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Background End-stage renal disease (ESRD) is the final stage during the development of renal failure. Depression is the most common psychiatric disorder in patients with ESRD, which in turn aggravates the progression of renal failure, however, its underlying mechanism remains unclear. This study aimed to reveal the pathogenesis and to discover novel peripheral biomarkers for ESRD patients with depression through metabolomic analysis. Methods Ultra-high-performance liquid chromatography coupled with mass spectrometry (UPLC-MS) was used to explore changes of serum metabolites among healthy controls, ESRD patients with or without depression. The differential metabolites between groups were subjected to clustering analysis, pathway analysis, receiver operating characteristic (ROC) curve analysis. Results A total of 57 significant serum differential metabolites were identified between ESRD patients with or without depression, which were involved in 19 metabolic pathways, such as energy metabolism, glycerolipid metabolism, and glutamate-centered metabolism. Moreover, the area under the ROC curve of gentisic acid, uric acid, 5-hydroxytryptamine, 2-phosphoglyceric acid, leucyl-phenylalanine, propenyl carnitine, naloxone, pregnenolone, 6-thioxanthene 5'-monophosphate, hydroxyl ansoprazole, zileuton O-glucuronide, cabergoline, PA(34:2), PG(36:1), probucol and their combination was greater than 0.90. Conclusions Inflammation, oxidative stress and energy metabolism abnormalities, glycerolipid metabolism, and glutamate-centered metabolism are associated with the pathogenesis of ESRD with depression, which may be promising targets for therapy. Furthermore, the identified differential metabolites may serve as biomarkers for the diagnosis of ESRD patients with depression.

Tibetan hulless barley (“qingke” in Chinese) is a valuable food in Tibet. Purple qingke (PQK) and black qingke (BQK), two special pigmented types of hulless barley, have traditionally been widely cultivated and consumed in Tibet for thousands of years. The composition and contents of anthocyanins of two cultivars are unknown. This study aimed to explore the composition and contents of anthocyanins of two cultivars and their antioxidant capacities. Six anthocyanins were identified by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry from barley, most of the anthocyanins were acylated by malonyl group. The total anthocyanin contents ranged from 141 to 2,304 μg/g in PQK and from 248.7 to 2902.9 μg/g in BQK. Furthermore, qingke has strong antioxidant activity against DPPH, ABTS˙+, and FRAP. Qingke may be useful for treating or preventing diseases caused by the overproduction of radicals.

Blackberry fruit (Rubus spp. L., Rosaceae) is a source of natural antioxidants for cosmetic products due to its abundance of phenolic compounds. The aim of the present study was to characterize blackberry extracts of the cultivars ‘Tupy’, ‘Guarani’, and ‘Xavante’ and to develop niosomes loaded with blackberry extracts for future applications in cosmetic formulations. After fruit collection at the Experimental Central Stations of the Federal University of Parana, Quatro Barras, Brazil, the extracts were solubilized in different solvents. The choice of extraction solvent and blackberry cultivar was established by analysis to evaluate the total phenolic content, radical scavenging capacity, and chemical characterization by ultra-performance liquid chromatography couple to quadrupole time-of-flight-mass spectrometry. These tests revealed that fruits of Rubus ‘Xavante’ had the highest potential to be used as an active cosmetic ingredient. Niosomes formulated with the acetone/water extract obtained from the fruit of this cultivar showed both good performance and encapsulation efficiency. The results indicate that this type of niosome can be considered a promising delivery system with the potential to improve skin absorption, in addition to the antioxidant, anti-aging, and UV protective activities of the polyphenol-rich blackberry extracts.

Background Neonatal acne occurs in the first few weeks after birth. Some lesions are more serious and leave scars. Maternal surface skin lipids (SSL) have a strong correlation with SSL of infants. The establishment of prediction rank model based on maternal SSL is essential to the prevention and treatment of neonatal acne. Method Surface skin lipids samples were collected from the mothers (M) of 56 neonatal acne patients and the mothers (HM) of 19 healthy infants. Surface skin lipids from the right forehead were collected using a noninvasive method. UPLC-QTOF-MS was applied to detect SSL. Partial least squares discriminant analysis and receiver operating characteristic (ROC) analysis were performed to screen and validate potential lipids. Random forest (RF) and ROC analysis were used to establish a prediction model and evaluate its accuracy. Results Sixteen altered potential lipids belonging to fatty acids, sphingomyelins, and glycerides were associated with M. M had less lipids than HM. Spearman's correlation of 16 lipids revealed 9 with high correlation. They were chosen as characteristic values of the RF prediction model. And the model showed an average accuracy of 98% in the validation set. Conclusion We have established an RF model for predicting neonatal acne and have shown that high skin barrier-related lipids were markers for predicting neonatal acne.

