High-performance Thin-layer Chromatography Fingerprinting Research Articles

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.

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.

A simple and effective method for identification of Fraxini Cortex from different sources by multi-mode fingerprint combined with chemometrics.

Fraxini Cortex has a long history of being used as a medicinal plant in traditional Chinese medicine. However, it is challenging to differentiate and make quality evaluations for Fraxini Cortex from different origins due to their similarities in morphological features, as well as general chemical composition using traditional chemical analytical methods. In this study, a simple and effective method was developed to identify Fraxini Cortex from different origins by multi-mode fingerprint combined with chemometrics. Digital images of the high-performance thin-layer chromatography profiles were converted to grayscale intensity, and the common patterns of high-performance thin-layer chromatography fingerprints were generated with ChemPattern software. Authentication and quality assessment were analyzed by similarity analysis, hierarchical cluster analysis, principal component analysis, and multivariate analysis of variance. The ultra-high-performance liquid chromatography fingerprints were analyzed by similarity analysis, principal component analysis, and orthogonal partial least square-discriminant analysis. When combined with chemometrics, high-performance thin-layer chromatography and ultra-high-performance liquid chromatography fingerprint provided a simple and effective method to evaluate the comprehensive quality of Fraxini Cortex, and to distinguish its two original medicinal materials (Fraxinus chinensis Roxb. and Fraxinus rhynchophylla Hance.) recorded in the Chinese Pharmacopeia and its three adulterants (Fraxinus mandschurica Rupr., Fraxinus pennsylvanica Marsh., and Juglans mandshurica Maxim.). A similar workflow may be applied to establish a differentiation method for other medicinal and economic plants.

High-performance thin-layer chromatography fingerprinting and antioxidant study of Mangifera indica mother tincture

Background Mangifera indica is widely used in the Homoeopathy systems of medicine. In Homoeopathy, Mang. ind. is one of the indicated medicines for passive haemorrhages, uterine, renal and gastric problems. Mangiferin is a xanthanoid present in leaves, stalks barks, peel and kernel of the mango plant. Mangiferin has antioxidant, antiviral, anticancer, antidiabetic, antiaging, immunomodulatory, hepatoprotective and analgesic properties. Therefore in the current study, we have tried to identify mangiferin and its antioxidant properties in Mang. ind. homoeopathic mother tincture. Objective The aim of the present study wais to identify Mangiferin, Total Phenol Content (TPC) and Total Flavonoid Content (TFC) in Mang. ind. homoeopathic mother tincture. Materials and Methods Mangiferin (A) was used as a primary reference standard and in sample In-house prepared mother tincture (B) and two different market samples (C and D) were used. An authentic sample of Mang. ind. bark was received from the Centre of Medicinal Plants Research in Homoeopathy, Emerald, Tamil Nadu, India for preparation of in house mother tincture. Results We have successfully identified Mangiferin in Mang. ind. in-house prepared mother tincture (B) and two market samples (C and D) by HPTLC. Total Phenolic Content (TPC) and Total Flavonoid Content (TFC) was determined by Ultraviolet–visible spectroscopy. Conclusion High Performance Thin Layer Chromatography study confirms the presence of Mangiferin whereas U. V spectroscopy proves to be a useful tool for in-vitro antioxidant assay study.

Comparative analysis of two Himalayan Aconitum species for their phytopharmaceutical properties

