Strategic enzymatic enantioselective desymmetrization of prochiral cyclohexa-2,5-dienones.

Asymmetric desymmetrization through the selective reduction of one double bond of prochiral 2,5-cyclohexadienones is highly challenging. A novel method has been developed for synthesizing chiral cyclohexenones by employing an ene-reductase (Bacillus subtilis YqjM) enzyme that belongs to the OYE family. Our strategy demonstrates high substrate scope and enantioselectivity towards substrates containing all-carbon as well as heteroatom (O, N)-containing quaternary centers. The mechanistic studies (kH/D = ∼1.8) indicate that hydride transfer is probably the rate-limiting step. Mutation of several active site residues did not affect the stereochemical outcomes. This work provides a convenient way of synthesizing various enantioselective γ,γ-disubstituted cyclohexanones using enzymes.

Silver Nanoparticles Induced Metabolic Perturbations in Pseudomonas aeruginosa: Evaluation Using the UPLC-QTof-MSE Platform.

Silver nanoparticles (AgNPs) have been widely utilized in various biomedical and antimicrobial technologies, displaying broad-spectrum activities against Gram-negative and Gram-positive bacteria including multidrug-resistant strains. However, the emergence of resistance to AgNPs upon repeated exposure and the survival of bacteria after initial exposure to antimicrobial agents pose a threat, as they may lead to the development of new resistant populations. To combat the early stages of antibacterial resistance, systematic analysis is essential to understand the immediate response of bacteria to antimicrobial agents. In this study, green-synthesized AgNPs with a diameter of approximately 14 nm were exposed toPseudomonas aeruginosaat three different inhibitory concentrations and at two different time intervals (1 and 4 h) to investigate the perturbations in the metabolome using liquid chromatography-high-resolution mass spectrometry. MetaboAnalyst 5.0 was employed for univariate and multivariate analysis, and the affected metabolic pathways were constructed using a variable important in projection scores above 1 from PLS-DA. The study revealed significant alterations in metabolites associated with cell wall synthesis, energy metabolism, nucleotide metabolism, the TCA cycle, and anaplerotic intermediates of the TCA cycle. Our investigation aimed to comprehensively understand the effects of green-synthesized AgNPs onP. aeruginosa metabolism, providing a more precise snapshot of the bacterium's physiological state through metabolomics approach.

LC-PDA-QTof-MS characterization of the stress degradation products of Zanubrutinib: A novel BTK inhibitor.

Zanubrutinib (ZAN) is an orally administered anti-cancer medication used for the treatment of Mantle cell lymphoma. Recently, it has also been approved by FDA for the treatment of chronic lymphocytic leukemia. Determination of impurities formed in drug substances/products as a result of manufacturing or storage forms an important aspect of drug life cycle management. The current study concentrated on understanding the stability of ZAN under various stress conditions as per the ICH Q1 (R2) guidelines. In total, ZAN produced thirteen degradation products under various hydrolytic (acid, base and neutral) and thermal stress conditions. The stress degradation products were separated by ultra-performance liquid chromatography, chemical structures of these products were characterized by MS/MS experiments combined with accurate mass measurements conducted on a LC-QTof-MS. The mechanism for the formation of these degradation products was also proposed. This study provides comprehensive information on the inherent stability of ZAN which will be useful in the drug development and manufacturing processes.

Multiplatform Metabolomics to Understand the Imidacloprid-Induced Toxicity in Drosophila.