The preventive and therapeutic effects of dark tea fermented by Eurotium cristatum (DTE) in glucose metabolism have been demonstrated. However, few studies have investigated comprehensive changes in the chemical composition and activity in DTE before and after fermentation. In this study, the metabolic profiling of raw samples and fermented samples was determined by ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS). Furthermore, a systematic analytical strategy combining global metabolomics and the spectrum-effect relationship based on α-glucosidase inhibition was employed for screening discriminant metabolites. As a result, 15 discriminant metabolites were identified in DTE samples. Among them, 10 metabolites (4 fatty acids, 1 dyphylline derivative, 3 lysophosphatidylcholines, and 2 triterpenes) increased in relative contents and the contents of the other 5 polyphenol metabolites decreased after fermentation. These metabolites were critical constituents possibly associated with DTE's hypoglycemic activity, which also might be suitable as quality evaluation indicators. This study provided a worthy insight into the exploration of representative active constituents or quality indicators of DTE.

The current data report describes the predictive identification of phytochemical constituents in the bioactive extract of Ipomoea mauritiana (IM) whole plant. For several formulations this plant was commonly used as ‘Vidari’ for Ayurvedic medicine. Traditionally, IM tubers are used to alleviate spinal cord pain, improve breast milk, as a tonic, increase sperm count and treating jaundice. The methanol extract can potentially scavenge free radicals and possess antibacterial activity that could be correlated with its chemical composition. So it is crucial to identify the major compounds of IM. An ultra-performance liquid chromatography coupled electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF/MS) method was adopted to detect the flavonoids, saponins, alkaloids, terpenoids in IM methanol extract. Data presented here is related to a published work Antioxidant and antibacterial activity of Ipomoea mauritiana Jacq.: A traditionally used medicinal plant in Bangladesh (Alam et al., 2020). Secondary metabolites were analyzed by the comparison of the mass fragmentation arrangements with Waters UNIFI library that enables for positive identification of the compounds based on the spectral match.

Aminoglycosides are a class of broad-spectrum antibiotics with several clinical uses. Owing to the ototoxicity and nephrotoxicity of aminoglycosides, therapeutic drug monitoring is required. This study aimed to devise a high-throughput method for identification and quantitative determination of aminoglycoside antibiotics in human plasma samples using ultra-performance liquid chromatography–quadrupole time-of-flight-mass spectrometry (UPLC–Q-ToF-MS). Plasma samples (100 µL) spiked with five aminoglycosides (streptomycin, spectinomycin, amikacin, kanamycin, and gentamycin) and an internal standard (ribostamycin) were diluted and centrifuged in aqueous formic acid and acetonitrile. The clear supernatant extract was evaporated and reconstituted in the mobile phase, of which 4 µL was subjected to UPLC–Q-ToF-MS. Prominent peaks were observed for the drugs within 3 min. The recoveries of five aminoglycosides from plasma samples were 92.6–120%. The regression equations showed excellent linearity (0.9999 ≥ r2 ≥ 0.9987) within the range of 1.0–100 µg/mL, and detection limits of 0.5–2.0 µg/mL. The coefficients of the intra- and inter-day variations for five drugs were less than 11.8%, while the accuracy of quantitation was in the range of 89–111%. In this study, a novel method was presented for identification and determination of aminoglycosides in human plasma samples using UPLC–Q-ToF-MS analysis. This method can be applied to high-throughput analysis used for clinical and environmental purposes.

The metabolism and pharmacokinetics of gefitinib (Iressa®, N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholino-propoxy)quinazolin-4-amine), a selective thymidylate kinase inhibitor for the epidermal growth factor receptor (EGFR), was studied after IV and PO administration to male C57BL6 mice at 10 and 50 mg/kg respectively.The pharmacokinetics and metabolism of gefitinib were investigated using a range of rapid UHPLC-MS and UHPLC-IM-HRMS methods, using both reversed-phase (RP) and hydrophilic interaction liquid chromatography (HILIC), to rapidly determine the drugs pharmacokinetics and metabolic fate.Rapid oral absorption resulted in peak plasma concentrations at 1 h of ca. 7 µg/mL, that declined with a half-life of 3.8 h (2.6 h for the IV route), and providing an estimated oral bioavailability of 53%. Gefitinib itself was the major circulating drug-related compound in plasma extracts, with a total of 11 metabolites identified.The urinary profiles determined using both HILIC and RP-UPLC-IM-MS detected gefitinib and 10 metabolites or 15 metabolites respectively including the detection of a number of novel glucuronide conjugates.Despite rapid, sub 5 min, LC profiling methods being employed metabolite coverage was shown to be high and compared well with that of previous studies. The metabolism and pharmacokinetics of gefitinib (Iressa®, N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholino-propoxy)quinazolin-4-amine), a selective thymidylate kinase inhibitor for the epidermal growth factor receptor (EGFR), was studied after IV and PO administration to male C57BL6 mice at 10 and 50 mg/kg respectively. The pharmacokinetics and metabolism of gefitinib were investigated using a range of rapid UHPLC-MS and UHPLC-IM-HRMS methods, using both reversed-phase (RP) and hydrophilic interaction liquid chromatography (HILIC), to rapidly determine the drugs pharmacokinetics and metabolic fate. Rapid oral absorption resulted in peak plasma concentrations at 1 h of ca. 7 µg/mL, that declined with a half-life of 3.8 h (2.6 h for the IV route), and providing an estimated oral bioavailability of 53%. Gefitinib itself was the major circulating drug-related compound in plasma extracts, with a total of 11 metabolites identified. The urinary profiles determined using both HILIC and RP-UPLC-IM-MS detected gefitinib and 10 metabolites or 15 metabolites respectively including the detection of a number of novel glucuronide conjugates. Despite rapid, sub 5 min, LC profiling methods being employed metabolite coverage was shown to be high and compared well with that of previous studies.

