Gas Chromatography-mass Spectrometry Studies Research Articles

Phytochemical Analysis, Cytotoxic, Antioxidant, and Anti-coccidian Activities of Vitis vinifera Leaves Grown in Saudi Arabia

Background: Natural products are essential for medicinal purposes in developing countries. Traditional and contemporary medicine utilized plant products as alternative treatments for parasitic diseases. Objectives: The purpose of the investigation is to analyze and assess the bioactive compounds in Vitis vinifera leaf extracts, as well as their effect on coccidiosis. Materials and methods: Several analytical techniques were utilized, such as infrared spectroscopy (IR) and gas chromatography-mass spectrometry (GC-MS). Additionally, we assessed the total phenol, tannin, flavonoid examination, and cytotoxicity of the plant extracts using an MTT assay and UV-visible spectrophotometry (UV-Vis). Moreover, the samples’ levels of antioxidant activity (measured with DPPH and ABTS) were analyzed. Results: The leaf extract's GC-MS study identified many bioactive elements linked to plant secondary metabolites. Further, the total phenol, tannin, and flavonoid were 160.217 ± 0.652, 65.872 ± 0.263, and 34.0463 ± 0.545 (mg TAE/g DW), respectively. It also contains these extracts with a high antioxidant and DPPH radical scavenging activity. This reflects the number of bioactive chemicals that are present in these extracts. Conclusions: The findings of this research discuss techniques in phytochemical analysis of Vitis vinifera leaf extracts and the identification of biologically active components that support and justify the utilization of grape leaves in folk medicine in developing countries.

Phytochemical Screening, Antioxidant, Cytotoxic, Analgesic, Antidiarrheal Activity and GC-MS Analysis of the Leaves of Stachyphrynium Placentarium

The goal of the present study is to analyze the phytochemicals, antioxidants, cytotoxic, analgesic and anti-diarrheal properties of the leaves of Stachyphrynium placentarium methanol extract as well as its dichloromethane soluble fraction using GC-MS technique. The leaves of the S. placentarium were extracted with methanol and designated as SPM. Phytochemicals investigation exhibited the presence of several important secondary metabolites. Antioxidants such as total phenolic content, flavonoids content, total antioxidant, reducing power capacity and DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay SPM revealed non-significant antioxidant properties. The cytotoxicity study of the extract showed significant cytotoxicity (LC50 35.1093 µg/mL) compared to standard vincristine sulfate (LC50 0.482 µg/mL). Analgesic study of S. placentarium by hot plate model indicates both the doses exhibited significant central analgesic activity compared with the control. The maximum analgesic activity was observed at 90 min. SPM (500 mg/kg) is 60.21 %. SPM250 also showed a significant percentage of elongation or latency from 30 to 150 min 57.00, 59.64, 59.88, 58.07 and 53.71 %, respectively 30 min time interval. In the acetic acid writhing test, S. placentarium significantly outperformed the control and standard (SPM250 - 52.27 %, SPM500 - 34.09 and STD - 27.27 %), showing a significant increase in peripheral analgesic activity (p < 0.001). Antidiarrheal activity of SPM showed that SPM250 and SPM500 substantially decreased the feces in castor oil-induced diarrhea (p < 0.05). According to the study, the incidence and severity of diarrhea are significantly reduced by SPM500. Four chemicals are found in the dichloromethane-soluble portion of the methanol extract of S. placentarium leaves, according to GC-MS studies such as Octadecane, 6-methyl- (1.326 %), Dodecane (1.02 %), 1-Iodo-2-methylundecane (0.47 %), tetradecanoic acid 12-methyl, methyl ester (72.64 %). Tetradecanoic acid exhibits anti-inflammatory, anticancer, 5-reductase inhibitory, antioxidant, hemolytic and antifibrinolytic properties. HIGHLIGHTS The phytochemicals, antioxidants, cytotoxic, analgesic and anti-diarrheal properties of Stachyphrynium placentarium methanol extract (SPM) leaves. The dichloromethane soluble fraction is examined using Gas chromatography–mass spectrometry (GC-MS) and identified Octadecane, 6-methyl-, Dodecane, 1-Iodo-2-methylundecane, tetradecanoic acid 12-methyl, methyl ester. The brine shrimp lethality test exhibited that SPM has significant cytotoxicity (LC50 35.1093 µg/mL) compared to vincristine sulphate (LC50 0.482µg/mL), the extract showed significant cytotoxicity (LC50 35.1093 µg/mL). S. placentarium at both dosages (250 and 500 mg) provided significant central analgesia compared to the control in a hot plate model. In the acetic acid writhing test, S. placentarium showed significantly higher peripheral analgesic action (SPM250 - 52.27, SPM500 - 34.09 and STD - 27.27 %) compared to control and standard. In castor oil-induced diarrhoea, SPM250 and SPM500 (250 and 500 mg) showed significantly reduced faeces production (p < 0.05). GRAPHICAL ABSTRACT

