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Plasma from patients with Parkinson’s disease (PD) is a valuable source of information indicating altered metabolites associated with the risk or progression of the disease. Neurotoxicity of dopaminergic neurons, which is triggered by aggregation of α-synuclein, is the main pathogenic feature of PD. However, a growing body of scientific reports indicates that metabolic changes may precede and directly contribute to neurodegeneration. Identification and characterization of the abnormal metabolic pattern in patients’ plasma are therefore crucial for the search for potential PD biomarkers. The aims of the present study were (1) to identify metabolic alterations in plasma metabolome in subjects with PD as compared with the controls; (2) to find new potential markers, some correlations among them; (3) to identify metabolic pathways relevant to the pathophysiology of PD. Plasma samples from patients with PD (n = 25) and control group (n = 12) were collected and the gas chromatography-time-of-flight-mass spectrometry GC-TOFMS-based metabolomics approach was used to evaluate the metabolic changes based on the identified 14 metabolites with significantly altered levels using univariate and multivariate statistical analysis. The panel, including 6 metabolites (L-3-methoxytyrosine, aconitic acid, L-methionine, 13-docosenamide, hippuric acid, 9,12-octadecadienoic acid), was identified to discriminate PD from controls with the area under the curve (AUC) of 0.975, with an accuracy of 92%. We also used statistical criteria to identify the significantly altered level of metabolites. The metabolic pathways involved were associated with linoleic acid metabolism, mitochondrial electron transport chain, glycerolipid metabolism, and bile acid biosynthesis. These abnormal metabolic changes in the plasma of patients with PD were mainly related to the amino acid metabolism, TCA cycle metabolism, and mitochondrial function.

The present study aimed to investigate the anti-proliferative effect of the ionic liquid-Graviola fruit (IL-GFE) extract on colon adenocarcinoma (HT29) cell lines and their kinetics behaviour to assess the Graviola fruit potential as a therapeutic alternative in cancer treatment. The phytoconstituents content of IL-GFE was identified using GC-TOFMS apparatus and measured its cytotoxicity on HT29 by tetrazolium bromide. Then the cytokinetic behaviour of the treated HT29 cells with IL-GFE was illustrated using the cells' growth curve. Besides, the cell cycle phase perturbation for the treated HT29 was applied using a flow cytometry technique. Qualitative identification of phytoconstituents of IL-GFE showed that Graviola fruit contains acetogenins, alkaloids, flavonoids, tannins and saponins compounds. IL-GF extract displayed a cytotoxicity effect on HT29 cells with the IC50 value of 10.56 µg/mL, while Taxol showed an IC50 value of 1.22 µg/mL. IL-GFE also decreased the cell generation number from 3.93 to 2.96 generations compared to Taxol-treated cells 2.01 generations. The microscope observation of the HT29 cells treated with the crude IL-GFE displayed loss of density and cell detachment. The extract's growth inhibition was related to the cell cycle arrest at the G0/G1 phase. IL-GFE inhibited colon adenocarcinoma HT29 cells' proliferation and affected their kinetic behaviour by lowering cell viability, inducing apoptosis, and arresting the cell cycle at the G0/G1 phase. ABSTRAK: Kajian ini bertujuan untuk mengkaji kesan anti-proliferatif ekstrak buah-ion Graviola (IL-GFE) pada garis sel adenokarsinoma kolon (HT29) dan tingkah laku kinetik mereka untuk menilai potensi buah Graviola sebagai alternatif terapi untuk barah rawatan. Kandungan fitokonstituen IL-GFE dikenal pasti menggunakan alat GC-TOFMS dan mengukur sitotoksisitasnya pada HT29 oleh tetrazolium bromida. Kemudian tingkah laku sitokinetik sel HT29 yang dirawat dengan IL-GFE digambarkan menggunakan keluk pertumbuhan sel. Selain itu, gangguan fasa kitaran sel untuk HT29 yang dirawat diaplikasikan menggunakan teknik sitometri aliran. Pengenalpastian kualitatif fitokonstituen IL-GFE menunjukkan bahawa buah Graviola mengandungi asetogenin, alkaloid, flavonoid, tanin dan sebatian saponin. Ekstrak IL-GF memperlihatkan kesan sitotoksisiti pada sel HT29 dengan nilai IC50 10.56 µg/mL, sementara Taxol menunjukkan nilai IC50 1.22 µg/mL. IL-GFE juga menurunkan jumlah penjanaan sel dari 3.93 hingga 2.96 generasi berbanding sel yang dirawat Taxol 2.01 generasi. Pemerhatian mikroskop sel HT29 yang dirawat dengan IL-GFE kasar menunjukkan kehilangan ketumpatan dan detasmen sel. Perencatan pertumbuhan ekstrak berkaitan dengan penangkapan kitaran sel pada fasa G0/G1. IL-GFE menghalang percambahan sel HT29 adenokarsinoma kolon dan mempengaruhi tingkah laku kinetik mereka dengan menurunkan daya maju sel, mendorong apoptosis, dan menghentikan kitaran sel pada fasa G0/G1.