IntroductionBhutanese Sowa Rigpa Medicine (BSM) uses two species of Aconitum in 25 important multi-ingredient formulations. The formulations are used for treating chronic disoders, parasitic and microbial infections, inflammatory diseases, bilious fever, or high fever related to bile disorders, including colds and flu. Some Aconitum species are toxic, and there is a need for stringent quality control measures. Materials and methodsAconitum orochryseum Stapf and Aconitum laciniatum (Bruehl) Stapf, were collected from Lingzhi, which is geographically located under Thimphu district, Bhutan, between the altitudes of 4000–4900 meters above sea level (masl), between July and September. Their comparative pharmacognostic properties were evaluated by: 1) using physicochemical parameters, such as foreign matter and moisture contents, total ash, acid-insoluble ash and alcohol-soluble extracts values and loss on drying of individual crude drugs; 2) developing a comparative High-Performance Thin Layer Chromatography (HPTLC) fingerprint, at Menjong Sorig Pharmaceuticals Corporation Limited (MSCPL), - Bhutan and 3) conducting a literature review to identify the nature and toxicity of alkaloids present in two Aconitum species. ResultsTwo Aconitum species possess distinct macros- and micros-copic characteristic features (including colour, taste, smell, appearance, and shape). The physicochemical analysis and the HPTLC fingerprinting of their methanol extracts yielded adequate data to differentiate and validate these two species’ identities, before adding them to 25 essential traditional medicine drugs (ETMDs) or polyherbal formulations in BSM. A literature review revealed that the two Aconitum species contained two distinct types of alkaloids. ConclusionsTwo Aconitum species could be differentiated based on their botanical features, macros- and micros-copic characteristics, physicochemical values and HPTLC fingerprint profiles. These pharmacognostic properties can be used as quality control parameters to confirm the two species' identity, thereby ensuring the quality and safety of BSM.

Standardization of Rasagandhi mezhugu

Background: Rasagandhi mezhugu (RGM) is the higher order Siddha formulation comprising of mercury, arsenical compounds, minerals and herbs and is indicated for chronic diseases like Megarogam, Kiranthi, Puttrunoi, etc. Objectives: The objective of the present investigation was to standardize the formulation of RGM. Materials and Methods: The organoleptic characters such as color, appearance, taste, and odor were noted. RGM was screened for moisture content, total ash value, water-soluble ash, acid-insoluble ash, alcohol-soluble extractive value, water-soluble extractive value, microbial load, and specific pathogen to estimate the quality of the study drug. Results: The outcomes of the physiochemical analysis of RGM were found to be within standard limits. RGM is free from specific pathogens such as Escherichia coli, Salmonella, Pseudomonas, and Staphylococcus aureus. The overall safety regarding heavy-metal content was assured by quantifying the metal content using inductively coupled plasma optical emission spectroscopy. The high-performance thin-layer chromatography fingerprint of RGM was generated. Conclusion: The results attained from the study could be used as a reference for setting limits for the quality assurance and quality control of Rasagandhi mezhugu.

Pharmacognostical and phytochemical studies of Atibala (Abutilon indicum [Linn.] sweet) fruit.

Abutilon indicum (Linn.) Sweet (Malvaceae), generally called as "Atibala" is a plant of high medicinal importance. The plant possesses several beneficial effects such as cooling, laxative, digestive, analgesic, anti-inflammatory, astringent, diuretic, expectorant, antihelmintic, aphrodisiac, and demulcent which is widely used in the Ayurveda system of medicine. The current study is aimed to establish the macroscopy, powder microscopy and physicochemical analysis of A. indicum fruits. The Pharmacognostical studies on A. indicum fruits, including parameters such as morphological evaluation, powder microscopy, ash values, foreign organic matter, extractive value, phytochemical, fluorescence studies, and high-performance thin-layer chromatography fingerprint profile, are established in the current study. Fruit powder microscopy has shown diagnostic characteristics such as stellate hairs of different sizes, testa, lignified endocarp, and palisade cells. The loss on drying value of fruit powder was 7.7% w/w. The total ash values of the drug were found to be 10.5% and acid insoluble ash 2.4% w/w with respect to the air-dried crude drug. Water-soluble and alcohol-soluble extractives were found to be 9.64% w/w and 9.04% w/w, respectively. Phytochemical characterization of aqueous, alcoholic extracts of A. indicum fruit revealed the presence of proteins, carbohydrates, phenols, flavonoids, saponins, and steroids. The powder microscopical and phytochemical studies observed in this study can serve as a valuable resource for the authentication of A. indicum fruits.

Comparative HPTLC fingerprinting profile and GC-MS analysis of raw and purified Guggulu (Commiphora wightii. Arn. Bhand) by Ayurvedic purification method.

