High-performance Liquid Chromatography-quadrupole Time-of-flight Mass Spectrometry Research Articles

Ethyl Acetate Fraction of Hedyotis diffusa Willd Induces Apoptosis via JNK/Nur77 Pathway in Hepatocellular Carcinoma Cells.

Background Hepatocellular carcinoma (HCC) is characterized by poor diagnosis and high mortality. Novel and efficient therapeutic agents are urgently needed for the treatment. Hedyotis diffusa Willd (HDW) is used to treat cancers, especially HCC in China. Purpose The study aimed to identify the main anti-HCC extract in HDW and to explore the mechanism of the active extract. Materials and Methods The high-performance liquid chromatography-quadrupole-time of flight mass spectrometry (HPLC-QTOF-MS) method was used for the simultaneous determination of main compounds in the ethyl acetate fraction of HDW (EHDW). The toxicity test of different HDW fractions was carried out on larvae at 2 day-post-fertilization (dpf) for 72 h. The in vivo anti-HCC effect of different HDW fractions was evaluated on a zebrafish tumor model by immersion administration. The antiproliferative effect of HDW fractions was determined with MTT assay, as well as hematoxylin and eosin (HE) staining assay. Hoechst 33258 staining was used to observe changes in nucleus morphology. Flow cytometry analysis was used to investigate apoptosis induction. Western blot analysis was used to examine apoptosis-related proteins, and key proteins in JNK/Nur77 signaling pathway. SP600125 was served to validate the apoptotic mechanism. Results EHDW showed the strongest tumor cell growth inhibitory effect on zebrafish tumor model. Further study revealed that EHDW induced apoptosis in zebrafish tumor model and in cultured Hep3B cells. Meanwhile, it has been shown that the levels of BCL2-associated X (Bax), cytochrome c (cyto c), cleaved-caspase 3, and poly-ADP-ribose polymerase (PARP) cells were upregulated. In contrast, the level of antiapoptotic B cell lymphoma-2 (Bcl-2) was downregulated in Hep3B cells. Additionally, EHDW activated JNK/Nur77 pathway by increasing the levels of p-JNK(Thr183/Tyr185) and p-Nur77(Ser351). Further study showed that blockage of JNK by SP600125 reversed EHDW-induced JNK/Nur77 pathway and the downstream apoptotic proteins. Conclusion In conclusion, EHDW exerted the anti-HCC effect, which may be attributed to the activation of JNK/Nur77 pathway. This study supported the rationale of HDW as an HCC therapeutic agent.

LC-MS Metabolite Profiling and the Hypoglycemic Activity of Morus alba L. Extracts.

Morus alba L. is used in traditional Chinese medicine for its anti-diabetic activity; however, the part of the hypoglycemic activity and related active metabolites are still not fully clarified. In this study, the metabolites in the M. alba roots, leaves, twigs, and fruits extracts (70% ethanol extracts) were systematically identified, and their hypoglycemic activity was evaluated by the high-fat diet/streptozotocin-induced 2 diabetes mellitus (T2D) mouse model. A total of 60 high-level compounds, including 16 polyphenols, 43 flavonoids, and one quinic acid, were identified by high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS) combined with the fragmentation pathways of standards and the self-established database. Among them, 23 metabolites were reported for the first time from this plant. In contrast to the extracts of M. alba leaves and fruits, the extracts of roots and twigs displayed significant hypoglycemic activity The glycemia was significantly reduced from 32.08 ± 1.27 to 20.88 ± 1.82 mmol/L and from 33.32 ± 1.98 to 24.74 ± 1.02 mmol/L, respectively, after 4 weeks of treatment with roots and twigs extracts. Compound 46 (morusin), which is a high-level component identified from the extracts of M. alba roots, also displayed significant activity in decreasing the blood glucose level of T2D mice reduced from 31.45 ± 1.23 to 23.45 ± 2.13 mmol/L. In addition, the extracts of roots and twigs displayed significant activity in reducing postprandial glycemia. This work marks the first comparison of the metabolites and hypoglycemic activity of M. alba roots, leaves, twigs, and fruits extracts, and provides a foundation for further development of M. alba extracts as anti-diabetic drugs.