Trichoderma harzianum TIND02 upregulates the expression of pathogenesis-related genes and enzymes and enhances gray blight resistance in tea

The gray blight incited by Pestalotiopsis and allied genera is a prevalent disease affecting tea cultivation, and managing it with Trichoderma spp. is an alternative to synthetic fungicides. Plants modify their arsenal system against pathogens when they are exposed to Trichoderma spp., which produces proteins and enzymes associated with pathogenesis. Understanding the expression pattern of defense-related markers will help in developing gray blight resistance tea cultivars. Thus, this study intended to induce resistance against gray blight in tea by Trichoderma harzianum TIND02. For this, a total of eight Trichoderma isolates originated from organic tea rhizospheres were characterized and evaluated for their efficacy. Dual culture test revealed isolate TIND02 as the most potential candidate with 74.6% inhibitory activity against gray blight pathogen Pseudopestalotiopsis theae. Molecular characterization based on ITS and tef-1 alpha genes confirmed isolate TIND02 as T. harzianum. Scanning electron microscopic study showed the mycoparasitic nature of T. harzianum TIND02 (TH-TIND02) to Ps. theae. The ethyl acetate extract of TH-TIND02 at 100 and 200 μg mL−1 showed potential inhibitory activity (>69.9%) against Ps. theae which confirmed the presence of higher volatile metabolites. Gas chromatography–Mass spectrometry study revealed that ethyl acetate extract of TH-TIND02 was composed of 21 major and minor volatile organic compounds with acetamide, 2, 2, 2-trifluoro-N, N-bis trimethyIsilyl–C (94.74%) as a major component. The isolate also produced chitinase, cellulase, β-1, 3 glucanase, and protease hydrolytic enzymes. Nursery experiments revealed that 2% and 5% doses (2 × 106 CFU mL−1) of TH-TIND02 significantly reduced respective 65.0% and 70.0% disease severity over control with improved plant growth. Besides, expressions of defense-related enzymes (chitinase, pHenolics, peroxidase, phenylalanine ammonia lyase, β-1, 3-glucanase, and polyphenol oxidase) and pathogenesis-related genes (chitinase and β-1, 3-glucanase) due to TH-TIND02 were determined. The secretion of defense-related enzymes was highly upregulated in plants applied with TH-TIND02 followed by Ps. theae inoculation compared to controls. The RT-qPCR analysis showed that the expression of both genes in co-inoculated plants was two-fold higher than in control after 21-day post incubation. These results suggest that TH-TIND02 application reduced gray blight severity by elevated enzyme activity and overexpressed pathogenesis-related genes in tea plants which offer for its eco-friendly and sustainable use as a bio-fungicide in tea gardens.

GC-MS Analysis and In-silico Molecular Docking of Phytoconstituents of Sansevieria roxburghiana against IL-1β and TNF-α for Anti-Rheumatic Activity

Background: Rheumatoid arthritis (RA) is recognized as a long-term inflammatory form of arthritis. This inflammation becomes long-lasting, and the synovial tissue thickens as a result of increased cell activity, protein production, and other variables in the joint, causing discomfort, redness, and warmth. It can also harm the joints of the hands, wrists, fingers, elbows, shoulders, toes, spinal column, and knees. Due to the autoimmune response, there is an aberrant production of inflammatory mediators such as Tumour necrosis factor- alpha (TNF-a), Interleukins (IL-1, IL-6, IL-7, IL-15, IL-17, IL-18, and IL-23), Granulocyte-macrophage colony-stimulating factor (GM-CSF), and Interferon (IFN)-ϒ. By inhibiting their production, those inflammatory mediators will play a pivotal role in the therapy of RA. Purpose of the Study: The goal of the current study was to explore the anti-rheumatic activity of Sansevieria roxburghiana using the computational docking method. Research Rationale: Administration of Non-steroidal anti-inflammatory drugs, steroidal medicaments, and Disease-modifying anti-rheumatoid therapies creates substantial adverse effects in people suffering with RA. To overcome the burdens and toxicity of drug products herbal medicines were taken into the field of research. Materials and Methods: For this, 14 bioactive compounds from ethanolic extract of S. roxburghiana leaves were identified using the GC-MS (Gas chromatography-mass spectrometry) study and utilized as a binding compound (ligand) for biological target interactions. The crystallographic geometries of two targeted molecules, Interleukin-1β (IL-1β) and Tumour necrosis factor-alpha (TNF-α), have been retrieved from the PDB database. Methotrexate, a well-known medication for IL-1β and TNF-α inhibitors, was chosen as the reference for the comparative analysis. Computational docking was performed using the Autodock4 (version 1.5.6) choice based on the score functions. Results: The results displayed the binding energy as follow -10.13, -9.77, -8.98 and -8.67 Kcal/mol of 2-isopropyl-5 methyl cyclohexyl 3-(1-(4-chlorophenyl)-3-oxobutyl)-C, 3-chloro-5-cholestene, stigmasterol, and cycloartanol against IL-1 β (PDB ID: 4GAI) and standard methotrexate was -8.14 Kcal/mol. For TNF-α (PDB ID: 5M2J) the binding energy is -8.94, -8.85, -8.46, and -8.24 Kcal/mol of cycloartanol, stigmasterol, 2-isopropyl-5 methyl cyclohexyl 3-(1-(4-chlorophenyl)-3-oxobutyl)-C and 3-chloro-5-cholestene by with comparing methotrexate (-8.89 Kcal/mol). Conclusion: Hence, it was indicated that the phytoconstituents of S. roxburghiana could serve as a potential approach for designing future antirheumatic drugs.