Guggulu is an oleo gum resin obtained from the plant Commiphora wightii (Arn.) Bhand., used in Ayurved ic medicines for various ailments like anti-inflammatory conditions, hyperlipidemia, thyroid disorders etc. Guggulsterones E & Z are responsible for these broad ranges of pharmacological actions. It is recommended to do Shodhana (purification) before incorporating it into medicinal formulations. Sahasrayoga, an Ayurvedic text, emphasizes the purification of Guggulu in a particular media, which is a long run practice in Kerala. To compare the physicochemical and phytochemical parameters, quantitative estimation of Guggulsterone E & Z using high performance thin layer chromatography (HPTLC) and qualitative gas chromatography-mass spectrometry (GC-MS) analysis of Guggulu before and after purification. Shodhana of Guggulu was performed in Water boiled with crushed fresh leaves of Neem (Azadirachta indica) and fresh rhizome of turmeric (Curcuma longa) using a special equipment (Dolayantra). Preliminary physicochemical and phytochemical evaluation, quantification of Guggulsterones E & Z using HPTLC and GC-MS analysis of raw and purified Guggulu were performed. Phytochemical evaluation of metabolites revealed marked variations. The mean concentrations of Guggulsterone E & Z showed significant differences before and after purification (p<0.01). On GC-MS analysis, it was found that few new compounds were added in the purified Guggulu. Therapeutic efficacy of Guggulu might have enhanced after traditional purification.

Coriandrum sativum attenuates microglia mediated neuroinflammation and MPTP-induced behavioral and oxidative changes in Parkinson's disease mouse model.

Coriandrum sativum Linn. (family: Umbelliferae; C. sativum), is a potential herb widely used as a spice and traditional medicine. In the present work, the effects of C. sativum fruit extract (CSE), against lipopolysaccharide (LPS)-stimulated BV-2 microglia-mediated neuroinflammation in vitro and 1-methyl-4 phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) animal model in vivo was investigated. LPS-stimulated increase in nitric oxide (NO), inducible NO synthase, cyclooxygenase-2, interleukin-6 and tumor necrosis factor-alpha were significantly (p < 0.05 ~ p < 0.001) inhibited by CSE (25, 50 and 100 μg/mL) in BV-2 microglial cells. Further, CSE inhibited the LPS-induced nuclear factor of kappa-beta activation and IκB-α phosphorylation in BV-2 microglia. In vivo studies, CSE (100, 200 and 300 mg/kg) ameliorated the MPTP (25 mg/kg, i.p.)-induced changes in locomotor, cognitive and behavior functions evaluated by rotarod, passive avoidance and open field test significantly (p < 0.05 ~ p < 0.001). The MPTP-induced changes in brain oxidative enzyme levels such as superoxide dismutase, catalase, and lipid peroxidation were significantly (p < 0.01 and p < 0.001 at 200 and 300 mg/kg, respectively) restored with CSE treatment. High-performance thin-layer chromatography fingerprinting analysis of CSE exhibited several distinctive peaks with quercetin and kaempferol-3O-glucoside as identifiable compounds. In conclusion, our study indicated that CSE attenuated neuroinflammatory processes in LPS-stimulated microglia in vitro and restored the MPTP-induced behavioral deficits and brain oxidative enzyme status in vivo proving its therapeutic potential in the treatment of neuroinflammatory and oxidative stress-mediated neurodegeneration seen in PD.

HPTLC Fingerprints of Alcoholic Extracts of Phyllanthus species collected from Various Parts of Eastern Ghats in Andhra Pradesh

The genus Phyllanthus L. consists of 750 to 850 species distributed in tropical and subtropical around the world while 51 species in India. Most of them have been used in traditional medicine since ancient times. Phyllanthus spp.. like P. amarus, P. debilis, P. fraternus, P.urinaria and P. virgatus are called bhumyamalaki, used in Ayurveda. More than 514 compounds have been isolated from Phyllanthus, the majority of which are lignins, triterpenoids, flavonoids, and tannins. Lignins like phyllanthin and hypophyllanthin have been shown hepatoprotective activity and tannins exhibit various biological activities. Corilagin, geranin, and gallic acid are another three most prevalent compounds in this genus. P.amarus shows high concentration of Phyllanthin than P. fraternus, P. virgatus, P. maderaspatensis, P. urinaria, and P. debilis. Our aim is toestablish physical constants and fingerprint profile of Phyllanthus species collected from various parts of Eastern Ghats in Andhra Pradesh using high performance thin layer chromatography (HPTLC) technique. It is one of the fast growing techniques, used to analyze the phytochemical constituents. Leaves of thirteen Phyllanthus species were used to prepare methanolic extracts. Extracted samples used for carrying out HPTLC. Sample was spotted on TLC plate as triplicate and developed the plate with the mobile phase as Toluene: Ethyl acetate: Methanol (7: 2: 1). Dried TLC plate was then scanned under the UV at 366nm. All the Phyllanthus spp.. exhibits significant fractions of phyllanthin, niranthin, quercetin and phenolic compounds in alcoholic extracts. It can be used as a diagnostic tool for the correct identification of potential Phyllanthus spp.. and it is useful as a phytochemical marker and also a good estimator of genetic variability in the plant populations.