Neonicotinoids, the class of insecticides used for crop protection, are subjected to vigilance due to their pernicious impacts. Imidacloprid (IMD) is one of the most representative insecticides of the neonicotinoid family, which has shown unfriendly consequences for non-target species. Metabolomics, a multidisciplinary approach, is being used in toxicological research to understand the metabolic responses to toxicant exposure by utilizing modern analytical techniques. Yet, no solitary analytical technique can cover the broad metabolite spectrum, but a multi-technique metabolomics platform can aid in analyzing the majority of the metabolites. In the present study, an effort has been made to identify the differential metabolites in Drosophila after exposure to IMD at 2.5 and 25 ng/mL using liquid chromatography-high-resolution mass spectrometry (LC-HRMS), gas chromatography-MS (GC-MS), and NMR-based untargeted metabolomics. Multivariate pattern recognition analysis helped in identifying/recognizing 19 (LC-HRMS), 7 (GC-MS), and 13 (NMR) differential metabolites mainly belonging to the category of amino acids, sugars, fatty acids, and organic acids. The pathway analysis of differential metabolites predominantly showed impact on aminoacyl-tRNA biosynthesis, amino acid metabolism, and glycerophospholipid metabolism. Among these, arginine and proline metabolism was observed to be the common metabolic pathway perturbed in Drosophila due to IMD exposure. The multiplatform metabolomics based on LC-HRMS, GC-MS, and NMR analysis with an advanced level of statistical analysis can provide insights into potential perturbations in the metabolome of IMD-exposed Drosophila.

A green deep eutectic solvent based dispersive liquid-liquid microextraction for the quantitative analysis of 21 polychlorinated biphenyl metabolites in food of animal origin using injector port silylation-gas chromatography-tandem mass spectrometry.

An analytical method was developed for the quantitative determination of 21 polychlorinated biphenyls (PCBs) metabolites (17 were -OH, 1 -MeO, and 3 were MeSO2) in foods of animal origin using deep eutectic solvent (DES) based dispersive liquid-liquid microextraction followed by injector port silylation-gas chromatography-tandem mass spectrometry. The type of DES (thymol: camphor, 1:1 molar ratio) and optimum volume of DES (300 µL), pH (7.0), and disperser solvent (acetonitrile) were optimized to attain the maximum extraction efficiency. The limit of detection, limit of quantification, and percent recovery were found to be in the range of 0.12-0.23 ng/mL, 0.40-0.76 ng/mL, and 80.1-111.4%, respectively. The expanded uncertainty was observed to be in the range of 7.2-22.8% for the targeted analytes. The proposed method was applied to real food samples (milk, meat, fish, and egg) and the levels were found to be in the range of 0.64-32.14 ng/g. This is first of its kind method using green solvent based method for the analysis of PCB metabolites (-OH, MeO, and MeSO2) and will find extensive application in routine testing for foods of animal origin.

Development of liquid chromatography-triple quadrupole mass spectrometric method for the quantitative determination of a novel adjuvant, Imidazoquinoline gallamide in aluminum hydroxide gel-Imidazoquinoline gallamide and COVAXIN.

Imidazoquinoline gallamide is a toll-like receptor 7/8 agonist, belongs to the imidazoquinoline class, has the potential to activate antigen-presenting cells, and enhances immune response, primarily Th1 response. The COVAXIN is a whole virion inactivated Coronavirus disease 2019 vaccine formulated with this novel adjuvant called, aluminum hydroxide gel Imidazoquinoline gallamide, wherein, Imidazoquinoline gallamide is chemisorbed onto aluminum hydroxide gel. Herein, an analytical method based on liquid chromatography-tandem mass spectrometry was developed to identify and quantify Imidazoquinoline gallamide in aluminum hydroxide gel Imidazoquinoline gallamide and COVAXIN. The multiple reaction monitoring transitions were optimized for Imidazoquinoline gallamide quantification are [M+H]+ ions with 512.24→343.19 m/z (quantifier ion) and 512.24→360.22 m/z (qualifier ion). The developed method was validated as per the international conference on harmonization quality2 revison1 guidelines. The method was linear in the range of 0.025-10 µg/mL with a coefficient of determination of 0.9985 and the limit of quantification is 0.025 µg/mL. The accuracy was in the range of 82-121 % and intra- and inter-day precision was less than 7.1% and 5.39%, respectively. The expanded uncertainty results are 9.2% for Imidazoquinoline gallamide in the sample. The validated method was successfully applied to evaluate Imidazoquinoline gallamide concentration in every batch of COVAXIN.

Chemical profiling and simultaneous quantification of major bioactive constituents from Cocculus hirsutus by UPLC-QqQ-MS.