Detections of Chemicals and Migratory Plastics in Peritoneal Dialysis Fluids

Background: Peritoneal dialysis (PD) is an important modality of renal replacement therapy (RRT). Peritonitis and ultrafiltration failure are complications that have a long-term impact on PD patients. Besides touch contamination, procedural errors and clinical reasons of peritonitis, contaminants, and constituents of peritoneal dialysis fluids (PDFs) have been implicated in causing peritonitis and ultrafiltration failure. This study was aimed to test the PDFs in India for the presence of migratory plastics. Materials and Methods: PDFs from the two manufacturers in India were tested using liquid chromatography mass spectrometry (LCMS) and gas chromatography mass spectrometry (GCMS) with headspace analysis (volatile compounds) and pyrolysis of plastics (polymer compounds). The storage conditions and handling were uniform. Results: The results revealed impurities of acetate compounds and aldehyde derivatives of glucose degradation products (GDPs) with contaminants and leachable plastics. There were high levels of GDP derivative in the form of 5-hydroxymethylfurfural compounds (5-HMF). The analysis revealed the presence of plastic softeners in very high concentrations. Conclusion: The study unmasks the presence of chemicals and GDPs that can be implicated in pathogenesis of sterile peritonitis and ultrafiltration failure. The study demonstrated the presence of leachable plastics. In conclusion, LCMS and GCMS studies can be used to test PDFs for unwanted chemicals prior to human use.

Biomarkers for assessment of the polychemotherapy results in patients with adrenocortical cancer based on gas chromatography-mass spectrometry studies of urine steroid profiles

Background: The effectiveness of polychemotherapy (PCT) for adrenocortical cancer (ACC) is assessed by imaging tests with the RECIST 1.1 criteria. However, the presence of subclinical tumor foci does not allow for an objective measurement of the true tumor burden. As shown previously, postoperative assessment of the steroid metabolome by gas chromatography-mass spectrometry (GCMS) in ACC patients makes it possible to identify early signs of adrenal steroidogenesis abnormalities and of the recurrence of adrenocortical carcinoma.
 Aim: To identify biomarkers of response to PCT by GCMS study of the urine steroid profile in ACC patients after surgical resection of the tumor.
 Materials and methods: Urine steroid profiles were studied by GCMS (Shimadzu GCMS-TQ8050 gas chromatography-mass spectrometer) in 30 ACC patients (stages II, III and IV) after surgery and first line (combination of etoposide, doxorubicin and cisplatin with daily mitotane) and second line (gemcitabine combined with capecitabine and mitotane) PCT. The control group included 25 patients with hormonally inactive adenomas.
 Results: The response to PCT according to RECIST 1.1 criteria was obtained in 23 patients (Group 1, responders) and in 7 patients ACC progressed under PCT (Group 2, non-responders). In the responders, the urinary excretion of etiocholanolone, pregnanediol and pregnanetriol was lower than in the control group. The non-responders had higher urinary excretion of androgens, progestogens and tetrahydro-11-deoxycortisol (THS), compared to the responders and the control group. The patients with ACC progression under PCT had an increase in 3β,16,20-pregnenetriol (3β,16,20-dP3) levels and a decrease of the 3α,16,20-dP3/3β,16,20-dP3 ratio, compared to those in the PCT responders. The threshold values for urinary excretion of dehydroepiandrosterone (DHEA, ≤ 469 mcg/24h; AUC = 1.0), THS (≤ 223 mcg/24h; AUC = 1.0), and 3β,16,20-dP3 (≤ 130 mcg/24h; AUC = 0.986), as well as the 3α,16,20-dP3/3β,16,20-dP3 ratio (≥ 2.13; AUC = 1.0) had 100% sensitivity and specificity for the assessment of the PCT effectiveness.
 Conclusion: Different urine steroid profiles were obtained by GCMS in the ACC patients after PCT with and without treatment response. The 100% sensitivity and specificity of the threshold values for urinary excretion of DHEA, THS, 3β,16,20-dP3 and the 3α,16,20-dP3/3β,16,20-dP3 ratio for the assessment of PCT results indicate the potential to use these parameters as biomarkers of response or progression of the disease in the monitoring of PCT effects in ACC patients.

