Rapid quality assessment and traceability of ginger powder from Northeast India and Indian market based on near infrared spectroscopic fingerprinting.

INTRODUCTION: Ginger (Zingiber officinale Rosc.) varies widely due to varying concentrations of phytochemicals and geographical origin. Rapid non-invasive quality and traceability assessment techniques ensure a sustainable value chain. OBJECTIVE: The objective of this study is the development of suitable machine learning models to estimate the concentration of 6-gingerol and check traceability based on the spectral fingerprints of dried ginger samples collected from Northeast India and the Indian market using near-infrared spectrometry. METHODS: Samples from the market and Northeast India underwent High Performance Liquid Chromatographic analysis for 6-gingerol content estimation. Near infrared (NIR) Spectrometer acquired spectral data. Quality prediction utilized partial least square regression (PLSR), while fingerprint-based traceability identification employed principal component analysis and t-distributed stochastic neighbor embedding (t-SNE). Model performance was assessed using RMSE and R2 values across selective wavelengths and spectral fingerprints. RESULTS: The standard normal variate pretreated spectral data over the wavelength region of 1,100-1,250 nm and 1,325-1,550 nm showed the optimal calibration model with root mean square error of calibration and R2 C (coefficient of determination for calibration) values of 0.87 and 0.897 respectively. A lower value (0.24) of root mean square error of prediction and a higher value (0.973) of R2 P (coefficient of determination for prediction) indicated the effectiveness of the developed model. t-SNE performed better clustering of samples based on geographical location, which was independent of gingerol content. CONCLUSION: The developed NIR spectroscopic model for Indian ginger samples predicts the 6-gingerol content and provides geographical traceability-based identification to ensure a sustainable value chain, which can promote efficiency, cost-effectiveness, consumer confidence, sustainable sourcing, traceability, and data-driven decision-making.

Ultra-performance liquid chromatography-Quadrupole time-of-flight tandem mass spectrometry-based metabolite profiling, quality evaluation, and marker analysis of Trachyspermum ammi (L.) Sprague by high-performance thin-layer chromatography.

Trachyspermum ammi (L.) Sprague (Apiaceae), commonly known as "Ajwain" is distributed throughout India. Ajwain fruits contain fiber, carbohydrates, phenolic acids, flavonoids, and tannins. The fruits also yield a small amount of essential oil, with Thymol as the principal constituent. Ajwain has various pharmacological activities like anti-leishmanial, antimicrobial, cytotoxic, antispasmodic, nematocidal, and anthelmintic. The fruits are of high therapeutic value; thus, it becomes quite essential to evaluate the quality of Trachyspermum ammi (L.) Sprague to authenticate and ensure its therapeutic and nutritional properties. The ethyl acetate fraction of Trachyspermum ammi (L.) Sprague fruits exhibited the highest total phenolic and flavonoid content values of 149.55 ± 1.19 mg rutin equivalent and 682.85 ± 3.68 mg gallic acid equivalent, respectively. Metabolite profiling of the ethyl acetate fraction using ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry analysis resulted in identifying 19 phytomolecules. A validated high-performance thin-layer chromatography method was developed to quantify standard phytomolecules in the ethyl acetate fraction. The highest and lowest percentages of phytomarker were found to be caffeic acid (5.51% ± 0.16% w/w) and gallic acid (1.29% ± 0.09% w/w), respectively. This validated rapid, accurate, and precise method for standardization of Trachyspermum ammi (L.) Sprague will be beneficial for its quality evaluation as well as the derived products.

Quality evaluation of different black rice varieties of northeastern region of India.