Eleven compounds were identified during profiling of polyphenols by UPLC-QTOF/MS. In abundance was quercetin-3-O-α- l -arabinofuranoside in M. malabathricum ethanolic leaves extract while 6-hydroxykaempferol-3-O-glucoside was present in the leaves extract of M. decenfidum (its rare variety). TPC and TFC were significantly higher in M. decemfidum extract than M. malabathricum extract. During DPPH, FRAF and β-carotene bleaching assays, M. decemfidum extract exhibited greater antioxidant activity compared to M. malabathricum extract. Effect of M. malabathricum and M. decemfidum extracts on viability of MDA-MB-231 cell at concentrations 6.25–100 μg/mL were evaluated for 24, 48 and 72 h. After 48 and 72 h treatment, M. malabathricum and M. decemfidum leaves extracts exhibited significant activity in inhibiting MDA-MB-231 cancer cell line with M. malabathricum extract being more cytotoxic. M. malabathricum and M. imbricatum serves as potential daily dietary source of natural phenolics and to improve chemotherapeutic effectiveness.

Objective : The purpose of this study was to observe the difference of extraction method (both conventional and non-conventional) based on metabolite profile using UPLC-QToF-MS/MS system from Peperomia pellucida (L.) Kunth. Methods: Dried samples were extracted using the conventional maceration method and the optimum ofionic liquid-based microwave-assisted extraction(IL-MAE) methods.Metabolite profiling was performed using UPLC-QToF-MS/MS system with some modifications adjusted to the instrument condition. The data was analyzed using Masslynx 4.1 software. Results: based on the results, there were differences on profiling metabolite from both conventional and non-conventional extraction methods that was extraction method using organic solvent and ionic liquid solvent ([BMIM]BF 4 at the optimum condition.The extract obtained using ILMAE method had a peak depth with a well-separated Rt (retention time) value ranging from 0.5 to 7.5 min which means that the extracted compound was from polar to nonpolar properties. The extract obtained using maceration method, the peak spread on a separate Rt ranges from 2.5 to 7.5 min. Also, both extracts obtained have different area under curve (AUC) values with amount total of 22285 (IL-MAE) and 12679 (maceration), respectively, and showed the IL-MAE twice as large as that of the maceration. Conclusion: based on the results, conventional and non-conventional extraction method showed differences in metabolite profiling based on Rt value and mass spectrum m/z of each peak. Key words: Peperomia pellucida (L) Kunth, Hyphenated system, Ionic liquid- based microwave-assisted extraction, Maceration, Metabolite profiling.

The OPLS-DA, the UPLC–QTOF/MS and chemometric approaches were adopted to discriminate the different varieties belonging to three pumpkin species. Furthermore, this study included a comparison of the phenolic metabolites, antioxidant (DPPH, FRAP and ABTS activity), in vitro α-glucosidase and α-amylase activities, and cell cytotoxicity of the leaves of three varieties of pumpkin species (Cucurbita moschata, C. maxima, and C. pepo). Leaves of all three varieties of pumpkin species contained 23 phenolic metabolites. The OPLS-DA, the UPLC–QTOF/MS and chemometric approach showed that genistin was the candidate biomarker responsible for the separation of butternut squash (C. maxima) from the other two species, while 7-methylquercetin-3-galactoside-6″-rhamnoside-3‴-rhamnoside separated the Caserta squash (C. pepo) from the other two pumpkin species. Leaves of Green Hubbard (C. maxima) showed the highest antioxidant capacity (FRAP and ABTS activities), while butternut squash (C. moschata) showed the highest DPPH scavenging activity. Leaf extracts of butternut squash (C. moschata) and Green Hubbard (C. maxima) showed higher inhibition of α-glucosidase activity (IC50 18.22 μg mL−1) and α-amylase (IC50 18.11 μg mL−1) activities respectively, with no cytotoxic effect detected. The phenolic metabolites identified herein for the leaves of each variety of pumpkin species can be potential control markers for further standardisation. Leaves of Green Hubbard (C. maxima) provides functional properties for health benefits.