Acacia Seyal gum (ASG), also known as gum Arabic, is an antioxidant-rich soluble fiber. ASG has been reported to have many biological activities, including anticancer, antidiabetic, antiulcer, and immunomodulatory activity. Extraction of bioactive compounds from ASG is commonly performed using conventional extraction methods. However, these techniques have certain limitation in terms of extraction time, energy, and solvent requirements. Ultrasound-assisted extraction (UAE) could be used as an alternative technique to extract bioactive compounds in less time, at low temperature, and with less energy and solvent requirements. In this study, the UAE extraction of ASG was optimized using response surface methodology (RSM). A face-centered central composite design (FCCCD) was used to monitor the effect of different independent factors of ultrasound operation (sonication time, temperature, and solvent ratio) on ASG extraction yield. In addition, screening and characterization of phytochemicals in 60% ethanol ASG extract was carried out using Raman microscopy, Fourier transform infrared spectroscopy (FTIR), and gas chromatography time-of-flight mass spectroscopy (GC-TOFMS) analysis. The results indicated that, under optimal conditions (extraction time 45 min, extraction temperature 40 °C, and solid–liquid ratio of 1:25 g/mL), the yield of ASG was 75.87% ± 0.10. This yield was reasonably close to the predicted yield of 75.39% suggested by the design of experiment. The ANOVA revealed that the model was highly significant due to the low probability value (p &lt, 0.0001). Raman spectrum fingerprint detected polysaccharides, such as galactose and glucose, and protein like lysine and proline, while FTIR spectrum revealed the presence of functional groups peaks value of alkanes, aldehydes, aliphatic amines, and phenol. GC-TOFMS spectroscopic detected the presence of strong d-galactopyranose, carotenoid, and lycopene antioxidant compounds. In conclusion, this study demonstrated that the UAE technique is an efficient method to achieve a high yield of ASG extracts. The selected model is adequate to optimize the extraction of several chemical compounds reported in this study.

Acacia Seyal gum (ASG), also known as gum Arabic, is an antioxidant-rich soluble fiber. ASG has been reported to have many biological activities, including anticancer, antidiabetic, antiulcer, and immunomodulatory activity. Extraction of bioactive compounds from ASG is commonly performed using conventional extraction methods. However, these techniques have certain limitation in terms of extraction time, energy, and solvent requirements. Ultrasound-assisted extraction (UAE) could be used as an alternative technique to extract bioactive compounds in less time, at low temperature, and with less energy and solvent requirements. In this study, the UAE extraction of ASG was optimized using response surface methodology (RSM). A face-centered central composite design (FCCCD) was used to monitor the effect of different independent factors of ultrasound operation (sonication time, temperature, and solvent ratio) on ASG extraction yield. In addition, screening and characterization of phytochemicals in 60% ethanol ASG extract was carried out using Raman microscopy, Fourier transform infrared spectroscopy (FTIR), and gas chromatography time-of-flight mass spectroscopy (GC-TOFMS) analysis. The results indicated that, under optimal conditions (extraction time 45 min, extraction temperature 40 °C, and solid–liquid ratio of 1:25 g/mL), the yield of ASG was 75.87% ± 0.10. This yield was reasonably close to the predicted yield of 75.39% suggested by the design of experiment. The ANOVA revealed that the model was highly significant due to the low probability value (p < 0.0001). Raman spectrum fingerprint detected polysaccharides, such as galactose and glucose, and protein like lysine and proline, while FTIR spectrum revealed the presence of functional groups peaks value of alkanes, aldehydes, aliphatic amines, and phenol. GC-TOFMS spectroscopic detected the presence of strong d-galactopyranose, carotenoid, and lycopene antioxidant compounds. In conclusion, this study demonstrated that the UAE technique is an efficient method to achieve a high yield of ASG extracts. The selected model is adequate to optimize the extraction of several chemical compounds reported in this study.

Depression is the most common mental illness, affecting approximately 4.4% of the global population. Despite many available treatments, some patients exhibit treatment-resistant depression. Thus, the need to develop new and alternative treatments cannot be overstated. Adenosine receptor antagonists have emerged as a promising new class of antidepressants. The current study investigates a novel dual A1/A2A adenosine receptor antagonist, namely 2-(3,4-dihydroxybenzylidene)-4-methoxy-2,3-dihydro-1H-inden-1-one (1a), for antidepressant capabilities by determining its metabolic profiles and comparing them to those of two reference compounds (imipramine and KW-6002). The metabolic profiles were obtained by treating male Sprague-Dawley rats with 1a and the reference compounds and subjecting them to the forced swim test. Serum and brain material was consequently collected from the animals following euthanasia, after which the metabolites were extracted and analyzed through untargeted metabolomics using both 1H-NMR and GC-TOFMS. The current study provides insight into compound 1a’s metabolic profile. The metabolic profile of 1a was similar to those of the reference compounds. They potentially exhibit their antidepressive capabilities via downstream effects on amino acid and lipid metabolism.

