TLC Profiles Research Articles

Microscopic Characters, Phytochemical Screening Focus on Alkaloid and Total Phenolic Content of Uncaria gambir Roxb. and Uncaria sclerophylla Roxb. Leaves

Objective: Uncaria gambir Roxb. and Uncaria sclerophylla Roxb. are medicinal plants widely grown in Indonesia. Genus Uncaria is well known for catechin content that has potent antioxidant activity. Uncaria also was reported containing alkaloid that has antihypertensive action. This study aims to provide microscopic characters and phytochemical profiles associated with the phenolic and alkaloid compounds contained in these two species of Uncaria. Methods: Microscopic characteristics were performed on fresh leaf samples using a microscope. Presence of alkaloids from ethanol and chloroform extract of U. gambir and U. sclerophylla leaf were performed using Dragendorff reagents. Thin Layer Chromatography profile was developed in the mobile phase that consisted of acetone: chloroform (8:2). Total Phenolic Content was determined according to the Folin-Ciocalteu method using microplate reader spectroscopically at λ = 765 nm. Results: Microscopy of U. gambir leaf revealed the presence of vascular bundle fragment, oil fragments, parenchyma cells, midrib transverse-section and lower epidermis with paracytic stomata. Whereas in U. sclerophylla can be observed midrib transverse-section and lower epidermis unicellular with trichomes. Both U. gambir and U. sclerophylla positively contain alkaloids. Chloroform extract showed the presence of alkaloids while ethanolic extracts showed negative results. The TLC profile shows the presence of catechin compound in the ethanolic extract, whereas in the chloroform extract does not show the presence of catechin. The total phenolic content of U. gambir and U. sclerophylla leaf ethanolic extract and chloroform extract was 7.309; 5.734 and 0.437; 0.161 mg of gallic acid equivalents each gram sample, respectively. Conclusion: Microscopic characteristics, TLC and phytochemicals obtained can be used for the authentication of Uncaria gambir Roxb. and Uncaria sclerophylla Roxb leaf.

Review on Plants Fingerprinting and Tlc Fingerprinting of Two Aqueous Herbal Products

As herbal medicines have become commercialized, the safety, quality and efficacy of medicinal plants and herbal products have become of great concern. In view of this a review on Plant fingerprinting was carried out using systematic review and TLC fingerprinting was also carried out on two herbal products using simple Thin Layer Chromatography. The results revealed Morphological, Chemical and genetic methods are the major types studied. The High Performance Thin Layer Chromatography (HPTLC) is the most common type to chemical method utilized and most of the studies on Plants fingerprinting were carried out in Asia. Comparing the TLC profile of the two herbal preparations reveals that the two products are basically the same

Phytochemical studies and tlc profile of madhuca longifolia L. Bark with different solvents

Phytochemical studies and tlc profile of madhuca longifolia L. Bark with different solvents

Effect of compound Musizin isolated from Rhamnus wightii Wight and Arn on the immature stages of filarial vector mosquito Culex quinquefasciatus Say (Diptera: Culicidae) and its non-target studies

Abstract The present study was aimed to check the activity of Rhamnus wightii Wight and Arn against Culex quinquefasciatus (Say). Eggs and larvae of Cx. quinquefasciatus were exposed to different concentrations (0.5, 1.0, 1.5 and 2.0 ppm) of R. wightii crude extracts prepared using DMSO. The active ethyl acetate extract was packed in column chromatography and eluted. Totally 10 fractions were obtained based on the TLC profile. Among these fractions, fraction 2 obtained from Hexane:Chloroform (1:1) presented promising larvicidal activity compared to the other fractions. This fraction was identified as Musizin. Musizin caused 70% larvicidal activity with LC50 and LC90 values of 1.62 and 4.51 ppm respectively at 24 h. The larvae exhibited restless movement and convulsion followed by death; in control the larvae presented normal movement. Musizin presented 83% ovicidal activity against Cx. quinquefasciatus eggs after 120 h post-treatment at 2 ppm. Musizin treated eggs were shrunken, and most of the eggs did not hatch. Musizin was tested for toxicity against non-target organism Poecilia reticulata, and no toxicity was found against this organism. Musizin treated larvae were subjected to histopathological studies, and the results revealed severe damage to the larval midgut cells; in control there was no damage in the midgut cells of larvae. The present study revealed that the isolated compound Musizin could be used as a natural pesticide to control vector mosquitoes.