Unveiling the phytochemical profiles, selective bioactivity potential, and molecular docking study of bioactive compounds with target proteins using optimized bark extracts of Grewia asiatica L.

The current study aimed to explore the phytochemicals, estimate the anti-inflammatory, antioxidant, antimicrobial, and anticancer potentials of polyphenol enriched methanolic bark extract of Grewia asiatica (PEMGa), and perform molecular docking studies with identified phytocompounds. Here, we utilized response surface methodology (RSM) with a Box Behnken design (BBD) to enhance the phenolic content of the methanolic extract from G. asiatica bark by optimizing three crucial parameters like extraction time (h), temperature of extraction (°C), and solvent ratio (% v/v of methanol: water). Phytochemical quantification of PEMGa illustrated that the bark part contains a good amount of therapeutically potent phytochemical groups like phenolics, flavonoids, and tannins. Additionally, through HPLC analysis, eight compounds belonging to the phenolic and flavonoid groups have been quantified from PEMGa. By GC-MS (Gas chromatography–mass spectrometry) analysis of PEMGa, eleven phytochemicals were identified. Moreover, employing in-vitro bioactivity studies, PEMGa has unveiled its remarkable potential as an anti-inflammatory, antimicrobial, antioxidant, and anticancer agent. During the in-silico molecular docking study, out of eleven phytocompounds identified by the GC-MS study, three compounds, namely, γ-sitosterol, stigmasterol, and campesterol, formed the most stable docked complex with anti-inflammatory, antimicrobial, and antioxidant target proteins. The present study provides justification for the bark part of G. asiatica as a rich source of potent phytochemicals, and its potential against oxidative stresses, inflammations, cancer, and microbial infections.

Composition of Impurities of Germanium Tetrafluoride: A Gas Chromatography–Mass Spectrometry Study

Composition of Impurities of Germanium Tetrafluoride: A Gas Chromatography–Mass Spectrometry Study

The GC-MS Study of the Ayurvedic Formulation “Dhanwantharam Thailam” Used for Rheumatism

ABSTRACT Background: The scientific validity of contemporary and alternative medicinal practices, such as Ayurveda and Siddha, holds significant importance in today’s context. Aim and Objective: The present study employs gas chromatography-mass spectrometry (GC-MS) analysis to investigate a pain-relieving Ayurvedic oil, Dhanwantharam Thailam, aiming to establish correlations between its medicinal activity and the biomolecules it contains. Materials and Methods: Procured sample from a reputable Ayurvedic vendor in Chennai, Dhanwantharam Thailam underwent GC-MS analysis using standard procedures. Results and Discussion: The resulting profile revealed the presence of crucial molecules like oleic acid, dodecanoic acid, 1,2,3-propanetriyl ester, ethenyl ester, and 9,12-octadecadienoyl chloride (Z, Z), aligning with the medicinal properties attributed to Dhanwantharam Thailam. Conclusion: The identification of these biomolecules supports the role of Dhanwantharam Thailamas an effective pain-relieving oil.

Phytochemical Characterization of Anisomeles indica (L.) Kuntze Extract Using GC-MS: Evaluation of Antioxidant and Antimicrobial Properties

indica (L.) Kuntze leaves were tested to explore their chemo profiling through gas chromatography-mass spectrometry (GC-MS). The samples were also analyzed to evaluate antioxidant and antimicrobial activities. The antioxidant efficacy of the plant extract was evaluated using the FRAP, DPPH, and ABTS methodologies. The results revealed that extract had a high concentration of flavonoids and phenols and exhibited the highest antioxidant properties reflected by the inhibition of free radical DPPH (30.87 ± 0.001%). The plant extracts showed antimicrobial activities against Pseudomonas aeruginosa (gram-negative) and Staphylococcus aureus (gram-positive) bacteria with 10 and 9 mm inhibition zones, respectively. The GC-MS studies exhibited the presence of 25 compounds. The major constituents are desogestrel (100%), n-hexadecanoic acid (8.59%), 9, 12-octadecadienoic acid (Z, Z) (8.5%), gamma.-sitosterol (7.57%), and palmitic acid, TMS derivative (3.31%). The GC-MS analysis exhibited that leaves consist of bioactive compounds, such as alkaloids, terpenoids, phenolics, and fatty acids.