INTRODUCTION: Black rice (Oryza sativa L.), which is rich in polyphenols and flavonoids, is indigenous to Northeast India, specifically Manipur, and traditionally consumed for its protective effects on human health. Due to its economic value, it is crucial to evaluate the quality of different black rice varieties to authenticate their therapeutic and nutritional properties. OBJECTIVE: We aimed to evaluate the quality of pre- and post-marketed black rice samples by a validated high-performance thin layer chromatography method and determine variations of total phenolics and total flavonoids with antioxidant potential. MATERIAL AND METHODS: The ferulic acid, gallic acid, quercetin, and caffeic acid contents of three black rice varieties-Poireiton, Amubi, and Sempak-along with two marketed samples of Amubi from Manipur, India, were quantified based on standards. Antioxidant potential was measured by the 2,2-diphenyl-1-picryl-hydrazyl hydrate free radical scavenging assay. RESULTS: The highest and lowest relative biomarker contents were found in hydroalcoholic extracts of Amubi [caffeic acid (1.43% w/w), ferulic acid (1.15% w/w), quercetin (0.6% w/w), and gallic acid (0.39% w/w)] and the marketed sample Var. Amubi from Kakching District, respectively. Pearson's correlation coefficient of antioxidant potential with phenolic and flavonoid content showed a moderate to strong correlation for all samples. CONCLUSION: This validated, rapid, accurate standardization method for black rice varieties will be beneficial for the quality evaluation of black rice and its derived products. It will also be helpful to authenticate the nutritional benefits for the consumers.

6-Gingerol contents of several ginger varieties of Northeast India and correlation of their antioxidant activity in respect to phenolics and flavonoids contents.

INTRODUCTION: Ginger constitutes the rhizome part of the plant Zingiber officinale from the Zingiberaceae family. A large number of ginger varieties with high sensorial and functional quality are found in Northeast India. Hence, phytopharmacological screening of different ginger varieties is essential that will serve as a guideline in applied research to develop high-end products and improve economical margins. OBJECTIVE: To determine the variation in total phenolics content (TPC), total flavonoids content (TFC), and antioxidant activities and correlate that with 6-gingerol contents of different ginger varieties collected from Northeast India using Pearson's correlation analysis. MATERIALS AND METHODS: The TPC and TFC values were determined using standard methods. Antioxidant activities were measured using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radical scavenging assays, while reversed-phase high-performance liquid chromatography (RP-HPLC) analysis was utilised for quantitative determination of 6-gingerol content. RESULTS: The result revealed that ginger variety 6 (GV6) contains the highest 6-gingerol content and TPC value showing maximum antioxidant activity, followed by GV5, GV4, GV9, GV3, GV2, GV8, GV1, and GV7. The findings also suggested that the antioxidant activity has much better correlations with TPC as compared with TFC values. Pearson's correlation analysis showed a significant correlation between 6-gingerol contents and TPC values. CONCLUSION: This work underlines the importance of ginger varieties from Northeast India as a source of natural antioxidants with health benefits.

A reversed-phase ultra-fast liquid chromatography-photodiode array detector (RP-UFLC-PDA) method for simultaneous estimation of ayapanin and umbelliferone in Ayapana triplinervis Vahl.

A rapid validated ultra-fast liquid chromatography-photodiode array detector (UFLC-PDA) method was developed to identify and quantify ayapanin (AY) and umbelliferone (UM) simultaneously in Ayapana triplinervis Vahl methanolic extract. The method was validated for linearity, limit of detection (LOD; 3:1σ/S), limit of quantification (LOQ; 10:1σ/S), precision, accuracy, specificity and robustness. The response was linear with a good correlation between concentration and mean peak area through a correlation coefficient of 0.9996, y = 7025.7x - 2269.8 and 0.9997, y = y = 16,262x - 946 with LOD of 6.256 ± 0.52 and 3.325 ± 0.36, and LOQ of 18.838 ± 0.18 and 8.870 ± 0.85 for AY (0.67% w/w) and UM (0.18% w/w), respectively. The relative standard deviation (%) of precision and recovery of AY and UM was <2.0%. The proposed method was simple, accurate, specific, precise and reproducible.

Quality Related Safety Evaluation of a South African Traditional Formulation (PHELA®) as Novel Anti-Biofilm Candidate.