AKTIVITAS ANTIBAKTERI FRAKSI SAPONIN DARI KULIT BATANG TUMBUHAN KASTURI (Mangifera casturi)

This research to study antibacterial activity (E.coli and S.aureus) of saponin fractions from stem bark of Mangifera casturi. Powdered of stem bark of M.casturi (512,67 g) were extracted with methanol. Methanol extract (32.57 g) was partitionated with n-hexane to give saponin extract. Saponin extract (5.98 g) was partitionated by Vacuum Liquid Chromatography (VLC) over silica gel to give 39 fractions were grouped into four major fractions A-D by combining fractions with similar TLC profile. Every fractions studied antibacterial activity with E. coli and S. Aureus with difusi methode. Fraction A most active then other fraction with inhibition diameter 10.3 ± 0.5 mm (E.coli) and 10.8 ± 0.3 mm (S.aureus).

Preliminary Study of Hyptis pectinata (L.) Poit Extract Biotransformation by Aspergillus niger

One alternative approach to increase the content of bioactive compounds is fermentation. Hyptis pectinata (L.) Poit is a plant that can be found in tropical area and potentially as anticancer, anti-inflammatory, insect repellant, antiviral and antioxidant. In this research, efforts have been made to increase bioactive plant capacity of Hyptis pectinata (L.) Poit through submerged fermentation using Aspergillus niger. The study was performed by adding methanol extract of Hyptis pectinata (L.) Poit on two conditions, that was added at the beginning of fermentation and while entering a phase of death. Aspergillus niger growth rate in both conditions was observed by determining the dry weight of cells every 24 hours. The transformation profil of extract was observed after 24 hours of extract addition in early death phase by the TLC method. The results show that the addition of Hyptis pectinata (L.) Poit extract at log phase triggers the cells to growth faster, whereas the addition at the early death phase precisely accelerates cell death. TLC profile shows the emergence of new compounds suspected as the products of transformation of Hyptis pectinata (L.) Poit extract on day 8 after addition of extract.

TLC Profile and Activity Test of Secondary Metabolites Aspergillus flavus “In-Habiting” Queen Termite’s Nest Macrotermes gilvus on Enriched Media

Antibiotic are secondary metabolites yielded by microbe especially fungus. Previous research succesfull screened four kinds of fungus that live in termites queen’s nest, one of them was Aspergillus flavus. Furthermore, Alen et al (2016g) states that this fungus last to produce metabolite compounds on SDA media which only exist in first and second subculture extract, omit gradually for the next culture. It was presumed happen caused a different habitat to grow. So it is necessary to enrich the media with queen termite nest to get back the initial metabolites. The enrichment was done using four different media concentrations (0.25; 0,50; 0.75 and 1 grams of nest/mL media). The results show that enrichment of 1 gram of nest/mL media provides the most optimum fungus growht. The third subculture of Aspergillus flavus is cultured on enriched media which will become the fourth subculture, this fungus cultured until tenth subculture, do extraction and fracination to each culture. Based on TLC profile analysis, the initial metabolite not yet formed until tenth subculture, but forms six new stain terpenoid compounds. The result of columns chromatography obtained 10 sub-faction. Activity test was done by diffusion method to 12 test bacteria and 3 test fungus. Spot 2,5,6 (Rf 0.84; 0,36; 0,26) inhibit the growth of Pseudomonas aeruginosa ATCC 27853, spot 3,4 (Rf 0.74; 0,52) inhibit the growth of Micrococcus luteus ATCC 10240. The use of enriched media affect formation of secondary metabolites Aspergillus flavus. Keyword : Secondary metabolites, Aspergillus flavus, Macrotermes gilvus Hagen., Enriched media, TLC Profiles, Activity Assay