A Comprehensive Study on Chemical and Biological Investigation of Thymus Brachychilus Jalas: A Rich Source of Ursolic and Oleanolic Acids

The significance of Thymus species in the scientific community is growing steadily due to their extensive utilization in traditional medicine, food industry, and pharmaceutical sector, owing to their abundance in essential oil and phytochemical content, rendering them commercially significant species. The current work focuses on conducting a comprehensive analysis of the ethanol extract and essential oil derived from the root and aerial portions of Thymus brachychilus Jalas, an endemic species that has not been previously investigated. Additionally, a novel GC-MS (gas chromatography–mass spectrometry) technique was developed to quantify the levels of triterpenoids, which are frequently found in many plants, particularly those belonging to the Lamiaceae family. The approach was then used to assess the triterpenoid content of the species. While the phenolic content of the species was determined by LC-MS/MS (liquid chromatography–tandem mass spectrometry), the chemical composition of triterpenoid, essential oil and flavor (aroma) of the plant was determined by GC-MS. Eucalyptol was the primary ingredient in both the essential oil and the flavor, accounting for 11.05% and 12.35%, respectively. In the DPPH and ABTS radical scavenging techniques, the root ethanol extract exhibited the greatest antioxidant activity with IC50 values of 26.70 ± 0.23 and 19.16 ± 0.11 µg/mL, respectively. There was a high level of urease (59.54 ± 1.67% at 100 µg/mL) inhibitory activity observed in the root ethanol extract in enzyme inhibition assays, as well as angiotensin (94.80 ± 0.56%), elastase (40.19 ± 0.39%), and collagenase (48.26 ± 0.12%) inhibition in the aerial ethanol extract. Moreover, the MCF-7 cell line for breast cancer had a strong cytotoxic impact when exposed to the essential oil of the species (vitality%: 1.45 ± 0.01 at 200 µg/mL. The LC-MS/MS and GC-MS studies revealed that the roots had a significant concentration of rosmarinic acid (15,801 µg analyte/g extract), but the aerial portions had a substantial quantity of ursolic acid (152,374 µg analyte/g extract). The plant exhibits potential in the food, cosmetic, and pharmaceutical sectors on account of its elevated levels of rosmarinic and ursolic acid, superior antioxidant capacity, and angiotensin and urease inhibitory effect.

MalbacR: A Package for Standardized Implementation of Batch Correction Methods for Omics Data.

Mass spectrometry is a powerful tool for identifying and analyzing biomolecules such as metabolites and lipids in complex biological samples. Liquid chromatography and gas chromatography mass spectrometry studies quite commonly involve large numbers of samples, which can require significant time for sample preparation and analyses. To accommodate such studies, the samples are commonly split into batches. Inevitably, variations in sample handling, temperature fluctuation, imprecise timing, column degradation, and other factors result in systematic errors or biases of the measured abundances between the batches. Numerous methods are available via R packages to assist with batch correction for omics data; however, since these methods were developed by different research teams, the algorithms are available in separate R packages, each with different data input and output formats. We introduce the malbacR package, which consolidates 11 common batch effect correction methods for omics data into one place so users can easily implement and compare the following: pareto scaling, power scaling, range scaling, ComBat, EigenMS, NOMIS, RUV-random, QC-RLSC, WaveICA2.0, TIGER, and SERRF. The malbacR package standardizes data input and output formats across these batch correction methods. The package works in conjunction with the pmartR package, allowing users to seamlessly include the batch effect correction in a pmartR workflow without needing any additional data manipulation.

Lipid extraction and analysis of microalgae strain pectinodesmus PHM3 for biodiesel production

The current study is focused on the lipid extract of microalgae; Pectinodesmus strain PHM3 and its general analysis in terms of chemical contents. Combinations of both chemical and mechanistic approaches were applied to obtain the maximum yield of lipids which was recorded to be 23% per gram through continuous agitation using Folch solution. The extraction methods used in this study included: Bligh and Dyers method, Continuous agitation method, Extraction using Soxhlet and Acid base extraction method. Lipid quantification of ethanol and Folch solution lipid extract was performed through gravimetric methods and qualification was done through Fourier Transmission Infrared Spectroscopy (FTIR) and Gas Chromatographymass spectrometry (GC-MS). Phytochemical analysis identified other compounds in ethanol extract and the results confirmed the presence of steroids, coumarins, tannins, phenols and carbohydrates. Transesterification of lipids showed 7% per gram dry weight yield of Pectinodesmus PHM3. GC-MS studies of extracted biodiesel suggested that 72% of biofuels was in the form of dipropyl ether, ethyl butyl ethers, methyl butyl ether and propyl butyl ether. Lipid processing of acid-base extract showed that oily nature of lipid shifted to a more precipitated form which is a common observation when mixture of lipids is converted to phosphatides.