A South African traditional formulation, PHELA®, is consumed by the traditional people for severe chest problems with coughing, diarrhea, oral ulcers etc. The present study focused on establishing the anti-infective properties of a safe and standardized poly-herbal formulation through a series of criteria and specifications.

Lagenaria siceraria and it's bioactive constituents in carbonic anhydrase inhibition: A bioactivity guided LC-MS/MS approach.

INTRODUCTION: Lagenaria siceraria, is a popular food plant among Indians, contains a large number of phenolic compounds with several medicinal benefits, mentioned in Indian System of Medicine (ISM). OBJECTIVES: To investigate the carbonic anhydrase inhibitory potential and inhibitory mechanism of the most potent fraction of L. siceraria fruits. MATERIALS AND METHODS: The extract and fraction of dried fruit of L. siceraria screened for their in vitro carbonic anhydrase II (bCA II) inhibitory activity. The active fraction was purified by using flash chromatography. The bioactive compounds were identified and quantified through liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS) and reverse-phase high-performance liquid chromatography (RP-HPLC). Finally, the underlying carbonic anhydrase inhibitory mechanism of the compounds was explained by enzyme kinetics and molecular docking study. RESULTS: The LC-QTOF-MS based identification of the most active fraction revealed the presence of phenolic compounds. The results of the enzyme inhibition assay revealed that coniferyl alcohol, ferulic acid and p-Coumaric acid inhibited bCA II activity [half maximal inhibitory concentration (IC50 ) value range of 80 to 250 µM) in a dose dependent manner. The kinetics study of enzyme inhibition revealed that p-Coumaric acid binds to the enzyme competitively whereas the non-competitive type of inhibition was observed for ferulic acid and coniferyl alcohol. The molecular docking study explored the interaction mechanism of phenolic compounds at the active site of bCA II. CONCLUSION: The present research led us to conclude that, the phenolic compounds from L. siceraria serve as major contributors for carbonic anhydrase inhibition, which could play a useful role in the management of oedema, hypertension, obesity and related metabolic disorders.

Immunoprotective potential of Ayurvedic herb Kalmegh (Andrographis paniculata) against respiratory viral infections - LC-MS/MS and network pharmacology analysis.

INTRODUCTION: Immunity boosting has emerged as a global strategy to fight the SARS-CoV-2 pandemic situation. In India, AYUSH systems of medicine have been promoted as an immune-protection strategy. Andrographis paniculata (Burm. F) Nees (AP) mentioned in Ayurveda has been widely used for treating sore throat, flu, and upper respiratory tract infections which may provide possible novel therapeutic approaches, exclusively targeting SARS-CoV-2 and its pathways. OBJECTIVES: The present work uses liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics and combination synergy analysis based on network pharmacology to mine multimode evidence to understand the possible mechanism of action, diseases association, protein-protein interaction and major pathways involved therein. MATERIAL AND METHODS: Metabolite profiling was performed by Agilent QTOF LC-MS/MS system. Network pharmacology analysis was performed by using functional annotation analysis based on databases like Binding DB, STRING, DAVID and KEGG for further data mining. Further combination synergy was evaluated using "neighbourhood approach" and networks were constructed through Cytoscape 3.2.1. RESULTS: The molecules from kalmegh provides immune-protection and anti-viral response via involving different pathways, like toll-like receptor pathway, PI3/AKT pathway and MAP kinase pathways against COVID-19 infection. The KEGG analysis showed that in a vast majority of the most enriched pathways, AP were associated with viral infections and upper respiratory tract infections. CONCLUSIONS: The results suggest a synergy between andrographolide and other molecules identified as safe and efficacious anti-inflammatory agent having effects on upper respiratory tract infections and can significantly decrease the production of cytokines and pro-inflammatory factors in viral infections.

Optimized piperine-phospholipid complex with enhanced bioavailability and hepatoprotective activity.