Comparative Evaluation of Thin-layer Chromatography/ High-performance Thin-layer Chromatography Fingerprint Profiles of Successive Extracts in Development of Standardization Protocol for Ayurvedic, Siddha or Unani Formulations

Aim: Trikatu churna is an important and unique polyherbal formulation used for various therapeutic indications. The World Health Organization (WHO) has emphasized the need to ensure the quality of medicinal plant products and herbal formulations by using any modern controlled techniques. High-performance thin-layer chromatography (HPTLC) is a frequently used technique to develop the chromatographic fingerprints. Therefore, the present study was carried out for quality evaluation of Trikatu churna through the development of standardization protocol with comparative HPTLC fingerprint profiles of successive extracts besides routine analysis. Materials and methods: Successive extracts of various solvents of polarity in increasing order through soxhlet apparatus for Trikatu churna and its authenticated raw materials were prepared; comparative chromatographic fingerprint profiles developed along with the evaluation of qualitative and quantitative tests parameters have been carried out by using the Ultraviolet (UV)–visible spectrophotometer, flame photometer, and atomic absorption spectrophotometer. Results: The following test parameters physicochemical parameters, evaluation of qualitative, quantitative phytochemicals such as total tannins, sugars and phenols, micronutrients such as sodium (Na), potassium (K); safety parameters such as microbial load, heavy metals and aflatoxins; comparative TLC and HPTLC fingerprint profiles of successive extracts of ingredients in increasing the polarity order of the solvents (n–hexane to alcohol) with respect to that of consecutive extracts of the final polyherbal formulation have been carried out for the quality evaluation. Conclusion: Trikatu churna was evaluated for its identity and purity by the systematic analysis of the above parameters for raw botanical ingredients and formulation in the course of development of standardization protocol for quality control.

Fluorescence Detection of Phytoconstituents in Hydroalcoholic Extract of Ipomoea aquatica Forssk. by Thin Layer Chromatographic Analysis

Introduction: The medicinal plant water spinach is applicable for many therapeutic activities like anti cancer, anti oxidant, etc. the plant is used for purification of metals and waters of different categories. The benefit of hydroalcoholic extract in this study is non toxic, preservative and high yield of extract from the herbal powder. Thin layer chromatographic analysis is one of the best analytical methods for the separation and identification of phytoconstituents from the crude extract. The spots separated on the stationary phase could be detected by using the non destructive fluorescence method. The detection will be done at shorter and longer wavelength in UV cabinet. Aim and Objective: The aim was to separate and identify the various active metabolites present in the crude extract of the herb of interest using Rf value by thin layer chromatographic analysis. Method: The thin layer chromatographic parameters were set by trial and error method. Ethyl acetate: Methanol: Water (10.0:1.35:1.0 v/v/v) for alkaloids, Ethyl acetate: Pyridine: Water (5.0:1.0:4.0 v/v/v) for glycosides, Toluene: Acetone: Formic acid (4.5:4.5:1.0 v/v/v) for tannins and Toluene: Ethyl acetate (9.3:0.7 v/v) for triterpenoids were the mobile phases for the individual constituents. The constituents were identified at 365nm. Results and Discussion: Nearly 24 various constituents of alkaloids, glycosides, flavonoids, triterpenoids and tannins were separated using respective mobile phases through Rf value. Conclusion: The mentioned TLC profile for the plant of interest will hopefully help the researchers in herbal field to develop more herbal medicines requiring the particular constituent or its extract.