A novel normal-phase UPLC-QToF MS method was developed for sphingolipids analysis, which was then applied to separate and identify seven classes of sphingolipids in plasma. The time-consuming of this novel method was much shorter than previous 2D LC–MS method, while the sensitivity, repeatability and resolution for sphingolipids analysis were better or comparable. Besides, the differences of lipids in human plasma among diabetes mellitus patients, diabetes mellitus associated with atherosclerosis patients and control subjects were also compared. Finally, 12 sphingolipids species were identified as the amounts of which in diabetes mellitus associated with atherosclerosis patients were significantly higher than them in diabetes mellitus patients (VIP >1.0, p value 1.5). As the isomers of GalCs and GluCs were successfully separated, it was found that five GluCs, rather than GalCs, were included in these 12 potential biomarkers. These results suggest the need to separate GalCs from GluCs, and also indicated that there may be some extra activation pathways of diabetes mellitus associated with atherosclerosis, different from the atherosclerosis pathway and the diabetes mellitus pathway, causing cardiovascular disease.

To improve tea leaves' utility as a functional crop, it is necessary to understand the distribution and alteration of bioactive secondary metabolites during different growth stages. In the present study, secondary metabolites changes in C. sinensis leaves were investigated according to growth stage using ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC–QTof MS) with multivariate analysis. Through principal component analysis of the metabolites, patterns could be distinguished from samples harvested between April and August. On the loading plot, significant changes in the contents of metabolites were found during growth, and two gallotannins (4 and 10), three flavan-3-ols (6, 7, and 17), two flavonols (14 and 16), and two theaflavins (29 and 31) were evaluated as growth markers among thirty-one isolated metabolites. In particular, the flavonols and theaflavins increased gradually, whereas gallotannins and flavan-3-ols decreased continuously depending on the growth stage. A possible pathway could be deduced using metabolomics analysis, and information regarding physiological characterization and optimal harvesting time was obtained.

Accurate assays of plant antioxidants and other phytochemicals require efficient extraction conditions and enable rigorous assessments of crop varieties and production systems. This study assessed the extraction of phytochemicals and antioxidants from conventionally or organically grown red and golden beets (Beta vulgaris L.), using twenty solvent (S1-S20) mixtures containing water, methanol, and ethanol alone or with acids (ascorbic, formic, acetic). Red beetroot extracted with methanol with or without acid had the highest betanin content (2791.0 μg/g and 8222.3 μg/g of fresh weight [FW], respectively) and golden beetroot extracted with methanol/ascorbic acid/water had the highest vulgaxanthin I (193.7 μg/g and 15.0 μg/g of FW, respectively). The radical-scavenging activity and total phenolics in beetroot extracts reflected the different extraction efficiency of each solvent. UHPLC-QTOF-MS was used to identify twenty-seven phytochemicals, including 23 betalains, 2 amino acids, and 2 phenolic acids. Chemometric approaches discriminated the beet varieties and different extracts within one variety based on the composition and abundance of the key phytochemicals. The red beetroot extracted with aqueous ethanol with or without acid (S5, S7, S8, S9), and golden beetroot extracted with methanol-containing solvents (S15 for conventionally and S20 for organically) had the highest levels of phytochemicals, suggesting that these conditions efficiently extract key phytochemicals.

Four previously undescribed ginsenosides, along with five known analogues were isolated from wild ginseng by a UPLC-QTOF-MS-guided fractionation procedure. Their structures were elucidated on the basis of spectroscopic and spectrometric data (1D and 2D NMR, HR-ESI-MS). The isolated compounds could significantly inhibit the cigarette smoke extract (CSE)-induced inflammatory reaction in A549 cells. The HDAC2 pathway might be involved in the protective effect against the CSE-mediated inflammatory response in A549 cells.

Ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC–QToF–MS) analysis of dextromethorphan (DXM) and its metabolites—dextrorphan, 3-methoxymorphinan (3-MEM) and 3-hydroxymorphinan—in skeletal remains of rats exposed to DXM under different dosing patterns is described. Rats (n = 20) received DXM in one of four dosing patterns: acute (ACU1 or ACU2—100 or 200 mg/kg, i.p.; n = 5, respectively) or repeated (REP1 or REP2—3 doses of 25 or 50 mg/kg, i.p., 30 min apart; n = 5, respectively). Drug-free animals (n = 5) served as negative controls. Following euthanasia, the animals decomposed to skeleton outdoors. Bones were sorted by animal and skeletal element (vertebra, femur, pelvis, tibia, rib and skull), washed, air-dried and pulverized prior to dynamic methanolic drug extraction, filtration/pass-through extraction and analysis by UPLC–QToF–MS in positive electrospray ionization mode. Analyte levels (expressed as mass-normalized response ratios, RR/m) differed significantly between ACU1 and ACU2 (Mann–Whitney (MW), P

Diverse peanut varieties are widely cultivated in China. However, few studies have investigated the effects of germination on the phenolic profiles and antioxidant activities of specific Chinese peanut cultivars. Therefore, this study was conducted to evaluate the effects of germination on total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity, and phenolic profiles of seven peanut cultivars in China. The TPC, TFC, and antioxidant activities were determined by spectrophotometry, while phenolic profiles were analyzed by using ultra-high performance liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry (UPLC-QTOF-MS). The results found that germination significantly increased TPC, TFC, and antioxidant activity. Antioxidant activity was found to be closely related to TPC in germinated peanut extracts, which indicates that phenolics are the main contributors of antioxidants in germinated peanuts. In addition, germination induced significant changes in polyphenolic profiles. In the analyzed samples, 36 phenolic compounds were identified in which most were flavonoids. Overall, these findings highlight that germinated peanuts can be a good natural source of natural antioxidants for human consumption and functional food development.