GC-MS Studies on Nitric Oxide Autoxidation and S-Nitrosothiol Hydrolysis to Nitrite in pH-Neutral Aqueous Buffers: Definite Results Using 15N and 18O Isotopes.

Nitrite (O=N-O-, NO2-) and nitrate (O=N(O)-O-, NO3-) are ubiquitous in nature. In aerated aqueous solutions, nitrite is considered the major autoxidation product of nitric oxide (●NO). ●NO is an environmental gas but is also endogenously produced from the amino acid L-arginine by the catalytic action of ●NO synthases. It is considered that the autoxidation of ●NO in aqueous solutions and in O2-containing gas phase proceeds via different neutral (e.g., O=N-O-N=O) and radical (e.g., ONOO●) intermediates. In aqueous buffers, endogenous S-nitrosothiols (thionitrites, RSNO) from thiols (RSH) such as L-cysteine (i.e., S-nitroso-L-cysteine, CysSNO) and cysteine-containing peptides such as glutathione (GSH) (i.e., S-nitrosoglutathione, GSNO) may be formed during the autoxidation of ●NO in the presence of thiols and dioxygen (e.g., GSH + O=N-O-N=O → GSNO + O=N-O- + H+; pKaHONO, 3.24). The reaction products of thionitrites in aerated aqueous solutions may be different from those of ●NO. This work describes in vitro GC-MS studies on the reactions of unlabeled (14NO2-) and labeled nitrite (15NO2-) and RSNO (RS15NO, RS15N18O) performed in pH-neutral aqueous buffers of phosphate or tris(hydroxyethylamine) prepared in unlabeled (H216O) or labeled H2O (H218O). Unlabeled and stable-isotope-labeled nitrite and nitrate species were measured by gas chromatography-mass spectrometry (GC-MS) after derivatization with pentafluorobenzyl bromide and negative-ion chemical ionization. The study provides strong indication for the formation of O=N-O-N=O as an intermediate of ●NO autoxidation in pH-neutral aqueous buffers. In high molar excess, HgCl2 accelerates and increases RSNO hydrolysis to nitrite, thereby incorporating 18O from H218O into the SNO group. In aqueous buffers prepared in H218O, synthetic peroxynitrite (ONOO-) decomposes to nitrite without 18O incorporation, indicating water-independent decomposition of peroxynitrite to nitrite. Use of RS15NO and H218O in combination with GC-MS allows generation of definite results and elucidation of reaction mechanisms of oxidation of ●NO and hydrolysis of RSNO.

Untargeted GC-MS and FT-NIR study of the effect of 14 processing methods on the volatile components of Polygonatum kingianum.

Polygonatum kingianum is a traditional medicinal plant, and processing has significantly impacts its quality. Therefore, untargeted gas chromatography-mass spectrometry (GC-MS) and Fourier transform-near-infrared spectroscopy (FT-NIR) were used to analyze the 14 processing methods commonly used in the Chinese market.It is dedicated to analyzing the causes of major volatile metabolite changes and identifying signature volatile components for each processing method. The untargeted GC-MS technique identified a total of 333 metabolites. The relative content accounted for sugars (43%), acids (20%), amino acids (18%), nucleotides (6%), and esters (3%). The multiple steaming and roasting samples contained more sugars, nucleotides, esters and flavonoids but fewer amino acids. The sugars are predominantly monosaccharides or small molecular sugars, mainly due to polysaccharides depolymerization. The heat treatment reduces the amino acid content significantly, and the multiple steaming and roasting methods are not conducive to accumulating amino acids. The multiple steaming and roasting samples showed significant differences, as seen from principal component analysis (PCA) and hierarchical cluster analysis (HCA) based on GC-MS and FT-NIR. The partial least squares discriminant analysis (PLS-DA) based on FT-NIR can achieve 96.43% identification rate for the processed samples. This study can provide some references and options for consumers, producers, and researchers.

Inhibition of Autophagy Increases Cell Death in HeLa Cells through Usnic Acid Isolated from Lichens.

