Performance Liquid Chromatography-quadrupole Time Research Articles

Determination of twelve neonicotinoid pesticides in chili using an improved QuEChERS method with UPLC-Q-TOF/MS

In this study, a QuEChERS method based on citrate was developed and utilized for the analysis of twelve neonicotinoid pesticides in fresh red chilies, fresh green chilies, and dried chilies, coupled with ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS). In the sample preparation, acetonitrile containing 1% formic acid was used as the extraction solvent. Anhydrous sodium sulfate replaced the traditional anhydrous magnesium sulfate for water removal, effectively eliminating the issues of salt caking. Graphitized carbon black, octadecyl silica, and primary secondary amine were used as cleaning agents. The method showed good sensitivity, with the limits of quantification below 0.03 mg/kg for fresh chilies and below 0.15 mg/kg for dried chilies. Values of matrix effects ranged from −19.5% to 8.4%, and the recovery was 86.9% - 105.2%. The analytical method provided an effective tool for the high throughput detection of neonicotinoid pesticide residues in multiple chili matrices.

A breakthrough in periodontitis treatment: Revealing the pharmacodynamic substances and mechanisms of Kouqiangjie formula

Ethnopharmacology relevancePeriodontitis, a complex inflammatory disease, significantly affects people's lives. Traditional Chinese multi-herbal formulas, composed of various herbs, exhibit their therapeutic efficacy holistically. Kouqiangjie Formula (KQJF), comprising 12 herbs including Rhizoma smilacis glabrae, Polygonatum sibiricum Delar. ex Redoute, Taraxacum mongolicum Hand.-Mazz, etc., has been clinically proven to effectively treat periodontitis. However, the potential active substances conferring these effects and their mechanisms of action remain unclear. Aim of the studyThe current investigation endeavours to utilize Ultra Performance Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry (UPLC-Q-TOF-MS), network pharmacology, and in vivo animal experiment confirmation to explore the plausible bioactive compounds and operational mechanisms underpinning KQJF's therapeutic impact on periodontitis. Materials and methodsUsing the UPLC-Q-TOF-MS technique, we deciphered the chemical constituents of KQJF. Network pharmacology was employed to earmark key bioactive elements, forecast principal targets, and operational pathways which were later substantiated through molecular docking. Experimental validations were carried out in a periodontitis animal model using a range of techniques, including micro-CT, H&E staining, qRT-PCR, and protein blotting procedures, providing comprehensive verification of our initial assumptions. ResultsUtilizing UPLC-Q-TOF-MS, we characterized 87 individual chemical constituents in KQJF. Network pharmacology revealed that 14 components, including senkyunolide A, glycycoumarin, licoflavonol, glycyrin, senkyunolide I, and senkyunolide H, form the key therapeutic basis of KQJF in targeting periodontitis. Significant targets and pathways were discerned as AKT1, MMP9, JUN, PTGS2, CASP3, TLR4, IL1β, BCL2, PPARG, and pathways such as the TNF signaling pathway, NF-κB signaling pathway, osteoclast differentiation, and Wnt signaling pathway. Molecular docking demonstrated robust binding activity between these crucial targets and the key active ingredients. In vivo experimentation corroborated that, compared with the model group, KQJF significantly ameliorated symptoms and micro-CT imaging parameters of periodontitis in the rat model, down-regulating the expression of AKT1, MMP9, JUN, PTGS2, CASP3, TLR4, and IL1β, while up-regulating the expression of BCL2 and PPARG. ConclusionIn summary, this study has pioneered a comprehensive exploration of the potential therapeutic constituents, targets, and mechanisms of KQJF for periodontitis treatment, adopting a synergistic strategy of “chemical component analysis-network pharmacology screening-in vivo animal experiment validation”. This provides experimental evidence for the clinical application of KQJF and further in-depth research. Additionally, it presents an effective strategy for the research of other Chinese herbal formulations.

