Encapsulation of Cymbopogon khasiana × Cymbopogon pendulus Essential Oil (CKP-25) in Chitosan Nanoemulsion as a Green and Novel Strategy for Mitigation of Fungal Association and Aflatoxin B1 Contamination in Food System.

The present study deals with the encapsulation of Cymbopogon khasiana × Cymbopogon pendulus essential oil (CKP-25-EO) into a chitosan nanoemulsion and efficacy assessment for inhibition of fungal inhabitation and aflatoxin B1 (AFB1) contamination in Syzygium cumini seeds with emphasis on cellular and molecular mechanism of action. DLS, AFM, SEM, FTIR, and XRD analyses revealed the encapsulation of CKP-25-EO in chitosan with controlled delivery. The CKP-25-Ne displayed enhanced antifungal (0.08 µL/mL), antiaflatoxigenic (0.07 µL/mL), and antioxidant activities (IC50 DPPH = 6.94 µL/mL, IC50 ABTS = 5.40 µL/mL) in comparison to the free EO. Impediment in cellular ergosterol, methylglyoxal biosynthesis, and in silico molecular modeling of CKP-25-Ne validated the cellular and molecular mechanism of antifungal and antiaflatoxigenic activity. The CKP-25-Ne showed in situ efficacy for inhibition of lipid peroxidation and AFB1 secretion in stored S. cumini seeds without altering the sensory profile. Moreover, the higher mammalian safety profile strengthens the application of CKP-25-Ne as a safe green nano-preservative against fungal association, and hazardous AFB1 contamination in food, agriculture, and pharmaceutical industries.

Identification and quantification of phytochemicals of Chamaecostus cuspidatus (Nees & Mart.) C.D.Specht & D.W.Stev and Cheilocostus speciosus (J. Koenig) C.D.Specht by LC-MS techniques and their in-vitro anti-adipogenic screening.

Chamaecostus cuspidatus (Nees & Mart.) C.D.Specht & D.W.Stev and Cheilocostus speciosus (J.Koenig) C.D.Specht contain bioactive compounds that possess many pharmacological activities including antidiabetic and hypolipidemic. These plants are used to treat diabetes by herbal healers. Considering the traditional use of C. cuspidatus and C. speciosus, the present study is designed to perform qualitative and quantitative analysis as well as in-vitro anti-adipogenesis against 3T3-L1 cells to ensure efficacy. A total of thirty-eight compounds were identified using HPLC-QTOF-MS/MS. Quantification of ten bioactive compounds among identified compounds was performed by UPLC-QqQLIT-MS/MS. The quantification method was validated according to ICH guidelines (International conference on harmonization guidelines). Quantification of bioactive compounds of different organs of C. cuspidatus and C. speciosus showed remarkable differences in the content. Microscopic and ORO absorbance confirmed the antiadipogenic potential of leaves (L-02), roots (R-02) of C. cuspidatus and leaves of C. speciosus (L-01) in 3T3-L1 cells.

LCMS-DNP based dereplication of Araucaria cunninghamii Mudie gum-resin: identification of new cytotoxic labdane diterpene.

As a part of natural defense, plants initiate the secretion of gum containing numerous pharmacologically active essential metabolites. A fraction of such gum-resin from Araucaria cunninghamii Mudie, when screened against human cancer cell lines, was found to be active. Further, it was subjected to an LCMS-DNP (Dictionary of Natural Products) based dereplication study followed by a detailed phytochemical investigation to obtain pure metabolites. Also, the gum resin of A. cunninghamii was found to be a rich source of abietanes and labdanes. The LCMS-DNP-based dereplication study identified many known metabolites, which were isolated for the first time from this plant as well as a new labdane diterpenoid (9). The compounds were characterized via spectroscopic techniques, which were subsequently compared with the already existing literature data. The metabolites were screened against seven human cancer cell lines. The anticancer activity was further supported by molecular docking studies.

Scientific Appraisal and Therapeutic Properties of Plants Utilized for Veterinary Care in Poonch District of Jammu and Kashmir, India.