Pharmacognostic, phytochemical, physicochemical and TLC profile study Mace (Aril) of Myristica malabarica Lamk. (Myristicaceae)

The plant Myristica malabarica Lamk. is traditionally used as a medicine and spices in food . It is belonging to family Myristicaceae. The plant is native to India and endangered trees are mostly found in western ghat. Extracted with various solvents by successive soxhlet hot extraction processs with increasing order of polarity on phytochemical investigation. The extract has shown alkaloids, saponin, tannin and flavones glycosides. It has important medicinal uses like Ayurvedic Medicines. It is traditionally used as anticancer, antiInflammatory, anti-Oxidant, Sedative hypnotics, Antimicrobial, Antifertility, Hepatoprotective and cytotoxicity. The chemical constituents such as Malabaricones, Malabaricanol, Isoflavones are isolated .Myristica Fragrans also known as fragnant Nutmeg or true Nutmeg. The present study i.e. Pharmacognostic, Phytochemical, Physicochemical and TLC Profile Study of Mace (Aril) Of Myristica malabarica Lamk. is helpful in the characterization of the crude drug. Physiochemical and phyto-chemical analysis of mace confirm the quality and purity of plant and its identification. The information collected is useful for further pharmacological and therapeutical evaluation of mace (Aril) Of Myristica malabarica Lamk. and anthology of quality control of crude drug.

Toxic effects of Solanum xanthocarpum Sch &Wendle against Helicoverpa armigera (Hub.), Culex quinquefasciatus (Say.) and Eisenia fetida (Savigny, 1826).

Many commercially available agro and household chemicals are used as pesticides, repellents, and growth inhibitors against insect pests. The repeated uses of these chemicals against insect pests have caused the development of resistance in them; they also cause ill effects on nontarget organisms. The present study was aimed to evaluate the antifeedant, larvicidal, pupicidal, and biochemical effects of the solvent extracts of Solanum xanthocarpum against third instar larvae of Helicoverpa armigera. Hexane, chloroform and ethyl acetate extracts were subjected to phytochemical analysis. The results revealed the presence of terpenoids, flavonoid, and quinone. Maximum antifeedant activity of 72.30% was recorded in chloroform extract followed by hexane (69.02%) and ethyl acetate (57.40%) extracts against H. armigera. Chloroform extracts of S. xanthocarpum showed more than 60% larvicidal and pupicidal activity against H. armigera. The effective chloroform extract was fractionated with increasing polarity of solvent system (hexane, chloroform, and ethyl acetate extracts). Based on the TLC profile, nine major fractions were isolated. The fourth fraction showed higher antifeedant, larvicidal, and pupicidal activity against H. armigera. The effective fraction reduced the hemolymph and gut protein concentration in a concentration-dependent manner (r 2 0.99). The effective fraction 4 showed 100% larvicidal activity at 500ppm concentration with LC50 value of 227.95ppm. The fourth fraction did not show any toxic symptom or mortality of earthworm. Based on these results, this effective fraction could be used in the development of a pesticide formulation to control insect.

Symbiotic bacteria associated with puffer fish Gastrophysus spadiceus and evaluation of their antimicrobial activities.

The present study reports the diversity of culturable bacteria associated with the puffer fish Gastrophysus spadiceus. During the study, a total of 31 strains affiliated to the genera Pseudomonas, Janthinobacterium, Rahnella, and Psychrobacter were isolated from liver, intestines, and flesh of G. spadiceus. These strains exhibited a diverse range of metabolites as indicated by the HPLC and TLC profiles of the chemical extracts of their fermentation products. Some of these crude extracts showed strong antimicrobial activities against pathogenic bacterial strains. In addition, few crude extracts exhibit insecticidal activity against Artemia salina.

