Non-thermal plasma modulated l-tyrosine self-assemblies: a potential avenue for fabrication of supramolecular self-assembled biomaterials.

Aromatic amino acids (AAs) have garnered particular interest due to their pivotal roles in numerous biological processes and disorders. Variations in AA self-assembly not only affect protein structures and functions, but their non-covalent interactions such as hydrogen bonding, van der Waals forces, and π-π stacking, yield versatile assemblies vital in bio-inspired material fabrication. Tyrosine (Tyr), a non-essential aromatic amino acid, holds multifaceted significance in the body as a protein building block, neurotransmitter precursor, thyroid hormone contributor, and melanin synthesis regulator. The proficiency of Cold Atmospheric Plasma (CAP) in generating a spectrum of reactive oxygen and nitrogen species has spurred innovative research avenues in the studies of biomolecular components, including its potential for targeted cancer cell ablation and biomolecule modification. In this work, we have assessed the chemical as well as the structural changes in Tyrosine-derived self-assembled structures arising from the CAP-induced reactive species. For a comprehensive understanding of the mechanism, different treatment times, feed gases, and the role of solvent acidification are compared using various spectroscopic and microscopic techniques. LC-ESI-QQQ mass spectra unveiled the emergence of oxygenated and nitro derivatives of l-tyrosine following its interaction with CAP-derived ROS/RNS. SEM and TEM images demonstrated an enhanced surface size of self-assembled structures and the formation of novel nanomaterial-shaped assemblies following CAP treatment. Overall, this study aims to explore CAP's interaction with a single-amino acid, hypothesizing the creation of novel supramolecular structures and scrutinizing CAP-instigated transformations in l-tyrosine self-assembled structures, potentially advancing biomimetic-attributed nanomaterial fabrication which might present a novel frontier in the field of designing functional biomaterials.

Phytochemically analysed extract of Ageratina adenophora (Sprengel) R.M.King & H. Rob. initiates caspase 3-dependant apoptosis in colorectal cancer cell: A synergistic approach with chemotherapeutic drugs.

ETHNOPHARMACOLOGICAL RELEVANCE: Ageratina adenophora (Sprengel) R.M.King & H.Rob. has been used as traditional indigenous medicine all across the globe for its diverse therapeutic applications such as anticancer, analgesic, antipyretic, thermogenic, antiseptic, antimicrobial as well as astringent. The various ethnic groups of India use plant parts to treat cuts and wounds, venomous insect bites, skin lesions, blisters, scabies and other skin irritations, gastritis and indigestion problems, cough, stomach ache and dysentery. The Portuguese traditionally extract the juice from the plant and use it for cancer, diabetes, liver disorder, gallbladder and stomach ailments. Nigerian healers use different parts of the plant to treat diabetes, fever and inflammation. AIM OF THE STUDY: The aim of this study is to investigate the cytotoxic potential of A. adenophora hydroalcoholic leaves extract (AHL) on Colorectal cancer (CRC) cell lines (HCT-116, HCT-15 and HT-29), synergistic potential with chemotherapeutic drugs 5FU and Cisplatin as well as reactive oxygen species (ROS) generation, based on the sample collected from Mao district of Manipur, India. Identification of bioactive phytocompounds in AHL was also performed by HRLCMS. METHODS: The AHL was evaluated for its cytotoxic as well as antiproliferative activities by 3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide (MTT) assay, clonogenic and cell migration assays. The total phenolic content (TPC) and total flavonoid content (TFC) were quantified by Folin-ciocalteu and Aluminium chloride assays respectively. Caspase 3 activation was evaluated using Caspase-3 Assay Kit. Apoptosis detection by flow cytometry was carried out using annexin V-FITC/PI apoptosis detection kit. The apoptotic cells were also visualized by Giemsa and 4',6-Diamidino-2-phenylindole (DAPI) staining. The intracellular Reactive oxygen species (ROS) generation was also evaluated using fluorescent probe 2',7'-dichlorodihydrofluorescein di-acetate (H2DCFDA) in flow cytometry. The combination effects of AHL with chemotherapeutic drugs 5FU and Cisplatin were also evaluated. The identification of phytochemical constituents of AHL were analysed by HR-LCMS. RESULTS: The AHL induced cytotoxic activity significantly in HCT-116 with IC50 of 65.65 ± 2.10 µg/mL, but non-cancerous cell HeK-293 was least cytotoxic. Colony formation and cell migration were inhibited in a dose and time dependent manner. The cell morphology upon AHL treatment was significantly altered with apoptotic features. The extract was rich in total phenolic (82.09 ± 0.35mgGAE/g) and total flavonoid (58.31 ± 0.55 mgQAE/g) contents. AHL induced apoptosis as detected by AnnexinV/PI, via activation of caspase 3 and elevated production of Reactive oxygen species (ROS). AHL in combination with 5FU and Cisplatin acts synergistically and potentiates the therapeutic properties of the extract. Sesquiterpenes, phenolic as well as flavonoid derivatives with anticancer properties were detected in AHL by HRLCMS, and these phytoconstituents may be attributed for anticancer property of AHL. CONCLUSION: The present study evaluates the effectiveness of AHL against Colorectal cancer cell lines. AHL is cytotoxic and induces apoptosis in HCT-116 cells by caspase 3 activation and increased ROS production that can be attributed to sesquiterpenoids. Thus, the plant A. adenophora has therapeutic potential for Colorectal cancer and can be further exploited for developing anticancer drug.