The importance of traditional and indigenous knowledge is acknowledged on a worldwide scale for its coexistence principles and sustainable use techniques. In view of this, the present study is an attempt to document the ethno-veterinary plants used by the tribal communities of Western Himalaya. This study also provides the scientific validation of herbal medicines used in ethno-veterinary practices through a reverse pharmacological approach. A total of 59 informants were selected through a non-probability sampling method. Detailed information on the medicinal plants used in ethno-veterinary practices along with their habits and habitats, part/s used, remedy preparation methods, additives/ingredients used during preparation and administration, dosages administered, and route of administration was collected. Data was analyzed for the Relative Frequency of Citations (RFC), Use Values (UV), Informant Consensus Factor (ICF), and Jaccard Index (JI). Further, a reverse pharmacological approach was used for scientific validations of the documented herbal knowledge of plant species. During the study, 56 plant species belonging to 54 genera and 39 families were documented. Asteraceae was the dominant family followed by Lamiaceae, Amaranthaceae and Fabaceae. Life forms were dominated by herbaceous species and leaves were the most common plant parts used. The highest Relative Frequency of Citations (RFC) and Use Values (UV) were recorded for Brassica rapa L. (Brassicaceae). The Pearson correlation coefficient between RFC and UV shows a strong positive correlation between the proportion of uses of a plant species within a sample of informants and the number of times that a particular use of a plant species was mentioned by the informant. Studies of the biological activity of ethno-veterinary plants can provide clues of promising leads for the isolation and identification of useful compounds that may be developed into pharmaceuticals for human welfare.

A Coumarin-Donepezil Hybrid as a Blood-Brain Barrier Permeable Dual Cholinesterase Inhibitor: Isolation, Synthetic Modifications, and Biological Evaluation of Natural Coumarins.

Plants have immensely contributed to the drug discovery for neurodegenerative diseases. Herein, we undertook the phytochemical investigation of Nardostachys jatamansi (D.Don) DC. rhizomes followed by semisynthetic modifications to discover cholinesterase (ChE) and beta-site amyloid precursor protein cleaving enzyme 1 (BACE-1) inhibitors. The 8-acetyl-7-hydroxycoumarin isolated from the bioactive extract moderately inhibits acetylcholinesterase (AChE) and BACE-1 with IC50 values of 22.1 and 17.7 µM, respectively. The semisynthetic trifluoromethyl substituted coumarin chalcone display a 5-fold improvement in BACE-1 inhibition (IC50 3.3 µM). Another semisynthetic derivative, a coumarin-donepezil hybrid, exhibits dual inhibition of both ChEs with IC50 values of 1.22 and 3.09 µM, respectively. Molecular modeling and enzyme kinetics revealed that the coumarin-donepezil hybrid is a non-competitive inhibitor of AChE. It crosses the blood-brain barrier and also inhibits Aß self-aggregation. The results presented herein warrant a detailed investigation of the coumarin-donepezil hybrid in preclinical models of Alzheimer's disease.

Genus Rauvolfia: A review of its ethnopharmacology, phytochemistry, quality control/quality assurance, pharmacological activities and clinical evidence.

ETHNOPHARMACOLOGICAL RELEVANCE: The plants are from the genus Rauvolfia Plum. ex L. (Apocynaceae), which is represented by 74 species with many synonyms, and distributed worldwide, especially in the Asian, and African continents. Traditionally, some of them are used for the treatment of various disorders related to the central nervous system (CNS), cardiovascular diseases (CVD), and as an antidote due to the presence of monoterpene indole alkaloids (MIAs) such as ajmaline (144), ajmalicine (164) serpentine (182), yohimbine (190) and reserpine (214). AIM: The present review provides comprehensive summarization and critical analysis of the traditional to modern applications of Rauvolfia species, and the major focus was to include traditional uses, phytochemistry, quality control, pharmacological properties, as well as clinical evidence that may be useful in the drug discovery process. MATERIALS AND METHODS: Information related to traditional uses, chemical constituents, separation techniques/analytical methods, and pharmacological properties of the genus Rauvolfia were obtained using electronic databases such as Web of Science, Scopus, SciFinder, PubMed, PubChem, ChemSpider, and Google Scholar between the years 1949-2021. The scientific name of the species and its synonyms were checked with the information of The Plant List. RESULTS: A total of seventeen Rauvolfia species have been traditionally explored for various therapeutic applications, out of which the roots of R. serpentina and R. vomitoria are used most commonly for the treatment of many diseases. About 287 alkaloids, seven terpenoids, nine flavonoids, and four phenolic acids have been reported in different parts of the forty-three species. Quality control (QC)/quality assurance (QA) of extracts/herbal formulations of Rauvolfia species was analyzed by qualitative and quantitative methods based on the major MIAs such as compounds 144, 164, 182, 190, and 214 using HPTLC, HPLC, and HPLC-MS. The various extracts of different plant parts of thirteen Rauvolfia species are explored for their pharmacological properties such as antimicrobial, antioxidant, antiprotozoal, antitrypanosomal, antipsychotic, cardioprotective, cholinesterase inhibitory, and hepatoprotective. Of which, clinical trials of herbal formulations/extracts of R. serpentina and MIAs have been reported for CVD, CNS, antihypertensive therapy, antidiabetic effects, and psoriasis therapy, while the extracts and phytoconstituents of remaining Rauvolfia species are predominantly significant, owning them to be additional attention for further investigation under clinical trials and QC/QA. CONCLUSION: The present communication has provided a comprehensive, systematic, and critically analyzed vision into the traditional uses, phytochemistry, and modern therapeutic applications of the genus Rauvolfia are validated by scientific evidence. In addition, different plant parts from this genus, especially raw and finished herbal products of the roots of R. serpentina have been demonstrated for the QC/QA.