Comparative Metabolomics Revealed Differences in Alkaloids Metabolism Between Morus nigra L. and Morus. alba L. at Different Growth Stages

Mulberry (Morus spp.), a flowering plant in the Moraceae family, is known for the edible fruit, and mulberry leaves have various applications. This experiment utilized ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) and Progenesis QI software to rapidly identify secondary metabolites in the leaves of Morus nigra L. and Morus alba L.. Orthogonal partial least squares discriminant analysis (OPLS-DA), principal component analysis (PCA) and Hierarchical clustering analysis (HCA) were used to examine the dynamic changes and stage specificity of metabolites, particularly alkaloids, in M. nigra and M. alba leaves during seven primary growth stages. In results, the leaf samples from different growth stages uncovered a total of 182 metabolites from 13 distinct categories. M. nigra had significantly higher levels of metabolites than those in M. alba, according to differences in leaf metabolite accumulation patterns. Multiple statistical analyses revealed that the metabolome of M. nigra and M. alba leaves exhibited a significant interspecific and developmental specific accumulation patterns, with most DNJ alkaloids having the highest content in M. nigra leaves. In different growth stages, the metabolites of M. nigra leaves showed significant stage specificity. During the development stage of small fruit stage, obvious changes occurred in the chemical composition of M. nigra leaves, with a significant increase in DNJ alkaloid metabolites. Overall, this study provides a theoretical reference for the comprehensive development and sustainable utilization of M. nigra resources in Xinjiang.

Characterization of key bitter compounds in Idesia polycarpa var. vestita Diels fruit by sensory-guided fractionation

Idesia polycarpa var. vestita Diels (I. vestita) has become a promising oil crop due to its easily digestible and highly nutritious fruit oil. However, the intense bitter taste of its fruit greatly limits its development and promotion in the food industry. Herein, five key bitter compounds from I. vestita fruit were isolated by sensory-guided fractionation and characterized using ultra-high performance liquid chromatography-quadrupole time of flight-mass spectrometer and nuclear magnetic resonance. The bitter taste of the identified compounds was subsequently validated by threshold tests and computational molecular docking. The bitterness threshold in water of idesin was the lowest (12.051 mg/L), and all bitter substances spontaneously bound to the bitter receptors hTAS2R16 and hTAS2R14, with a stronger affinity for the latter (approximately −6.5 – −9.0 kcal/mol). This is the first systematic study of bitter compounds in I. vestita fruit, providing a scientific basis for revealing the mechanism of bitterness formation and bitterness control.

Electrooxidation of amoxicillin in aqueous solution with graphite electrodes: Optimization of degradation and deciphering of byproducts using HRMS

Contaminants of emerging concern (CECs) such as antibiotics have become a matter of worry in aquatic environments worldwide. Their presence in the environment has been increasing due to the inability of conventional wastewater and water treatments to annihilate them. Hence, attempts have been made to remove CECs using electrochemical oxidation (EO). Present study employed the low cost, active carbon based graphite sheet electrodes as anode and cathode to oxidize and degrade Amoxicillin (AMOX)- a β-lactum thiazolidine antibiotic. Optimization studies found pH 9, 45 mA cm−2, 81 cm2 electrode surface area, 6 mM electrolyte concentration and 60 min treatment time to be optimal for AMOX removal. Studies with varying concentrations of AMOX (20 mg L−1, 30 mg L−1 and 40 mg L−1) found that increase in concentrations of AMOX require higher current densities and treatment time for better TOC removal. High performance liquid chromatography photo diode array (HPLC-PDA) studies found 94% removal for 40 mg L−1 of AMOX at optimal conditions with 90% COD and 46% TOC removal. High resolution mass spectrometry (HRMS) studies using Ultra performance liquid chromatography-quadrupole time of flight-mass spectrometry (UPLC-Q-ToF-MS) identified major degradation mechanisms to be hydroxylation, β-lactum ring cleavage, breakage of thiazolidine ring chain from the aromatic ring and piperazinyl ring formation. The final byproducts of AMOX oxidation were carboxylic acids.

