Ultra-performance Liquid Chromatography Coupled With Quadrupole Time-of-flight Mass Spectrometry Research Articles

Investigating the mechanism of enhanced medicinal effects of Terminalia chebula fruit after processing based on intestinal flora and metabolomics

Background and objectiveTerminalia chebula is a classical medicine for the treatment of lingering dysentery, and both raw and processed T. chebula can alleviate ulcerative colitis (UC). The therapeutic efficacy of T. chebula is enhanced after processing, but the mechanism that processing improves this efficacy is still unknown. We investigated the medicinal effects of raw and processed T. chebula on dextran sulfate sodium (DSS)-induced UC model rats using intestinal flora and metabolomics analyses, in order to elucidate the mechanism by which processing enhances the therapeutic effect. MethodsThe major constituents of raw and processed T. chebula were detected by high-performance liquid chromatography (HPLC). UC model was replicated using the DSS method, and then UC rats were administered raw and processed T. chebula. The general physical signs, disease activity index (DAI) scores, colon histopathological morphology, and the expressions of inflammatory cytokines were used to evaluate the therapeutic effect of T. chebula. In addition, 16 s rRNA sequencing and gas chromatography-mass spectrometry (GC–MS) were used to characterize the intestinal flora and contents of short-chain fatty acids (SCFAs). Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was utilized to identify the nontargeted fecal metabolites. ResultsRaw and processed T. chebula significantly improved the general physical signs and colon inflammatory symptoms and decreased DAI scores of UC rats. Both raw and processed T. chebula mitigated intestinal flora disorders in UC rats, increasing probiotic bacteria, including Lactobacillus and Romboutsia. However, the effect of processed T. chebula was more pronounced. Moreover, the levels of SCFAs of DSS-induced UC rats were restored after drug administration, and the processed T. chebula had a better regulatory effect than raw T. chebula. In the fecal nontargeted metabolomics analysis, differential metabolites such as lipids and amino acids were identified. The processed T. chebula can regulate purine metabolism and other pathways to improve UC, and the levels of the disordered metabolites gradually approached those of the control group. ConclusionRaw and processed T. chebula had the capacity to mitigate DSS-induced UC by rebalancing the intestinal flora, restoring the contents of SCFAs, and regulating fecal metabolites, while processed T. chebula showed preferable effects.

Evaluating the Ability of External Electric Fields to Accelerate Reactions in Solution.

This study investigates the catalytic effects of external electric fields (EEFs) on two reactions in solution: the Menshutkin reaction and the Chapman rearrangement. Utilizing a scanning tunneling microscope-based break-junction (STM-BJ) setup and monitoring reaction rates through high-performance liquid chromatography connected to a UV detector (HPLC-UV) and ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-q-ToF-MS), we observed no rate enhancement for either reaction under ambient conditions. Density functional theory (DFT) calculations indicate that electric field-induced changes in reactant orientation and the minimization of activation energy are crucial factors in determining the efficacy of EEF-driven catalysis. Our findings suggest that the current experimental setups and field strengths are insufficient to catalyze these reactions, underscoring the importance of these criteria in assessing the reaction candidates.

Development of an integrated strategy for comprehensive characterization of Sinomenii Caulis extract and metabolites in rats based on UPLC/Q-TOF-MS

Sinomenii Caulis (SC), a commonly used traditional Chinese medicine for its therapeutic effects on rheumatoid arthritis, contains rich chemical components. At present, most studies mainly focus on sinomenine, with little research on other alkaloids. In this study, a comprehensive profile of compounds in SC extract, and biological samples of rats (including bile, urine, feces, and plasma) after oral administration of SC extract was conducted via ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS). The fragmentation patterns and potential biotransformation pathways of six main types of alkaloids in SC were summarized, and the corresponding characteristic product ions, relative ion intensity, and neutral losses were obtained to achieve rapid classification and identification of complex components of SC from in vitro to in vivo. As a result, a total of 114 alkaloid compounds were identified, including 12 benzyl alkaloids, 4 isoquinolone alkaloids, 32 aporphine alkaloids, 28 protoberberine alkaloids, 34 morphinan alkaloids and 4 organic amine alkaloids. After administration of SC extract to rats, a total of 324 prototypes and metabolites were identified from rat plasma, urine, feces and bile, including 81 aporphines, 95 protoberberines, 117 morphinans and 31 benzylisoquinolines. The main types of metabolites were demethylation, hydrogenation, dehydrogenation, aldehydation, oxidation, methylation, sulfate esterification, glucuronidation, glucose conjugation, glycine conjugation, acetylation, and dihydroxylation. In summary, this integrated strategy provides an additional approach for the incomplete identification caused by compound diversity and low abundance, laying the foundation for the discovery of new bioactive compounds of SC against rheumatoid arthritis.

