Thin Layer Chromatography Fingerprinting Research Articles

Comparative bioactivity evaluation of secondary metabolites from the leaves, stem and flowers of Tithonia diversifolia

Natural products have been extensively described as important in the process of drug.discovery and development. Drugs based on natural products are used in the treatment of infectious diseases, cancer, lipid disorders and immunomodulation. These drugs illustrate the importance of natural products as sources of innovative therapeutic agents. Recent studies point towards therapeutic potential of large number of natural products that originate from microbes and plants. These type of natural products plays a crucial role to shape the future of drug discovery process. Numerous studies indicate the use of Tithonia diversifolia (TD) as folk medicine to cure diseases like malaria, jaundice, hepatitis and dermatological problems. The present study is focused on identifying and comparing the bioactivity of different extracts from TD. Bioactive extracts of the leaves, stem, flowers were subjected to quantitative and qualitative phytochemical analysis. Qualitative analysis of TD extracts showed the presence of terpenoids, flavanoids, alkaloids, phenolics as the major class of compounds. High-Performance Thin-Layer Chromatography (HPTLC) fingerprinting analysis was carried to establish a quantitative profiling of the extracts. GC–MS of the extracts indicated the presence of Hexadecanoic acid, 2,3- dihydrobenzofuran, Squalene, dotriacontane, 1- hexacosanol, D- tagatose as the major phytocompounds present in the TD Hexane and ethyl acetate extract. The tested Tithonia diversifolia extracts showed significant anti-oxidant and anti-bacterial activities indicating therapeutic potential for TD.

Determination of quality standards and evaluation of In vitro Antibacterial activity of Kanchanara Guggulu: An ayurvedic formulation

Introduction: Ayurveda is one of the most renowned Indian traditional health-care systems and has been adopted globally in recent decades. In Ayurveda Kanchanara guggulu (KG) tablet is one of the Indian traditional medicines used in the treatment of various indications such as tonsilitis, cyst, abdominal lump, and fistula-in-ano. Methods: In the present study, KG tablet was prepared and standardized on basis of physical, physico-chemical properties, high-performance thin-layer chromatography (HPTLC) fingerprint, and phytochemical analysis. Heavy metal test of lead, arsenic, mercury, and cadmium was carried out to confirm the permissible limit as per the AYUSH and WHO standards. The antibacterial activity of the KG tablet was evaluated on two human pathogenic bacteria Escherichia coli and Staphylococcus aureus. Results: The heavy metal limit was found below the limit of quantification and within the allowable limit as per the AYUSH and WHO standards. The tablet showed comparable antimicrobial activity against both strains. The tablets were found to be having satisfactory quality parameters such as hardness, disintegration test, and friability values as per the pharmacopeia standards. Conclusion: The tested parameters based on the present study can be used as a reference approach for quality control of KG formulation. The HPTLC study was found to be an effective tool for quality control of herbal formulation to check the inter-batch batch consistency of the formulation. The antimicrobial activity against human pathogens substantiates its traditional uses.

HPTLC Fingerprinting Profile of Vacha (Acorus calamus Linn.)

Introduction: Vacha (Acorus calamus Linn.) commonly called Sweet flag is a significant herb with well-recognized therapeutic potential. Choorna (powder) of its rhizome is the commonly used dosage form according to Ayurveda. High-performance thin-layer chromatography (HPTLC) fingerprinting of rhizome powder of Vacha was carried out to estimate the active constituents in the dosage form choorna. ß-asarone, which is the major phytoconstituent of rhizome, was used as standard.Materials and Methods: HPTLC plate 5 × 10 cm, pre-coated with silica gel 60 F254 TLC plates, was used as stationary phase. Toluene: ethyl acetate: acetic acid (90:10:2) was used as mobile phase. Visualization was done under ultraviolet light 254 nn and 366 nm.Results and Discussion: HPTLC chromatogram showed the presence of 3 comparable peaks at 254 nm and 2 comparable peaks at 366 nm, in methanolic extract of rhizome powder and standard.Conclusion: This approach creates profile analysis of the plant rhizome and can be helpful for quality control and further researches