Cocculus hirsutus is a widely used herb in traditional systems of medicine for the treatment of various diseases. In the present study, five alkaloids (1-5), two flavonoids (6-7), one triterpenoid (8), and three steroids (9-10) were isolated from the roots of Cocculus hirsutus and further crude extract was analyzed by LC-Q-Tof-MS/MS in positive ionization mode leading to the identification of ten metabolites through comparison of exact molecular masses from their MS/MS spectra, mass fragmentation studies and with literature data. In addition, a method was developed and validated for the quantification of four bio-active compounds [Sinococuline (1), Magnoflorine (2), (E)-N-feruloyltyramine (3), and 20-Hydroxyecdysone (10)] using UPLC-QqQ-MS in multiple reaction monitoring (MRM) mode for the first time. The method has shown good linearity with correlation coefficients (r2) higher than 0.9916 for all four compounds. The intra- and inter-day precision were in the range of 0.3-6.1% and from 0.7% to 8.8%, respectively. The matrix effects of all the four analytes were found in the range of 94.7 ± 2.8-112.7 ± 3.7%. Overall, our study provides a reliable and rapid approach by hyphenated LC-MS/MS using high-resolution mass spectrometers for identification and quantification of bioactive constituents from the root extracts of Cocculus hirsutus.

Chemometric assisted natural DES based VA-DLLME-LC-MS/MS method for the quantitative determination of Garcinol in biofluids/tissues: A practical application to pharmacokinetics and biodistribution studies.

Garcinol (GAR) is a polyisoprenylated benzophenone obtained from Garcinia indica used as anti-oxidant and anti-inflammatory in traditional medicine and due to these activities, it possesses anticancer properties. It is considered to be a next generation epigenetic drug. A green solvent based analytical method which is efficient, sophisticated, and highly enriched has been developed for the quantitative analysis of GAR in biological samples (plasma, liver, kidney and spleen) with the use of deep eutectic solvent (DES) for its extraction. A series of 23 DESs were synthesized and out of which, Thymol (Th)-Terpeniol (T), 2:1 molar ratio with a more hydrophobic environment and high interaction efficiency between GAR and DES was identified for the better extraction from mice plasma and tissue samples. The Design of Experiment approaches like placket-burmann design and central composite design were used to optimize the method conditions. The method validation characteristics, such as limit of detection (0.193-0.237 ng/mL), limit of quantification (0.644-0.697 ng/mL), lower limit of quantification (0.5 ng/mL), broad range of linearity with R2 (0.9994-0.9997) with a percent recovery not less than 87% was observed, which are well within the acceptance criteria for a bioanalytical method. The enrichment factor is upto 53-60 folds, with high extraction efficiency (89-97%). The measurement uncertainty was estimated with an expanded uncertainty ranged between 10.9%-19.0%. The method developed and validated was effectively applied to examine the pharmacokinetic and biodistribution patterns for GAR in mice.

Comprehensive metabolomic analysis of Mangifera indica leaves using UPLC-ESI-Q-TOF-MSE for cell differentiation: An in vitro and in vivo study.

The comprehensive metabolic profiling was performed in the leaf extracts of Mangifera indica and assessed for their significant therapeutic application in tissue engineering and regenerative medicine in both in vitro and in vivo studies. About 147 compounds were identified in the ethyl acetate and methanol extracts of M. indica using MS/MS fragmentation analysis and the selected compounds were quantified using LC-QqQ-MS analysis. The in vitro cytotoxic activity showed that the M. indica extracts enhance the proliferation of mouse myoblast cells in concentration-dependent manner. As well, the extracts of M. indica induce the myotube formation by generating oxidative stress in the C2C12 cells was confirmed. The western blot analysis clearly showed that the M. indica induce myogenic differentiation by upregulating the myogenic marker proteins such as PI3K, Akt, mTOR, MyoG, and MyoD. The in vivo studies showed that the extracts expedites the acute wound repair by formation of crust, wound closure and improves the blood perfusion towards the wound area. Together, the leaves of M. indica can be used as excellent therapeutic agent for tissue repair and wound healing applications.

Microbial degradation of organochlorine pesticide: 2,4-Dichlorophenoxyacetic acid by axenic and mixed consortium.

