Anuja Bhardwaj

HPTLC Based Chemometrics of Medicinal Mushrooms

The current study presents application of chemometrics to high performance thin layer chromatography (HPTLC) fingerprints of Indian medicinal mushrooms, Cordyceps sinensis (CS) and Ganoderma lucidum (GL). These mushrooms possess numerous bio-active constituents that endow them with several medicinal properties. Five CS extracts using different compositions of water and ethanol and nine sequentially extracted GL extracts were prepared employing the accelerated solvent extraction technique. A rapid and convenient HPTLC method was developed for generating phytochemical profiles and simultaneous detection of antioxidant compounds: ascorbic acid, hesperidin, gallic acid, rutin, and quercetin. The presence of these known and some of selective unknown zones visualized at 254 nm or 366 nm were confirmed by HPTLC–ESI/MS (HPTLC–Electronspray Ionization/Mass Spectrometry). The most probable m/z value of selective unknown HPTLC zones was searched against public databases to find putative compounds. Chemometric methods such as cluster analysis and principal component analysis (PCA) were performed on HPTLC datasets acquired during HPTLC scanning at 254 nm and 366 nm. Data matrices consisting of maximum peak heights of HPTLC zones at a specific Rf (retention factor) values were used as input variables. Also, the quantitative patterns of metabolites across the extracts were represented using HeatMap. This study illustrated HPTLC assisted chemometric analysis of Cordyceps sinensis and Ganoderma lucidum.

Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Inhibits Candida Biofilms: A Metabolomic Approach

This article presents a comparative gas chromatography (GC)-mass spectrometry (MS)-based metabolomic analysis of mycelia and fruiting bodies of the medicinal mushroom Ganoderma lucidum. Three aqueous extracts-mycelia, fruiting bodies, and a mixture of them-and their sequential fractions (methanolic and ethyl acetate), prepared using an accelerated solvent extractor, were characterized by GC-MS to determine volatile organic compounds and by high-performance thin-layer chromatography to quantify ascorbic acid, a potent antioxidant. In addition, these extracts and fractions were assessed against Candida albicans and C. glabrata biofilms via the XTT reduction assay, and their antioxidant potential was evaluated. Application of chemometrics (hierarchical cluster analysis and principal component analysis) to GC data revealed variability in volatile organic compound profiles among G. lucidum extracts and fractions. The mycelial aqueous extract demonstrated higher anti-Candida activity and ascorbic acid content among all the extracts and fractions. Thus, this study illustrates the preventive effect of G. lucidum against C. albicans and C. glabrata biofilms along with its nutritional value.

Screening of Indian Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes): A UPC2-SQD-MS Approach.

Oriental medicinal mushroom Ganoderma lucidum has been widely used for the promotion of health and longevity owing to its various bioactive constituents. Therefore, comprehending metabolomics of different G. lucidum parts could be of paramount importance for investigating their pharmacological properties. Ultra-performance convergence chromatography (UPC2) along with mass spectrometry (MS) is an emerging technique that has not yet been applied for metabolite profiling of G. lucidum. This study has been undertaken to establish metabolomics of the aqueous extracts of mycelium (GLM), fruiting body (GLF), and their mixture (GLMF) using ultra-performance convergence chromatography single quadrupole mass spectrometry (UPC2-SQD-MS). Aqueous extracts of G. lucidum prepared using an accelerated solvent extraction technique have been characterized for their mycochemical activities in terms of total flavonoid content, 1,1-diphenyl-2-picryl-hydrazyl scavenging activity, and ferric ion reducing antioxidant power. The UPC2-SQD-MS technique has been used for the first time for metabolite profiling of G. lucidum on a Princeton Diol column (4.6 × 250 mm; 5 µm) using supercritical CO2 (solvent) and 20 mM ammonium acetate in methanol (co-solvent). In the present study, UPC2-SQD-MS was found to be a rapid, efficient, and high-throughput analytical technique, whose coupling to principal component analysis (PCA) and phytochemical evaluation could be used as a powerful tool for elucidating metabolite diversity between mycelium and fruiting body of G. lucidum. PCA showed a clear distinction in the metabolite compositions of the samples. Mycochemical studies revealed that overall GLF possessed better antioxidant properties among the aqueous extracts of G. lucidum.

Nanoformulations: A Novel Approach Against Hypoxia

Abstract Global emergence of nanotechnology has led to the development of revolutionary commercial products from machines to medicines. It encompasses innovative biotechnology processes, the synthesis of new drugs and their targeted delivery, regenerative medicine, neuromorphic engineering, and developing a sustainable environment. It is an advanced technique that employs purposive and precise manipulation of atoms and molecules in order to use the unique properties of matter that could materialize at the nanoscale. Because of the potential technological applications, nanotechnology is developing rapidly with more than 300 products already in the market. Considering the current prevalence and advances in the field of nanotechnology, this chapter presents the basics of this nanoscaled technology and nanoformulations. It focuses on the use of nanoformulations concerning hypoxia-related diseases, including the research carried in context to high-altitude (hypobaric) hypoxia maladies. The chapter also highlights the prospective nanoformulations in managing the high-altitude induced pathophysiologies.

Characterization of nucleobases in sea buckthorn leaves: An HPTLC approach

ABSTRACT The present study aims to establish a high-performance thin layer chromatography (HPTLC)-based comparative analysis, directed toward characterization of nucleobases in aqueous and alcoholic extracts of sea buckthorn leaves from three different varieties: Hippophae salicifolia, Hippophae rhamnoides mongolica, and Hippophae rhamnoides turkestanica. The alcoholic and aqueous leaf extracts from these sea buckthorn varieties were prepared using accelerated solvent extraction technique. A novel HPTLC method for separating and identifying six nucleobases, namely, guanosine, guanine, cytosine, adenine, uracil, and thymine were adopted. HPTLC analysis indicated the presence of one or more of these nucleobases in a total of six leaf extracts evaluated, their quantities varying from 0.23 to 7.76 µg nucleobase per mg of extract. Though a typical trend could not be observed in the values obtained, the extracts were found to be considerably rich with respect to nucleobase contents. The results acquired from HPTLC were subsequently validated by hyphenation with mass spectrometry and also by applying chemometric tools in form of heat maps, hierarchical cluster dendrograms, and principal component analysis. The presence of nucleobases in the leaf extracts was confirmed by HPLC as well but HPTLC proved to be a better approach for characterization of nucleobases in plant extracts, than high performance liquid chromatography (HPLC). GRAPHICAL ABSTRACT