Physicochemical and Phytochemical Evaluation of Siddha formulation Saaranai chooranam

Background: Siddha system of medicine depends largely on herbal for the treatment of diseases which was practiced at village levels and now becomes an important medicine in worldwide, According to the Akasthiyar -2000-Part– III textbook, Saaranai Chooranam is a herbal drug that is indicated for Raththa kothippu (Systemic Hypertension) Saaranai is the Tamil name for Trianthema portulacastrum, which belongs to the family Aizoaceae. Aim & Objective: This study primarily aims to evaluate the Physicochemical and Phytochemical evaluation of Saaranai Chooranam, Preliminary Phytochemical analysis such as High Performed Thin Layered Chromatography and Powder Microscopy of Saaranai Chooranam. Methodology: The Physicochemical analysis of Saaranai Chooranam is carried out using standard procedures. Results: Preliminary Phytochemical screening found, the presence of phytochemicals such as Proteins, Terpenoids, Alkaloids, Carbohydrates, and Tannins. High-performance thin-layer chromatography fingerprinting revealed the presence of many phytochemicals with different Rf values and densitometric scans of the plates showed numerous bands and peaks. The Powder Microscopy reveals the presence ofTracheidal fibre, Vessel with bordered pits, Calcium oxalate crystal, Stone cell, Group of sclereids with brownish content, Paracytic stomata and surrounding subsidiary cells, Rosette Calcium oxalate crystal, Sclereid with narrow lumen, Thick walled cells, Tracheidal fibre and Trichome. Physicochemical analysis revealed the values of total ash (17.86%), acid-insoluble ash (1.34%), water-soluble ash (13.62), sulphated ash (25.72%), pH (6.05 in 4% alcohol solution), volatile oil (0.5%), foaming index (111.11%) and swelling index (4ml). Conclusion: This study is an effort to explore the different Physico and Phytochemical compounds of Saaranai Chooranam effective in the management of Raththa kothippu (Systemic hypertension).

Antioxidant Capacity, Total Phenolic, and Flavonoid Content from Java Tea (Orthosiphon aristatus) Extracts

Orthosiphon aristatus, known as kumis kucing in Indonesia, has some biological activity, i.e., as an anti-inflammatory, fever, rheumatism, diabetes, and kidney stone. This plant is also known for being rich in phenolic compounds such as flavonoids. In this study, we would like to compare the antioxidant activity, total phenolics, and flavonoid content from the stems and leaves of Orthosiphon aristatus extracted with water and ethanol by maceration. The yield stem and leaves of the water extract are 12.98-31.28%, and ethanol extract 3.58 - 9.82% (dry basis). The stem and leaves water extract contains a phenolic of 7.82-26.43 (mg GAE/g) and ethanol extract of 2.27-7.84 (mg GAE/g). Water extract Orthosiphon aristatus flavonoid content is 1.23-2.86 (mg EK/g), and ethanol extract is 1.09-3.95 (mg EK/g). The antioxidant activity of ethanol (IC50), extract from leaves was 123.91 (µg/mL) and from stems was 125.50 (µg/mL), while IC50 of water extract from leaves was 114.70 (µg/mL) and from stems was 125.03 (µg/mL). The correlation test result of active compounds using linear regression is phenolic compounds with antioxidants stronger than flavonoids. The qualitative thin-layer chromatography fingerprint analysis showed that ethanol and water extract contained natural sinensetin compounds.

Deciphering the impact and mechanism of Trikatu, a spices-based formulation on alcoholic liver disease employing network pharmacology analysis and in vivo validation.