Piper species is one of the most widely consumed spices for culinary purposes. Piperine (PIP) present in Piper species has a wide range of therapeutic activity including hepatoprotection. However, the major biological limitation of PIP is its low bioavailability after oral administration. Purpose of the study was to prepare an optimized and adequately characterized PIP-phospholipid complex (PPC) as a delivery system to overcome these limitations and to investigate the pharmacokinetics and hepato-protectivity of the formulation in the animal model. Response surface methodology was adopted to optimize the process parameters for PPC preparation. FT-IR, DTA, PXRD, SEM, molecular docking etc. were used for characterization. Solubility, log P, dissolution efficiency and in vivo pharmacokinetics were also investigated. PPC showed enhanced hepatoprotective potential as compared to pure PIP at the same dose level (25 and 50 mg/kg). PPC restored the levels of serum marker and antioxidant enzymes. PPC also increased the bioavailability of PIP in rat serum by 10.40-fold in comparison with pure PIP at the same dose level and enhanced the elimination half-life (t1/2 el) from 0.477 ± 1.76 to 9.80 ± 1.98 h. Results concluded that PPC enhanced the hepatoprotection of PIP which may be due to the improved bioavailability and pharmacokinetics of PIP in rats.

LC-QTOF-MS-based metabolite profiling and evaluation of α-glucosidase inhibitory kinetics of Coccinia grandis fruit.

Coccinia grandis is an important food crop of the Cucurbitaceae family, widely used for culinary purposes in India. It is reported to possess hypoglycemic, hypolipidemic and antioxidant activities. The current study was aimed to explore the inhibition kinetics as well as major constituents of the active fraction of C. grandis against α-glucosidase. The kinetic study was performed through spectrophotometric assay, with p-nitrophenyl-α-d-glucopyranoside as a substrate with varying concentrations. An in vitro antioxidant study was performed by DPPH assay. In addition, UPLC-QTOF-MS analysis was carried out for metabolite profiling of the bioactive fraction of C. grandis. The results showed that the difference between the α-glucosidase inhibitory activity of the ethyl acetate fraction of C. grandis (EFCG) (IC50 2.43 ± 0.27 mg/ml), and standard inhibitor, acarbose (2.08 ± 0.19 mg/ml), was not statistically significant at a P-value of 0.05. The enzyme kinetics confirmed the inhibition mode in a mixed manner. The EFCG also showed the highest antioxidant activity (101.74 ± 1.95 µg/ml) among all of the fractions. A significant correlation between antioxidant and α-glucosidase inhibitory activity of EFCG was observed. The LC-QTOF-MS study of the EFCG putatively identified 35 metabolites, which may be responsible for its antioxidant and α-glucosidase inhibitory properties. Thus, C. grandis fruits can serve as a functional food to address diabetes-related disorders associated with α-glucosidase.

Safety assessment of selected medicinal food plants used in Ayurveda through CYP450 enzyme inhibition study.

BACKGROUND: Andrographis paniculata, Bacopa monnieri and Centella asiatica are mentioned in Ayurveda for the management of neurodegenerative disorders. These plants and their phytomolecules, such as andrographolide, bacoside A and asiaticoside, were studied for their inhibition potential on pooled CYP450 as well as human CYP3A4, CYP2D6, CYP2C9 and CYP1A2 by CYP-CO complex assay and fluorogenic assay respectively followed by IC50 determination. Quantification of bioactive compounds present in the extracts was done by RP-HPLC. Heavy metal content in the selected medicinal plants was determined by atomic absorption spectroscopy. RESULT: CYP-CO complex assay indicated significantly less inhibition potential than standard inhibitor (P < 0.05 and above). A. paniculata showed highest inhibitory activity against CYP3A4 and CYP2D6 (IC50 = 63.06 ± 1.35 µg mL-1 ; 88.80 ± 3.32 µg mL-1 ), whereas C. asiatica and B. monnieri showed least inhibitory activity against CYP1A2 (IC50 = 288.83 ± 1.61 µg mL-1 ) and CYP2C9 (184.68 ± 3.79 µg mL-1 ), respectively. In all cases the extract showed higher inhibition than the single bioactive compounds. The heavy metals content in the plant extracts were within the permissible limits. CONCLUSION: The findings suggested that selected food plants and bioactive compounds contributed negligible interaction potential with CYP isozymes and may not possess any harmful effect with regard to their therapeutic application. © 2016 Society of Chemical Industry.