The protective effects of peptides from Chinese baijiu on AAPH-induced oxidative stress in HepG2 cells via Nrf2-mediated signaling pathway

Antioxidant peptides have been widely reported. However, only a few reports have been published examining the antioxidant peptides derived from Chinese baijiu. In this study, 6 novel peptides derived from Chinese baijiu were identified successfully using high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (HPLC-QTOF-MS) with a concentration of 0.835–24.540 µg/L. The underlying molecular mechanisms were investigated, and their cytoprotective effects were examined against 2,2’-azobis (2-methylpropanimidamidine) dihydrochloride (AAPH)-induced oxidative stress in HepG2 cells. The results showed that these peptides exerted protective effects by suppressing reactive oxygen species (ROS) generation, preventing malondialdehyde (MDA) formation, and upregulating cellular antioxidant enzyme activities (SOD, CAT, and GSH-Px) in a dose-dependent manner. Further experiments proved that these peptides exerted antioxidant effects via Nrf2/ARE-mediated signaling pathway by promoting Nrf2 nuclear translocation, inhibiting ubiquitination, and enhancing transcription capacity of Nrf2 in HepG2 cells. These findings provide the molecular basis for the effects of antioxidant peptides derived from Chinese baijiu, which is important for a deeper understanding of the relationship between human health and moderate drinking.

Screening and identifying cucurbitacins and cucurbitacin glycosides in Cucumis sativus using high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry combined with in-source fragmentation and alkali adduct ions.

Cucumber, as a popular fruit and vegetable, has tremendously contributed to providing a sufficient and high-quality food supply. However, the cucumber plant metabolites, which may possess potential benefits for human health, were rarely reported. In addition, rapid detection of these metabolites from the complex biological matrix of cucumber samples is a tremendous challenge. A rapid detection method was established to systematically screen cucurbitacins and cucurbitacin glycosides in cucumber plants by combining high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC-Q-TOF-MS) with in-source fragmentation (ISF). Moreover, the alkali cations, including acetic acid, 0.1% LiCl, 0.1% NH4 Cl, 0.1% NaCl, and 0.1% KCl, were added to the mobile phase for improving the ion response. The fragmentation pathways of seven cucurbitacins and cucurbitacin glycosides were primarily investigated. The characteristic ISF ions at m/z 501.3211 and 503.3367 were identified and employed to screen 40 cucurbitacins and cucurbitacinglycosides from the complex biological matrix. Their structures were identified by their tandem mass spectrometry (MS/MS) spectra and fragmentation pathways of references. Finally, the metabolic distribution and network of cucurbitacins and cucurbitacin glycosides in cucumber plants were also proposed. This work marks the first systematic and comprehensive study of the metabolites in cucumber plants using HPLC-Q-TOF-MS technology, providing a template for screening and identifying the triterpenoids from other plant-derived medicines or food.

Ultraviolet-coupled advanced oxidation processes for anti-COVID-19 drugs treatment: Degradation mechanisms, transformation products and toxicity evolution.

Remdesivir (RDV), dexamethasone (DEX) and hydroxychloroquine (HCQ) were widely used in the treatment of COVID-19 pneumonia, possibly causing environmental risks and drug-resistance viruses. This study elucidated the degradation mechanisms and potential toxicity risks of the three anti-COVID-19 drugs by UV and ultraviolet-coupled advanced oxidation processes (UV/AOPs). All the drugs could be degraded by more than 98% within 3 min under the following optimal conditions: pH of 5.0 and drug-to-oxidant (H2O2) molar ratio of 1:200. Combined with density functional theory (DFT) analysis and high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS), twenty-four transformation products (TPs) were detected and the main degradation pathways were investigated. Based on bacterial luminescence inhibition test and the peak-area evolution of TPs, RDV and HCQ showed an obvious toxicity-increase region when TPs were generated in large quantities, while the toxicity of DEX continued to decline during degradation processes. By QSAR predictions, the main contributors to the toxicity evolution during the UV/AOPs were predicted. Halogen-containing TPs showed significantly higher toxicity than other TPs, and thus the chlorine-containing structure in HCQ presented the potential toxicity. Appropriate reaction parameters and adequate reaction time for the UV/AOPs could eliminate the toxicity of TPs and ensure environmental safety. This study could play a positive role in the treatment of anti-COVID-19 drugs and their environmental hazard assessment.