Trikatu Churna (TC) comprising Zingiber officinale rhizome, Piper longum, and Piper nigrum fruit, is effective in treating liver diseases and has high nutraceutical values. However, the efficacy of TC in treating alcoholic liver disease (ALD) and its mechanism remain largely unknown. This study evaluated the hepatoprotective effects of different doses of TC as well as to identify the bioactive components and determine their mechanism of action against ethanol-induced ALD. A compound-target network analysis model of TC was established to identify its potential bioactive compounds and pathways that might regulate its hepatoprotective effects. Further, in-vivo studies were performed to validate the potential of TC (200 mg/kg and 400 mg/kg b.w.) in the treatment and management of ALD. The study revealed that both the dosages of TC demonstrate significant (p > 0.0001) hepatoprotective effects by improving body weight, total bilirubin, serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), serum alkaline phosphate (ALP), total cholesterol, total protein, globulin, albumin, and liver morphology. The High-performance thin-layer chromatography (HPTLC) fingerprinting of TC showed the presence of piperine. Network pharmacology identifies the role of TC in regulating various signaling processes including Advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE), Hypoxia-inducible factors (HIF-1), Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-Kappa B), and Phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling to exert its anti-inflammatory, antioxidant and anti-apoptotic role in managing ALD. Based on the bioinformatics analysis, some of the key targets of TC were found to be Prostaglandin-Endoperoxide Synthase 2 (PTGS2) or Cyclooxygenase-2 (COX-2), Sirtuin 1 (SRT1), and caspase-3. These effects may serve as a novel therapeutic option for the treatment of ALD. These preclinical validation studies for the ethnopharmacological potential of TC in ALD treatment further paved the way for researchers to perform next-level translational and clinical studies. Further, in-depth experimental studies for the validation of these bioinformatics-based results will give a clearer picture of mechanisms.

Bioactive composition and TLC profile data on PAX herbal health tea and PAX herbal diatea

Background: The phytoconstituents of herbal drugs are largely influenced by the quality control system used during and post-production processes including the handling of such products. Thin Layer Chromatography is one of such quality control parameters that demonstrate uniqueness and uniformity between various substances, serving as an identity for such substances. Pax Herbal Health tea (PHT) and Pax Herbal Diatea (PDT) are polyherbal drugs, PHT is used as a tonic for general wellness, while, PDT is used in the management and treatment of diabetes. This study evaluated the different phytoconstituents present and developed Thin-layer chromatography (TLC) fingerprint profiles for PHT and PDT to serve as quality control checks during the production for consistency and market uniqueness after production.Material and Methods: Qualitative phytochemical and chromatographic analyses were carried out using standard methods.Results: The phyto-screening revealed the presence of Alkaloid, Flavonoid, Tanin, Terpenoids, Reducing sugar, Steroid, and Cardiac glycoside in PHT while, Saponin, Tanin, Steroids, Reducing sugar, Flavonoid, and Terpenoids were observed in PDT. The TLC finger-print chromatograms of PHT after development with n-Hexane:Ethyl acetate (3:2) showed four distinct components under ultraviolet light at 365 nm, and three spots when sprayed with 20% methanolic sulphuric under visible light, while, PDT developed in n-Hexane:ethyl acetate:methanol (2.5:2:0.5) revealed three fluorescent components at 365 nm and four components after sulphuric acid treatment.Conclusion: From this present study, identity cards have been designed for PHT and PDT through bioactive composition and TLC profiles which can be used in accessing the quality and consistency of the herbal drugs. Â

Antimicrobial activity and Quality Control Parameters of Talicati Vatakam - A Classical Siddha Drug

Tāḷicāti vaṭakam (TSV), a polyherbal siddha drug was chosen, it was screened for antimicrobial study and also subjected to standardization parameters. The ingredients were procured, authenticated and prepared the drug as per standard operating procedure. The ethanolic extract of TSV was screened for nine bacteria and two fungi. Then the drug was investigated for the phytochemical profile, physicochemical parameters, thin layer chromatographic photo documentation (TLC), high-performance thin-layer chromatography (HPTLC) finger print profile. Antimicrobial assay revealed inhibitory activity against all test pathogens. TLC under UV showed 8 bands at short wavelength, 13 bands at long wavelength; 8 bands showed post derivatization with vanillin Sulphuric acid reagent. The present investigation concluded that the siddha herbal preparation of TSV have great potential on antimicrobial against pathogens. This siddha formulation can be used to prevent the bacterial and fungal infections and the standards could be used for quality control of the drug.

