Yulin Feng

The metabolic change of serum lysophosphatidylcholines involved in the lipid lowering effect of triterpenes from Alismatis rhizoma on high-fat diet induced hyperlipidemia mice

ETHNOPHARMACOLOGICAL RELEVANCE Alismatis rhizoma (AR), a Traditional Chinese Medicine with lipid-regulating properties, is usually used to treat hyperlipidemia. Lysophosphatidylcholines (Lyso PCs) play a crucial role in lipid metabolism disorders. In this study, the triterpene fraction purified from boiling water extract of AR was evaluated for its lipid lowering activity using mice with high-fat diet (HFD) induced hyperlipidemia. The metabolic changes of individual Lyso PCs treated with the triterpene fraction were detected by ultra-high performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometer (UHPLC-QTRAP-MS/MS). MATERIALS AND METHODS HFD induced hyperlipidemia mice were administrated with triterpene and non-triterpene fractions at doses of 180, 360 and 720 mg/kg body weight/day for 4 weeks, respectively. Total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and atherogenic Index (AI) in mice serum were measured. The chemical components in the lipid-lowering fraction were characterized by ultra-high performance liquid chromatography-quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS/MS). The changes of Lyso PC in the serum of mice treated with the lipid-lowering fraction were quantified by UHPLC-QTRAP-MS/MS. RESULTS A total of 18 alisol derivatives were identified in the triterpene fraction. The hyperlipidemia mice treated with the triterpene fraction showed a significant decrease in serum TC, LDL-C and AI after continuous consumption of HFD for 4 weeks. The results also showed that 27 serum Lyso PCs in mice fed with HFD were down-regulated, and 19 were up-regulated. The abnormal serum level of Lyso PCs associated with hyperlipidemia was intervened by the alisol derivatives, with increase of unsaturated Lyso PCs and decrease of saturated ones. CONCLUSIONS The study demonstrated for the first time that triterpenes from the AR extract can lower serum lipid level in HFD induced hyperlipidemia mice. These metabolism changes of Lyso PCs could further improve our understanding of the potential mechanism of lipid lowering effect of AR.

A novel dereplication strategy for the identification of two new trace compounds in the extract of Gastrodia elata using UHPLC/Q-TOF-MS/MS

An ultra performance liquid chromatography (UHPLC) coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS/MS) was used in the structural determination of natural compounds in Gastrodia elata. A total of 64 compounds were identified or tentatively characterized. The strategy used for characterization was comparing their retention time and fragmentation behaviors with those of the reference standards, or investigating their accurate mass measurements and characteristic fragmentation patterns followed by low-energy collision dissociation tandem mass spectrometry (CID-MS/MS). Phenolic conjugates mainly underwent consecutive losses of gastrodin residues and combined losses of H2O and CO2 from their citric acid units under negative MS/MS conditions. According to these rules, we have successfully characterized fifteen potential novel compounds. To confirm the reliability of this strategy, two targeted unknown trace parishins were obtained from G. elata by LC/MS-guided isolation. Based on the analysis of data from NMR spectroscopy and other techniques, the two unknown parishins were identified as 2-[4-O-(β-d-glucopyranosyl)benzyl]-3-methyl-citrate (parishin J) and 1,2-di-[4-O-(β-d-glucopyranosyl)benzyl]-3-methyl-citrate (parishin K), respectively. The fully established structures were consistent with the MS-oriented structural elucidation. This study expanded our knowledge on parishins in Gastrodia species, and the proposed strategy was proven efficient and reliable in the discovery of new minor compounds from herbal extracts.

Ionic liquid-based ultrasound-assisted extraction and aqueous two-phase system for analysis of caffeoylquinic acids from Flos Lonicerae Japonicae

In this work, an ionic liquid-based ultrasonic-assisted extraction (ILUAE) method was developed to extract caffeoylquinic acids (CQAs) from Flos Lonicerae Japonicae (FLJ). ILUAE parameters were optimized by response surface methodology, including IL concentration, ultrasonic time, and liquid-solid ratio. Optimized ILUAE approach gained the highest extraction yields of 28.53, 18.21, 3.84mg/g for 3-O-caffeoylquinic acid (C1), 3,5-di-O-caffeoylquinic acid (C2), 3,4-di-O-caffeoylquinic acid (C3), respectively. C1-C3 are the three most abundant CQAs compounds in FLJ. The method showed comparable extraction yield and shorter extraction time compared with conventional extraction techniques. Subsequently, an aqueous two-phase system (ATPS) was applied in extraction solutions. Two trace CQAs, 5-O-caffeoylquinic acid (C4) and 4,5-di-O-caffeoylquinic acid (C5), were significantly enriched with signal to noise values increasing from less than 10 to higher than 1475. The results indicated that ILUAE and ATPS are efficient and environmentally-friendly sample extraction and enrichment techniques for CQAs from herbal medicines.

The rapid discovery and identification of physalins in the calyx of Physalis alkekengi L.var.franchetii (Mast.) Makino using ultra-high performance liquid chromatography-quadrupole time of flight tandem mass spectrometry together with a novel three-step data mining strategy.

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.

Metabolites profiling of Pulsatilla saponin D in rat by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS/MS).