The Western Ghats, India, is a hotspot for lichen diversity. However, the pharmacological importance of lichen-associated metabolites remains untapped. This study aimed to evaluate the cytotoxic potential of lichens of this region. For this, sixteen macrolichens were collected and identified from two locations in the Western Ghats. The acetone extract of Usnea cornuta (UC2A) showed significant cytotoxicity towards multiple human cancer cell lines. Interestingly, co-treatment with chloroquine (CQ), an autophagy inhibitor, increased the cytotoxic potential of the UC2A extract. A gas chromatography mass spectrometry (GCMS) study revealed usnic acid (UA), atraric acid and barbatic acid as the dominant cytotoxic compounds in the UC2A extract. Further, UA was purified and identified from the UC2A extract and evaluated for cytotoxicity in HeLa cells. The monodansyl cadaverine and mitotracker red double staining revealed the autophagy-inducing activities of UA, and the inhibition of autophagy was confirmed via CQ treatment. Autophagy inhibition increased the cytotoxicity of UA by 12-16% in a concentration-dependent manner. It also increased lipid peroxidation, ROS levels and mitochondrial depolarization and decreased glutathione availability. A decrease in zeta potential and a 40% increase in caspase 3/7 activity were also noted after CQ treatment of UA-treated cells. Thus, cytotoxicity of UA can be increased by inhibiting autophagy.

In silico molecular docking approach and in vitro cytotoxic, antioxidant, antimicrobial and anti-inflammatory activity of Ixora brachiata Roxb

The Ixora brachiata Roxb. methanolic leaf extract yielded significant in vitro free radical scavenging activity against DPPH (60% of inhibition obtained in 100 μg/mL) and ABTS (62% of inhibition obtained in 100 μg/mL) radicals, compared to ascorbic acid positive control. The antibacterial activity was modest when compared with antibiotics controls. The extract was effective, especially against Escherichia coli and Staphylococcus aureus, yielding 5 ± 0.9 (mm) and 4 ± 0.5 (mm), respectively, at a 10 mg/mL concentration. MTT assay showed an IC50 value of 51 μg/mL. The plant extract was rich in antioxidants, and anthraquinones were detected. When compared to aspirin (100–800 μg/mL) which had much greater anti-inflammatory (HRBC stabilisation) activity of 45–84%, the methanolic extract of I. brachiata was found to possess 10–45% activity in the same concentration range of 100–800 μg/mL. Since aspirin is a pure compound (and a routinely employed anti-inflammatory standard) and the plant extract contains multiple compounds with differential anti-inflammatory activities, the activity of aspirin may have been much higher. Qualitative and quantitative analyses showed the presence of different classes of primary and secondary metabolites in the leaf extract. The GC-MS (Gas chromatography-mass spectrometry) study in the plant extract showed that ∼22 different volatile compounds were present. The in vitro cytotoxic potential of the extract was assessed in MCF-7 breast cancer cell lines, and the bioactive compounds β-sitosterol, γ-sitosterol, δ-tocopherol and squalene, which were identified through GC-MS, were further assessed for binding to salient receptor proteins which are routinely involved in breast cancer pathophysiology, like PI3K, MMP-9, mTOR and ERβ.

GC-MS Studies on the Conversion and Derivatization of γ-Glutamyl Peptides to Pyroglutamate (5-Oxo-Proline) Methyl Ester Pentafluoropropione Amide Derivatives.

Glutathione (γ-L-glutamyl-L-cysteinyl-glycine, γ-Glu-Cys-Gly) is the most abundant intra-cellular dicarboxylic tripeptide with multiple physiological roles. In biological samples, glutathione exists in its reduced form GSH and in two stable oxidized forms, i.e., in its symmetric disulfide form GSSG and as S-glutathionyl residue in proteins. S-Glutathionylation is a post-translational modification, which is involved in several pathophysiological processes, including oxidative stress. The GSH-to-GSSG molar ratio is widely used as a measure of oxidative stress. γ-Glutamyl is the most characteristic structural moiety of GSH. We performed gas chromatography-mass spectrometry (GC-MS) studies for the development of a highly specific qualitative and quantitative method for γ-glutamyl peptides. We discovered intra-molecular conversion of GSH, GSSG, γ-Glu-Cys and of ophthalmic acid (OPH; γ-glutamyl-α-amino-n-butyryl-glycine) to pyroglutamate (pGlu; 5-oxo-proline, also known as pidolic acid) during their derivatization with 2 M HCl/CH3OH (60 min, 80 °C). For GC-MS analysis, the methyl esters (Me) were further derivatized with pentafluoropropionic (PFP) anhydride in ethyl acetate (1:4, v/v; 30 min, 65 °C) to their PFP derivatives. At longer reaction times, pGlu is hydrolyzed to Glu. Internal standards were prepared by derivatizing GSH, GSSG, γ-Glu-Cys and OPH in 2 M HCl/CD3OD. Quantification of the Me-PFP derivative of pGlu was performed in the electron-capture negative-ion chemical ionization (ECNICI) mode by selected-ion monitoring (SIM) of the mass-to-charge (m/z) ions 269 for unlabeled pGlu (d0Me-PFP-pGlu) and m/z 272 for the in situ prepared deuterium-labeled pGlu (d3Me-PFP-pGlu). Although not inherent to the analysis of small peptides, the present GC-MS method is useful to study several biochemical aspects of GSH. Using pentafluorobenzyl bromide (PFB-Br) as the derivatization reagent, we found that synthetic pGlu is converted in aqueous acetone (60 min, 50 °C) into its pentafluorobenzyl (PFB) ester (PFB-pGlu). This derivatization procedure is useful for the GC-MS analysis of free pGlu in the ECNICI mode. Quantitative analysis of PFB-pGlu by GC-MS requires the use of stable-isotope labeled analogs of pGlu as an internal standard.