High Performance Thin Layer Chromatography Fingerprinting Analysis of Piper betle L. Leaves

Introduction: Many modern medicines used today based on plants and plant products. Piper betle is generally known as the betle vine, it is an important medicinal and recreational plant. High performance thin layer chromatography (HPTLC) is an advanced powerful analytical method with more separation power, high performance and superior reproducibility than classic thin layer chromatography (TLC). A chromatographic fingerprint of a plant extract is a chromatographic pattern of some common chemical constituents of pharmacologically active and/or chemical characteristics. Chromatographic fingerprints are useful in authentication and identification of plant.&#x0D; Objectives: Objectives of present research was to establish HPTLC fingerprinting of methanolic extract of Piper betle L. leaves.&#x0D; Materials and Methods: Methanolic extract of Piper betle leaves was prepared using soxhlet apparatus. HPTLC studies were performed using a CAMAG HPTLC system equipped with automatic TLC sampler-4 (ATS 4), TLC scanner 4, and vision CATS 3.0 software.&#x0D; Results: The study revealed the presence of alkaloids with Rf value 0.65, flavonoids with Rf values 0.19, 0.29, 0.72, 0.95., and phenolic compound with Rf value 0.7.&#x0D; Conclusion: The HPTLC fingerprinting profile developed for the methanolic extract of Piper betle L. leaves will help in proper identification of the plant.Piper betle

High-performance thin-layer chromatography fingerprinting and anti-inflammatory and antinociceptive activities of Pyracantha coccinea M.Roem.: A laboratory-based study.

This study aimed to investigate the high-performance thin-layer chromatography (HPTLC) fingerprinting and in-vivo anti-inflammatory and antinociceptive activities of different leaf extracts (ethanolic extract, n-hexane, chloroform, and ethyl acetate) of Pyracantha coccinea M.Roem. plant. A total of one hundred and twenty-four Wistar rats for anti-inflammatory and antinociceptive tests (carrageenan and formalin tests, respectively) were treated with two doses of the ethanolic extract (100 and 300 mg/kg), two doses of other plant fractions (30 and 100 mg/kg), Diclofenac (25 mg/kg) as the positive control, and normal saline as the negative control group, by oral gavage route. HPTLC fingerprinting is used for assay of terpenoids, flavonoids, alkaloids, and antioxidant activity. Treatment of the animal with the ethanolic extract at doses of 100 and 300 mg/kg, both ethyl acetate and chloroform fractions at the dose of 30 mg/kg and 100 mg/kg decreased the pain score in the formalin test and paw edema caused by carrageenan relative to control group significantly. Moreover, these extracts reported the highest amounts of flavonoid contents. In conclusion, phytochemicals present in Pyracantha coccinea M.Roem. leaves have anti-inflammatory and antinociceptive activities. Future studies are needed to identify the compounds with the anti-inflammatory and antinociceptive potential present in the plant.

HPTLC fingerprinting, photopigments, and nutrient analysis of Selinum tenuifolium along the altitudinal gradient