The presence of 2,4-dichlorophenoxyacetic acid (2,4-D), an organochlorine herbicide, in the environment has raised public concern as it poses hazard to both humans and the ecosystem. Three potential strains having the capability to degrade 2,4-D were isolated from on site agricultural soil and identified as Arthrobacter sp. SVMIICT25, Sphingomonas sp. SVMIICT11 and Stenotrophomonas sp. SVMIICT13. Over 12 days of incubation, 81-90% of 100 mg/L of 2,4-D degradation was observed at 2% inoculum. A shorter lag phase with 80% of degradation efficiency was observed within 5 days when the inoculum size was increased to 10%. Six microbial consortia were prepared by combining the isolates along with in-house strains, Bacillus sp. and Pseudomonas sp. Consortia R3 (Arthrobacter sp. + Sphingomonas sp.), operated with 10% of inoculum, showed 85-90% degradation within 4 days and 98-100% in 9 days. Further, targeted exo-metabolite analysis confirmed the presence and catabolism of intermediate 2,4-dichlorophenol and 4-chlorophenol compounds.

Experimental design of non-ionic hydrophobic DES-DLLME coupled with injector port silylation-GC-MS/MS for the quantitative determination of 13 bisphenols in food samples.

Non-ionic hydrophobic deep eutectic solvent based DLLME method with thymol and camphor (1:1 mole ratio) as extraction solvent and acetonitrile as disperser solvent was developed for the determination of 13 bisphenols (BPs) in food commodities by auto IPS-GC-MS/MS. Further, the influential parameters pH, vortex time, and volume of NIHDES for extraction of 13 BPs were optimized by employing the design of experiment. The LOQ and recoveries were observed to be 0.1 ng mL-1, and 77.8-109.2%, respectively. The expanded uncertainty was found to be 4.8-18.0% for the developed and validated method, which was applied to food commodities to determine 13 BPs and their levels were in the range of 0.01-1.24 ng mL-1 in liquid foods (milk, water, and beverages) and 0.29-3.25 ng g-1 in meat samples (goat, chicken, and fish). The present green analytical method is highly suitable for the rapid determination of 13 BPs in food commodities for routine analysis.

Evaluating chemicals of emerging concern in the Ganga River at the two major cities Prayagraj and Varanasi through validated analytical approaches.

Evaluating environmental water quality means to assess and protect the environment against unfriendly impacts from various organic impurities emerging from industrial emissions and those released during harvesting. Potential risks related with release of polycyclic aromatic hydrocarbons (PAHs), pesticides and pharmaceuticals (PhAcs), and personal care products (PCPs) into the environment have turned into an increasingly serious issue in ecological safety. Monitoring helps in control of chemicals and ecological status compliance to safeguard specific water uses, for example, drinking water abstraction. A longitudinal review was carried out for 55 different persistent organic pollutants (POPs) for the Ganga River which passes through the urban areas of Prayagraj and Varanasi, India, through validated analytical approaches and measurement uncertainty (MU) estimation to assess their potential use for routine analysis. Furthermore, environmental risk assessment (ERA) carried out in the present study has revealed risk quotient (RQ) higher than 1 in a portion of the aquatic bodies. Using a conservative RQ strategy, POPs were assessed for having extensive risks under acute and chronic exposure, proposing that there is currently critical ecological risk identified with these compounds present in the Ganga River. In general, these outcomes demonstrate a significant contribution for focusing on measures and feasible techniques to minimize the unfavorable effects of contaminants on the aquatic environment.

Chemical profiling of marine seaweed Halimeda gracilis using UPLC-ESI-Q-TOF-MSE and evaluation of anticancer activity targeting PI3K/AKT and intrinsic apoptosis signaling pathway.