Discovery, isolation and structural identification of alkaloid glycosides in six traditional Chinese medicine such as Coptis chinensis

Alkaloids are important active ingredients occurring in many traditional Chinese medicines, and alkaloid glycosides are one of their existence forms. The introduction of saccharide units improves the water solubility of alkaloid glycosides thus presenting better biological activity.Because of the low content in plants, alkaloid glycosides have been not comprehensively studied. In this study, ultrahigh performance liquid chromatography-quadrupole time of flight-tandem mass spectrometry(UPLC-QTOF-MS/MS) was employed to identify and analyze the alkaloid glycosides in Coptis chinensis, Phellodendron chinense, Menispermum dauricum, Sinomenium acutum, Tinospora sagittata and Stephania tetrandra. The results showed that except Tinospora sagittata, the other five herbal medicines contained alkaloid glycosides. Furthermore, the alkaloid glycosides in each herbal medicine were identified based on UV absorption spectra, quasimolecular ion peaks in MS, fragment ions information in the MS/MS, and previous literature reports. A total of 42 alkaloid glycosides were identified. More alkaloid glycosides were identified in C. chinensis and Menispermum dauricum, and eleven in C. chinensis were potential new compounds. Furthermore, the alkaloid glycosides in the water extract of C. chinensis were coarsely se-parated by macroporous adsorption resin, purified by column chromatography with D151 cation exchange resin, ODS and MCI, combined with semi-preparative high performance liquid chromatography. Two new alkaloid glycosides were obtained, and their structures were identified by mass spectrometry and NMR data as(S)-7-hydroxy-1-(p-hydroxybenzyl)-2,2-N,N-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-O-β-D-glucopyranoside and(S)-N-methyltetrahydropalmatubine-9-O-β-D-glucopyranoside, respectively. This study is of great significance for enriching the information about the chemical composition and the in-depth development of C. chinensis. Meanwhile, it can provide a reference for rapid identification and isolation of alkaloid glycosides from other Chinese herbal medicines.

6-Oxofurostane and (iso)Spirostane Types of Saponins in Smilax sieboldii: UHPLC-QToF-MS/MS and GNPS-Molecular Networking Approach for the Rapid Dereplication and Biodistribution of Specialized Metabolites.

Identifying novel phytochemical secondary metabolites following classical pharmacognostic investigations is tedious and often involves repetitive chromatographic efforts. During the past decade, Ultra-High Performance Liquid Chromatography-Quadrupole Time of Flight-Tandem Mass Spectrometry (UHPLC-QToF-MS/MS), in combination with molecular networking, has been successfully demonstrated for the rapid dereplication of novel natural products in complex mixtures. As a logical application of such innovative tools in botanical research, more than 40 unique 3-oxy-, 3, 6-dioxy-, and 3, 6, 27-trioxy-steroidal saponins were identified in aerial parts and rhizomes of botanically verified Smilax sieboldii. Tandem mass diagnostic fragmentation patterns of aglycones, diosgenin, sarsasapogenin/tigogenin, or laxogenin were critical to establishing the unique nodes belonging to six groups of nineteen unknown steroidal saponins identified in S. sieboldii. Mass fragmentation analysis resulted in the identification of 6-hydroxy sapogenins, believed to be key precursors in the biogenesis of characteristic smilaxins and sieboldins, along with other saponins identified within S. sieboldii. These analytes' relative biodistribution and characteristic molecular networking profiles were established by analyzing the leaf, stem, and root/rhizome of S. sieboldii. Deducing such profiles is anticipated to aid the overall product integrity of botanical dietary supplements while avoiding tedious pharmacognostic investigations and helping identify exogenous components within the finished products.

Determination of multiple mycotoxins in chili powder using cold-induced liquid–liquid extraction and Fe3O4@MWCNTs-NH2 coupled with UPLC-Q-TOF/MS

Food matrix interference is still a big challenge in analyzing multiclass mycotoxins. Herein, a novel cold-induced liquid–liquid extraction-magnetic solid phase extraction (CI-LLE-MSPE) coupled with ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) method was explored for the simultaneous determination of multiple mycotoxins in chili powders. Fe3O4@MWCNTs-NH2 nanomaterials were prepared and characterized, and the factors affecting the MSPE process were investigated. Based on this, the CI-LLE-MSPE-UPLC-Q-TOF/MS method was established for determining ten mycotoxins in chili powders. The proffered technique eliminated the matrix interference effectively and demonstrated strong linearity (0.5–500 µg/kg, R2 ≥ 0.999), high sensitivity (limit of quantification was 0.5–1.5 µg/kg), and the recovery was 70.6%-111.7%. The extraction process is simpler than conventional methods, as the adsorbent can be separated using magnets, and reusable adsorbents are beneficial in reducing costs. In addition, the method can provide a valuable reference for pretreatment procedures for other complex matrices.