Photodegradation of bisphenol A (BPA) in coastal aquaculture waters: Influencing factors, products, and pathways

Bisphenol A (BPA), an endocrine-disrupting contaminant, is ubiquitous in the environment due to its presence in plastics, wastewater, and agricultural runoff. This study investigated the photodegradation behavior of BPA in coastal aquaculture waters near Qingdao, China. Lower salinity promoted BPA photodegradation, while higher salinity has an inhibitory effect, suggesting slower degradation in seawater compared to ultrapure water. Triplet-excited dissolved organic matter (3DOM*) was identified as the primary mediator of BPA degradation, with additional contributions from hydroxyl radicals (•OH), singlet oxygen (1O2), and halogen radicals (HRS). Alepocephalidae aquaculture water exhibited the fastest degradation rate, likely due to its high DOM and nitrate/nitrite (NO3−/NO2−) content, which are sources of 3DOM* and •OH. A positive correlation existed between NO3−/NO2− concentration and the BPA degradation rate. Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) analysis identified the primary BPA photodegradation products, formed mainly through oxidative degradation, hydroxyl substitution, nitration, and chlorination pathways. Elucidating these photodegradation mechanisms provides valuable insights into the environmental fate and potential ecological risks of BPA in aquaculture environments. This knowledge can inform strategies for marine environmental protection and the development of sustainable practices.

Study on the material basis and immunological enhancement activity of dangdi oral liquid

Studies have shown that a lot of traditional Chinese medicines could improve the immunity of the body. Dangdi oral liquid (DDO) was mainly composed of Angelica sinensis (Oliv.) Diels (Danggui), Rehmannia glutinosa Libosch. (Dihuang), Achyranthes bidentata Bl. (Niuxi), Glycyrrhiza uralensis Fisch. (Gancao). In this study, the rapid ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) method was used to identify the potentially effective compounds of DDO. Then the immune activity of DDO was measured by lymphocyte proliferation, macrophage phagocytic function, NK cell activity, delayed type hypersensitivity reaction, hemolytic plaque number, sIgA content and immune organ index. The results showed that a total of 51 compounds were identified. In addition, DDO could significantly promote the lymphocyte proliferation, improve macrophage phagocytic ability, NK cell activity, hemolytic plaque number, sIgA content and immune organ index compared with control group, and the medium dose possessed the best efficacy (P＜0.05). These results indicated that DDO could enhance the immunity of mice.

Rapid identification of chemical constituents and dynamic metabolic profile of Shenqi-Tiaoshen formula in rat plasma based on UPLC-Q-TOF/MSE

Shenqi-Tiaoshen formula (SQTSF) is a traditional Chinese medicine (TCM) prescription that has been employed in the treatment of chronic obstructive pulmonary disease (COPD). Clinical practice has demonstrated that SQTSF is an effective prescription for stable COPD. However, owing to the complexity of TCM prescription, there is a lack of in-depth understanding of the chemical components of SQTSF and its in vivo metabolism studies. In this study, a comprehensive analytical strategy based on ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was established to identify the chemical components, the absorbed components, and the metabolites of SQTSF given by gavage in rats, and analyze their dynamic changes. As a result, 86 chemical components of SQTSF were characterized, which were mainly categorized into flavonoids, saponins, organic acids, terpenoids, etc. Among them, 13 compounds were confirmed unambiguously by reference standards. Furthermore, 20 prototype components and 46 metabolites were detected in rat plasma at different time points. It was found that one prototype component and thirteen metabolites could be detected during the entire 24 h, indicating that these compounds were slowly eliminated and thus accumulated in vivo over a prolonged duration. Interestingly, the phenomenon that three prototype components and fourteen metabolites reappeared after a period of disappearance from the plasma was found. It was also observed that different prototype components may generate the same metabolite. The metabolic processes of SQTSF in rats mainly included oxidation, reduction, hydration, demethylation, deglycosylation, methylation, acetylation, glucuronidation, glutathionylation, and associated combination reactions. Overall, the present study identified the chemical components of SQTSF and their dynamic metabolic profile in rat plasma, which provided a systematic and applicable strategy for screening and characterization of the prototype components and metabolites of TCM compound preparations.