Marine seaweeds are predominant for nutraceuticals and have been food source since ancient times and are currently being used in Chinese medicines and Japanese traditional medicines. In the present study, the chemical profile analysis of Halimeda gracilis was performed using UPLC-ESI-Q-TOF-MSE analysis and assessed for its anticancer activity. A cursory investigation of the total ion chromatograms of the both methanol (MHG) and ethyl acetate (EAHG) of H. gracilis extracts reveals that both extracts have different kind of metabolites including phenols and its derivatives, diterpenes, cinnamic acids derivatives etc. The in vitro anticancer activity of EAHG and MHG exhibiting significant activity and induced apoptosis against skin cancer cells by generating excessive ROS, damaging the mitochondrial membrane and nuclear components. Further, the western blot analysis showed that the EAHG and MHG downregulates the oncoproteins PI3K, AKT, p-AKT, and BcL2 and upregulates the apoptotic proteins Bax, Cyto C, p21, p53, Caspase 9 and Caspase 3. To best of our knowledge, this is the first report which implies the UPLC-ESI-Q-TOF-MSE based chemical profiling of H. gracilis and can be used for the treatment of cancer.

Bombax ceiba calyx displays antihyperglycemic activity via improving insulin secretion and sensitivity: Identification of bioactive phytometabolomes by UPLC-QTof-MS/MS.

Vegetables are considered good food for the management of hyperglycemia. Bombax ceiba L. (family: Bombacaceae) calyces are part of traditional vegetables. This study evaluated its usefulness on various parameters responsible for the development of hyperglycemia and conducted phytometabolomic analysis to identify phytochemicals responsible for the observed activities. It was found that the aqueous methanol extract of its calyces (B. ceiba calyx extract, BCE) reduced (12.4%) significantly (p < 0.05) the development of sucrose-induced postprandial hyperglycemic load in rats. In-vitro studies revealed that BCE improved glucose-stimulated insulin secretory activity in MIN6 cells plausibly by decreasing ADP/ATP ratio. BCE also augmented concentration-dependent (5 µg, 10 µg, and 20 µg) increase in glucose uptake in hyperglycemic L6 myotubes both by non-insulin-dependent manner (35%, 68%, and 132%, respectively) and insulin-dependent manner (42%, 59%, and 172%, respectively). The insulin-stimulated GLUT4 translocation was compromised (34%) significantly (p < 0.05) under hyperglycemic condition; however, it was improved by 23% and 72% (p < 0.001) when L6 myotubes were primed with 10 and 20 µg of BCE, respectively. Hyperglycemia aggravated reactive oxygen species (ROS) generation in L6 myotubes. The ROS generation was significantly (p < 0.001) reduced by priming myotubes with BCE before challenging myotubes to hyperglycemic environment, possibly by preserving cellular antioxidant enzymes catalase, glutathione peroxidase, and reduced glutathione levels. Phytometabolomic analysis disclosed a number of phytochemicals present in B. ceiba calyces known to display these activities. This is the first study reporting antihyperglycemic activity in B. ceiba calyces, its mechanisms of action, and phytometabolomic profile applying UPLC-QTof-MS/MS technique. PRACTICAL APPLICATION: B. ceiba calyces are part of traditional vegetables. Our study finds that B. ceiba calyces contain phytochemicals possessing antihyperglycemic, insulin secretory, insulin sensitization properties, and potentials for preserving hyperglycemia-induced vitiations in cellular antioxidant defense. These observations provide foundation for exploring further possibilities of B. ceiba calyces to become valuable dietary inclusion in the diet of people suffering from metabolic disorders.

Analytical developments of p-hydroxy prenylamine reference material for dope control research: Characterization and purity assessment.