Effect of Organic Manure and Moisture Regimes on Soil Physiochemical Properties, Microbial Biomass Cmic:Nmic:Pmic Turnover and Yield of Mustard Grains in Arid Climate.

(1) Background: Arid conditions occur due to climate abnormality in the different biogeography regions of the world. The aim of this research is to investigate the stoichiometry of manure and moisture regimes on soil properties, microbial biomass C:N:P turnover, and the grain yield of mustard crops under stress in arid conditions; (2) Methods: The field experiment was carried out for 2 years at the farms of the agriculture college of SKN, Jobner (SKRAU Bikaner, Rajasthan). The effects of organic manure, moisture regimes, and saline water treatment on soil properties, such as the soil microbial biomass build-up, loss, turnover, and recycling of carbon (Cmic), nitrogen (Nmic), and phosphorus (Pmic) in the mustard crop were investigated. The twenty-seven treatments studied are described as follows: organic manures (control, FYM @ 10 t ha-1 and vermicompost @ 5 t ha-1), moisture regimes (0.4, 0.6, and 0.8 IW/CPE ratio), and saline irrigation water (control, 6, 12 dSm-1); (3) Results: Our findings indicate that vermicompost @ 5 t ha-1 significantly increases moisture retention and the available water in soil at 33 kPa and 1500 kPa. The microbial biomass build-up of Cmic increases by 43.13% over the control and 14.36% over the FYM. Similarly, the soil microbial biomass of Nmic, and Pmic also increase considerably. The SHC of the soil is enhanced by the application of farmyard fertilizer and vermicompost. The BD and pH decrease significantly, while the SHC, OC, CEC, and ECe of the soil increase significantly. The build-up, losses, and fluxes of the soil microbial biomass of Cmic, Nmic, and Pmic increase significantly, and the turnover rate decreases under vermicompost @ 5 t ha-1. A significant increase in grain yield was observed. Irrigation with a 0.8 IW/CPE moisture regime significantly decreases the pH of the SHC; (4) Conclusions: We hypothesized the interactive outcomes of the moisture regime and found that organic manure significantly influenced grain and stover yield. The treatments of quality irrigation water and the addition of organic manure are efficient enough to improve soil properties, water holding capacity, and soil microbial biomass C:N:P in stress climatic conditions.

Rapid determination and optimisation of berberine from Himalayan Berberis lycium by soxhlet apparatus using CCD-RSM and its quality control as a potential candidate for COVID-19.

SARS-CoV-2 (or COVID-19) has become a global risk and scientists are attempting to investigate antiviral vaccine. Berberis are important plants due to the presence of bioactive phytochemicals, especially berberine from the protoberberine group of benzylisoquinoline and recent studies have shown its potential in treating COVID-19. B. lycium Royle growing in subtropical regions of Asia had wide applications in Indian system of medicine. Rapid determination and novel optimisation method for berberine extraction has been developed by Soxhlet extraction utilising central composite design-response surface methodology (CCD-RSM). Berberine was detected by high-performance liquid chromatography (HPLC), and the highest yield (13.39%) was obtained by maintaining optimal extraction conditions i.e., extraction time (7.28 hrs), ethyl alcohol (52.21%) and solvent to sample ratio (21.78 v/w). Investigation of two geographic regions (Ramnagar and Srinagar) showed high berberine content in lower altitude. This novel optimisation technique has placed berberine as a potential candidate for developing pharmaceutical products for human health care.

Catharanthus roseus (L.) G. Don: A review of its ethnobotany, phytochemistry, ethnopharmacology and toxicities.