Distilled liquors are important products, both culturally and economically. Chemically, as a complex mixture, distilled liquor comprises various chemical compounds in addition to ethanol. However, the chemical components of distilled liquors are still insufficiently understood and compositional differences and similarities of distilled liquors from different cultures have never been compared. For the first time, both volatile organic compounds (VOCs) and non-VOCs in distilled liquors were profiled using mass spectrometry-based metabolomic approaches. A total of 879 VOCs and 268 non-VOCs were detected in 24 distilled liquors including six typical Chinese baijiu and 18 typical Western liquors. Principal component analysis and a correlation network revealed important insights into the compositional differences and similarities of the distilled liquors that were assessed. Ethyl esters, a few benzene derivatives, and alcohols were shared by most distilled liquors assessed, suggesting their important contribution to the common flavor and mouthfeel of distilled liquors. Sugars and esters formed by fatty alcohol differ significantly between the assessed Chinese baijiu and Western liquors, and are potential marker compounds that could be used for their discrimination. Factors contributing to the differences in chemical composition are proposed. Our results improve our understanding of the chemical components of distilled liquors, which may contribute to more rigorous quality control of alcoholic beverages.

Although many studies have been conducted on the components present in human breast milk (HM), research on the differences of chemical metabolites between HM, bovine milk (BM) and formula milk (FM) is limited. This study was to explore the chemical diversity of HM, BM and FM by metabolomic approaches. GC-TOFMS and UPLC-QTOFMS were applied to investigate the metabolic compositions in 30 HM samples, 20 FM samples and 20 BM samples. Metabolite profiling identified that most of the non-esterified fatty acids, which reflected the hydrolysis of triglycerides, were much more abundant in HM than those in FM and BM, except for palmitic acid and stearic acid. The levels of tricarboxylic acid (TCA) intermediates were much higher in FM and BM than those in HM. Each type of milk also showed its unique composition of free amino acids and free carbohydrates. In conclusion, higher levels of non-esterified saturated fatty acids with aliphatic tails

Schizophrenia is a severe mental disorder that affects 0.5–1% of the population worldwide. Current diagnostic methods are based on psychiatric interviews, which are subjective in nature. The lack of disease biomarkers to support objective laboratory tests has been a long-standing bottleneck in the clinical diagnosis and evaluation of schizophrenia. Here we report a global metabolic profiling study involving 112 schizophrenic patients and 110 healthy subjects, who were divided into a training set and a test set, designed to identify metabolite markers. A panel of serum markers consisting of glycerate, eicosenoic acid, β-hydroxybutyrate, pyruvate and cystine was identified as an effective diagnostic tool, achieving an area under the receiver operating characteristic curve (AUC) of 0.945 in the training samples (62 patients and 62 controls) and 0.895 in the test samples (50 patients and 48 controls). Furthermore, a composite panel by the addition of urine β-hydroxybutyrate to the serum panel achieved a more satisfactory accuracy, which reached an AUC of 1 in both the training set and the test set. Multiple fatty acids and ketone bodies were found significantly (P

Species of Tagetes, which belong to the family Asteraceae show different characteristics including, bloom size, shape, and color; plant size; and leaf shape. In this study, we determined the differences in primary metabolites and carotenoid yields among six cultivars from two Tagetes species, T. erecta and T. patula. In total, we detected seven carotenoids in the examined cultivars: violaxanthin, lutein, zeaxanthin, α-carotene, β-carotene, 9-cis-β-carotene, and 13-cis-β-carotene. In all the cultivars, lutein was the most abundant carotenoid. Furthermore, the contents of each carotenoid in flowers varied depending on the cultivar. Principal component analysis (PCA) facilitated metabolic discrimination between Tagetes cultivars, with the exception of Inca Yellow and Discovery Orange. Moreover, PCA and orthogonal projection to latent structure-discriminant analysis (OPLS-DA) results provided a clear discrimination between T. erecta and T. patula. Primary metabolites, including xylose, citric acid, valine, glycine, and galactose were the main components facilitating separation of the species. Positive relationships were apparent between carbon-rich metabolites, including those of the TCA cycle and sugar metabolism, and carotenoids.