Degradation of OBS (Sodium p-Perfluorous Nonenoxybenzenesulfonate) as a Novel Per- and Polyfluoroalkyl Substance by UV/Persulfate and UV/Sulfite: Fluorinated Intermediates and Treatability in Fluoroprotein Foam.

Sodium p-perfluorous nonenoxybenzenesulfonate (OBS) is a novel fluorosurfactant used as the alternative to perfluorooctanesulfonic acid (PFOS) in several applications such as fire-fighting foams and chemical enhanced oil recovery ("EOR") in China, with the annual production capacity of about 3,500 t. Here, for the first time, we investigated the degradability of OBS under the conditions of UV/persulfate (UV/PS) and UV/sulfite (UV/SF) as typical redox processes. A higher reaction rate (1.05 min-1) and total organic carbon (TOC) reduction (46.9%) but a low defluorination rate (27.6%) along with the formation of a series of fluorinated intermediates were found in UV/PS, while a high defluorination rate (87.7%) was realized in UV/SF. In particular, a nontargeted workflow using high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC/Q-TOF-MS) was established to detect fluorinated intermediates. Combined with the theoretical calculation, the distinctive degradation pathways in both oxidation and reduction processes were proposed. The degradation mechanism of OBS in UV/SF was proposed to be H/F exchange and subsequent HF elimination. Furthermore, the diluted OBS-based fluoroprotein (FP) foam was used to investigate the degradation of OBS, which confirms the treatability using the redox approach. This work provides insights into the degradability of OBS, fluorinated intermediate search, and proper treatment of related contamination.

Determination of free and bound phenolics in northern highbush blueberries by a validated HPLC/QTOF methodology

Blueberry consumption has increased in recent years, partially due to the health benefits attributed to their phenolic content. This study aimed to optimize and validate a high-performance liquid chromatography/quadrupole-time of flight mass spectrometry (HPLC/QTOF-MS) methodology for the determination of free (i.e., solvent extractable) and bound (i.e., covalently bound to the plant cellular structures) phenolics in northern highbush blueberries. A total of 31 phenolics, including anthocyanins, flavonols, flavan-3-ols, and phenolic acids, were targeted in the positive electrospray ionization mde [ESI (+)], including MS/MS confirmation of their product ions. Free phenolics determination was validated in terms of sensitivity, reproducibility, linearity, and accuracy, and 21 free phenolic compounds were found above their limits of detection among six commercial blueberry cultivars, which were quantified with correction for the matrix effect. ‘Elliott’ presented the highest free phenolic and anthocyanin contents (3640 and 3430 mg kg−1 fresh weight, respectively). Malvidin-3-O-galactoside was the most abundant free phenolic for ‘Duke’, ‘Draper’, ‘Elliott’, and ‘Last Call’ while chlorogenic acid predominated for ‘Bluecrop’ and ‘Calypso’. Syringetin-3-O-glucoside was the most abundant bound phenolic compound among the six blueberry cultivars.

Discrepancies in the German Pharmacopoeia procedure for quality control of Quillaja saponin extracts.