Metabolites profiling of protein enriched oyster mushroom (Pleurotus ostreatus (Jacq.) P. Kumm.) grown on oil palm empty fruit bunch substrate

This study aimed to compare the nutritional composition and metabolites profiles of the Pleurotus ostreatus oyster mushroom grown on oil palm empty fruit bunch (EFB) and rubber wood sawdust (RWS) substrates. The mushrooms were prepared for proximate analysis, beta-glucan content, and metabolites profiling using ultra high performance liquid chromatography-quadrupole time of flights mass spectrometer (UHPLC-QToF). Proximate analysis showed significantly higher crude protein, and beta-glucan contents (p < 0.05) were recovered from the mushrooms grown on EFB substrate as compared to those grown on RWS. Metabolite profiling of hot water extracts showed that three lipids (PE (17:1/0:0), PE (16:1/0:0), and hydroxyoctadecadienoic), two amino acids (acetyl-leucine, and threonic acid), and one polysaccharide (4-O-beta-d-galactopyranosyl-beta-d-xylopyranose) were present at a relatively higher level (maximum fold change >10) in mushrooms grown on EFB. Our results suggest that the metabolites of the mushroom cultivated on EFB substrate were altered. The innovative use of palm oil waste as mushroom cultivation substrate exerts the concept of waste-to-wealth, promoting Sustainable Palm Oil Initiative (SPOI), and enhancing food security through the production of high-protein mushrooms.

Mitochondrial Metabolomics of Sym1-Depleted Yeast Cells Revealed Them to Be Lysine Auxotroph.

Metabolomics has expanded from cellular to subcellular level to elucidate subcellular compartmentalization. By applying isolated mitochondria to metabolome analysis, the hallmark of mitochondrial metabolites has been unraveled, showing compartment-specific distribution and regulation of metabolites. This method was employed in this work to study a mitochondrial inner membrane protein Sym1, whose human ortholog MPV17 is related to mitochondria DNA depletion syndrome. Gas chromatography-mass spectrometry-based metabolic profiling was combined with targeted liquid chromatography-mass spectrometry analysis to cover more metabolites. Furthermore, we applied a workflow employing ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry with a powerful chemometrics platform, focusing on only significantly changed metabolites. This workflow highly reduced the complexity of acquired data without losing metabolites of interest. Consequently, forty-one novel metabolites were identified in addition to the combined method, of which two metabolites, 4-guanidinobutanal and 4-guanidinobutanoate, were identified for the first time in Saccharomyces cerevisiae. With compartment-specific metabolomics, we identified sym1Δ cells as lysine auxotroph. The highly reduced carbamoyl-aspartate and orotic acid indicate a potential role of the mitochondrial inner membrane protein Sym1 in pyrimidine metabolism.

Facile synthesis of triazine-based porous organic polymer for the extraction and determination of nitrofuran metabolites residues from meat samples with ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry

In this work, a novel triazine-based porous organic polymers, TAPT-BPDD, was firstly synthesized by a facile method at room temperature. After characterized by FT-IR, FE-SEM, XRPD, TGA, and nitrogen-sorption experiments, TAPT-BPDD was applied as solid-phase extraction (SPE) adsorbent for the extraction of four trace nitrofuran metabolites (NFMs) from meat samples. The key parameters including the adsorbent dosage, sample pH, type and volume of eluents, type of washing solvents were evaluated in the extraction process. Combined with ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS/MS) analysis, good linear relationship (1–50 µg·kg−1, R2>0.9925) and low limits of detection (LODs, 0.05–0.56 µg·kg−1) were obtained under the optimal conditions. When spiked at different level, the recoveries were in the range of 72.7–111.6%. The adsorption isothermal model and extraction selectivity of TAPT-BPDD were also studied in detail. The results showed that TAPT-BPDD was a kind of promising SPE adsorbent for the enrichment of organics in food samples.

