyses of the 58 chemical components in three Zicao species including Lithospermum erythrorhizon, Arnebia euchroma and Arnebia guttata samples were conducted using the established analytical method, which can be instructive for future utilization of Zicao and its related medicinal products.

•Fragmentation pathways and rules of physalins were obtained.•Diagnostic ions for rapid screening of physalins were determined.•Diagnostic ions for classification of physalins were determined.•A novel three-step data mining strategy for analysis of MS/MS data was established.•A UHPLC–QTOF-MS/MS method was established for efficient identification of physalins.Physalins, uniquely discovered from genus physalis, showed significant bioactivities in many aspects. It is therefore very important for the exploration of natural resources rich of physalins. However, there is no efficient approach for rapid discovery and identification of this class of compounds due to their structural complexity. To address the issue, the fragmentation pathways and correspondingly fragmentation rules of physalins in negative MS/MS mode were thoroughly investigated in this study using seven physalin standards. As a result, diagnostic ions for the rapid screening of physalins and classification of different types of physalins were determined based on their MS/MS fragmentation patterns. On top of that, an integrated approach using UHPLC–QTOF-MS/MS together with a novel three-step data mining strategy was developed for the systematic analysis of physalins in complex samples. Consequently, 46 physalins including 20 novel ones were efficiently discovered and identified from the crude extracts of Ph. alkekengi calyx. The present study laid a foundation for future study of different parts of Ph. alkekengi and other physalis species with regard to rapid discovery of novel physalins. In addition, this study provided a base for establishing a quality control method of the raw materials of Ph. alkekengi according to the profile of physalins.

•74 lyso PCs were recognized and characterized in mice serum using LC-QTOF-MS/MS.•59 lyso PCs were quantitated in hyperlipidaemic mice fed turmeric.•Preventive effects of turmeric on HFD-induced mice were elucidated.•A targeted metabolomic approach for the analysis of serum lyso PC was established.Consumption of turmeric diets demonstrated preventive effects on high-fat diet (HFD) induced hyperlipidaemia. However, the preventive effects on serum lysophosphatidylcholines (Lyso PCs) have not yet been studied. For this study, an approach for characterization of Lyso PCs in mice serum using LC-QTOF-MS/MS was developed. The preventive effects of turmeric diets were evaluated on quantitative changes of individual Lyso PCs in mice serum by LC-QTRAP-MS/MS. As a result, 74 Lyso PCs were characterized, among which 59 Lyso PCs were quantified. The results indicated that significant changes occurred for the Lyso PCs with higher degree of unsaturation in the fatty acid chain. This is the first report regarding a targeted metabolomic approach for analyses of serum Lyso PCs and its application to interpretation of preventive effects of turmeric on mice hyperlipidaemia. These metabolic changes and potential biomarkers might serve as scientific evidence for future diagnosis and therapeutic intervention of hyperlipidaemia.