This study focused on the evaluation of Quillaja saponin extracts with the additional quality designation DAB-which means the abbreviation of the German Pharmacopoeia (Deutsches Arzneibuch). This label suggests that Quillaja saponin extracts marked in this way are of pharmacopoeial quality and thus stand out from other Quillaja saponin extracts. The DAB ninth edition listed Quillaia saponin as a reagent. According to DAB, its quality must be checked by thin-layer chromatography (TLC), and three closely spaced zones in a defined retention factor (Rf) interval specify the saponin reagent. All the Quillaja saponin extracts obtained from different manufacturers and labeled as DAB quality complied with the TLC test. However, the analysis with high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (HPLC-Q-ToF-MS) clearly showed additionally an intense peak pattern of Madhuca saponins in all measured samples. The TLC test for Mahua seed cake, which is the press residue from Madhuca longifolia, surprisingly showed the same three closely spaced zones in the defined Rf interval. The three zones could be identified as Mi-saponins from Madhuca after scraping and extracting them from the stationary phase of the TLC plate and subsequent measurement by HPLC-Q-ToF-MS. Therefore, the specification of the saponin reagent in DAB characterizes erroneously Madhuca saponins that are not listed as a saponin plant source for the saponin reagent.

Identification of the Trace Components in BopuzongJian and Macleaya cordata Extract Using LC-MS Combined with a Screening Method.

Bopu powder® and Sangrovit® were developed from Macleaya cordata and are widely used in agriculture and animal husbandry, but their impurities have been rarely reported in the literature. Impurity analysis is of great importance to the quality and safety of veterinary drugs. In this study, high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS) combined with a screening method was used to screen and characterize the impurities in Bopu powder® and Sangrovit®. A total of 58 impurities were screened from Bopu powder® and Sangrovit® using the screening strategies, of which 39 were identified by their accurate m/z value, characteristic MS/MS data, and fragmentation pathways of references. This established method was used for impurity analysis for the first time and proved to be a useful and rapid tool to screen and identify the impurities of Bopu powder® and Sangrovit®, especially for those at trace levels in a complex sample. In addition, this study marks the first comprehensive research into impurities in these two products and has great significance for the systematic detection of impurities in other plant-derived drugs.

Effects of Citri Reticulatae Pericarpium and grapefruit juice on the pharmacokinetics of omeprazole in rats.

The effects of Citri Reticulatae Pericarpium (CRP) and grapefruit juice (GFJ) on the pharmacokinetics of omeprazole were investigated in this study. Sprague-Dawley rats were pretreated with CRP decoction or GFJ for 28 consecutive days. After a single intragastric administration of 6.0mg/kg, the concentration of omeprazole in the plasma was determined by high-performance liquid chromatography (HPLC), and the pharmacokinetic parameters were calculated by Kinetica software 5.0. A high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (HPLC-Q-TOF-MS) method was established to identify the chemical components in CRP decoction and GFJ. The results showed that the AUCt-∞ was significantly increased when coadministrated with CRP. The AUC0-t and AUC0-∞ was remarkably increased; the Cl was decreased when coadministrated with GFJ. A total of 31 and 28 bioactive compounds were identified in the CRP decoction and GFJ, respectively. Flavonoids and furanocoumarins, including hesperidin, hesperetin, naringenin, sinensetin, tangeretin, nobiletin, and 6',7'-dihydroxybergamottin, were simultaneously identified in CRP decoction and GFJ. This study indicates that the increased bioavailability of omeprazole may be due to the inhibition of hepatic cytochrome P450 enzymes, and the systemic exposure should be monitored when concomitant administration with CRP and GFJ. PRACTICAL APPLICATIONS: Citri Reticulatae Pericarpium (CRP) has been widely consumed as a daily condiment, functional food, and a traditional Chinese medicine. Omeprazole, primary metabolized by CYP450 enzymes, was usually coadministered with CRP for the treatment of gastrointestinal disease. Studies have confirmed that much fruit juices, including grapefruit juice, may affect drug metabolism enzymes. CRP and grapefruit (Citrus paradisi Macf.) belong to the genus Citrus and family Rutaceae with different species. Therefore, the pharmacokinetic interaction of CRP decoction and grapefruit juice with omeprazole is worthy of attention. The results of this study can provide basic pharmacological data support for the safe and effective clinical use of omeprazole. It can also provide a theoretical basis for the development of new functional products and daily application of CRP.