Phytochemical screening, Gas chromatography: Mass spectrometry and antidiabetic properties of aqueous extract of ginger (Zingiber officinale) in Alloxan induced diabetic Wistar rats

The study investigates the phytochemistry, Gas-Chromatography–Mass Spectrometry (GC-MS), atomic Absorbtion spectroscopy (AAS) and antidiabetic activities of aqueous ginger (Zingiber officinale) extract in diabetic alloxan-induced Wistar rats. Qualitative phytochemical analysis, GC-MS, AAS and antidiabetic properties of the aqueous ginger extract were determined using standard procedures. Phytochemical analysis of the aqueous ginger extract shows that the extract contains tannins, flavonoids, saponins, alkaloids, simple phenolic, glycosides, carbohydrates, reducing sugar and steroids. The GC-MS study of the aqueous Zingiber officinale extract revealed the presence of eleven different compounds with 1,3-Cyclohexadiene, 5-(1,5-dimethy 4-hexenyl)-2-methyl-, [S-(R*,S*)]- been the most abundant with peak area of 33.98%. The AAS analysis shows that ginger contain: Ca, Na, Fe, K, P and Zn minerals. Twenty five (25) Wistar rats were grouped into 5 groups and investigated for antidiabetic study for a period of 15 days. Group A and B animals were normal and negative control respectively. Group B rats were induced with alloxan and not treated with drugs or extract. Animals in other groups (C, D and E rats) were diabetic and treated with standard drug (glibenclamide with concentration of 10 mg/kg), 200 and 400 mg/kg body weight of aqueous ginger extract respectively. The group of rats treated with 10, 200 and 400 mg/kg body weight of glibenclamide and aqueous ginger extract showed significant reduction (p

POTENTIAL ANTIOXIDANT EFFICACY OF THE SECONDARY METABOLITES ISOLATED FROM CALOCYBE INDICA (VAR.APK2): AN EDIBLE MACROFUNGI

Objective: Mushrooms have been valued as traditional sources of natural bioactive compounds for many centuries and targeted as promising therapeutic agents. The bioactive mycomolecules of mushrooms are reported to have antioxidant, antitumor, antidiabetic, anti-inflammatory, and antimicrobial activity, which are the important medicinal targets in terms of drug discovery today. Hence, an attempt was made in the present study, to evaluate the immunomodulatory and oxidative process of secondary metabolites from the milky mushroom Calocybe indica (P&C) var.APK2 using radical scavenging assays. Methods: The fruiting bodies of milky mushrooms were found to produce an array of mycomolecules such as phenols, flavonoids, alkaloids, tannins, terpenoids, steroids, and saponins in their methanolic extract which was confirmed using Fourier-transform infrared spectrophotometer (FT-IR) analysis and standard phytochemical studies; hence, chromatography fractions of these mushroom seem greatly promising biological activities including antioxidants. Results: The functional analysis of the secondary metabolites of these macrofungi was evaluated by the separation of potential fractions using preparative thin-layer chromatography (TLC) that revealed seven distinct bands with Rf values of 0.14, 0.26, 0.31, 0.42, 0.52, 0.70, and 0.82; the antioxidant activity was determined through TLC in situ bio autography. The quenching property of metabolite compound which was ranging from 19% to 77.9% and the half effective concentration values of 2,2-diphenyl-1-picrylhydrazyl and hydroxyl radical scavenging activity was recorded as 64.26 μg/ml and 54.5 μg/ml sample concentration, respectively. The active mycomolecules of C. indica from the TLC was, further, confirmed using Gas Chromatography–Mass Spectrometry studies. Conclusion: The present investigation of the study revealed that the antioxidant efficacy of edible milky mushroom may be further proceeded for in vivo studies for novel drug discovery.