Prenylamine was initially used for the treatment of angina pectoris and later on withdrawn from the market in 1988 due to cardiac arrhythmias concern. The major phase I metabolite of prenylamine is p-hydroxy prenylamine that has a chiral center in the structure. Even though p-hydroxy prenylamine was synthesized earlier, it lacked complete analytical developments for chiral high-performance liquid chromatography (HPLC) separation. However, p-hydroxy prenylamine reference material is not commercially available. The innovation of this manuscript is the development and validation of a chiral HPLC separation method and more extensive characterization of the reference material than previously reported method. Therefore, it was hypothesized to develop and validate normal phase HPLC method for p-hydroxy prenylamine reference material. p-Hydroxy prenylamine was synthesized in two batches and characterized successfully using 13 C NMR, 1 H NMR, high-resolution mass spectrometry (HRMS), Fourier transform infrared spectroscopy (FT-IR), and thermogravimetric analysis (TGA). A normal phase chiral HPLC method was developed to analyze the p-hydroxy prenylamine purity. Separation of the p-hydroxy prenylamine enantiomers were achieved using ultra-high-performance liquid chromatography (UHPLC) on a ChiralCel ODH column at wavelength of 220 nm. The developed method was validated in terms of its linearity, accuracy, precision, and robustness for purification, purity assessment, and stability studies. Proton and carbon peaks were confirmed by nuclear magnetic resonance (NMR) analysis. Functional groups were confirmed by FT-IR. Loss on drying was 0.3% and 0.6% for Batches 1 and 2, respectively. The purity of the developed reference material for Batches 1 and 2 was found to be 99.59% and 100%, respectively. Therefore, the synthesized batches of p-hydroxy prenylamine can be used in dope testing as reference material.

Using bioanalytical tools to detect and track organic micropollutants in the Ganga River near two major cities.

Major rivers in India are subject to ongoing impacts from urban drain discharges, most of which contain high levels of domestic and industrial wastewater and stormwater. The aim of the present study was to determine the levels of bioactive organic micropollutants at the discharge points of major urban drains in comparison to upstream and downstream sites. To achieve this, we employed a panel of in vitro bioanalytical tools to quantify estrogenic, androgenic, progestogenic, glucocorticoid and peroxisome proliferator-like activity in water extracts collected from two Indian cities in the Ganga Basin. Cytotoxicity of the water extracts in a human-derived cell line and the potential to cause oxidative stress in a fish cell line were also investigated. We found high levels of activity for all endpoints in samples directly receiving urban drain discharge and low levels at sites upstream from drain discharges. Estrogenicity was detected at levels equivalent to 10 ng/L 17ß-estradiol, representing a high likelihood of biomarker effects in fish. Sites located downstream from drain discharges exhibited low to intermediate activity in all assays. This study demonstrates the importance of managing urban drain discharges and the utility of applying bioanalytical tools to assess water quality.

Phytometabolomic analysis of boiled rhizome of Nymphaea nouchali (Burm. f.) using UPLC-Q-TOF-MSE, LC-QqQ-MS & GC-MS and evaluation of antihyperglycemic and antioxidant activities.

Phytometabolomic analysis of Nymphaea nouchali (Burm. F.) boiled rhizome was carried out utilizing UPLC-Q-TOF-MSE, LC-QqQ-MS and GC-MS techniques and evaluated for antihyperglycemic and antioxidative stress potentials. Metabolomic analysis revealed presence of multiple antidiabetic and antioxidant compounds. Boiled rhizome powder exhibited potent antihyperglycemic activity against sugar-induced postprandial hyperglycemia in rats plausibly due to the presence of intestinal α-glucosidase inhibitory and augmenting cellular glucose uptake activities. It also prevented hyperglycemia-induced hemoglobin and insulin glycation. Rhizome displayed potent reducing power, effectively scavenged various reactive oxygen species. It displayed antioxidative stress potential in assuaging H2O2 induced erythrocyte hemolysis and antioxidant activity by inhibiting membrane lipid peroxidation. Boiled rhizome was also found to preserve the loss of cellular antioxidants under H2O2 induced oxidative stress and disturbances caused to mitochondrial membrane potential. This is the first research reporting boiled N. nouchali rhizome as an ideal food material to manage the cause of hyperglycemia and resultant oxidative stress.

Development of an analytical method for the quantitative determination of multi-class nutrients in different food matrices by solid-phase extraction and liquid chromatography-tandem mass spectrometry using design of experiments.

A simple and rapid analytical method based on solid-phase extraction (SPE) followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the aid of design of experiments (DOE) approach was developed and validated for simultaneous determination of multi-class nutrients (water- and fat-soluble vitamins and flavonoids) in various food matrices (vegetables, fruits, and cereals). The factors affecting the SPE method were optimized using DOE tools. The separation was achieved in 13 min using the C18 column by gradient LC programme. The SPE-LC-MS/MS method was validated in terms of limit of detection (1.29-29.17 ng/g), linearity range (25-1000 ng/g), coefficient of determination (0.993-0.999) and recovery (72.53-104.24%) for multi-class nutrients in different food samples. Inter- and intra-day precision were evaluated and found to be within acceptable range. The developed method finds extensive application in the routine analysis of multi-class nutrients in various food matrices.