Comparisons of the anti-inflammatory, antiviral, and hemostatic activities and chemical profiles of raw and charred Schizonepetae Spica

Ethnopharmacological relevanceA common view in traditional Chinese medicine (TCM) theory is that “processing can alter the efficacy of crude drugs”. The clinical usage of some processed products may have already changed greatly over time during the development of modern scientific analysis. Therefore, the view of “processing can alter the efficacy of crude drugs” should be confirmed by comparative studies. Schizonepetae Spica (SS), a Chinese medicinal herb, is the dried spike of Schizonepeta tenuifolia Briq. It is available in two forms: raw products and charred products (Schizonepetae Spica Carbonisata, SSC; raw SS processed by stir-frying until carbonization). Raw SS is commonly used to treat TCM symptoms that resemble common cold, fever, respiratory tract infection and allergic dermatitis, while SSC has long been used as a remedy for TCM symptoms that resemble bloody stool and metrorrhagia. Aim of the studyWe aimed to examine whether stir-fry processing alters the anti-inflammatory, antiviral and hemostatic activities of SS and explore the chemical profile behind the potential changes in medicinal properties caused by stir-fry processing. Materials and methodsWe used cell models to examine the anti-inflammatory and antiviral effects of raw SS and SSC. The bleeding time of the tail bleeding model and clotting time of the capillary method in mice were used to compare the hemostasis properties of raw SS and SSC. The chemical profiles of SS and SSC were compared using a method combining gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF-MS) analysis. ResultsThe anti-inflammatory effects of SSC were less potent than those of raw SS. Both raw SS and SSC effectively inhibited viral infection in a dose-dependent manner, with IC50 values of 96.30 and 9.73 μg/mL and selectivity index (SI) values of >1.56 and 7.78, respectively. Interestingly, SSC showed more potent antiviral activities than raw SS. Intragastric administration of raw SS and SSC to mice demonstrated that the hemostatic effects of SSC were more potent than those of raw SS. By comparing the volatile chemical profiles of SSC, we found that twenty-nine constituents disappeared and that fifty-four new constituents were formed while the relative contents of five other components decreased and three other components increased. Additionally, the nonvolatile chemical profiles of raw SS and SSC differed, with thirty-two lower peaks and seven higher peaks in SSC than in SS. ConclusionOur study showed that raw SS and SSC support traditional practice for the clinical applications of these two products except for raw SS used for the treatment of viral infection. It is a fascinating challenge to form SSCs with both traditional hemostatic activities and antiviral properties after stir-fry processing. In addition, the volatile and nonvolatile chemical constituents of raw SS changed dramatically during processing. Further studies are warranted to explore whether the change in chemical constituents is in accordance with the purpose of processing.

A comprehensive study of three species of Paeonia stem and leaf phytochemicals, and their antioxidant activities

Ethnopharmacological relevancePaeonia plants have been widely used as traditional Chinese medicinal materials for more than 2,000 years in the treatment of cardiovascular, extravasated blood and female genital diseases; paeoniflorin and paeonol have been implicated as the plants’ primary active ingredients. Aim of the studyPrevious studies have been singularly focused on the chemical constituents and content variation of the Paeonia roots in the advancement of traditional Chinese medicine, with the plants’ stems and leaves considered useless. This study aims to explore the chemical constituents, content variation, and antioxidant capacity in Paeonia stems and leaves for the future utilization of traditional Chinese medicine, given that current practices of digging and trade endanger Paeonia in the wild. Materials and methodsHerein, secondary metabolites from the stems and leaves from six developmental stages of the annual growth cycle of Paeonia ostii T. Hong & J. X. Zhang, P. ‘Hexie’, and P. lactiflora Pall. were qualitatively and quantitatively analyzed via high-performance liquid chromatography with a diode array detector (HPLC-DAD) and high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). Antioxidant capacity at each stage was also evaluated by various free radical scavenging assays. ResultsA total of 24 metabolites were detected and identified, including 5 monoterpene glycosides, 4 tannins, 5 phenols, 9 flavonoids, and paeonol. Excepting paeonol and the phenols, the levels of each metabolite category were significantly higher in the leaves than the stems during all developmental stages. The paeoniflorin content in the P. ostii leaves was the highest during the first developmental stage and higher than the standards of the Chinese Pharmacopoeia, suggesting it to be the optimal harvesting stage for medicinal uses. Notably, the antioxidant capacity of the leaves was significantly greater than in the stems, particularly for the leaves of P. ‘Hexie’. ConclusionOur study indicates that the leaves of P. ‘Hexie’ have the potential to be a worthy medicinal substitute to Paeonia roots due to their high monoterpene glycosides, phenols, and flavonoids as well as their strong antioxidant capacity. Further, this study provides a theoretical basis for the development and utilization of non-root Paeonia plant sections as medicinal plant resources.