ETHNOPHARMACOLOGICAL RELEVANCE: Catharanthus roseus (L.) G. Don is a well known medicinal plant belonging to family Apocynaceae that have been traditionally used as medicine since ancient times. C. roseus is a well-recognized herbal medicine due to its anticancer bisindole alkaloids (vinblastine (111), vincristine (112) and vindesine (121)). In the Ayurvedic system of medicine, different parts of C. roseus are used in folklore herbal medicine for treatment of many types of cancer, diabetes, stomach disorders, kidney, liver and cardiovascular diseases. AIM OF THE STUDY: The main idea behind this communication is to update comprehensively and analyze critically the traditional applications, phytochemistry, pharmacological activities, and toxicity of various extracts and isolated compounds from C. roseus. MATERIALS AND METHODS: The presented data covers scientific works on C. roseus published across the world between 1967 and 2021 was searched from various international publishing houses using search engines as well as several traditional texts like Ayurveda and relevant books. Collected data from different sources was comprehensively summarized/analyzed for ethnomedicinal uses, phytochemistry, analytical chemistry, biological activities and toxicity studies of C. roseus. RESULTS AND DISCUSSION: C. roseus has a wide range of applications in the traditional system of medicine especially in cancer and diabetes. During phytochemical investigation, total of 344 compounds including monoterpene indole alkaloids (MIAs) (110), bisindole alkaloids (35), flavonoids (34), phenolic acids (9) and volatile constituents (156) have been reported in the various extracts and fractions of different plant parts of C. roseus. The extracts and isolated compounds of C. roseus have to exhibit many pharmacological activities such as anticancer/cytotoxic, antidiabetic, antimicrobial, antioxidant, larvicidal and pupicidal. The comparative toxicity of extracts and bioactive compounds investigated in dose dependent manner. The investigation of toxicity showed that the both extracts and isolated compounds are safe to a certain limit beyond that they cause adverse effects. CONCLUSION: This review is a comprehensive, critically analyzed summarization of sufficient baseline information of selected topics in one place undertaken till date on C. roseus for future works and drug discovery. The phytochemical investigation including biosynthetic pathways showed that the MIAs and bisindole alkaloids are major and characteristic class of compounds in this plant. The present data confirm that the extracts/fractions and their isolated alkaloids especially vinblastine (111) and vincristine (112) have a potent anticancer/cytotoxic and antidiabetic property and there is a need for further study with particular attention to the mechanisms of anticancer activity. In biosynthesis pathways of alkaloids especially bisindole alkaloids, some enzymes and rearrangement are unexposed therefore it is required to draw special attention. It also focuses on attracting the attention of scientific communities about the widespread biological activities of this species for its better utilization prospects in the near future.

Ethnobotany of the Himalayas: Safeguarding Medical Practices and Traditional Uses of Kashmir Regions.

The present study was carried out to enlist the medicinal plants used by the local inhabitants of developing countries such as India, and the district of Kupwara of the Kashmir Himalaya has been targeted. Our research is one of the first study focusing on the statistical evaluation of the cross-cultural analysis between three different communities i.e., Dard, Kashmiri and Gujjar, of the study area. Sampling was carried out in eight villages in 2017 to 2020, and data were collected from 102 informants based on walking transects, to collect plant specimens, and semi-structured interviews. The medical usages of all collected taxa were grouped into 15 disease categories and 81 biomedical ailments. In this study, we documented around 107 plant taxa belonging to 52 families from the local inhabitants of the Kashmir Himalaya, which regulate the livelihood of the people and support cultural ecosystem services. Asteraceae, Rosaceae, Lamiaceae, Malvaceae, Ranunculaceae, Poaceae, Solanaceae, Polygonaceae, Plantaginaceae and Brassicaceae are the top most dominant families. Herbaceous groups of plants were more common than trees and shrubs, and 71.96% of herb taxa were employed as medicine. Liliaceae, Caprifoliaceae and Portulacaceae (FUV = 0.24 each) have the highest family use value (FUV). The most prominent family was Asteraceae (seven genera, nine taxa), followed by Rosaceae and Lamiaceae (six genera, six taxa each). Persicaria Mill., Rheum L., Aconitum L. and Artemisia L. were prominent genera. Valeriana jatamansi Jones ex Roxb. (47UR), Fritillaria cirrhosa D. Don (45UR), Arisaema jacquemontii Blume (37UR), Asparagus racemosus Willd. (36UR) and Rumex acetosa L. (35UR) were the most important plant taxa with reference to use-reports. The ethnomedicinal applications of Aesculus indica Wall. ex Cambess., Solanum pseudocapsicum L., Ranunculus hirtellus Royle and Cormus domestica (L.) Spach plant taxa are reported here for the first time from the Himalayan Kashmiri people. We recommend further research on ethnopharmacological application of these newly recorded ethnobotanical plants. The medical usage of the plant was limited to different parts of the plant. In terms of the usage percentage, whole plant (26.17%), leaves (24.30%) and roots (19.63%) were found to have the highest utilization. The powder form (40.19%) was the most frequently employed method of drug/medicine preparation, followed by the utilization of extracted juice and/or other extracts (22.43%). The ICF values range from 0.85 to 1.00. Their use to remedy parasitic problems (PAR) and insect bites (IB) (ICF = 1.0 each) had the maximum consensus mentioned by the informants, although the number of taxa employed under this category was very limited. The different plant taxa used for the treatment of the gastrointestinal problems (GAS) was the most prominent disease category (262 URs, 16.19%, 25 taxa, ICF = 0.90). About 65% of the plant taxa studied is indigenous to the Asia or Himalayan regions, and around 35% is found to be exotic in nature. A strong positive correlation was found between age, gender, educational qualification and medicinal plant knowledge. No significant association was between people of different communities interviewed in terms of medical knowledge of the plants, p = 0.347 (>0.05) and χ2 = 2.120. No significant difference was found between the number of species documented concerning gender as p = 0.347 (>0.05) and χ2 =0.885. This study provides the comprehensive status of ethnomedicinal knowledge among three different communities of the study area. This study provided an impetus in discovering the baseline primary data for molecules which would help in drug discovery and management of various diseases, apart from conserving the genepool of plants in the investigated area.