Osteonecrosis of the femoral head (ONFH), one of the widespread orthopedic diseases with a decrease in bloodstream to the femoral head, is frequently accompanied by cellular death, trabecula fracture, and collapse of the articular surface. The exactly pathological mechanism of ONFH remains to explore and further identify. The aim was to identify the global urinary metabolic profiling of ONFH and to detect biomarkers of ONFH. Urine samples were collected from 26 ONFH patients and 26 healthy people. Ultra-performance liquid chromatography–quadrupole time of flight tandem mass spectrometry (UPLC-QTOF/MS) in combination with multivariate statistical analysis was developed and performed to identify the global urinary metabolic profiling of ONFH. The urinary metabolic profiling of ONFH group was significantly separated from the control group by multivariate statistical analysis. 33 distinctly differential metabolites were detected between the ONFH patients and healthy people. Sulfate, urea, Deoxycholic acid and PE(14:0/14:1(9Z)) were screened as the potential biomarkers of ONFH. In addition, the up/down-regulation of sulfur metabolism, cysteine and methionine metabolism, glycerophospholipid metabolism, and histidine metabolism were clearly be associated with the ONFH pathogenic progress. Our results suggested that metabolomics could serve as a promising approach for identifying the diagnostic biomarkers and elucidating the pathological mechanism of ONFH.

About 3.0 million people living under a typical tropical savannah climate in the Brazilian Federal District (FD) have faced an unprecedented water crisis. Considering the need for indirect reuse of wastewater for public supply, this work aimed to investigate FD water sources regarding the presence and risks of three contaminants of emerging concern: caffeine, carbamazepine, and atrazine. Samples from two current water sources (Descoberto and Santa-Maria Lakes) and two future water sources of the FD (Paranoá and Corumbá Lakes) were analyzed by solid-phase extraction followed by liquid chromatography coupled to hybrid quadrupole-time-of-flight mass spectrometry (UPLC-QTOF/MS). Method precision and accuracy were satisfactory and limits of quantification ranged from 0.37 to 0.54 ng/L. Higher concentrations were observed for caffeine in the future water sources (39 to 180 ng/L) followed by carbamazepine (5.4 to 25 ng/L) and atrazine (3.9 to 15 ng/L). The less-impacted water sources, in current use in the FD, present caffeine concentrations ranging from 4.8 to 32 ng/L and atrazine levels varying between 2.4 and 5.5 ng/L. Carbamazepine was not detected in these reservoirs. Environmental risk assessment indicates a possible risk for carbamazepine and atrazine, evidencing the need for further studies. No human health risk was depicted within the results.

Severe sepsis (SS) remains among the leading causes of death in both developed and developing countries. There is an urgent need for accurate biomarkers for early diagnosis of SS. Non-targeted mass-spectrometry was used to characterize sensitive and economical peripheral biomarker(s) associated with the serum metabolome from SS patients. In this study, serum samples were obtained from healthy controls and age-matched patients with SS. Multivariate statistical analysis was performed, and the metabolites identified were studied in relation to subsequent intervention procedures by receiver operating characteristic curve analysis. Metabolic differences among SS and control subjects were identified based on orthogonal signal correction-partial least squares discriminant analysis. Sphingosine, 5-methylcytidine, 3-dehydrocarnitine, 4-acetamido-2-aminobutanoic acid and phenyllactic acid in the SS subjects were significantly different from the controls. Three metabolites comprising sphingosine, 5-methylcytidine and 3-dehydrocarnitine were selected as candidate biomarkers and validated in separate, independent patient cohorts. These metabolite markers hold great potential for further development of SS early diagnostic markers. These findings suggest mass-spectrometry serum metabolomics may possess great potential for early diagnosis of SS patients.

As one of the most troublesome complications in patients with chronic renal disease, the etiology of uremic pruritus remains unknown, and the current therapeutic approaches are limited and unsatisfactory. To identify potential biomarkers for improving diagnosis and treatment and obtain a better understanding of the pathogenesis of uremic pruritus, we compared serum metabolome profiles of severe uremic pruritus (HUP) patients with mild uremic pruritus (LUP) patients using ultraperformance liquid chromatography-quadruple time-of-flight mass spectrometry (UPLC-QTOF MS). Partial least squares discriminant analysis (PLS-DA) showed that the metabolic profiles of HUP patients are distinguishable from those of LUP patients. Combining multivariate with univariate analysis, 22 significantly different metabolites between HUP and LUP patients were identified. Nine of the 22 metabolites in combination were characterized by a maximum area-under-receiver operating characteristic curve (AUC = 0.899) with a sensitivity of 85.1% and a specificity of 83.0% distinguishing HUP and LUP. Our results indicate that serum metabolome profiling might serve as a promising approach for the diagnosis of uremic pruritus and that the identified biomarkers may improve the understanding of pathophysiology of this disorder. Because the 9 metabolites were phospholipids, uremic toxins, and steroids, further studies may reveal their possible role in the pathogenesis of uremic pruritus.