Paris polyphylla Sm. characterized extract infused ointment accelerates diabetic wound healing in In-vivo model.

ETHNOPHARMACOLOGICAL RELEVANCE: The dried rhizome of Paris polyphylla Sm. is extensively used by traditional healers in India, China, and Vietnam to treat skin inflammation, cut wounds, uterine and traumatic bleeding, and cancer. AIM OF THE STUDY: The traditional use of P. polyphylla rhizomes for treating wounds and bleeding has been reported previously. However, the potential of P. polyphylla in the treatment of diabetic wounds has not yet been explored. Our present study focused on the investigation of the wound-healing activity of P. polyphylla infused ointment in streptozotocin (STZ)-induced diabetic rats to validate the traditional claim. MATERIALS AND METHODS: Hydroalcoholic extract of the dried rhizomes of P. polyphylla were quantified by validated and optimized HPTLC (High-performance thin layer chromatography) method for Paris saponin VII, Dioscin and Polyphyllin V. The extract was used to prepare P. polyphylla ointments (5 and 10%). P. polyphylla ointment was subjected to physiochemical analysis and skin irritation test. Thirty STZ-induced diabetic adult male Wistar albino rats were divided into five groups (n = 6) and a circular excision wound was created. P. polyphylla ointment, ointment base (OB), and standard (STD) (Povidone Iodine 10%) were administered topically. The wound area of all groups were recorded every six days and compared with that of control. The epithelization period of each group was recorded. On day 18, the histopathological study of skin tissues of all groups was performed using hematoxylin and eosin (H&E) and Mallory's trichrome (MT). RESULTS: Marker analysis and quantification of phytomolecules in hydroalcoholic extract ofP. Polyphylla were found to be of paris saponin VII (3.28 ± 0.08% w/w), dioscin (1.94 ± 0.12% w/w), and polyphyllin V (1.87 ± 0.84% w/w). A physiochemical study of P. polyphylla ointment showed that the prepared ointment was within an acceptable range and was not irritable to the skin. Daily topical administration of 10% P. polyphylla ointment (PP10) for 18 days completely healed the STZ-induced diabetic wounds. On day 18, the 5% P. polyphylla ointment (PP5) showed 99.1 ± 2.9% wound closure, while that of the standard and control was 78.4 ± 7.3% and 18.5 ± 5.9%, respectively. The epithelialization period of PP10 was 18 days, whereas that of the control was 28 days. Histopathological analysis of the progression of PP10 and PP5 wounds showed a decrease in inflammatory cells, regenerated epithelial layer, keratosis layer, hair follicles, fibroblasts, and collagen. Upon collagen intensity quantification of MT stained sections, an increase in collagen density of PP10 and PP5 treated groups was observed, showing accelerated wound healing potential of P. polyphylla extract in diabetic wounds compared to the standard ointment. CONCLUSION: This study suggested the potential of P. polyphylla rhizomes derived formulation to treat diabetic wounds, although the plant is traditionally used to treat normal wounds. The results indicate the validation of traditional claim, which has been explored commercially in industrial aspect.

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.

Exploration of anti-diabetic activity and metabolite profiling of Bruguiera cylindrica (l.) Bl.-in vivo anti-diabetic activity, exploration of molecular mechanism, and network pharmacological analysis.