A Systematic Survey, Taxonomic Study and Conservation of Genus Salacia (Celastraceae) in North Central Province of Sri Lanka

Genus Salacia is a climbing shrub comprised of nearly 200 species out of which, five have been reported in Sri Lanka. Salacia species are important components in many Ayurveda preparations and traditional medicine. Since S. reticultata and S. oblonga possess scientifically proven therapeutic benefits for an array of burning health issues, people tend to over exploit them from the wild to fulfill the existing demand. As a result, these two have become endangered in Sri Lanka. Therefore, recording existing populations in different agro-ecological regions with their abundance has become one of the key issues in conservation and sustainable use of these plants. Moreover, these species exhibit morphological plasticity and documented vegetative characters to differentiate one from the other are scares. The present study was launched to carry out a systematic survey and taxonomic study of different populations of Salacia species in the North Central province with the view of confirming taxonomic position of existing populations, documenting vegetative characters as there are overlapping morphological characters and flowering is seasonal and identifying unique populations for conservation. In the Systematic survey 28 localities out of which 15 are new records in the North Central Province where Salacia species were growing naturally were found. Representative samples were collected from twenty locations and identified, subsequently authenticated and used for the morphological study. Ninety morphological and anatomical characters of each population were scored by using standard methods. Methods for leaf clearing and wood maceration suitable for Genus Salacia were developed. A preliminary screening of phytochemicals was done using TLC. Leaf and stem extracts were obtained by cold extraction and Soxhlet method. TLC profiles were recorded under UV, using Anisaldehyde-sulphuric acid and Iodine vapour. Taxonomic relationships among collected samples were analyzed. Out of the ninety characters studied, fifty four could be identified as clear characters for species demarcation. Based on the study, eight samples could be clearly identified as S. reticulata, six as S. oblonga but the remaining six displayed a mixture of morphological characters of S. chinensis, S. reticulata and S. oblonga. The population collected from Ritigala (a new locality) showed deviation in characters from all others and hence need to draw the attention of conservationists. Keywords: S. reticulata, S. oblonga, Taxonomic relationship

Toxicity of Ag Nanoparticles Synthesized Using Stearic Acid from Catharanthus roseus Leaf Extract Against Earias vittella and Mosquito Vectors (Culex quinquefasciatus and Aedes aegypti)

Currently, many plant extracts have been employed to green synthesize metal nanoparticles toxic to insect pests and vectors. However, scarce efforts focused on the main molecules responsible of reduction and stabilization processes. In this study, we assessed the larvicidal activity of hexane, dichloromethane (DCM), acetone and aqueous extracts of Catharanthus roseus on the insect pest Earias vittella. Among them, DCM extract exhibited maximum larvicidal activity against E. vittella. Therefore, it was fractionated using a silica gel column. Based on the TLC profile, 7 fractions were isolated. Fraction 5 showed the highest larvicidal activity (81.33%) when tested at 1000 ppm. Fraction 5 showed a single spot on TLC and was subjected to UV–Vis spectroscopy, H1NMR, FTIR and mass spectroscopy. Based on the spectral data, we isolated stearic acid, which was further used as a reducing agent to synthesize Ag nanoparticles. The formation of Ag nanoparticles was recognized by the presence of broad peak at 416 nm corresponding to the surface plasmon resonance of Ag nanocomposites in UV–Vis spectroscopy. Ag nanoparticles were spherical in shape with size ranging from 35 to 55 nm. The nanoparticles showed significant antifeedant (87.13%) and larvicidal (93.77%) activities against E. vittella when tested at 200 ppm and exhibited LC50 values of 45.46 ppm in antifeedant experiments and 25.12 ppm in larvicidal assays on E. vittella. The nanoparticles showed acute toxicity against mosquitoes Culex quinquefasciatus and Aedes aegypti with LC50 values lower than 40 ppm on both vectors, showing their promising potential to develop newer and effective biopesticides to control arthropod pests.