Standardization of polyherbal Siddha formulation aavarai poovarisi chooranam – anti-diabetic drug by pharmaceutical analytical techniques

Aavarai Poovarisi Chooranam is a Siddha poly herbal formulation plays an important role in treating Diabetes alone or in combination. Chooranam is applied to the powder made by a single or a mixture of herbal ingredients. The drugs required for making Aavarai Poovarisi Chooranam were collected and authenticated by experts and prepared as per siddha literature. It was then subjected to preliminary phyto-chemical analysis, physico-chemical, high performance thin layer chromatographic finger printing was done through CAMAG software, biochemical analysis, heavy metal analysis, pesticide residue, aflatoxin and microbial load analysis prior to toxicology and pharmacology testing. The obtained results were within normal limits in this study, which is briefly described in this paper.

HPTLC fingerprinting reveals leaf and roots phytochemical variability in developmental stages of Withania somnifera

Background: Withania somnifera has been the foundation of numerous contemporary medicines today for treatment of various diseases. However, phytochemical variability profile is lacking in the root and leaf organs. Aim: This study was aimed to elucidate the phytochemical profile in seedling, vegetative and reproductive stages of leaf and roots in W. somnifera using High performance thin layer chromatography (HPTLC) fingerprinting. Methods: The HPTLC with dual wavelength UV (254/366 nm) was used for the analysis of phytochemical variation in leaf and root methanolic extracts of W. somnifera. Results: HPTLC chromatograms in 366 nm, 254 nm, and normal day light, seedling, vegetative and reproductive stages leaf and root exhibited bands variability. At 450 nm leaf HPTLC fingerprints in seedling stage showed five peaks with Rf values in the range of 0.42 to 0.90, vegetative stage with nine peaks of Rf values in range of 0.10 to 0.89, and reproductive stage with six peaks of Rf values in range of 0.30 to 0.92. In root showed six peaks with Rf values in range of 0.00 to 0.82 in seedling stage, ten peaks with Rf values in range of 0.00 to 0.96 in vegetative stage, seven peaks with Rf values in range of 0.01 to 0.91 in reproductive stage. In roots, higher peak areas were found at the reproductive stage (3.33%) compared to vegetative stage (1.65%). Conclusion: The comparative HPTLC fingerprints of leaves and roots in W. somnifera presented phytochemical variability in leaf and root that varied with developmental stages.

Development of Synergy-Based Combination of Methanolic Extract of Andrographis paniculata and Berberis aristata Against E. coli and S. aureus.

This study evaluates the antibacterial activity and phytochemical characterizations of Andrographis paniculata extract (APE) and Berberis aristata extract (BAE). The stem of Andrographis paniculata (AP) and root of Berberis aristata (BA) were extracted with methanol. The results confirmed that APE and BAE possess high phenolic and flavonoid content. The antioxidant activity of the APE and BAE showed an elevated potential to scavenge DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals with IC50 of 95.03μg/mL and 256.26μg/mL, respectively. A total of 35 and 32 metabolites in APE and BAE, respectively, were identified through mass spectrometry analysis, whereas 17 and 12 metabolites in APE and BAE, respectively, were detected through high-performance thin-layer chromatography (HPTLC) fingerprinting profiling. Antibacterial activity of the extracts was performed by the well diffusion and microdilution method, and the findings showed that APE and BAE had antibacterial activities against E. coli and S. aureus. The growth curve and time-kill study showed that the extracts had a bacteriostatic effect. A combination study with the standard drug was carried out using the microdilution checkerboard method in which most of the combinations showed synergistic interactions. The findings of this study have shown that APE and BAE are good sources of antibacterial compounds and can be used for treating infectious diseases caused by E. coli and S. aureus.