INTRODUCTION: The Bruguiera cylindrica L. is a mangrove plant that is typically found in coastal areas of Asia, including India. It has been known for its medicinal properties, which have been utilized for generations. For example, in Thailand, it has been used to treat wounds and diarrhoea, while in India, it has been effective in addressing diabetes, ulcers, and other health issues. This particular study sought to investigate the potential of B. cylindrica bark extract in reducing the symptoms of diabetes in rats. METHODS: In this study, we examined the potential of B. cylindrica bark extract as an inhibitor of α-amylase and α-glucosidase enzymes in vitro. We also evaluated the effects of the extract and Metformin on rats fed high-fat diets and measured their lipid profiles and biochemical parameters. Furthermore, we conducted a network pharmacology analysis to identify proteins and pathways involved in the amelioration of diabetes. RESULTS: Through metabolite profiling, we identified 58 compounds in B. cylindrica hydroalcoholic extract. These compounds include alkaloids, phenolics, flavonoids, and fatty acids. The extract was found to have a dose-dependent inhibition activity against α-amylase and α-glucosidase, with IC50 values similar to acarbose. In rats, oral administration of 200-400 mg/kg of B. cylindrica led to reduced blood glucose levels and normalized serum biochemical parameters. CONCLUSIONS: Bruguiera cylindrica bark may reduce blood sugar levels in rats with diabetes. The study found metabolites that interact with protein targets associated with different types of diabetes.

Chemo-profiling and exploring therapeutic potential of Momordica dioica Roxb. ex Willd. for managing metabolic related disorders: In-vitro studies, and docking based approach.

ETHNOPHARMACOLOGICAL RELEVANCE: Momordica dioica Roxb. ex Willd. (M. dioica Roxb.) a nutritious and therapeutic property rich crop of Cucurbitaceae plant family. In various folklore medicine including Ayurveda fruits are used to treat several metabolic related disorders i.e., hyperglycemia, hyperlipidemia, diabetes, obesity etc. Furthermore, traditionally it is used to treat fever, inflammation, ulcer, skin diseases, haemorrhoids, hypertension and also employed as cardioprotective, hepatoprotective, analgesic, diuretic. AIM OF THE STUDY: This study focuses to explore the therapeutic potential of Momordica dioica Roxb. ex Willd. through in-vitro and in-silico approach for managing hyperlipidemia, hyperglycemia and related metabolic disorders along with its phytochemical profiling for quality evaluation and validation of traditional claim. MATERIALS AND METHODS: The present study was carried out on hydroalcohol extract of dried leaf and fruit of Momordica dioica. In-vitro antioxidant potential using DPPH and Nitric oxide scavenging assay along with in-vitro enzyme inhibitory potential against α-amylase, α-glucosidase, and pancreatic lipase enzymes was studied. The bioactive metabolites were identified from the most potent bioactive extract by analysis with LC-QTOF-MS and also studied their role to lessen the metabolic related disorder through in-silico approaches. RESULTS: The results confirmed that the fruit extract is more active to possess antioxidant and prominent enzyme inhibition potential compared to the leaf. Sixteen identified metabolites in M. dioica Roxb. fruits may be responsible for the therapeutic potential related to metabolic related disorder. The in-silico study of the identified phytomolecules against α-amylase, α-glucosidase and pancreatic lipase showed significant docking scores ranging from -9.8 to -5.5, -8.3 to -4.8 and -8.3 to -6 respectively. CONCLUSION: The current study illustrated that M. dioica Roxb., a traditionally important plant is potential against metabolic related disorders. Phytocomponents present in the fruit extract may be responsible for antioxidant as well as the enzymes' inhibitory potential. Thus, fruits of M. dioica Roxb. will be useful as alternative therapeutics for treatment of hyperlipidemia, hyperglycemia and related metabolic disorders.

Investigation on Anti-diabetic Efficacy of a Cucurbitaceae Food Plant from the North-East Region of India: Exploring the Molecular Mechanism through Modulation of Oxidative Stress and Glycosylated Hemoglobin (HbA1c).