Identification of plant-based multitargeted leads for Alzheimer's disease: In-vitro and in-vivo validation of Woodfordia fruticosa (L.) Kurz.

BACKGROUND: Alzheimer's disease (AD) is a complex neurodegenerative disease with no availability of disease-modifying therapeutics. The complex etiology and recent failures in clinical trials indicate the need for multitargeted agents. PURPOSE: The present study aims to discover new plant-based multitargeted anti-AD leads. METHODS: A library of plant extracts was screened for inhibition of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and beta-site amyloid precursor protein cleaving enzyme 1 (BACE-1). The secondary metabolites of active extracts were also tested, followed by enzyme-kinetics and molecular modeling to understand the mechanism of inhibition. The most active extract was investigated for in-vivo anti-dementia activity in behavioral mice models. RESULTS: Among the library of 105 extracts, Woodfordia fruticosa (SBE-80) and Bergenia ciliata (SBE-65) extracts displayed significant inhibition of all three enzymes. Gallic acid, one of the constituents of both plants, shows moderate inhibition of AChE and BACE-1. Catechin-3-O-gallate (CG), another constituent of SBE-65, inhibits EeAChE, rHuAChE, and eqBChE with IC50's of 29.9, 1.77, and 8.4 µM, respectively; along with a mild-inhibition of BACE-1. Ellagic acid, the constituent of SBE-80, inhibits BACE-1 with an IC50 value of 16 µM. The W. fruticosa extract SBE-80 at the dose of 25 mg/kg QD × 9 (PO) displayed memory-enhancing activity in Morris Water Maze and Passive Avoidance Test in Swiss albino mice. Treatment with SBE-80 also inhibits AChE in-vivo; whereas, a non-significant decrease in the serum TBARS was observed. CONCLUSION: W. fruticosa is identified for the first time as an anti-AD lead candidate. The in-vitro and in-vivo data presented herein and the documented safety profile of W. fruticosa indicate its strong potential for preclinical development as a botanical drug for dementia/AD.

Andrographis paniculata (Burm.f.) Nees: Traditional uses, phytochemistry, pharmacological properties and quality control/quality assurance.