Alzheimer's disease (AD) is a progressive neurodegenerative disease with neither definitive pathogenesis nor effective treatment method so far. Huang-Lian-Jie-Du-Tang (HLJDT) is a classic formula of traditional Chinese medicine (TCM) proven to have ameliorative effects on learning and memory deficits of dementia. Morris water maze (MWM) test and pathology analysis have demonstrated that HLJDT could ameliorate learning and memory deficits in AD mouse model, which may act via its anti-neuroinflammation properties. According to our previous studies, an UPLC-QTOF/MS-based metabolomics approach was performed to explore the potential mechanisms of HLJDT on preventing AD. As a result, a total of 23 potential metabolites (VIP1, |Pcorr|0.58, CUFjk excludes 0, P 0.05) contributing to AD progress were identified. The metabolic pathway analysis with MetPA revealed that glycerophospholipid metabolism, sphingolipid metabolism, arachidonic acid metabolism, linoleic acid metabolism and tryptophan metabolism were disturbed in mouse model of AD. After HLJDT treatment, 14 metabolites were restored back to the control-like levels.

Antifungal hordatines and their precursors, hydroxycinnamoylagmatines, were analyzed in dormant barley seeds and beer sample by ultra-performance liquid chromatography (UPLC) coupled to quadrupole time-of-flight mass spectrometry (QTOF-MS) operating in MSE-mode. Three hordatine basic structures, namely A, B and C, were found in barley and beer samples. All of these hordatines had analogous structures containing either one (hordatines A1, B1 and C1) or two (hordatines A2, B2 and C2) hydroxylated agmatine residues. All hordatines and hydroxycinnamoylagmatines were also present as sugar conjugates. Furthermore, N6 –methylated form of hordatines A, B and C, as well as their precursors, coumaroyl-N6-methylagmatine and feruloyl-N6-methylagmatine, were tentatively identified from barley samples. In addition, a number of other phenolamides, namely hydroxycinnamic acids conjugated to spermidine, were identified from barley and beer.

In this study, response surface methodology (RSM) and two artificial neural networks (ANN), multi-layer perceptron (MLP) and radial basis function (RBF), were used for the first time to model and optimize the extraction conditions of phenolic compounds in kidney beans in order to compare and establish effective prediction models. A total of 40 experiments were performed to screen for variables. The highest amount of polyphenols (917.2 mg GAE/100 g DW) and the strongest antioxidant activity (56.03 μmol Trolox/g DW) were obtained under optimal extraction conditions (70 min extraction time, 53% acetone, and 37 mL/g solvent-to-solid ratio). Response values were close to the predicted values with prediction accuracy as RBF > MLP > RSM, indicating that ANN model has higher prediction accuracy than RSM. Furthermore, UPLC-QTOF-MS analysis of extracts under the optimal extraction conditions showed that there were 25 antioxidant compounds detected, 13 of which were proposed from kidney beans for the first time.

Chrysanthemum cinerariifolium (C. cinerariifolium) is an empirically proven medicinal plant that has anticancer activities. This study aimed to profile metabolites, and cytotoxic activity of root, leaf, stem, and root extracts C. cinerariifolium on T47D cells and to determine the correlation of metabolite content with cytotoxic activity. The metabolite profile was carried out using UPLC-QToF-MS / MS, and cytotoxic activity was carried out using the MTT method. The results obtained in the form of a chromatogram were processed with the application Masslynk so that a metabolite profile data obtained. The data is then analyzed statistically using Principal Component Analysis (PCA). The results obtained on metabolite profiling showed that there were differences in metabolite profiles in the roots, stems, leaves, and chrysanthemums. The characteristic compounds in the flower section are D - (-) - Morphine and in the leaf part of genistein and N - [(5-Chloro-1,2,3-thiadiazol-4-yl) methyl] -1- (2-isopropyl-4 -methyl-1,3-thiazol-5-yl) -N-methyl ethanolamine. The major compounds in the roots, stems, and leaves are Orphenadrine with successive percentages of 9.11%, 10.16%, and 3.24%, and the major compounds in the flower section are D - (-) - Morphine with a percentage of 10, 86%. Furthermore, the results of the cytotoxic activity test showed differences in anticancer cytotoxic activity in the parts of the flower, leaf, stem, and root of C. cinerariifolium. Besides, there is a relationship between the metabolite content and anticancer cytotoxic activity of each part of C. cinerariifolium. The higher the level of orphenadrine compounds in the plant, the higher the potential for anticancer. Keywords: C. cinerariifolium, UPLC-QToF-MS/MS, T47D cells.