Purification and characterization of pure curcumin, desmethoxycurcumin, and bisdemethoxycurcumin from North-East India Lakadong turmeric (Curcuma longa).

Lakadong turmeric has been outlined for its high content of curcuminoids across the globe. Three significant molecular markers are widely present in turmeric viz, curcumin, desmethoxycurcumin, and bisdemethoxycurcumin, and they are present very high amount in Lakadong turmeric. Curcuminoids have been reported for structural and spectrum similarity of 3 to 4 nm (432, 434, and 436 nm, respectively). Current purification methods are based on recrystallisation where it is difficult to get highly pure material and preparative methods associated with tedious separation with high cost. Lakadong turmeric has not been explored commercially since long time. No reports are available in the literature with highly pure reference materials with in-depth characterization data and purity assessment. Curcumin, desmethoxycurcumin, and bisdemethoxycurcumin were characterized using different analytical techniques viz, UV-Visible Spectroscopy, Fourier Transform Infrared Spectroscopy (FT-IR), Differential Scanning Calorimetry (DSC), Thermogravimetric Analysis (TGA), Proton Nuclear Magnetic Resonance (1HNMR), Carbon-13 Nuclear Magnetic Resonance (13CNMR), High-Resolution Mass Spectrometry (HR-MS) and Inductive Coupled Plasma Mass Spectrometry (ICP-MS). Purified 3 markers has shown High-Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD) purity more than 99.5%. DSC the melting peaks of curcumin, desmethoxycurcumin and bisdemethoxycurcumin were observed at 168 °C, 165 °C, and 210 °C, respectively. These plant-based markers have high commercial potential as reference material for routine Quality Assurance and Quality Control (QAQC) in herbal industries.

Phyto-metabolomics of phlogacanthus thyrsiformis by using LC-ESI-QTOF-MS/MS and GC/QTOF-MS: Evaluation of antioxidant and enzyme inhibition potential of extracts.

Phlogacanthus thyrsiformis (P. thyrsiformis) is a non-conventional edible plant that has been used as a vegetable and as a traditional medicine to treat various diseases. This non-conventional edible plant is widespread in India, Yunnan-Chinese provinces, and Vietnam. Despite this potential claim, the phytochemical investigation of plants remains incomplete. In the present study, the chemical profile analysis of P. thyrsiformis leaves extracts was performed using LC-ESI-QTOF-MS/MS and GC/QTOF-MS and assessed for its antioxidants and enzyme inhibitory potential. A cursory examination of the phytometabolites of different solvents extracts revealed chloroform extract has different metabolites, including phenols, terpenes, glycosides and alkaloids. 113 and 138 metabolites, including primary and secondary metabolites, were identified using LC-ESI-QTOF-MS/MS and GC/QTOF-MS, respectively. Total phenolic content was observed highest in chloroform extract (60.59 ± 1.25 mg GAE/g). Maximum antioxidant activity was found to be in chloroform extract, which was identified by DPPH (0.88 ± 0.29 mg/mL), ABTS (0.54 ± 0.1 mg/mL) and FRAP assay (26.46 ± 1.65 mg AA/g extract). Ethyl acetate extract have the highest α-amylase (0.09 ± 0.11 mg/mL) and α-glucosidase (0.088 ± 0.13 mg/mL) enzyme inhibition potential. For all the detected molecules, docking and molecular dynamics studies were performed. To the best of our knowledge, this is the first report that discusses LC-ESI-QTOF-MS/MS and GC/QTOF-MS-based metabolites profiling of P. thyrsiformis that can help to treat various diseases.

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.

Synthesis, characterization, method development, and validation of nor-ethylmorphine hydrochloride reference material using established analytical techniques for dope control analysis.

Ethylmorphine is permitted internationally for therapeutic purposes where morphine is not indicated across the globe. Nor-ethylmorphine a major metabolite of ethylmorphine. To differentiate the intake of morphine from ethylmorphine, nor-ethylmorphine stable reference material is desirable. There is no available commercial source and no data for reference material context for this substance. Therefore, nor-ethylmorphine HCl was synthesized and characterized, and purity and potency were assessed using nuclear magnetic resonance (NMR), high-resolution mass spectrometry (HRMS), Fourier transform infrared spectroscopy (FT-IR), thermogravimetry (TGA), and high-performance liquid chromatography (HPLC). Purity and potency were found to be 98.29% and 96.40%, respectively, providing a fit for purpose reference material for doping control analysis in sports.

Liquid chromatography/electrospray ionisation tandem mass spectrometric study of sitagliptin and its stressed degradation products.

A sensitive UPLC positive ion electrospray tandem mass spectrometry method was developed and validated for identification of degradation products of sitagliptin formed during stress study. Six of the major degradants were identi-fied with the proposed method. The separation of sitagliptin and its degradation products was achieved on a Acquity BEH C-18 column (50×2.1 mm, 1.7 µm) using a gradient programme. The mobile phase consists of solvent A (10 mM ammonium formate, pH 6.4 adjusted with formic acid) and solvent B (acetonitrile) and quantitative evaluation was performed at 267 nm with a flow rate of 0.15 mL min(-1). Suitability of this method for the quantitative determination of the drugs was proved by validation in accordance with International Conference on Harmonization (ICH) guidelines and can be used for routine analysis of sitagliptin formulations in quality control.