Phytochemical studies and HPTLC fingerprint profiling of different extracts of Portulaca Quadrifida L.

The present study was designed to detect the class of compound by phytochemical evaluation and perform the high performance thin layer chromatography (HPTLC) fingerprint analysis of hexane extract of P. quadrifida (HEPQ), chloroform extract of P. quadrifida (CEPQ), ethyl acetate extract of P. quadrifida (EAEPQ) and ethanol extract of P. quadrifida (EEPQ). The successive extraction based on eluotropic solvent system was utilised for extraction. The preliminary phytochemical screening was carried out for all the extract. For the HPTLC fingerprint profiling the precoated Merck silica plates were used as stationary phase while, mobile phase utilised was Toluene: Chloroform: Ethanol (5:4:1 v/v/v). The scanning was done at White light, 254 nm and 366 nm. The presence of steroids, coumarins, flavonoids, glycosides, saponins, coumarins was found in all 4 extracts, except the terpenoids which were present in HEPQ and EAEPQ. The fingerprinting profile was successfully obtained for all four extracts of P. quadrifida. This profiling data can thus be utilized for quality control of the P. quadrifida both on qualitative and quantitative basis.

Enrichment of hydroalcoholic extract of Trigonella foenum-graecum L. seed with major components using TLC fingerprint, image analysis, and design of experiment studies

BACKGROUND: The seed of famous Indian spice Trigonella foenum-graecum L. (Methi) of family Fabaceae is the source of a large number of chemical components and used in a number of herbal formulations as an extract. For the separation of these chemical components from the complex plant matrix, the development of an effective extraction method is necessary. The present study aimed to identify major chemical constituents in the fingerprint developed by thin-layer chromatography (TLC) and to design an optimized extraction process for the enrichment of Methi seed extract with the selected constituents. METHODS: TLC fingerprint along with the image analysis was used to find the major components depending upon the area under the curve. The independent variables and their range were identified by one-factor-at-a-time experiments. The selected range of variables was further optimized by response surface methodology using the design of experiment (DoE) study to develop an extraction method for the maximum recovery of the selected components. The analysis of variance was utilized to check the fit of the developed model on the basis of quality assessment parameters. RESULTS: The information generated in this study recommends the use of reflux method with 70.57 mL/g of 80% ethanol for 31.19 min at 69.76°C for the maximum extraction of major components from seeds. CONCLUSION: The developed extraction method can be used for lab scale extractions in plant drug standardization and scale-up extractions in herbal industries for the manufacturing of Methi seed formulations.

Molecular docking analysis of estrogen receptor binding phytocomponents identified from the ethyl acetate extract of Salicornia herbacea (L).

It is of interest to evaluate the secondary metabolites using high performance thin layer chromatography (HPTLC) finger printing and Gas chromatography-Mass spectroscopy (GC-MS) in S. herbaceaextract. The powdered plant material extracted using different solvents were used for the qualitative analysis of alkaloids, flavonoids, terpenoids and saponins followed by HPTLC finger printing and GC-MS analysis. The components identified in the GC-MS were docked with estrogen receptor (ER) to identify the binding specificity of isolated compounds. The ethyl acetate extract of S. herbaceashowed the presence of high number of secondary metabolites when compared to other solvent system. The qualitative analysis of the plant material also showed the presence of carbohydrates, protein, amino acid, phenol, flavonoids, terpenoids, glycosides, saponins and steroids. The HPTLC finger printing analysis revealed the existence of alkaloid, flavonoid, terpenoid and saponin compounds and GC-MS. GC-MS was performed to identify the phytocomponents constituents in the extract. 8 phytocompounds were identified to analyse binding with ER. The binding affinity score (-6.8 kcal/mol) and interacting ER residues (28) the phyto compound di-n-octyl phthalate showed best docking score with ER α than the standard drugs lasofoxifene, and 4-hydroxytamoxifen. The binding affinity and number of interacting ER residues was -6.9 kcal/mol; 10 and -6.2; 11, respectively. The results identified the presence of ER antagonist in S. herbaceaand warrants further investigation to explore for treating ER regulated diseases.