Pulsatilla saponin D, an antitumor substance isolated from traditional Chinese herbal medicine Pulsatilla chinensis (Bge.) Regel, is a promising candidate for new drug development. The purpose of the present study is to establish a simple and practical strategy for the metabolite profiling of Pulsatilla saponin D in vivo. A total of 18 metabolites were identified in rat plasma, urine and feces samples based on MS and MS/MS data by using ESI-Q-TOF-MS/MS, and eight of them (M11-M18) were reported for the first time. The results indicated that deglycosylation, dehydrogenation, hydroxylation and sulfation were the major metabolic transformations of Pulsatilla saponin D in vivo. This study has improved our understanding of the metabolic fate of Pulsatilla saponin D in vivo, and the information gained from the current study is relevant to the pharmacological activity of Pulsatilla saponin D.

An integrated strategy for establishment of curcuminoid profile in turmeric using two LC–MS/MS platforms

Graphical abstract Figure. No Caption available. HighlightsA 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. ABSTRACT 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.

A robust platform based on ultra-high performance liquid chromatography Quadrupole time of flight tandem mass spectrometry with a two-step data mining strategy in the investigation, classification, and identification of chlorogenic acids in Ainsliaea fragrans Champ

The chlorogenic acids are the major bioactive constituents of the whole plant of Ainsliaea fragrans Champ. (Xingxiang Tuerfeng). These compounds are usually present as isomeric forms in Xingxiang Tuerfeng. Therefore, an efficient approach should be developed for the rapid discovery and identification of chlorogenic acids isomers through the fragmentation pathway and rules. In this study, the collision induced dissociation tandem mass spectrometry (CID-MS/MS) fragmentation routes of chlorogenic acids were systematically investigated by UHPLC-QTOF-MS/MS in the negative ion mode using eight chlorogenic acids standards. As a result, diagnostic product ions for rapid discovery and classification of chlorogenic acids isomers were determined according to their MS/MS fragmentation patterns and intensity analysis. Based on these findings, a novel two-step data mining strategy was established. The first key step was to screen different kinds of substitution and the skeleton of the quinic acid using the characteristic product ions and neutral losses. The second key step was to screen and classify different types of chlorogenic acids using their diagnostic product ions. It was apply to the rapid investigation, classification, and identification of chlorogenic acids. And the same carbon skeletons from a complex extract of Ainsliaea fragrans Champ. were effectively identified. 88 constituents, including 14 chlorogenic acids types, were rapidly discovered and identified, and in particular, 12 types of chlorogenic acids, including p-CoQC, FQA, BQC, CQA-Glu, CFQA, p-Co-CQC, di-p-CoQC, BCQA, di-CQA-Glu, PCQA, tri-QCA, and P-di-CQA, were first discovered in Ainsliaea fragrans Champ. In conclusion, UHPLC-QTOF-MS/MS method together with a systematic two-step data mining strategy was established as a feasible, effective, and rational technique for analyzing chlorogenic acids. Additionally, this study laid a foundation for the study of the active substances and quality control of Ainsliaea fragrans Champ.

Antiviral herbs--present and future.

Viral disease is a calamity which absolutely can not be ignored for human health. The emergence of drug resistance and spread of new virus will be the new challenge against viral disease. To find and develop new antivirus agents with properties of safety, significant effect and low toxicity is the pressing question facing humans today. Because of its advantages, including rich resources, low price, less adverse effect, Traditional Chinese medicine (TCM) have become the research focus in antiviral treatment. In recent years, there are numerous articles about the studies from separation of active ingredients to the antiviral mechanism. In this paper, the progress in experimental study was illustrated on the basis of active ingredients, species of virus, mechanism, clinical application. Obviously, TCM have obvious advantages in the treatment of virus infectious disease and has a broad prospect of application.

Comprehensive profiling and characterization of coumarins from roots, stems, leaves, branches, and seeds of Chimonanthus nitens Oliv. using ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry combined with modified mass defect filter

&NA; Chimonanthus nitens Oliv. (CNO), having been studied and developed as the tea beverages, tea raw materials and preparations, an unique species in China, have been extensively used for treating colds and influenza for centuries. In the present study, a method based on the modified mass defect filter (MDF) was firstly developed and validated for comprehensive profiling of coumarins in the different parts of CNO via ultra‐performance liquid chromatography tandem quadrupole time‐of‐flight mass spectrometry (UPLC‐QTOF/MS). The five‐point screening approach based on MDF and the visual isotopic ion technique could rapidly screen the interested precursor ions. The fragmentation behavior of coumarins was systematically investigated and a total of 42 coumarins including 27 potential new ones were unambiguously or tentatively identified in the CNO. Collectively, the results obtained in the present work may provide useful information for future utilization of CNO. Graphical abstract Figure. No caption available. HighlightsA method for the comprehensive characterization of coumarins was established.The five‐point screening approach was developed for screening coumarins.The fragmentation pathways of coumarins were investigated.42 quassinoids including 27 new ones were identified from Chimonanthus nitens Oliv.

Cell-based assays in combination with ultra-high performance liquid chromatography-quadrupole time of flight tandem mass spectrometry for screening bioactive capilliposide C metabolites generated by rat intestinal microflora.

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±1.55μg/mL.