The chemical constituents and antioxidant activity of the crude extracts of Mentha spicata were investigated. Phytochemical analysis indicated the presence of sugar, flavonoids and alkaloids in the crude extracts of Mentha spicata. GC-TOFMS (Gas Chromatography Time-of-Flight Mass Spectrometry) analysis indicated the presence of fatty acid methyl esters (hexa decane, hepta decane, octa decane) terpenoids, terpenoid alcohol, caryophyllene and glycosides. Total phenolic components of the crude extracts was found to be 27.26±0.62 mg/g gallic acid equivalent which was determined by Folin-Ciocalteu method. The DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity was found to increase with increasing concentrations and was found to be 54.84±0.57% with an IC50 value of 25.2µg/ml. The reported antioxidant activity may be due to the presence of flavonoids and fatty acid methyl esters which has the scavenging potential by reducing the free radicals.

A novel method for the residue analysis of wine spoilage compound 2,4,6-trichloroanisole is reported. Wine (60 ml) was extracted with 2 ml toluene in presence of 24 g MgSO4 and 6 g NaCl. Cleanup of the toluene phase by dispersive solid phase extraction with mixture of 100 mg CaCl2, 25 mg primary secondary amine and 50 mg MgSO4 was effective in minimising co-extractives and matrix effects. Time-of-flight and tandem mass spectrometric parameters were optimised to achieve linearity over 0.25–500 ng ml1 and method detection limit 0.0083 ng ml1 which is well below the odour threshold of 0.04 ng ml1 . Recoveries at 0.04, 0.2 and 0.8 ng ml1 were within 80–110% (±8%). The method was reproducible when tested for Argentinean wines with intra-laboratory Horwitz ratios being

The risks associated with a soil contaminated by polycyclic aromatic hydrocarbons (PAHs) are generally assessed by measuring individual PAHs in the soil and correlating the obtained amounts to known adverse biological effects of the PAHs. The validity of such a risk estimation is dependent on the presence of additional compounds, the availability of the compounds (including the PAHs), and the methods used to correlate the measured chemical data and biological effects. In the work underlying this thesis the availability, chemical composition and biological effects of PAHs in samples of soils from PAH-contaminated environments were examined. It can be concluded from the results presented in the included papers that the PAHs in the studied soils from industrial sites were not generally physically trapped in soil material, indicating that the availability of the PAHs was not restricted in this sense. However, the bioavailable fraction of the PAHs, as assessed by bioassays with the earthworm Eisenia Fetida, could not be assessed by a number of abiotic techniques (including: solid phase micro extraction, SPME; use of semi-permeable membrane devices, SPMDs; leaching with various solvent mixtures, leaching using additives, and sequential leaching) and it seems to be difficult to find a chemical method that can accurately assess the bioavailability of PAHs. Furthermore, it was shown that PAH-polluted samples may be extensively chemically characterized by GC-TOFMS using peak deconvolution, and over 900 components can be resolved in a single run. The chemical characterization also revealed that samples that appeared to be similar in terms of their PAH composition were heterogeneous in terms of their overall composition. Finally, single compounds from this large set of compounds, which correlated with different biological effects, could be identified using the multivariate technique partial least squares projections to latent structures (PLS). This indicates that PLS may provide a valid alternative to Effect Directed Analysis (EDA), an established method for finding single compounds that correlate to the toxicity of environmental samples. Thus, the instrumentation and data evaluation tools used in this thesis are clearly capable of providing a broad chemical characterization as well as linking the obtained chemical data to results from bioassays. However, the link between the chemical analyses and the biological tests could be improved as as an organic solvent that solubilised virtually all of the contaminants was used during the chemical analysis while the biological tests were performed in an aqueous solution with limited solubility for a number of compounds. Consequently the compounds probably have a different impact in the biological tests than their relative abundance in profiles obtained by standard chemical analyses suggests. The availability and bioavailability of contaminants in soil also has to be studied further, and such future studies should focus on the molecular interactions between the contaminants and different compartments of the soil. By doing so, detailed knowledge could be obtained which could be applied to a number of different contaminants and soil types. Such studies would generate the data needed for molecular-based modelling of availability and bioavailability, which would be a big step forward compared to current risk assessment practices.

Flavour research is a demanding domain in terms of analytical methodology as key odorants usually occur in trace amounts, often embedded in extracts containing volatile compounds at much higher concentrations. Since its early days, GC-MS has been a key tool in flavour laboratories enabling characterisation of thousands of volatile components in food products. However, as flavour chemists delve deeper into the understanding of flavour generation and delivery, there is a need for more powerful methodologies adapted to their specific needs. This paper will present two techniques that allow flavour separation and characterisation, namely GC-TOFMS and MS/MS. Moreover, APCI-MS, PTR-MS and REMPI-TOFMS will be discussed as they enable direct investigation of volatile compounds without any chromatographic step, thus studying release of flavour compounds during food processing or food consumption.