Deciphering the mysteries of Aconitum pendulum: Unique identification of various processed products and characteristic chemical markers

Aconitum pendulum, a member of the Aconitum genus within the Ranunculaceae family, is known for its remarkable anti-inflammatory, anti-tumor, and analgesic properties. However, the application of Aconitum pendulum in traditional medicine is often restricted by its inherent toxicity, necessitating specific processing methods to ensure safe usage. Tibetan medicine utilizes distinctive processing techniques such as the barley wine, Hezitang, and zanba frying systems to render the herb safe for medicinal use. Despite the known pharmacological importance, scant research exists on how these processing methods affect the phytochemical profile of Aconitum pendulum, indicating a necessity for comparative analysis of the resulting concoctions. In this study, we employed 1H nuclear magnetic resonance (1H NMR) spectroscopy and ultra-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) to explore the metabolomic variations among different processed products of Aconitum pendulum. Multivariate data analysis techniques, including pattern recognition, allowed us to investigate the intricacies of the compound profiles. To discover signature chemical markers that can differentiate the various processed products, we conducted paired t-tests and generated heat maps highlighting the detected differences. Our integrated analysis of the 1H NMR and UPLC-Q-TOF-MS data revealed distinct chemical signatures unique to each method of Aconitum pendulum processing. By employing chemometric analysis with variable importance in projection (VIP) scores greater than 1.0, and coupled with the statistical rigor of paired t-tests (P < 0.05), we successfully identified significant chemical markers in various processed products, including ferulic acid (FA), caffeic acid (CA), 3-acetylaconitine (AAC) and benzoylaconitine (BAC). Our findings provide a rapid and robust analysis of the chemical constituencies within raw and processed Aconitum pendulum. The discovery of potential chemical markers through an array of advanced pattern recognition techniques offers a substantial contribution to the pharmaceutical analysis. The results of this research pave the way for future comparative studies and facilitate the recognition of distinctively processed Aconitum pendulum products in traditional medicine.

An integrated strategy of UPLC-Q-TOF-MS analysis, network pharmacology, and molecular docking to explore the chemical constituents and mechanism of Zixue Powder against febrile seizures

Febrile seizures (FS) are the most common type of seizures for children. As a commonly used representative cold formula for resuscitation, Zixue Powder (ZP) has shown great efficacy for the treatment of FS in clinic, while its active ingredients and underlying mechanism remain largely unclear. This study aimed to preliminarily elucidate the material basis of ZP and the potential mechanism for the treatment of FS through ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), network pharmacology, and molecular docking. UPLC-Q-TOF-MS was firstly applied to characterize the ingredients in ZP, followed by network pharmacology to explore the potential bioactive ingredients and pathways of ZP against FS. Furthermore, molecular docking technique was employed to verify the binding affinity between the screened active ingredients and targets. As a result, 75 ingredients were identified, containing flavonoids, chromogenic ketones, triterpenes and their saponins, organic acids, etc. Through the current study, we focused on 13 potential active ingredients and 14 key potential anti-FS targets of ZP, such as IL6, STAT3, TNF, and MMP9. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that inflammatory response, EGFR tyrosine kinase inhibitor resistance, AGE-RAGE signaling pathway in diabetic complications, and neuroactive ligand-receptor interaction were the main anti-FS signaling pathways. Licochalcones A and B, 26-deoxycimicifugoside, and hederagenin were screened as the main potential active ingredients by molecular docking. In conclusion, this study provides an effective in-depth investigation of the chemical composition, potential bioactive components, and possible anti-FS mechanism of ZP, which lays the foundation for pharmacodynamic studies and clinical applications of ZP.

UPLC-QTOF-MS-based lipidomic study of wedelolactone in acute colitis mice induced by dextran sulfate sodium

Inflammatory bowel disease is a relapsing inflammatory disease seriously endanger human health. Wedelolactone (WED) is a major active ingredient from Eclipta prostrata (L.) L. and has shown anti-inflammatory effects. However, the mechanism of WED in treating inflammatory colitis remains unknown. We aimed to investigate the mechanisms of WED in treating ulcerative colitis through lipidomic study. Sixty male C57BL/6 mice were exposed to DSS to induce acute colitis. Disease progression was judged by the disease activity index (DAI) and pathological changes of colon tissue. An ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) method was performed for colon and plasma lipidomics analyses. Differential metabolites in the three groups were distinguished by univariate and multivariate analysis. WED exerted anti-inflammatory effects representing by body weight and DAI score. Three metabolites were identified in plasma and 20 in colon. According to pathway analysis, the effects of WED on colitis were associated with seven pathways. The glycerophospholipid metabolism and ether lipid metabolism were the primary pathways. The findings provide important insight of the mechanism of WED in treating DSS induced colitis through lipidomic perspective.