Effects of Different Storage Conditions on the Browning Degree, PPO Activity, and Content of Chemical Components in Fresh Lilium Bulbs (Liliumbrownii F.E.Brown var. viridulum Baker.)

Although Lilium brownii (L. brownii) bulbs are popular fresh vegetables, a series of quality problems still remain after harvest. In this study, fresh L. brownii bulbs were placed in the dark at 25, 4, and −20 °C and under light at 25 °C from 0 to 30 days; the chemical compositions were analyzed by ultraviolet spectrophotometry (UV) and high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). During the 30-day storage period, the browning degree increased over the storage time and with increasing temperature, but the contents of proteins and free amino acids decreased and were aggravated by light. The total polyphenol content increased until the 6th day at 25 °C (dark or light), but it did not significantly accumulate at −20 or 4 °C. The reducing sugar content showed a dynamic balance, but the total polysaccharide content decreased constantly in the four storage conditions. The polyphenol oxidase (PPO) activity increased with storage time and increasing temperature, while it was inhibited by light. The increase rates of malondialdehyde (MDA) content at −20 °C and light (25 °C) were higher than those at 4 and 25 °C. In addition, 12 secondary metabolites were identified, most of which accumulated during the storage period, for example, 1-O-feruloyl-3-O-β-D-glucopyranosylglycerol; 1,3-O-di-p-coumaroylglycerol; 1-O-feruloyl-3-O-p-coumaroylglycerol; and 1,2-O-diferuloylglycerol. The variations in nutrient levels had a low correlation with browning, but the variations in MDA, PPO, and secondary metabolite (phenolic acids) levels had a high correlation with browning. In conclusion, fresh L. brownii bulbs should be stored at a low temperature (4 °C) and in dark condition, and browning bulbs are excellent materials for secondary metabolite utilization.

Relationship between cardiotonic activity of Fuzi (Radix Aconiti Lateralis Preparata) and its fingerprint determined by liquid chromatography-mass spectrometry.

To investigate the relationship between the cardiotonic activity of Fuzi (Radix Aconiti Lateralis Preparata, RALP) and its fingerprint determined by liquid chromatography-mass spectrometry (LC-MS). First, the fingerprints of six processed products of RALP were established by high performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS) followed by analysis of the principal component of the relative peak area of its common peaks. Next, the scores of the first five principal components were used as input for an artificial neural network (ANN). Additionally, the therapeutic effect of RALP was assessed by measuring the hemodynamic indexes of heart failure model rats. Subsequently, fluorescence semi-quantitative polymerase chain reaction and an enzyme-linked immunosorbent assay kit were used to determine the effects of RALP-processed products on the serum levels of noradrenaline (NA), angiotensin-Ⅰ (Ang-Ⅰ), and the expression of β-norepinephrine receptor mRNA (β-NRm) in the rat cardiac tissues. P < 0.05 was used as the output of the ANN. Finally, a network was constructed to display the relationship between the LC-MS fingerprints and the cardiotonic activity of the RALP-processed products. Several types of RALPs can improve diastolic function, systolic function and heart rate. On the basis of the findings from the principal component analysis (PCA) of 16 common peaks of fingerprints of six RALP-processed products, it was revealed that the first five principal components may include 100% of the information of the original data. As observed from the multilayer perceptron neural network analysis, principal component 4 presented with the strongest effects on serum levels of NA and Ang-Ⅰ in rats, while principal component 1 exerted the greatest effect on β-NRm expression in cardiac tissue. The key findings obtained from this study indicated that the network constructed by the PCA-ANN may predict pharmacodynamic effects of the main ingredients of Traditional Chinese Medicine (TCM). This method may serve as a new approach to identify the relationship between LC-MS fingerprints and the pharmacodynamic effects of TCM ingredients.