Cytotoxicity of actinomycetes associated with the ascidian Eudistoma vannamei (Millar, 1977), endemic of northeastern coast of Brazil

Previous studies demonstrated that the crude extract of the ascidian Eudistoma vannamei, endemic from northeasttern Brazil, strongly hinders growth of tumor cells in vitro by inducing apoptosis due to tryptophan derivatives, which are commonly found in bacteria. This study presents a bioactivity-guided screening among actinomycetes, associated with E. vannamei, aiming at recognizing active principles with biological relevance. Twenty strains of actinomycetes, designated as EVA 0101 through 0120, were isolated from colonies of E. vannamei among which 11 were selected for cytotoxicity evaluation. The extracts from EVA 0102, 0103, 0106, 0109 and 0113 were the most active, and were further studied for IC50 determination and chemical analysis by 1H NMR. IC50 values obtained ranged from 3.62 µg mL-1 (for EVA 0109 in leukemia cells) to 84.65 µg/mL (for EVA 0106 in melanoma cells). All active extracts exhibited the same TLC and spectroscopic profiles, suggesting the presence of quinones and other related secondary metabolites. Furthermore, these strains were identified and compared based on their respective 16S rRNA sequences. The results herein identified the five strains as Micromonospora spp. while phylogenetic analysis suggests that they are possibly two different Micromonospora species producing the cytotoxic compounds.

Characterization and Comparison of Three Species of the Genus Salacia.

Some of the plant species of the genus Salacia are considered as an important anti-diabetic drug in ayurveda. Among them S. chinensis, S. fruticosa and S. oblonga possess great medicinal importance, mostly because of its promising anti-diabetic activity. The increasing demand of this drug, resulted in a huge decline in its availability and it is replaced by spurious ones mainly because of lack of adequate quality standards. There is now a felt need to develop a systematic approach for the authentication of these plants and to develop well-designed methodologies for its standardization. The present study focused on the Pharmacopoeial parameters like pharmacognostical characterization and preliminary phytochemical screening of these three species. which were found to be sufficient to evaluate the raw material and can also be used as reference standards for the quality control/quality assurance purposes. The pharmacognostic study revealed that there are specific diagnostic features for distinguishing these three species among themselves and also from other related species. The external morphology of these three species are also shows variation in their colour, odour and taste. Both the species of Salacia ie, S. chinensis is having bitter taste and S. fruticose is with astringent taste whereas S. oblonga has no characteristic taste of its own. The TLC profile showed similar pattern in S. chinensis and S. fruiticosa where as S. oblonga showed more band in the profile than that of the other two. This reveals that the S. chinensis and S. fruiticosa having similar compounds responsible for therapeutic efficacy, whereas S. oblonga has more chemical constituents.

Phytochemical analysis and differential in vitro cytotoxicity assessment of root extracts of Inula racemosa.

The root of Inula racemosa is known for its antifungal, hypolipdemic and antimicrobial properties in traditional Indian Ayurvedic and Chinese system of medicine. The biological efficacy of Inula species is mainly due to the presence sesquiterpene lactone (Isoalantolactone and Alantolactone), which are reported to be inducers of Nrf2 antioxidant pathway. The investigation of properties and efficacy of root extracts of I. racemosa and their comparison was done with a view to find most efficacious extract for use at cellular level (both normal and transformed). In the present study different extracts of root of I. racemosa (aqueous, ethanolic, and 50% aqueous-ethanolic) were prepared and compared for their antioxidant potential, reducing capacity, polyphenol content and flavonoid content. Our investigations suggested that the aqueous extract possess highest antioxidant capacity and reducing potential. The polyphenol content was found to be highest in aqueous extract in comparison with other two extracts. However, all the three extracts showed less flavonoid content. Further, the preliminary phytochemical screening of all the extracts revealed the presence of terpenoids, phytosterols and glycosides. The TLC profile of ethanolic and 50% aqueous-ethanolic extracts showed the presence of alantolactone while aqueous extracts did not exhibit its strong presence. This warrants the need of more stringent techniques for characterization of aqueous extract in future. The in vitro cell based toxicity assays revealed that the aqueous extract was less toxic to kidneys cells while ethanolic extract was toxic to cells even at low concentrations. Hence, the current investigations showed better efficacy of the aqueous extract with respect to other extracts and found to be promising for its future application at in vitro levels.