Evaluation of Biological Mechanisms of Quanduzhong Capsule for Treating Osteoporosis by Integrating Untargeted Metabolomics and Network Pharmacology

AbstractOsteoporosis (OP) is a metabolic disease characterized by bone formation and resorption disturbances. Quanduzhong Capsule (QDZC) is a common treatment for OP in China; however, the effective components and metabolites of the drug after oral administration remain largely unknown. This study aims to identify the active components, analyze the metabolite changes, and investigate the underlying mechanism against OP. In the study, ovariectomy-induced rat OP model was established, then treated with QDZC. Alendronate sodium tablets (ASTs) were used as a reference drug. The chemical constituents of QDZC were analyzed by UPLC-QTOF-MS (ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry) and network pharmacology. The metabolomics was used to analyze differences in serum metabolites of rats in different groups [Sham, Model, Model + QDZC, and Model + AST] at 4, 8, and 12 weeks. Body weight and bone mineral density (BMD) were assessed. Enzyme-linked immunosorbent assay was used to determine serum levels of Akt, p-Akt, ERK, and p-ERK. Our data suggested 86 different chemicals from QDZC, including nine core compounds. QDZC significantly regulated 25 biomarkers linked to arachidonic acid metabolism and unsaturated fatty acid biosynthesis, and promoted serum expression of Akt, p-Akt, ERK, and p-ERK. QDZC might act by activating PI3K-Akt and MAPK signaling pathways. In addition, QDZC may use arachidonic acid derivatives to inhibit osteoclast generation and bone resorption and enhance calcitriol formation to improve calcium absorption and increase bone mass.

Enhancing NKT cell-mediated immunity against hepatocellular carcinoma: Role of XYXD in promoting primary bile acid synthesis and improving gut microbiota

Ethnopharmacological relevance‘Xiayuxue decoction’ (XYXD) is a traditional Chinese medicine compound, composing of three natural medicines: Rheum officinale Baill., Prunus persica (L.) Batsch and Eupolyphaga sinensis Walker. It is derived from the famous traditional Chinese medical classics ‘Jingui Yaolue’ and has been used for thousands of years. In the Guidelines for the Diagnosis and Treatment of Primary liver Cancer issued by China's Health Commission, XYXD was applied in the treatment of primary liver cancer. Aim of the studyTo clarify the pharmacodynamic material basis and mechanism of XYXD in the treatment of hepatocellular carcinoma (HCC). Materials and methodsFirstly, the active components of XYXD and its distribution in vivo were identified by Ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Then, the effective components and mechanism of XYXD against HCC were explored by network pharmacology combined with cell experiments in vitro. Furthermore, the anti-HCC effect of XYXD was determined by animal experiments in vivo. Metagenomic sequencing was used to detect its effect in gut microbiota, and targeted metabolism was used to detect the changes of bile acids in the liver. Finally, the related targets of NKT cell immune function activation were detected by RT-qPCR and Elisa. ResultsA total of 113 active ingredients in XYXD were identified, and the distribution of active ingredients in blood, liver, tumor, cecum, intestinal contents and feces was clarified. The circulation process and active ingredient group of XYXD were preliminarily clarified. In addition, we found five anti-HCC active ingredients in XYXD through network pharmacology combined with cell experiments in vitro, among which aloe emodin had the most significant effect, and predicted the potential mechanism of XYXD against HCC through NKT cell pathway. Moreover, the inhibitory effect of XYXD on liver tumor growth was clarified by animal experiments in vivo. The mechanism was mainly to promote the production of bile salt hydrolase (BSH) by increasing the abundance of Bacteroides and Lactobacillus, BSH converts conjugated bile acids into primary bile acids, and reduces the conversion of primary bile acids to secondary bile acids by reducing the abundance of Eubacterium, thereby increasing the content of primary bile acids. Primary bile acids trigger NKT cells in the liver to produce interferon-γ to exert anti-HCC immune effects. ConclusionThis study found that the traditional Chinese herbal formula XYXD can trigger the immune effect of NKT cells against HCC by regulating the interaction between gut microbiota and bile acids.