BACKGROUND: The medicinal plants of the Cucurbitaceae family, such as Solena heterophylla Lour. fruits, have significant ethnobotanical value and are readily accessible in North East India. AIMS: We conducted a study on Solena heterophylla Lour. fruits to evaluate their anti-diabetic activity in vivo, standardize their HPTLC, and profile their metabolites using LC-QTOF-MS. We aimed to explore the molecular mechanism behind their effects on oxidative stress and glycosylated hemoglobin (HbA1c). METHODS: Firstly, the ethyl acetate fraction of Solena heterophylla Lour. fruits was standardized using Cucurbitacin B as a standard marker by conducting HPTLC evaluation. Next, we delved into analyzing metabolite profiling. In addition, the standardized fraction was utilized in an experimental study to investigate the molecular mechanism of action in an in vivo high-fat diet and a low dose of streptozotocin-induced diabetic model. RESULTS: We have reportedly identified 52 metabolites in the ethyl acetate fraction of Solena heterophylla (EASH). In the in vitro tests, it has been observed that this extract from plants possesses notable inhibitory properties against α-amylase and α-glucosidase. Solena heterophylla fruits with high levels of Cucurbitacin B (2.29% w/w) helped lower FBG levels in animals with EASH treatment. EASH treatment reduced HbA1c levels and normalized liver lipid peroxidation and antioxidant enzyme levels. SGOT, SGPT, and SALP serum enzyme levels also returned to normal. CONCLUSION: Based on the current evaluation, it was found that EASH exhibited encouraging hypoglycemic effects in diabetic rats induced by a low dose of STZ and high-fat diet, which warrants further investigation.

Combining LC-MS/MS profiles with network pharmacology to predict molecular mechanisms of the hyperlipidemic activity of Lagenaria siceraria stand.

ETHNOPHARMACOLOGICAL RELEVANCE: Lagenaria siceraria Stand. (Family: Cucurbitaceae), popularly known as bottle gourd, is traditionally used in Ayurvedic medicine as a food plant, especially in hypertension and obesity. AIM OF THE STUDY: Investigations were undertaken to assign novel lead combinations from this common food plant to multi-molecular modes of actions in the complex disease networks of obesity and hypertension. LC-MS/MS based metabolite screening, in-vivo high fat diet induced hyperlipidemia animal study and network pharmacology explorations of the mechanism of action for lipid lowering effects including a neighbourhood community approach for molecular combinations were performed. MATERIAL AND METHODS: Major chemical constituents of the fruits of LS (LSFE) were analysed by HPLC-DAD-MS/MS-QTOF. Wistar albino rats (n = 36), divided into 6 groups (n = 6) received either no treatment or a high-fat diet along with LSFE or Atorvastatin. Lipid profiles and biochemical parameters were evaluated. In silico cross-validated network analyses using different databases and Cytospace were applied. RESULTS: Profiling of LSFE revealed 18 major constituents: phenolic acids like p-Coumaric acid and Ferulic acid, the monolignolconferyl alcohol, the flavonoid glycosides hesperidin and apigenin-7-glucoside. Hyperlipidemic animals treated with LSFE (200 mg/kg, 400 mg/kg, 600 mg/kg) showed a significant improvement of their lipid profiles after 30 days of treatment. Network pharmacology analyses for the major 18 compounds revealed enrichment of the insulin and the ErbB signalling pathway. Novel target node combinations (e.g. AKR1C1, AGXT) including their connection to different pathways were identified in silico. CONCLUSIONS: The combined in vivo and bioinformatics analyses propose that lead compounds of LSFE act in combination on relevant targets of hyperlipidemia. Perturbations of the IRS→Akt→Foxo1 cascade are predicted which suggest further clinical investigation towards development of safe natural alternative to manage hyperlipidemia.

Quantification and Standardization of Andrographolide in Andrographis Paniculata Samples by Validated RP-HPLC and HPTLC Methods.