ETHNOPHARMACOLOGICAL RELEVANCE: Andrographis paniculata (Burm.f.) Nees is a medicinal herb of the Asian countries used in many traditional medicinal systems for the treatment of diarrhea, flu, leprosy, leptospirosis, malaria, rabies, upper respiratory infections, sinusitis, syphilis, tuberculosis and HIV/AIDS etc. AIM OF THE STUDY: This review aims to provide the comprehensive, accurate and authentic information on traditional uses, phytochemistry and pharmacological properties of various extracts/fractions as well as phytocostituents of A. paniculata. In addition, this review also aims to provide advance and sensitive analytical methods along with chemical markers used in the standardization of herbal products for quality control (QC)/quality assurance (QA). MATERIALS AND METHODS: All relevant publications were considered within the years 1983-2020. The publications were searched from Google Scholar, PubChem, Chemspider, PubMed, Elsevier, Wiley, Web of Science, China Knowledge Resource Integrated databases and ResearchGate using a combination of various relevant keywords. Besides, relevant published books and chapters were also considered those providing an overview of extant secondary literature related to traditional knowledge, phytochemistry, pharmacology and toxicity of the plant. RESULTS AND DISCUSSION: In this review, 344 compounds, including, terpenoid lactones, flavonoids, phenolic acids, triterpenes and volatile compounds were summarized out of which more than half of the compounds have no reported pharmacological activities yet. Terpenoid lactones and flavonoids are the major bioactive classes of compounds of A. paniculata which are responsible for pharmacological activities such as anticancer and antioxidant activities, respectively. Biosynthetic pathways and active sites for target proteins of both terpenoid lactones and flavonoids were considered. Analgesic, anticancer, antidiabetic, antifertility, antiinflammatory, antimalarial, antimicrobial, antioxidant, antipyretic, antiviral, antiretroviral, antivenom, cardioprotective, hepatoprotective, immunomodulatory and neuroprotective activities have been also reported. Andrographolide is a major characteristic active principle and responsible for most of the pharmacological activities. Therefore, andrographolide has been selected as a marker for the standardization of raw and marketed herbal products by TLC, HPTLC, HPLC, GC-MS, HPLC-MS and HPLC-MS/MS methods for QC/QA. CONCLUSIONS: Conclusive evidence showed that the pharmacological activities reported in crude extracts and chemical markers are supporting and provides confidence in the traditional use of A. paniculata as a herbal medicine. The andrographolide could be used as a chemical marker for the QC/QA of raw and A. paniculata derived herbal products. Lactone ring in terpenoid lactone is an active site for targeted proteins. More efforts should be focused on the identification of the chemical markers from A. paniculata to provide a practical basis for QC/QA. Several aspects such as the mechanism of therapeutic potential, molecular docking technology and multi-target network pharmacology are very important for drug discovery and needed more investigation and should be considered. This compilation may be helpful in further study and QC/QA.

In-vitro antitumor activity of compounds from Glycyrrhiza glabra against C6 glioma cancer cells: identification of natural lead for further evaluation.

A HP20 resin-based unique method was adopted to get an active fraction of the hydroalcoholic extract of G. glabra roots. The fraction showed potent cytotoxicity against cancer cell line and was further subjected to detailed phytochemical investigation to obtain ten biomarkers. The isolated compounds were also tested for the cytotoxicity against the C6 glioma cell line in vitro using MTT assay. Among the isolated compounds, glycyrrhetic acid (1), glabrol (6), and glabridin (9) exhibited significant cytotoxicity. The compounds showed a dose-dependent decrease in cell viability. The active compounds were subjected to molecular docking study against topoisomerase I and topoisomerase II to support the mechanism of antitumor activity.

Determination of bioactive compounds of Artemisia Spp. plant extracts by LC-MS/MS technique and their in-vitro anti-adipogenic activity screening.

Ultra Performance Liquid Chromatography coupled with hybrid triple quadrupole linear ion trap tandem mass spectrometry (UPLC-ESI-QqQLIT-MS/MS) method in multiple reaction monitoring (MRM) acquisition mode was developed and validated for identification and simultaneous determination of potential anti-diabetic and anti-malarial compounds in ethanolic extracts of different Artemisia species. The chromatographic separation was carried out on an Acquity BEH™ C18 column (1.7 µm, 2.1 × 50 mm) with 0.1 % (v/v) formic acid in water and acetonitrile as mobile phase under gradient condition in 6 min. The developed method was validated in terms of linearity, LOD, LOQ, precision, stability and recovery according to international conference on harmonization guidelines. The correlation coefficients of all the calibration curves were ≥0.9902 and recoveries ranged from 98.22 to 104.49% (RSD ≤2.18 %). Relative standard deviations of intra-day, inter-day precisions and stability were ≤ 1.04, 1.09 and 2.80 %, respectively. The quantitative results showed remarkable differences in the content of all the compounds in different Artemisia species. The quantitative values of each peak were summarized as mean ± SD. The statistical analysis for comparison of observed quantitative differences of each compound was done to show that they are statistically significant. In-vitro assessment of extracts of selected Artemisia species inhibited adipocyte differentiation in 3T3-L1 cells, hence it may have certain phytochemicals which are responsible for reducing obesity and related metabolic disorders.

Ecological niche modeling as a cumulative environmental impact assessment tool for biodiversity assessment and conservation planning: A case study of critically endangered plant Lagerstroemia minuticarpa in the Indian Eastern Himalaya.