Analysis of Intact Glycosidic Aroma Precursors in Grapes by High-Performance Liquid Chromatography with a Diode Array Detector.

Nowadays, the techniques for the analysis of glycosidic precursors in grapes involve changes in the glycoside structure or it is necessary the use of very expensive analytical techniques. In this study, we describe for the first time an approach to analyse intact glycosidic aroma precursors in grapes by high-performance liquid chromatography with a diode array detector (HPLC-DAD), a simple and cheap analytical technique that could be used in wineries. Briefly, the skin of Muscat of Alexandria grapes was extracted using a microwave and purified using solid-phase extraction combining Oasis MCX and LiChrolut EN cartridges. In total, 20 compounds were selected by HPLC-DAD at 195 nm and taking as a reference the spectrum of phenyl β-D-glucopyranoside, whose DAD spectrum showed a first shoulder from 190 to 230 nm and a second around 200–360 nm. After that, these glycosidic compounds were identified by High-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (HPLC-qTOF-MS). Disaccharides hexose pentose were the most abundant group observed with respect to the sugars and monoterpendiols the main aglycones found.

A Novel Chinese Honey from Amorpha fruticosa L.: Nutritional Composition and Antioxidant Capacity In Vitro.

False indigo (Amorpha fruticosa L., A. fruticosa) is the preferred tree indigenous for windbreak and sand control in Northwest China, while information on nutritional and bioactive characteristics of its honey is rare. Herein, 12 honey of Amorpha fruticosa L. (AFH) were sampled in Northwest China and the nutritional composition was determined. Sixteen mineral element and ten dominant polyphenols content were identified and quantified by ICP-MS (Inductively coupled plasma mass spectrometry) and HPLC-QTOF-MS (High performance liquid chromatography-Quadrupole time-of-flight mass spectrometry), respectively. Moreover, AFH demonstrated high levels of DPPH (1,1-Diphenyl-2-picrylhydrazyl) radical scavenging activity (IC50 100.41 ± 15.35 mg/mL), ferric reducing antioxidant power (2.04 ± 0.29 µmol FeSO4·7H2O/g), and ferrous ion-chelating activity (82.56 ± 16.01 mg Na2EDTA/kg), which were significantly associated with total phenolic contents (270.07 ± 27.15 mg GA/kg) and ascorbic acid contents (213.69 ± 27.87 mg/kg). The cell model verified that AFH exhibited dose-dependent preventive effects on pBR322 plasmid DNA and mouse lymphocyte DNA damage in response to oxidative stress. Taken together, our findings provide evidence for the future application of AFH as a potential antioxidant dietary in food industry.

Systematic identification metabolites of Hemerocallis citrina Borani by high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry combined with a screening method

Hemerocallis citrina Borani is an important crop and its flower buds are widely consumed in East Asian areas as a vegetable, as well as in traditional Chinese medicine, due to its health-promoting properties. Metabolites present in plant-derived foods or medicines are in part responsible for their desirable flavor profiles and health benefits. Nevertheless, detailed information about these compounds in H. citrina is scarce. Therefore, this study aimed to investigate the metabolites by high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). In this study, a total of 144 compounds, including 14 amides, 25 polyphenols, 44 flavonols, 35 anthraquinones, 15 naphthols, and 11 other components, were detected by the established screening method and were identified by their precise m/z values, characteristic tandem mass spectrometry (MS/MS) data and fragmentation pathways of references, 111 of which were reported in this plant for the first time. The distribution of identified ingredients in different parts of H. citrina was determined. Interestingly, colchicine, which had been reported as a toxic compound in the fresh flower buds in previous studies and various news reports, was not found. This work marks the first comprehensive study of metabolites from commercial flower buds and different parts of H. citrina.

Systematic Detection and Identification of Bioactive Ingredients from Citrus aurantium L. var. amara Using HPLC-Q-TOF-MS Combined with a Screening Method.