Development of a multiclass method to quantify phthalates, pharmaceuticals, and personal care products in river water using ultra-high performance liquid chromatography coupled with quadrupole hybrid Orbitrap mass spectrometry.

Rationale: The organic micropollutants such as phthalates, pharmaceuticals, and personal care products (PPPCPs) enter the surface water through various routes. The aim of this study is to develop a sensitive and efficient method to identify and quantify 26 PPPCPs found in river water with acceptable accuracy and precision using a liquid chromatograph hyphenated with quadrupole hybrid Orbitrap mass spectrometry (Q-Orbitrap-MS) in a single chromatographic run. Method: The organic micropollutants were extracted from river water by solid-phase extraction (SPE) using hydrophilic-lipophilic balance sorbent and analyzed using an ultra-high performance liquid chromatograph (UHPLC) equipped with C18 stationary phase for chromatographic separation. The targeted mass experiments were conducted in a Q-Orbitrap-MS system in positive and negative electrospray ionization mode. Results: The method was found to be linear in the concentration range of 1-125 ng/L with coefficient of determination lying in the range of 0.995-0.999. The method achieved limit of quantification in the range of 0.41-1.72 ng/L, and method recovery measured at three different concentrations was found to be in the range of 75-115%. Intra- and interday precision expressed as percent relative standard deviation was found to be <15%. Matrix effect was found to be in the range of 83.5-109.79%. The matrix match calibration was used for quantification of PPPCPs in river water sample. The method performance was evaluated by analyzing real samples collected from Ganga River, and the concentrations of 21 analytes were found to be in the range of 0.76-9.49 ng/L for pharmaceuticals, 1.49-8.67 ng/L for phthalates, and 0.9-7.58 ng/L for personal care products. Conclusions: The present method was found to be precise, sensitive, and rapid to determine 26 PPPCPs including phthalates in river water samples using SPE-UHPLC-Q-Orbitrap-MS.

Bombax ceiba (Linn.) calyxes ameliorate methylglyoxal-induced oxidative stress via modulation of RAGE expression: identification of active phytometabolites by GC-MS analysis.

Non-enzymatic reactions between proteins and methylglyoxal (MG) result in the formation of advanced glycation end products (AGEs). These AGEs play a vital role in the development of diabetic complications by stimulating oxidative stress and acting upon their receptor RAGE (Receptor for Advanced Glycation End products). This study examined the effect of aqueous methanol extract of Bombax ceiba L. calyxes (BCCE) on MG induced protein glycation and oxidative stress, followed by the identification of phytometabolites present in the calyxes using gas chromatography-mass spectrometry (GC-MS). The study revealed that priming of bovine serum albumin protein with the BCCE inhibited MG induced AGE formation in vitro and restrained AGE-induced RAGE up-regulation in HEK-293 cells. The BCCE significantly (p < 0.001) reduced the MG induced increase in reactive oxygen species (ROS), NADPH oxidase (NOX), and mitochondrial dysfunction. Improvements in the levels of antioxidant enzymes such as Mn and Cu/Zn-superoxide dismutase and glutathione reductase were also observed in HEK-293 cells. Furthermore, the decrease in primary cellular defense against AGEs, the glyoxalase 1 (Glo-1) activity, due to MG treatment was restored in BCCE treated cells. GC-MS analysis revealed the presence of antioxidant and antiglycation compounds such as myo-ionisitol, scopoletin, d-sedoheptulose, succinic acid, and xylitol in B. ceiba calyxes. The observed beneficial effect in our study might be attributed to the presence of these compounds in B. Ceiba calyxes. This is the first report presenting the antioxidant and antiglycation activities of B. ceiba calyxes and GC-MS analysis of active phytometabolites. These observations show that B. ceiba calyxes may become a potent and promising functional food to manage/control the development of diabetic complications.