Screening and Characterization of Lipid inclusions in Bacteria by Fluorescence Microscopy and Mass Spectrometry as a Source for Biofuel Production

Objectives: To screen and characterize lipid inclusions found mostly in the cytoplasm of soil bacteria as a source for biofuel production. Methods: In the present study, five bacterial isolates, from soil were observed under phase contrast microscope to study their morphology later stained with lipid specific dye, Nile red and the fluorescence intensity determined by Zen blue software. Lipid extraction was performed using chloroform: methanol(2:1) and quantified gravimetrically. The extracted lipids were further resolved by TLC with Triolein and Linoleic acid as standards. Lipid with highest yield was analysed by MALDI-TOF. Findings: The analysis has been of immense importance in selection of the highest neutral lipid accumulating isolate, where Rhodococcus pyridinivorans strain OB1 yielded the highest amount of lipid (40mg/100ml).TLC profile showed TAG spots correlating with standards. MALDI-TOF spectra showed many peaks corresponding to Mono and diacylglycerols, a predominant peak at m/z 907corresponding to 54:3 TAG. Novelty: This report is the first on the analysis of several soil bacterial strains for their lipid accumulating ability and has provided a thorough profile of lipid classes.

Toxicity and antifeedant activity of Caesalpinia bonduc (L.) Roxb. (Caesalpiniaceae) extracts and fractions against the cotton bollworm Helicoverpa armigera Hub. (Lepidoptera: Noctuidae)

Natural pesticides show a number of advantages if compared to synthetic pesticides, since they are cheap, biodegradable, and minimal harmful to humans and animals, livestock and wildlife. Furthermore, natural biopesticides are easily accessible by poor marginalized rural populations in developing countries. Here, crude extracts and fractions of Caesalpinia bonduc (L.) Roxb. (Caesalpiniaceae) were evaluated for their biotoxicity against the cotton bollworm, Helicoverpa armigera Hub. (Lepidoptera: Noctuidae). Maximum antifeedant, larvicidal and pupicidal activities were observed testing the chloroform extract of C. bonduc. Therefore, C. bonduc chloroform extract was fractionated using solvents of increasing polarity. Based on the TLC profiles, six fractions were isolated. All fractions were tested against H. armigera. Maximum antifeedant, larvicidal, pupicidal activities, as well as the highest treatment impact on larval-pupal duration were recorded testing fraction 3. Overall, the present study allowed us to consider the cheap and easy to obtain fraction 3 as a promising source of botanical products effective in the fight against agricultural pests, with special reference to the cotton bollworm, in an eco-friendly perspective.

Authentication of Piper betle L. folium and quantification of their antifungal-activity

The TLC profiles of intra- and inter-day precision for Piper betle L. (PBL) folium methanol extract was studied for their peak marker recognition and identification. The Numerical chromatographic parameters (NCPs) of the peak markers, the hierarchical clustering analysis (HCA) and the principal component analysis (PCA) were applied to authenticate the PBL. folium extract from other Piper species folium extract and to ensure the antifungal activity quality of the PBL essential oil. The spotted extract was developed with the mobile phase of toluene: ethyl acetate; 93:7, (v/v). The eluted plate was viewed with the TLC-Visualizer, scanned under absorption and fluorescent mode detection, and on each sample the in-situ UV spectra were recorded between 190 to 400 nm.The NCPs profiles of intra- and inter-day precision results offered multi-dimensional chromatogram fingerprints for better marker peak pattern recognition and identification. Using the r-value fingerprints data series generated with this method allowed more precise discrimination the PBL. from other Piper species compared to the marker peak area fingerprint method. The cosine pair comparison was a simple method for authentication of two different fingerprints. The ward linkage clustering and the pair cross-correlation comparison were better chemometric methods to determine the consistency peak area ratio between fingerprints. The first component PCA-loading values of peak marker area fingerprints were correlated linearly to both the bio-marker concentration as well as the antifungal activity. This relationship could be used to control the quality and pharmacological potency. This simple method was developed for the authentication and quantification of herbal medicine.