Assessment of Biological Activities of Fungal Endophytes Derived Bioactive Compounds Isolated from Amoora rohituka.

Fungal endophytes have remarkable potential to produce bioactive compounds with numerous pharmacological significance that are used in various disease management and human welfare. In the current study, a total of eight fungal endophytes were isolated from the leaf tissue of Amoora rohituka, and out of which ethyl acetate (EA) extract of Penicillium oxalicum was found to exhibit potential antioxidant activity against DPPH, nitric oxide, superoxide anion and hydroxyl free radicals with EC50 values of 178.30 ± 1.446, 75.79 ± 0.692, 169.28 ± 0.402 and 126.12 ± 0.636 µg/mL, respectively. The significant antioxidant activity of EA extract of P. oxalicum is validated through highest phenolic and flavonoid content, and the presence of unique bioactive components observed through high-performance thin layer chromatography (HPTLC) fingerprinting. Moreover, EA extract of P. oxalicum also displayed substantial anti-proliferative activity with IC50 values of 56.81 ± 0.617, 37.24 ± 1.26 and 260.627 ± 5.415 µg/mL against three cancer cells HuT-78, MDA-MB-231 and MCF-7, respectively. Furthermore, comparative HPTLC fingerprint analysis and antioxidant activity of P. oxalicum revealed that fungal endophyte P. oxalicum produces bioactive compounds in a host-dependent manner. Therefore, the present study signifies that fungal endophyte P. oxalicum associated with the leaf of A. rohituka could be a potential source of bioactive compounds with antioxidant and anticancer activity.

PHYTOCHEMICAL PROFILING, HPTLC FINGERPRINT AND ANTIBACTERIAL, ANTI-FUNGAL, AND ANTIOXIDANT PROPERTIES OF ESSENTIAL OILS EXTRACTED FROM CUMMINUM CYMINUM, ZINGIBER OFFICINALE, TRACHYSPERMUM AMMI, ALIPNIA GALANGA, CEDRUS DEODARA, AND ELETTARIA CARDAMOMUM

Objective: The objective of the study was to carry out the phytochemical profiling of essential oils (EOs) and evaluation of their anti-microbial activity. Methods: The EOs extracted from Cumminum cyminum, Zingiber officinale, Trachyspermum ammi, Alipnia galanga, Cedrus deodara, and Elettaria cardamomum using clavenger apparatus. Phytochemical analysis and high-performance thin layer chromatography (HPTLC) fingerprinting were carried out for the EO. The antibacterial and antifungal activity were evaluated using agar well-diffusion method against two bacterial strains, Escherichia coli, Staphylococcus aureus and two fungal strains, Candida albicans, and Aspergillus brasiliensis. Positive controls ciprofloxacin-30 mg, azithromycin-15 mg, and nystatin NS-50 mg were used. Antioxidant potential of the EOs was investigated by TLC-bioautography method using 1,1-diphenyl-2-picrylhydrazyl derivatization. Results: The phytochemical analysis reveals presence of various phytochemical such as steroids, terpenoids, and phenylpropanoids. The HPTLC fingerprint is found to be unique for each of the oil. The EO of Z. officinale and T. ammi showed strong antibacterial activity against S. aureus. The EOs of C. cyminum, Tachyspermum ommi and A. galanga displayed prominent antioxidant activity on TLC bioautography. The herbs Cuminum cynimun, T. ammi, C. deodara, and Ellateria cardamomum produce reasonable amount of essentials oil, which can be explored for useful their industrial applications. Conclusions: These EOs can be explored further for their antimicrobial activity. The HPTLC analysis along with derivatization with suitable chromogenic reagents can be a rapid and simple tool for quality control of various EOs.