Antinutrients and metabolomic compounds of Bambara groundnut (Vigna subterranean) as affected by traditional processing by smallholder farmers.

Bambara groundnut (BG) (Vigna subterranean) is an underutilized, indigenous crop in South Africa that has nutritional and associated health benefits. Decreasing the antinutrients in food sources can potentially increase the digestibility of proteins and mineral absorption. To determine the effect of traditional processing (cooking) on the antinutrient content and metabolome of this crop, BG was sampled from 12 rural farms in three districts of the Mpumalanga province, South Africa. The four main colors that were identified (cream, orange, brown, and purple) were pooled together according to the district they were obtained from. One-half of each color sample obtained from each of the three districts was dehulled, color sorted, milled, and subjected to subsequent antinutrient and metabolome analyses, while the other half was cooked, air-dried, and milled prior to analyses. Samples were analyzed for phytate and tannins (antinutrients) by hydrochloric acid extraction methods as well as metabolome constituents by ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). Phytate, tannins, as well as other metabolomic constituents, namely, catechin, epicatechin, procyanidin, as well as citric acid, were identified in all raw and cooked BG samples. The cooking process resulted in a significant decrease (p<0.05) in the phytate and tannin content as well as an increase in the health-associated phenolic compounds.

Rapid Analysis of 18 Flavonoids in Tea by Ultrahigh-Performance Liquid Chromatography Coupled with Quadrupole-Time of Flight Mass Spectrometry

A method was established for the determination of 18 flavonoids in tea by ultra-performance liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry (UPLC-Q-TOF/MS). The tea samples were extracted by 70% (V/V) methanol aqueous solution, and separation was achieved on a Kinetex F5 column (2.1 mm × 100 mm, 2.6 μm) with methanol and 0.1% formic acid in water as the mobile phase with a gradient elution. The samples were detected in TOF/MS and information-dependent acquisition (IDA)-MS/MS modes. The results showed that the relative standard deviations of 18 flavonoids were less than 5.0 ppm. The correlation coefficients (R2) of the linear equation were greater than 0.998 in the range of 0.10–200 ng/mL. The limits of detection were 0.0010–0.040 ppm, and the limits of quantification were 0.0020–0.10 ppm. The recoveries ranged from 73.8% to 107% at spiked levels of 0.0020–1.0 ppm, with relative standard deviations (RSDs) being less than 10%. The method was simple, specific, and reliable. It could be used for the rapid screening and quantitative analysis of flavonoids in tea.

Antihypertensive effects of Pleurospermum lindleyanum aqueous extract in spontaneously hypertensive rats.

Pleurospermum lindleyanum (Lipsky) B. Fedtsch is a perennial herb classified in the Apiaceae family, genus Pleurospermum, chiefly native to the Taxkorgan County, Xinjiang, China. In the Xinjiang Province, it is a well-known ethnic traditional herb, often addressed by its tribal name, Kurumuti. It grows in harsh conditions over 4000m above sea level, such as the Pamirs plateau. It is rich in flavonoids, coumarins, terpenoids, essential oil, substances that have been widely applied in the prevention and treatment of hypertension, diabetes, coronary heart disease, and cerebral thrombosis by local Tajik residents. The present study aimed to evaluate the antihypertensive effects of the Pleurospermum Lindleyanum aqueous extract (PLAE) in spontaneously hypertensive rats (SHRs). The Pleurospermum lindleyanum was collected from the Taxkorgan Tajik Autonomous County, Xinjiang, China. The main chemical composition of PLAE was identified using the ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). SHRs were treated by gavage with PLAE (equivalent to Pleurospermum lindleyanum 5 or 10g/kg/day) for 6 weeks, using Captopril (10mg/kg/day) as positive control. The systolic blood pressure (SBP), renal and cardiac morphology, plasma levels of angiotensin-converting enzyme (ACE), aldosterone (ALD), angiotensinⅡ (AngⅡ), superoxide dismutase (SOD), endothelin-1 (ET-1) and nitric oxide (NO) were measured. A total of 30 compounds were identified in PLAE. PLAE significantly attenuated the SBP of SHRs. The effects began after 3 weeks of administration and then became steady and long-lasting. Its potential mechanisms may be associated with the protective effects on renal and cardiac injury caused by hypertension, the decrease of plasma vasoconstrictors, such as ACE, ALD, AngⅡ, and ET-1 levels, the maintenance of NO/ET balance, the increase in plasma NO levels and SOD activity, thereby reducing oxidative stress. Pleurospermum lindleyanum can be suggested as a novel antihypertensive ethnic traditional herb, which lays the foundation for researching safe and effective antihypertensive herbal medicines.