An efficient method for qualitation and quantitation of multi-components of the herbal medicine Qingjin Yiqi Granules

Qingjin Yiqi Granules (QJYQ) is a Traditional Chinese Medicines (TCMs) prescription for the patients with post-COVID-19 condition. It is essential to carry out the quality evaluation of QJYQ. A comprehensive investigation was conducted by establishing deep-learning assisted mass defect filter (deep-learning MDF) mode for qualitative analysis, ultra-high performance liquid chromatography and scheduled multiple reaction monitoring method (UHPLC-sMRM) for precise quantitation to evaluate the quality of QJYQ. Firstly, a deep-learning MDF was used to classify and characterize the whole phytochemical components of QJYQ based on the mass spectrum (MS) data of ultra-high performance liquid chromatography quadrupole time of flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Secondly, the highly sensitive UHPLC-sMRM data-acquisition method was established to quantify the multi-ingredients of QJYQ. Totally, nine major types of phytochemical compounds in QJYQ were intelligently classified and 163 phytochemicals were initially identified. Furthermore, fifty components were rapidly quantified. The comprehensive evaluation strategy established in this study would provide an effective tool for accurately evaluating the quality of QJYQ as a whole.

Analysis of Intestinal Metabolites in SR-B1 Knockout Mice via Ultra-Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry.

Scavenger receptor class B type 1 (SR-B1), a multiligand membrane receptor, is expressed in a gradient along the gastrocolic axis. SR-B1 deficiency enhances lymphocyte proliferation and elevates inflammatory cytokine production in macrophages. However, whether SR-B1 affects intestinal metabolites is unclear. In this study, we detected metabolite changes in the intestinal tissue of SR-B1-/- mice, including amino acids and neurotransmitters, by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) and HPLC. We found that SR-B1-/- mice exhibited changes in intestinal lipid metabolites and metabolic pathways, including the glycerophospholipid, sphingolipid, linoleic acid, taurine, and hypotaurine metabolic pathways. SR-B1 deficiency influenced the contents of amino acids and neurotransmitters in all parts of the intestine; the contents of leucine (LEU), phenylalanine (PHE), tryptophan (TRP), and tyrosine (TYR) were affected in all parts of the intestine; and the contents of 3,4-dihydroxyphenylacetic acid (DOPAC) and dopamine (DA) were significantly decreased in both the colon and rectum. In summary, SR-B1 deficiency regulated intestinal lipids, amino acids, and neurotransmitter metabolism in mice.

Differences in Metabolomic Profiles by Birthplace in Mexican-Origin Hispanic Men Who Participated in a Weight Loss Lifestyle Intervention.

Birthplace, as a proxy for environmental exposures (e.g., diet), may influence metabolomic profiles and influence risk of cancer. This secondary analysis investigated metabolomic profile differences between foreign and U.S.-born Mexican-origin (MO) Hispanic men to shed light on potential mechanisms through which foreign- and U.S.-born individuals experience differences in cancer risk and risk factors. Plasma samples from MO Hispanic men (N = 42) who participated in a previous lifestyle intervention were collected pre-and post-intervention. Metabolomic profiles were characterized from samples using ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-QTOF). Models were visualized using supervised orthogonal projections to latent structures-discriminant analysis (OPLS-DA). Progenesis QI was used for peak integration and metabolite identification. Plasma metabolomic profiles differed between foreign- and U.S.-born pre-intervention (R2 = .65) and post-intervention (R2 = .62). Metabolomic profiles differed pre- versus post-intervention (R2 = .35 and R2 = .65) for the foreign- and U.S.-born group, respectively. Both endogenous metabolites and dietary components characterized differences between foreign- and U.S.-born participants pre- and post-intervention. Plasma metabolomic profiles from MO Hispanic men differed by birthplace. These results advance our understanding of relevant exposures that may affect cancer risk among MO Hispanic men born abroad or in the United States.