Ameliorative effect of the ethnomedicinal plant Blepharis maderaspatensis (L.) B.Heyne ex Roth on lipopolysaccharide-induced acute liver inflammation and oxidative stress in Wistar rat model

BackgroundBlepharis maderaspatensis is an ethnomedicinal plant used by the Mavilan and Koraga tribes of Kerala state, India for the treatment of liver diseases. Thus, the present study aims to evaluate the liver protective activity of defatted ethanolic extract of B. maderaspatensis on lipopolysaccharide-induced acute liver inflammation and oxidative stress in Wistar rat model. MethodsPreliminary phytochemical evaluation and high-performance thin-layer chromatography fingerprint validation were performed. The total phenolic and flavonoid contents were measured using the Folin-Ciocalteu method and the aluminium chloride method, respectively. The acute oral toxicity study was conducted in Swiss albino mice in accordance with OECD 423 guideline. Effect of defatted ethanolic extract of the whole plant of B. maderaspatensis (BmE) on liver inflammation was evaluated in LPS-induced Wistar rat model. The rats were treated with BmE (100, 250 and 500 mg/kg body weight) once daily for 7 days prior to the LPS (single dose i.p., 10 mg/kg b.w.) challenge. Liver tissue biochemicals (ALT, AST and ALP), and antioxidant status (SOD, CAT, GSH, MDA, NO and MPO) in euthanised experimental rats were carried out using commercial kits/standard procedures. Haematology (RBC, WBC, Hb and PLT; 4 and 24 h), and histopathology analysis were also performed. Serum IL-1β, IL-6, TNF-α and PGE2 production were determined by ELISA. The expressions of COX-2, iNOS, IL-1β, IL-6, TNF-α and NF-κB were analysed by qRT-PCR. Phosphorylation of IκB-α was measured by Western blotting. LC-MS analysis of BmE was also performed. ResultsThe LD50 value of BmE was found to be >5000 mg/kg b.w. BmE500 showed significant protection from LPS-induced liver injury as evidenced by reduced serum enzyme levels (AST, ALT and ALP; p≤ 0.001) and markedly improved the liver antioxidant status (MDA, GSH, SOD and CAT; p≤ 0.001), when compared to the LPS alone treated group. Haematology (24 h, RBC, WBC, Hb and PLT; p≤ 0.001) and histopathology also supported the above results. BmE500 pre-treatment significantly suppressed the LPS-induced expression of iNOS (p≤ 0.001) and COX-2 (p≤ 0.001), and the subsequent release of NO (p≤ 0.001) and PGE2 (p≤ 0.001). Moreover, BmE500 inhibited the gene expression of pro-inflammatory cytokines, including IL-1β (p≤ 0.001), IL-6 (p≤ 0.001), and TNF-α (p≤ 0.01), which is supported by ELISA results. In addition, BmE500 markedly attenuated the activation of transcription factor NF-κB (p≤ 0.001) as well as phosphorylation of IκBα, as evidenced from the qRT-PCR and Western blot analysis, respectively. The silymarin (100 mg/kg b.w.) drug standard also showed significant improvement in all parameters analysed. ConclusionResults of the present investigation suggest that BmE has a significant protective effect on LPS-induced acute liver inflammation via attenuating inflammatory reactions, evidenced by the inhibition of NF-κB signalling cascade, and also through its antioxidant effects. Thus, the pharmacological data generated provide experimental evidence that clearly justifies the use of B. maderaspatensis as a liver protective agent in tribal medicine.