Mining of kokumi peptides in chicken broth with peptidomics

Chicken soup is a high-quality traditional seasoning. In this study, 16,306 polypeptides in the Gushi chicken soup were analyzed by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), and two polypeptides with kokumi flavor (TPDFVR and TPELLH) were obtained by comparing the amino acid structure and performing electronic tongue analysis. The taste thresholds of TPDFVR and TPELLH are 0.4 and 0.2 mmol/L, respectively. Therefore, the taste time of the chicken soup can be prolonged in the mouth, and the taste of the chicken soup can be improved. Also, the electronic tongue experiment proved that the two polypeptides have a strong sour and salty taste. The experimental results revealed that the peptidomics method is quick and effective in discovering peptides associated with taste.

UPLC-QTOF-MSE-Based Metabolic Profile to Screening Candidates of Biomarkers of Dwarf-Cashew Clones Resistant and Susceptible to Anthracnose (Colletotrichum gloeosporioides (Penz) Penz. &amp; Sacc.)

Investigating specialized plant metabolites, traditionally referred to as ‘secondary metabolites’ present in leaf extracts of cashew trees (Anacardium occidentale) resistant and susceptible to anthracnose disease was carried out using metabolomics combined with chemometric tools. We used clones of dwarf-cashew with the following variations of characteristics: resistant and healthy (CCP 76, BRS 226, BRS 189), susceptible and healthy (BRS 265), and another clone also susceptible but affected by the disease (BRS 265). The UPLC-QTOF-MSE (ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry) analysis of the leaves of Anacardium clones allowed us annotation a total of 39 metabolites. The multiple orthogonal partial discriminant analysis of the least-squares (OPLS-DA) allowed twelve metabolites to be potential biomarkers of differentiation among the clones studied. Namely, the triene-(17:3)-anacardic acid found on CCP 76 and BRS 189 clones, respectively, is the main candidate for biomarker of resistance. While catechin, B-type procyanidin isomers, and procyanidin dimer monogallate identified in BRS 265 are significant potential biomarkers of susceptibility.

Rapid screening of 420 pesticide residues in fruits and vegetables using ultra high performance liquid chromatography combined with quadrupole-time of flight mass spectrometry

A new method for screening and identification 420 pesticide residues in fruits and vegetables by ultra-performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UPLC-Q-TOF/MS) were developed. The samples were extracted with acetonitrile/acetic acid (99:1, V/V), and clean-up by SinChERS-Nano (single-step, cheap, effective, rugged, safe, nano) column, determined by UPLC-Q-TOF/MS. The accurate mass database and MS/MS database which contains 420 pesticides were established, the automatic retrieval of detection results was carried on according to the accurate mass, retention time, isotope ratio, ion fragment information, and so on. Method verification was performed on leeks samples. The results showed that 420 pesticides had good linearity in the range of 0.1–100 μg/L, and the correlation coeffificients (R2) was greater than 0.990. The limits of detections (LODs) and limits of quantifications (LOQs) of 420 pesticides were in range of 0.05–2.0 and 0.1–5.0 μg/L, respectively. The average spike recoveries at 3 levels were 70.1 % to 119.7 %, and the relative standard deviations (RSD) were lower than 20 % (n = 6). With this method, a survey of pesticide residues was conducted for 110 samples of 10 different fruits and vegetables, which provided scientific data for ensuring pesticide residue safety of the fruits and vegetables consumed daily by the public. This method was simple, sensitive and accurate, and could be used for rapid screening of 420 pesticide residues in fruits and vegetables.