Optimization of Cordyceps sinensis fermentation Marsdenia tenacissima process and the differences of metabolites before and after fermentation

In this paper, we explored the interaction of factors which influenced the Cordyceps sinensis fermentation Marsdenia tenacissima (Roxb.) Wight et Arn, a Dai (a national minority of China) medicine, and the optimal fermentation conditions. The differences of C. sinensis metabolites in normal state (CN) and products of two-way liquid fermentation of C. sinensis and Marsdenia tenacissima (CM) and Marsdenia tenacissima (MT). The interactive effect of factors was analyzed and the best conditions are obtained through the box-behnken design (BBD) in response surface methodology (RSM). All metabolites were determined by ultra high performance liquid chromatography quadrupole time of flight mass spectrometer (UHPLC-Q-TOF-MS), analyzed and identified by metabonomics technology. Results showed that the optimum fermentation conditions were the concentration of raw medicinal materials is 160 g/L, the fermentation time is 6 days, the inoculation volume is 9.5%, the rotating speed is 170 rpm. 197 metabolites were identified in both positive ion and negative ion. 119 metabolites were significantly different between CN and CM. 43 metabolites were significantly different between CM and MT. Differential metabolic pathways were enriched. In conclusion, this paper optimizes the bidirectional fermentation process of M. tenacissima and C. sinensis through response surface methodology, and analyzes the changes of components from the level of metabonomics, so as to provide reference for exploring medicinal fungi fermentation of traditional Chinese medicine.

Analysis of chemical constituents of Sabia parviflora by ultrahigh performance liquid chromatography quadrupole time of flight tandem mass spectrometry

Effective and targeted identification of chemical components of the Chinese herbal medicine Sabia parviflora remains a major challenge. Herein, we used ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry to analyze the chemical composition of S.parviflora. Its chemical components were rapidly identified using the characteristic ion filtration method, which involves these steps: (1) summarize the characteristic ions based on similar skeletons and compounds with substitution patterns, and establish a database; (2) screen and classify different types of compounds in S. parviflora based on the characteristic ions; and (3) identify the compounds based on molecular weight, secondary fragments, and the database. In the present study, the characteristic ions in S. parviflora were grouped into five major classes. A total of 104 components were identified, including 12 potentially novel compounds. This rapid and accurate method provides an important basis for basic chemical research in S. parviflora.

Individual and synergistic effect of multi-frequency ultrasound and electro-infrared pretreatments on polyphenol accumulation and drying characteristics of edible roses

The freeze-dried (FD) edible roses with high content of bioactive substances and superior flavor have been favored by consumers. Nevertheless, the development of freeze-dried rose industry has been plagued by a long drying time and low efficiency. This study investigated the effects of ultrasonic pretreatment (UP) in multi-frequency modes and electro-infrared pretreatment (EIP) prior to FD on polyphenol accumulation and drying characteristics of roses. The mechanism was explored by the changes in microstructure, equivalent circuit parameters, and phenol identifications of rose. The results showed that the FD time of roses decreased by 26 % after ultrasonic-infrared sequential synergistic pretreatment (UP + EIP) due to the damage of cell membrane permeability from UP. The quality attributes of UP + EIP products including color, phenols, and antioxidant activity (DPPH and ABTS radical scavenging rates) remarkably improved. UP + EIP significantly (p < 0.05) increased the content of polyphenols, namely quercetin-3β-d-glucoside, phlorizin, procyanidin B2, gallicacid, and rutin in the FD roses quantified by ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-qTOF-MS/MS). Therefore, UP + EIP is an effective pretreatment method for shortening FD time and producing high-quality FD rose products with enhanced polyphenol content.