Cumulative environmental impact assessment (CEIA) at river basin level for hydroelectric projects is an evolving concept and has proved to be a useful tool to assess the cumulative impact of developmental projects on the natural ecosystems. However, the generality of CEIA studies is often contested because of methodological limitations, especially in the domain of biodiversity conservation and conservation planning. Ecological niche modeling (ENM) can be a useful tool in CEIA studies for conservation planning of threatened plants in hydroelectric project (HEP) areas. We elucidate this hypothesis taking the example of Lagerstroemia minuticarpa Debberm. ex P.C. Kanjilal, a critically endangered tree species in the Indian Eastern Himalaya. Standard ecological methods were employed to document occurrence records, estimate population size, and characterize habitats. ENM was used to estimate the species potential environmental niche and distribution areas. The possible impacts of HEPs on the potential habitats were predicted by overlaying the HEPs on the potential area map as well as using the conceptual network diagram. The study revealed that the species occupies an environmental niche characterized by humid to per-humid conditions, and is distributed mostly in the Lohit and Teesta basins. Potential areas of the species with high environmental suitability coincide with 19 HEPs, which point to a potential threat to the survival of the species. Network diagram indicated that project activities might deteriorate the habitats thereby affecting the population and regeneration of the species. Our study provides a framework for developing appropriate measures for species conservation and reintroduction at basin level using ENM.

Discovery and preclinical development of IIIM-160, a Bergenia ciliata-based anti-inflammatory and anti-arthritic botanical drug candidate.

OBJECTIVE: Bergenia ciliata (Haw.) Sternb. is used in the Indian traditional system of medicine to treat various ailments including rheumatism and to heal wounds. The objective of the present study was to perform a preclinical characterization of the B. ciliata-based botanical extract IIIM-160. METHODS: IIIM-160 was chemically standardized and analyzed for heavy metal content, aflatoxins, pesticides and microbial load. The in vitro and in vivo efficacies were determined in suitable models of inflammation, arthritis and nociception. An acute oral toxicity study was performed in Swiss albino mice. A suitable oral formulation was developed and characterized. RESULTS: Bergenin was found to be the major component (9.1% w/w) of IIIM-160. The botanical lead displayed inhibition of lipopolysaccharide-induced production of proinflammatory cytokines in THP-1 cells, with selectivity toward interleukin-6 (IL-6) and had an excellent safety-window. It showed anti-inflammatory, anti-arthritic and antinociceptive activity in animal models and was not toxic at oral doses up to 2 g/kg in Swiss-albino mice. The gastroretentive, sustained-release capsule formulation showed sustained-release of the bergenin over the period of 24 h, resulting in improved plasma-exposure of bergenin in Sprague-Dawley rats. CONCLUSION: The dual-activity of IL-6 inhibition and antinociception marks the suitability of IIIM-160 for treating rheumatoid arthritis. This study will serve as the benchmark for further research on this botanical formulation.

Assessment of chemical and genetic variability in Tanacetum gracile accessions collected from cold desert of Western Himalaya.

Genetic diversity is essential for survival and adaptation of high altitude plants such as those of Tanacetum genus, which are constantly exposed to environmental stress. We collected flowering shoots of ten accessions of Tanacetum gracile Hook.f. & Thomson (Asteraceae) (Tg 1-Tg 10), from different regions of cold desert of Western Himalaya. Chemical profile of the constituents, as inferred from GC-MS, exhibited considerable variability. Percentage yield of essential oil ranged from 0.2 to 0.75% (dry-weight basis) amongst different accessions. Tg 1 and Tg 6 were found to produce high yields of camphor (46%) and lavandulol (41%), respectively. Alpha-phellendrene, alpha-bisabool, p-cymene and chamazulene were the main oil components in other accessions. Genetic variability among the accessions was studied using RAPD markers as well as by sequencing and analyzing nuclear 18S rDNA, and plastid rbcL and matK loci. The polymorphic information content (PIC) of RAPD markers ranged from 0.18 to 0.5 and the analysis clustered the accessions into two major clades. The present study emphasized the importance of survey, collection, and conservation of naturally existing chemotypes of medicinal and aromatic plants, considering their potential use in aroma and pharmaceutical industry.

Development of Ultra Performance Liquid Chromatography Tandem Mass Spectrometry Method for Simultaneous Identification and Quantitation of Potential Osteogenic Phytochemicals in Butea monosperma.