In the present study, a chemical profiling approach based on ultra-performance liquid chromatography coupled with photodiode array detection and time-of-flight mass spectrometry (UPLC-PDA-TOF-MS) was proposed to rapidly evaluate the chemical consistency between traditional and dispensing granule decoctions of traditional medicine combinatorial formulae and validated using San-Ao-Tang (SAT) as a model combinatorial formula. SAT is an effective traditional Chinese medicine, which is usually used in treating asthma and other diseases of the respiratory system. Two decoctions were prepared: traditional decoction, which is a water extract of three mixed constituent herbs of SAT; and dispensing granule decoction, which is a mixed water extract of each individual herb of SAT. Batches of these two decoction samples were subjected to UPLC-PDA-TOF-MS analysis and the data sets of t(R)-m/z pairs, ion intensities and sample codes were processed with supervised orthogonal partial least squared discriminant analysis to holistically compare their differences. Once a clear classification trend was found in the score plot, further statistics were performed to generate points at the two ends of S, and the components that correlated to these ions were regarded as the most changed components during decoction of the combinatorial formula. The changed components were identified by comparing the mass/ultraviolet spectra and retention times with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of the known compounds published in the literature. Using the proposed approach, global chemical differences were found between traditional and dispensing granule decoctions, like ephedrine, pseudoephedrine, norpseudoephedrine, licorice saponine H2, licorice saponine G2 and amygdalin.

Metabolomics is likely an ideal tool to assess tobacco smoke exposure and the impact of cigarette smoke on human exposure and health. To assess reproducibility and feasibility of this by UPLC-QTOF-MS, three experiments were designed for the assessment of smokers' blood. Experiment I was an analysis of 8 smokers with 8 replicates. Experiment II was an analysis of 62 pooled quality control (QC) samples from 7 nonsmokers' plasma placed as every tenth sample among a study of 613 samples from 160 smokers. Finally, to examine the feasibility of metabolomic study in assessing smoke exposure, Experiment III consisted of 9 smokers and 10 nonsmokers' serum to evaluate differences in their global metabolome. There was minimal measurement and sample preparation variation in all experiments, although some caution is needed when analyzing specific parts of the chromatogram. When assessing QC samples in the large scale study, QC clustering indicated high stability, reproducibility, and consistency. Finally, in addition to the identification of nicotine metabolites as expected, there was a characteristic profile distinguishing smokers from nonsmokers. Metabolites selected from putative identifications were verified by MS/MS, showing the potential to identify metabolic phenotypes and new metabolites relating to cigarette smoke exposure and toxicity.

Recently, it has been a big issue to distinguish the dried roots of Cynanchum wilfordii and Cynanchum auriculatum in Korean herbal medicine market. Although C. wilfordii and C. auriculatum have similar morphology, the types and quantities of metabolites may differ depending on the species. Thus, in this study, UPLC-QTOF/MS based metabolomics was applied to discriminate the roots of C. wilfordii and C. auriculatum. In the optimal LC/MS conditions, 64 known metabolites were analyzed in the two species. PCA and PLS-DA of metabolic profile data was able to differentiate between C. wilfordii and C. auriculatum. Furthermore, OPLS-DA and S-plot were applied to find the potential biomarkers for the discrimination of C. wilfordii and C. auriculatum. Finally, 4 known and 10 unknown metabolites were determined as the biomarkers, and their repeatability and reliability were also validated. This indicated the metabolite profiling is a robust approach to find discriminating biomarkers of C. wilfordii and C. auriculatum.

Metabolite extraction methods have been shown to be a critical consideration for pharmacometabolomics studies and, as such, optimization and development of new extraction methods are crucial. In the current study, an organic solvent-free method, namely, pressurised hot water extraction (PHWE), was used to extract pharmacologically important metabolites from driedMoringa oleiferaleaves. Here, the temperature of the extraction solvent (pure water) was altered while keeping other factors constant using a homemade PHWE system. Samples extracted at different temperatures (50, 100, and 150°C) were assayed for antioxidant activities and the effect of the temperature on the extraction process was evaluated. The samples were further analysed by mass spectrometry to elucidate their metabolite compositions. Principal component analysis (PCA) evaluation of the UPLC-MS data showed distinctive differential metabolite patterns. Here, temperature changes during PHWE were shown to affect the levels of metabolites with known pharmacological activities, such as chlorogenic acids and flavonoids. Our overall findings suggest that, if not well optimised, the extraction temperature could compromise the “pharmacological potency” of the extracts. The use of MS in combination with PCA was furthermore shown to be an excellent approach to evaluate the quality and content of pharmacologically important extracts.