Pharmacognostic study of Calotropis procera (Aiton) W.T. Aiton root and stem

Calotropis procera (Aiton) W.T. Aiton, belonging to the family Apocynaceae is an important medicinal shrub. It is widely used in folklore and Ayurvedic system of medicine. Various parts of this plant were used to treat human diseases such as Pama (Scabies), Vicharchika (Eczema), Kasa (Cough), Mukhakrishnatwa (Hyperpigmentation of the face), Pleehodara (Splenomegaly) and preparation of Ayurvedic as well as traditional medicines like Arkalavana and Arkataila. The present communication deals with a detailed account of the pharmacognostic study of C. procera root and stem. The study includes macroscopy, microscopy, powder microscopy studies, physicochemical, phytochemical analysis, heavy metal tests, screening of microbiological parameters and development of High-Performance Thin Layer Chromatography (HPTLC) fingerprints profile of C. procera root and stem. HPTLC fingerprints profile of ethanolic extract was done by using mobile phase toluene: ethyl acetate: formic acid for root and stem.

Assessment of pharmacognostic specification of Cannabis sativa leaves in Thailand.

ABSTRACTLack of quality control can affect the safety, efficacy, and acceptability of herbal products that may lead to health problems. Cannabis sativa L. (Cannabaceae) has been widely used as an ethnomedicinal practice for its medicinal values. This study aims to establish pharmacognostic specifications of C. sativa as per standard procedures. Macroscopic-microscopic characteristics, physicochemical parameters, thin-layer chromatography (TLC) fingerprinting, and phytochemical screening of C. sativa leaves collected from various locations throughout Thailand were investigated. Leaves are palmate consists of seven leaflets with green color, margin is serrate with acuminate apex. Anomocytic stomata were found in the upper epidermis while unicellular and glandular trichomes with cystolith were found in the lower epidermis and the epidermis layer covered with cuticle. The physicochemical analysis revealed that the loss on drying (4.068 ± 0.084 %w/w) was within acceptable limits, total ash (14.360 ± 0.084%w/w), acid insoluble ash (2.726 ± 0.080%w/w), ethanol-soluble extractive (11.101 ± 0.223%w/w), water-soluble extractive (23.038 ± 0.306%w/w), and water content (7.523 ± 0.524%w/w). TLC fingerprint showed nine spots with Rf value 0.14, 019, 0.23, 0.29, 0.32, 0.45, 0.58, 0.70, and 0.76. Phytochemical screening of Cannabis leaves indicated the presence of phenolic compounds, flavonoids, alkaloids, diterpenes, triterpenes, and steroids. This study provided referential data for the accurate plant identity, and establishment of cannabis leaves monograph in Thailand.

Chromatographic Fingerprinting of an Ayurvedic Formulation by High-Performance Thin Layer Chromatography Method

In the present study, an attempt has been made to develop a high-performance thin layer chromatography fingerprinting of gallic acid, piperine and curcumin from an Ayurvedic formulation, Dekofcyn tablet. Chromatographic separation was achieved on the plate pre-coated with silica gel 60F254 as the stationary phase. The solvent system consisted of toluene:ethyl acetate:formic acid:glacial acetic acid in the ratio (5:3.5:1:1 v/v/v/v) respectively. The retardation factor value was calculated at 0.35 (±0.02), 0.66 (±0.02) and 0.89 (±0.02) for gallic acid, piperine and curcumin respectively. Densitometric analysis was carried out in the isoabsorptive mode at 295 nm. The regression analysis was carried out in the range of 100-700 ng/spot for gallic acid and piperine, 400-1600 ng/spot for curcumin and the regression coefficient for all three markers was found to be 0.99 respectively. Validation of the method was performed according to international conference on hominization guidelines Q2 (R1). This method can also be used to evaluate other formulations containing these markers as one of the constituents, thus conforming to the need for ensuring the quality of an Ayurvedic formulation.