Bitter orange, Citrus aurantium L. var. amara (CAVA), is an important crop and its flowers and fruits are widely used in China as a food spice, as well as in traditional Chinese medicine, due to its health-promoting properties. The secondary metabolites that are present in plant-derived foods or medicines are, in part, responsible for the health benefits and desirable flavor profiles. Nevertheless, detailed information about the bioactive ingredients in CAVA is scarce. Therefore, this study was aimed at exploring the phytochemicals of CAVA by high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). Here, a systematic screening method combined with HPLC-Q-TOF-MS was presented. This technique was used to systematically screen metabolites, primarily from the complex matrix of CAVA, and to identify these compounds by their exact masses, characteristic fragment ions, and fragmentation behaviors. A total of 295 metabolites were screened by the screening method and 89 phytochemicals were identified in the flowers, fruits, roots, leaves, and branches of CAVA. For the first time, 69 phytochemicals (flavonoids, alkaloids, terpenoids, etc.) were reported from CAVA. The results highlight the importance of CAVA as a source of secondary metabolites in the food, medicine, and nutraceutical industries.

Metabolomic profile perturbations of serum, lung, bronchoalveolar lavage fluid, spleen and feces in LPS-induced acute lung injury rats based on HPLC-ESI-QTOF-MS.

Acute lung injury (ALI) is a clinically common and serious disease, underscoring the urgent need for clarification of its pathogenesis. According to traditional Chinese medicine (TCM) theories on the "lung-spleen-intestine axis" and its correlation with ALI, a high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS) metabolomic platform was applied to identify biomarkers from five bio-samples of control and model rats challenged with intratracheally administered lipopolysaccharide (LPS) based on multivariate mathematical statistical analysis. As a result, 19, 24, 24, 15 and 29 altered metabolites were identified in serum, lung, bronchoalveolar lavage fluid (BALF), spleen and feces samples, respectively. Metabolic pathway analysis showed that linoleic acid, sphingolipid, glycerophospholipid and bile acid metabolism pathways were mainly altered by ALI. Additionally, ROC curves were applied to assess the specificity and sensitivity of the biomarkers. ALI characteristic metabolomic spectra were then established to differentiate the control from the model group with a similarity discriminative threshold of 0.7. Additionally, to compare the metabolomic profiles of the five bio-samples and establish metabolic similarities and differences among them, correlation analysis was conducted in order to delineate an objective law of endogenous linkage along the lung-spleen-intestine axis. Therefore, this study provides insights into the mechanisms involved in ALI from a metabolomics perspective, which can be applied in characterization of the mechanism and early disease detection. Graphical abstract.

Exogenous Melatonin and Abscisic Acid Expedite the Flavonoids Biosynthesis in Grape Berry of Vitis vinifera cv. Kyoho.

Grape polyphenols contributing to more than half of the global polyphenol market were well studied; however, how melatonin (MLT), a potential plant hormone, and abscisic acid (ABA) affects polyphenols profile is still poorly understood. To explore whether these hormones are involved in polyphenolic biosynthesis, grape (Vitis vinifera cv. Kyoho) was exposed to MLT, ABA, and NDGA (nordihydroguaiaretic acid, an ABA biosynthesis inhibitor) treatments, and 16 polyphenols were identified from grape extracts by high performance liquid chromatography quadrupole time of flight mass spectrometry (HPLC-Q-TOF-MS). Both exogenous MLT and ABA significantly enhanced the biosynthesis of each flavonol and flavanol component, especially catechin, which was almost increased double by 200 µM of MLT treatment. Furthermore, the expression of genes involved in flavonoid biosynthesis, including 4-coumaroyl-CoA synthase, chalcone synthase, flavonoid 3′-hydroxylase, anthocyanin 3′-methyltransferase, flavonol synthase, flavonoid-3-O-glucosyltransferase, and flavonoid 3′,5′-methyltransferase were highly up-regulated as well but were down-regulated by NDGA. The present study provided new insights for improving flavonoids accumulation in agricultural production and its underlying mechanism.