Andrographis paniculata (family Acanthaceae) is known as Kalmegh, one of the traditionally used important medicinal plant contains several biologically active phytochemical including andrographolide. A. paniculata is broadly used by healthcare practitioners in India and also used in different traditional medicinal system. In this study, the leaves of A. paniculata were collected from West Medinipur, East Medinipur, South 24 Parganas, Purulia and Hooghly district of West Bengal, India. This study aiming towards validation and development of a simple, precise and reproducible reverse-phase high-performance liquid chromatography (RP-HPLC) and high-performance thin layer chromatography (HPTLC) methods for quantification of andrographolide in A. paniculata extracts. The validated RP-HPLC and HPTLC study confirmed that different concentrations of andrographolide content present in the plant samples, which are collected from above different districts of West Bengal, India. The amounts of andrographolide were found to be 2.71% (w/w), 3.19% (w/w), 1.83% (w/w), 1.73% (w/w) and 2.94% (w/w) in RP-HPLC study and 2.13% (w/w), 2.51% (w/w), 1.01% (w/w), 1.25% (w/w) and 2.15% (w/w) in HPTLC study. This precise, reproducible, accurate and specific method can be used for the quantification of andrographolide in kalmegh, as per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines recommendations.

Hydroalcoholic root extracts of Houttuynia cordata (Thunb.) standardized by UPLC-Q-TOF-MS/MS promotes apoptosis in human hepatocarcinoma cell HepG2 via GSK-3ß/ß-catenin/PDL-1 axis.

Houttuynia cordata (Thunb.), an important medicinal plant of Northeast India, Korea, and China, is used to treat various ailments and for anticancer research. Knowing its traditional practices, we are interested in the mode-of-action of HCT on HepG2 to co-relate the traditional practice with modern drug therapeutics. UPLC-Q-ToF-Ms analysis of HCT reveals identification of 14 metabolites. Network pharmacology analysis of the 14 compounds showed interaction with 232 different targets with their potential involvement in hepatocellular carcinoma. Whole extracts impart cytotoxicity on variety of cell lines including HepG2. There was a significant morphological alteration in treated HepG2 cells due to impairment of cytoskeletal components like ß and γ- tubulin. Arrest at G1-S checkpoint was clearly indicated downregulation of Cyclin D1. The root extracts actuated apoptosis in HepG2 as evident from altered mitochondrial membrane potential, Annexin V- FITC, BrdU-PI, AO/EtBr assays, and modulations of apoptotic protein expression but without ROS generation. Whole extracts caused abrogation of epithelial to mesenchymal transition with repression of Snail, N-Cadherin, Vimentin, MMP-9, and upregulation of Pan-Cadherin. Pathway analysis found GSK-3ß in Wnt/ß-Catenin signaling cascade to be involved through Hepatocellular carcinoma (hsa05225) pathway. The GSK-3ß/ß-Catenin/PDL-1 signaling was found to be inhibited with the downregulation of pathway components. This was further confirmed by application of EGF, an inducer of the GSK-3ß/ß-Catenin pathway that neutralized the effect of Houttuynia cordata (Thunb.) root extract on the said pathway. Network pharmacology analysis also confirms the synergy network with botanical-bioactive-target-disease which showed Kaempferol to have the highest degree of association with the said pathway.

Quantification of cucurbitacin E in different varieties of melon (Cucumis melo L.) fruit through validated RP-HPLC method.

The different varieties of melons (Cucumis melo L.) have been used in various traditional systems of medicine for decades to treat different ailments, including inflammation, cancer, cardiovascular, diabetes, edema, etc. The present study was designed for the quantification of cucurbitacin E in five different varieties of melon fruit through a validated RP-HPLC method. A solvent system is being optimized with a 70:30 (v/v) ratio of acetonitrile: water (1% glacial acetic acid) at a 1 mL/min flow rate and scanning spectrum (λmax) of 230 nm. A calibration curve for standard cucurbitacin E was generated and found to be linear (1-100 µg/mL). The variation of cucurbitacin E content among five different varieties of melon fruits is 0.0129% w/w- 0.231% w/w. This precise and reproducible method may be beneficial in addressing the quality-related aspects of medicinal food plants of Cucurbitaceae and its derived products or formulations.