An ultra performance liquid chromatography coupled with hybrid triple-quadrupole linear ion trap tandem mass spectrometry (UPLC-ESI-QqQLIT-MS-MS) method in multiple reaction monitoring mode was developed for identification and simultaneous determination of potential osteogenic compounds in ethanol extracts of different plant parts of Butea monosperma collected from different geographical regions. The chromatographic separation was carried out on an Acquity UPLC CSH C18 column (1.7 µm, 2.1 × 100 mm) with 0.1% (v/v) formic acid in water and methanol as mobile phase under gradient conditions in 8 min. The developed method was validated according to the guidelines of international conference on harmonization. The correlation coefficients of all the calibration curves were ≥0.9995 and recoveries ranged from 95.2 to 105.8% (RSD ≤ 1.95%). Relative standard deviations of intra-day, inter-day precisions and stability were ≤1.74, 1.84 and 2.8%, respectively. The quantitative results showed remarkable differences in the content of all potential osteogenic compounds in different parts of the plant as well as samples from different geographical regions. Quantitative variations studied from principal component analysis indicated tentative markers for B. monosperma cultivars which can discriminate sample of different geographical regions.

Simultaneous quantitative determination of bioactive terpene indole alkaloids in ethanolic extracts of Catharanthus roseus (L.) G. Don by ultra high performance liquid chromatography-tandem mass spectrometry.

A rapid, sensitive and reproducible method using ultra-high-performance liquid chromatography coupled with electrospray ionization hybrid triple quadrupole-linear ion trap mass spectrometry (UHPLC-ESI-QqQLIT-MS/MS) in multiple reaction monitoring (MRM) mode was developed and validated for simultaneous quantitation of anticancer (vincristine, vinblastine, vindesine), antihypertensive (ajmaline, ajmalicine, reserpine), aphrodisiac (yohimbine), sedative (serpentine) agents, dietary supplement (vinpocetine, yohimbine) and precursor of vinblastine (vindoline) from crude extracts of Catharanthus roseus. The precursor to product ion transitions for these compounds were observed at m/z 327 → 144, 355 → 144, 754 → 355, 353 → 144, 349 → 317, 825 → 225, 811 → 224, 458 → 188, 351 → 280 and 609 → 195, respectively in positive ionization mode. Chromatographic separation of all targeted TIAs was performed on ACQUITY UPLC BEH™ C18 column (1.7 µm, 2.1 mm × 50 mm). The calibration curves were linear within the concentration range 0.5-1000 ng/mL and correlation coefficients (R2) were closer to 1. Limit of detection (LOD) and limit of quantitation (LOQ) ranged from 0.039-0.583 ng/mL and 0.118-1.767 ng/mL, respectively. The intra-day (0.23-2.71% RSD) and inter-day (0.40-2.90% RSD) precision, stability (0.69-3.45% RSD) and recovery (99.63-104.30% ±â€¯%RSD ≤ 3.03%) were acceptable indicating good accuracy of the developed method. The method was successfully applied in ethanolic extracts of 39 samples of C. roseus parts (leaf, stem and root) collected from five different locations in India. Serpentine was detected as one of the most abundant TIA. Principal component analysis (PCA) was able to successfully discriminate among C. roseus samples on the basis of content of targeted TIAs.

Development of a UHPLC-MS/MS method for the quantitation of bioactive compounds in Phyllanthus species and its herbal formulations.

Phyllanthus species are extensively used in traditional medicines for the treatment of hepatic diseases due to their bioactive hypophyllanthin and phyllanthin. This work describes the development and validation of an ultra high performance liquid chromatography with tandem mass spectrometry method in polarity switching multiple reaction monitoring mode for the simultaneous detection and quantitation of 23 compounds using ultra-performance liquid chromatography coupled with electrospray-hybrid triple quadrupole-linear ion trap mass spectrometer. The validated parameters showed good linearity (R2  ≥ 0.996), limit of detection (0.05-1.62 ng/mL), limit of quantitation (0.15-4.95 ng/mL), precisions (intra-day: RSD ≤ 2.11%), (inter-day: RSD ≤ 2.91%), stability (RSD ≤ 2.56%) and overall recovery (98.22-104.48%; RSD ≤ 2.93%). The validated method was successfully applied in ethanolic extracts of P. amarus, P. niruri, P. emblica, P. fraternus, fractions of P. amarus and their herbal formulations for quantitation. The maximum content of hypophyllanthin (29.40 mg/g) and phyllanthin (56.60 mg/g) was detected in ethyl acetate fraction of P. amarus. The total content of 23 compounds was abundant in the ethanolic extract of P. emblica fruit. Principal component analysis was used to differentiate the selected Phyllanthus species and their herbal formulations. The results indicated that the present method could be used for quality control of Phyllanthus species and its herbal formulations.