Identification of prototype compounds and their metabolites in rats’ serum from Xuefu Zhuyu Decoction by UPLC-Q-TOF/MS

ObjectiveAs a classic prescription in traditional Chinese medicine, Xuefu Zhuyu Decoction (XFZYD) has been widely used in the clinical treatment of cardiovascular and cerebrovascular diseases. In order to unveil the potentially effective compounds, a rapid ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) method was established to identify prototype compounds and their metabolites from XFZYD in rats’ serum. MethodsThe serum from rats after intragastric administration of XFZYD aqueous extract was analyzed by UPLC-Q-TOF/MS method. The prototype compounds and their metabolites were identified by comparison with the reference standards and tentatively characterized by comprehensively analyzing the retention time, MS data, characteristic MS fragmentation pattern and retrieving literatures. ResultsA total of 175 compounds (24 prototype compounds and 151 metabolites) were identified and tentatively characterized. The metabolic pathways of prototype compounds in vivo were also summarized, including glucuronidation, hydrolyzation, sulfation, demethylation, and hydroxylation, and so on. ConclusionIn this study, a UPLC-Q-TOF/MS technique was developed to analyze prototype compounds and their metabolites from XFZYD in serum, which would provide the evidence for further studying the effective compounds of XFZYD.

Characterization of phenolics and discovery of α-glucosidase inhibitors in Artemisia argyi leaves based on ultra-performance liquid chromatography-tandem mass spectrometry and relevance analysis

Artemisia argyi leaves (AAL) has been widely used as herbal medicine and food supplement and in China and other Asian countries. The aim of this work is to qualitative and quantitative characterization of phenolic compounds in AAL and screening of natural product inhibitors of α-glucosidase from AAL. Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) was employed to rapid and comprehensive identification of phenolic compounds in AAL, and a total of thirty-three phenolic compounds were identified. High performance liquid chromatography with diode array detection (HPLC-DAD) was established and validated to simultaneously determinate ten main bioactive phenolics compounds in different batches of AAL samples. Meanwhile, the inhibitory capacities of different batches of AAL samples on α-glucosidase were evaluated. Then, relevance analysis, including grey relational analysis and Pearson correlation analysis were employed to investigate the correlations between the contents of phenolic compounds and α-glucosidase inhibitory activities, and discover the α-glucosidase inhibitors in AAL. The relevance analysis results indicated that three phenolic compounds, 3-caffeoylquinic acid, 3,4-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid could be potential α-glucosidase inhibitors in AAL. Moreover, the α-glucosidase inhibitory activities of the three phenolic compounds were validated by in vitro and in vivo experiments. The possible inhibiting effect of the three phenolic compounds on α-glucosidase was also explored by molecular docking analysis, and the results indicated that the binding of the three α-glucosidase inhibitors to α-glucosidase mainly by hydrogen bonds, hydrophobic forces and ionic bonds. The present research provided a deep insight into phenolic compounds and α-glucosidase inhibitory activities of AAL, and discovered the α-glucosidase inhibitors in AAL.

A complementary chromatographic strategy for integrated components characterization of Imperatae Rhizoma based on convergence and liquid chromatography combined with mass spectrometry and molecular network

The complexity of natural ingredients and the diversity of preparations are the major obstacles to the quality evaluation of traditional Chinese medicines (TCMs). A more comprehensive characterization of herbal compounds using different types of chromatographic separation techniques and covering a diverse polarity range can help evaluate the quality of TCMs. In this study, we first proposed a comprehensive method for characterizing compounds derived from Imperatae Rhizoma by combining the complementary strengths of UPCC-QTOF-MS (ultra-performance convergence chromatography coupled with quadrupole-time of flight mass spectrometry) with UPLC-QTOF-MS (ultra-performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry). The method based on the UNIFI scientific platform significantly shortened the analysis time and enabled a more comprehensive characterization of known and unreported compounds. Meanwhile, a feature-based molecular network (FBMN) was established on the Global Natural Product Social (GNPS) to infer potential compounds by rapidly classifying and visualizing these components. A total of 62 compounds in Imperatae Rhizoma were jointly characterizedand classified into six types. In comparison, the UPCC-QTOF-MS technology individually characterized 17 components, including lactones, phenols, aldehydes, phenylpropanoids, and small polar organic acids. The UPLC-QTOF-MS technology characterized 16 compounds mainly phenylpropionic acids, flavonoid glycosides, and chromone glycosides. Furthermore, three types of characteristic compounds could be well aggregated into an FBMN approach. Five possible potential new compounds were detected through the supplementary identification of GNPS and the correlation analysis of vicinal known compounds. The strategy was first applied to Imperatae Rhizoma and facilitated the characterization of a large quantity of data to provide comprehensive chemical composition results. This approach can be easily extended to the study of the material basis of other herbs or preparations in order to improve the accuracy of herb quality evaluation.