Background Siwu decoction categorized formulae (SWDCF) are widely used for treating gynecological diseases. This study aims to elucidate the differences of bioactive constituents in SWDCF by ultra-high performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC - QTOF - MS /MS) and HPLC-DAD. Methods An efficient method based on UPLC - QTOF - MS /MS was developed for identifying the chemical profiles of SWDCF. HPLC-DAD method was used for quantifying seven chemical markers in SWDCF. Results Eighty four components were identified or characterized, including ten organic acids, thirty glycosides (monoterpene or iridoid or phenylpropanoids glycosides), fourteen lactones, eighteen flavonoids, and eleven alkaloids in the complex system. The datasets of tR-m/z pairs, ion intensities and sample codes were processed with supervised orthogonal partial least squared discriminant analysis to compare these decoction samples. After a clear classification was established, OPLS-DA was performed and 16 common components with relative quantity in SWDCF samples were determined. Gallic acid, protocatechuic acid, vanillic acid, caffeic acid, paeoniflorin, ferulic acid, and senkyunolide I were selected as the chemical markers to identify SWDCF by HPLC-DAD. Conclusion The chemical profiles with 84 components in SWDCF, including monoterpene glycosides, acetophenones, galloyl glucoses, even some isomers in the complex system were characterized by UPLC–QTOF–MS/MS.

Background . Currently available tests are insuffi cient to distinguish patients with epithelial ovarian cancer (EOC) from normal individuals. Metabolomics, a study of metabolic processes in biologic systems, has emerged as a key technology in the measurements of small molecular metabolites in tissues or biofl uids. Material and methods. To investigate the application of metabolomics on selecting EOC-associated biomarkers, 173 plasma specimens (80 newly diagnosed EOC patients and 93 normal individuals) were analyzed using ultra-performance liquid chromatography-quadrupole time-of- fl ight mass spectrometry (UPLC/QTOF/MS). A two-step strategy was performed to select EOC-associated biomarkers. The fi rst step was to select potential biomarkers in distinguishing 42 cancer patients from 58 normal controls through partial least-squares discriminant analysis (PLS-DA) and database searching, and the second step was to validate the discrimination performance of these biomarkers in a dataset contained 38 EOCs and 35 controls. Results . Eight candidate biomarkers were selected. The combination of these biomarkers resulted in the area of receiver operating characteristic curve (AUC) of 0.941, a sensitivity of 0.921, and a specifi city of 0.886 at the best cut-off point for detecting EOC. Discussion. Our fi ndings suggested that sharp differences in metabolic profi les exist between EOC patients and normal controls. The identifi ed eight metabolites associated with EOC may be served as novel biomarkers for diagnosis.

TCM has long been practised as an empirical system and retrieved tens of millions of lives from historically to currently [1]. It is considered as an ancient and classical paradigm of systems biology. In TCM, diagnosis and medication are based on “Syndrome” (“ZHENG” in Chinese Mandarin), which can be regarded as a profile of symptom combination, or clinical phenotypes, such as Cold or Hot Syndrome, and “Hot medication curing Cold Syndrome and Cold medication curing Hot Syndrome” is a standard therapeutic guide line. This classical systems medicine at the macro level has been validated and developed by its repeated clinical practice for thousands of years [2]. Hot and Cold medication are the four properties of Chinese medicinal herbs, precisely including cold, hot, warm and cool, which are also called the four natures or “four xing” in TCM. Cold-cool and warm-hot are two completely opposite categories of natures, whereas cold and cool or hot and warm differ in the degree. Chinese medicinal herbs with cold and cool nature can clear away heat, purge fire and eliminate toxic materials, which are mainly used for heat-syndrome; while with hot and warm nature have the actions of expelling cold, which are mainly used for coldsyndrome. The four natures are summarized mainly from the body's response. On the base of syndrome differentiation theory, only distinguish heat or cold nature of disease, and have a good understanding of the cold or hot property of TCM, so selectively apply corresponding medicinal herbs that you could achieve the desired effect. Herbal medicine has attracted much attention as a means of alternative therapy along with the orthodox medical system [3, 4]. In recent 30 years more and more reports on the four natures of TCM have appeared in the literature. To date several aspects of research such as the characteristics of thermodynamics, the changes of nervous system and the endocrine glands, energy metabolism, the systems biology analysis include genome, transcriptome, proteome, and metabolome are all supposed to explored the macro and micro framework on the four properties, among which metabonomics is the most novel tool [5]. It is a rapidly growing area of scientific research, which has been widely used in disease diagnosis, biomarker discovery, and research into the disease mechanisms [6-10]. Metabonomics aims at comparing the pattern of endogenous metabolites under defined temporal conditions as

Discriminating between two herbal medicines (Panax ginseng and Panax quinquefolius), with similar chemical and physical properties but different therapeutic effects, is a very serious and difficult problem. Differentiation between two processed ginseng genera is even more difficult because the characteristics of their appearance are very similar. An ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF MS)-based metabolomic technique was applied for the metabolite profiling of 40 processed P. ginseng and processed P. quinquefolius. Currently known biomarkers such as ginsenoside Rf and F11 have been used for the analysis using the UPLC-photodiode array detector. However, this method was not able to fully discriminate between the two processed ginseng genera. Thus, an optimized UPLC-QTOF-based metabolic profiling method was adapted for the analysis and evaluation of two processed ginseng genera. As a result, all known biomarkers were identified by the proposed metabolomics, and additional potential biomarkers were extracted from the huge amounts of global analysis data. Therefore, it is expected that such metabolomics techniques would be widely applied to the ginseng research field.