Ultraviolet (UV) light-induced geometrical isomerization of cinnamic acid containing molecules: A plausible, non-enzymatic approach to modify metabolite composition of plant extracts

• UV light can be used to enhance the cinnamic acid containing-metabolite composition of a plant. • LC-MS in combination with multivariate statistics can be used to probe for the presence of photo-induced metabolites in plant extracts. • UV-irradiated plant extracts have a coherent metabolite composition as compared to naturally growing plants. Cinnamic acid containing molecules are widely distributed in plants, and they are known to possess several pharmacological properties. Structurally, these molecules consist of C 6 C 3 skeletal backbone, with an olefin functional group on the C 3 part . Naturally, plants produce a broad spectrum of cinnamic acid derivatives with a trans configuration, however, cis isomers are formed due to excessive UV-light exposure, thereby increasing the already complex metabolomes of plants containing these compounds. In high sunlight intensive areas, the prevalence of the cis isomers of cinnamic acid containing molecules is becoming more prevalent and synonymous with extracts of many plants. It is therefore imperative to develop analytical and data handling strategies to account for the presence of these molecules, which can serve as a template for rational design of pharmacologically active plant extracts or smart photo-switchable drugs. Herein, we show that a combination of ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS) and principal component analysis (PCA) allows differentiation of plants extracts of which metabolome modification through UV-radiation has been carried out. The results of the current study show that geometrical isomerism is a feasible, non-enzymatic approach to modify metabolite composition of plants. This simple method can be used to create “super” extracts of plants containing structurally diverse metabolites to be evaluated for various pharmacological properties in the future.

UPLC-QToF-MS based fingerprinting of polyphenolic metabolites in the bark extract of Boehmeria rugulosa Wedd.

Boehmeria rugulosa Wedd. is an evergreen tree of Urticaceae family. Its bark has been extensively used in ethno-medicinal system for various ailments such as bone fracture, sprain, snakebite, and wound healing. Phyto-metabolites, which are considered as the principle components for biological activities, have been least explored for this plant. The present work investigated metabolite profiling of the stem bark of B. rugulosa in water extract using Ultra Performance Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry (UPLC-QToF-MS) technique coupled with the UNIFI platform. We identified, for the first time, 20 polyphenolic metabolites belonging to seven groups: caffeoylquinic acids, coumaroylquinic acids, flavan-3-ols, oligomeric flavonoids, caffeic acid derivatives, coumaric acid derivative, and flavone glycoside in the B. rugulosa extract. UNIFI informatics-coupled UPLC-QToF-MS platform aids in the quick identification and fragmentation pattern of metabolites, with higher degree of reproducibility. The present study provides a chemical and therapeutic basis for further exploration of B. rugulosa as a valuable source of phytochemicals that could be instrumental in deciphering its ethno-medicinal utility for various human diseases.

Identification of the chemical components of ethanol extract of Chenopodium ambrosioides and evaluation of their in vitro antioxidant and anti tumor activities

Purpose: To determine the characteristic chemical components of the ethanol extract of Chenopodium ambrosioides and evaluate their antioxidant and anti-tumor effects in vitro.&#x0D; Methods: The plant powder (5 g) was extracted with 1 L of 80 % ethanol at room temperature for 45 min, and then placed at 60 oC at varying microwave power and duration to obtain optimal extraction conditions. Characteristic chemical components were detected using ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS). Kaempferitrin was isolated from the 80 % ethanol extract using a D101 macroporous resin column, and its content was assessed by high performance liquid chromatography (HPLC). The antioxidant effect of kaempferitrin was evaluated by its ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radicals, while its anti-proliferation activity in human liver cancer cells SMMC-7721 was determined using cell counting kit-8 (CCK-8) reagent.&#x0D; Results: Three characteristic components of ethanol extract of C. ambrosioides were obtained, namely, kaempferitrin, kaempferol-3-O-apigenin-7-O-rhamnoside and kaempferol-3-O-acetylapigenin-7-O-rhamnoside. Kaempferitrin was shown to possess strong DPPH radical and moderate ABTS radical scavenging activities. Kaempferitrin significantly inhibited the proliferation of SMMC-7721 cells at doses of 4 and 8 μg/mL, with half-maximal concentration (IC50) of 0.38 μM (p &lt; 0.05).&#x0D; Conclusion: Kaempferitrin extracted from C. ambrosioides has antioxidant and anti-tumor activities. The results reported here indicate that C. ambrosioides may have potential use in herbal medicine practice.