This study aimed to explore phenolic acids, nutrient status, and content of photopigments and their role in the production of secondary metabolites in Selinum tenuifolium collected from different altitudes (2150 m; 2593 m; 3178 m, asl) in Chopta region of Uttarakhand, India. High-performance thin-layer chromatography (HPTLC) fingerprinting, nutrient analysis, and chlorophyll (Chl) content were assessed in various plant parts (leaves, stem, and roots) of S. tenuifolium. HPTLC analysis revealed a number of bands in the methanolic extracts of S. tenuifolium of which two with retention factor (Rf) values, 0.36 pertaining to gallic acid (GA) and 0.58 relating to ferulic acid (FA) indicating their presence in all three plant parts of S. tenuifolium. The elemental analysis (Fe2+, Zn2+, Na + K+ and, Ca2+) of different plant parts reflected the nutrient status of the plant, the total amount of nutrients being highest in stems of plants growing at 3178 m and lowest in roots of plants growing at 2150 m. The photopigments analysis of leaves of S. tenuifolium showed variations in their contents, being maximum at the highest altitude (3178 m) and minimum at the lowest altitude (2150 m). The correlation matrix suggested a strong positive correlation between elements like Na+, K+and Ca+2 with that of carotenoids, and Fe+2 and Zn+2 with that of chlorophyll a + b (Chl a + b). On the other hand, a negative correlation existed between photopigments and phenolic acids probably because the plant enjoys a good nutrient status, and therefore secondary metabolites that are usually triggered during stress need not be produced in high quantities.

In Vitro Anti Tubercular Activity and Physicochemical Standardization of Selected Medicinal Plant Extracts

Globally tuberculosis is a major health problem. The main question with the current tuberculosis treatment is adverse effects and drug resistance. The present study aimed to identify safe and effective alternative herbal drugs for the treatment of tuberculosis. This study involves the standardization of selected medicinal plant extracts and in vitro antitubercular activity. Based on the literature survey, four medicinal plant extracts were selected i.e. Glycyrrhiza glabra (Yastimadhu), Piper longum (Pippali), Curcuma longa (Haridra) and Adhatoda vasica (Vasaka). To evaluate the authenticity and quality control aspect, chromatographic fingerprinting and physicochemical analysis were carried out. Quantification of phytomarkers was analyzed by High performance liquid chromatography and in vitro anti-mycobacterial activity was performed by agar well diffusion method on Mycobacterium smegmatis. All necessary physicochemical parameters were performed and found to comply with specified limits. Extracts were authenticated by qualitative High-performance thin layer chromatography fingerprinting in comparison with suitable raw herbs and phytomarkers. Quantification by High performance liquid chromatography showed that Glycyrrhiza glabra contains 10 % Glycyrrhizin, Piper longum contains 7 % Piperine, Curcuma longa contains 82.41 % Curcumin and Adhatoda vasica contains 2.3 % Vasicine. In vitro activity of Glycyrrhiza glabra, Piper longum and Curcuma longa showed 20 mm, 16 mm and 14 mm of maximum Zone of inhibition respectively. Adhatoda vasica did not show any activity against Mycobacterium smegmatis. The present study demonstrates that the selected plant extracts have the potential to inhibit mycobacterial growth. Thus, these standardized herbal extracts could be used in the design and development of a polyherbal formulation for the treatment of primary stage tuberculosis.

Sugar Profiling of Honeys for Authentication and Detection of Adulterants Using High-Performance Thin Layer Chromatography.

Honey adulteration, where a range of sugar syrups is used to increase bulk volume, is a common problem that has significant negative impacts on the honey industry, both economically and from a consumer confidence perspective. This paper investigates High-Performance Thin Layer Chromatography (HPTLC) for the authentication and detection of sugar adulterants in honey. The sugar composition of various Australian honeys (Manuka, Jarrah, Marri, Karri, Peppermint and White Gum) was first determined to illustrate the variance depending on the floral origin. Two of the honeys (Manuka and Jarrah) were then artificially adulterated with six different sugar syrups (rice, corn, golden, treacle, glucose and maple syrup). The findings demonstrate that HPTLC sugar profiles, in combination with organic extract profiles, can easily detect the sugar adulterants. As major sugars found in honey, the quantification of fructose and glucose, and their concentration ratio can be used to authenticate the honeys. Quantifications of sucrose and maltose can be used to identify the type of syrup adulterant, in particular when used in combination with HPTLC fingerprinting of the organic honey extracts.

Phytochemical characteristics of bergamot oranges from the Ionian islands of Greece: A multi-analytical approach with emphasis in the distribution of neohesperidose flavanones

The present study describes the peculiar phytochemical characteristics of bergamots cultivated in distinct islands of the Ionian Sea. Ultrahigh-performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS) supported by 1 and 2D NMR spectroscopy was used for unambiguous metabolic profiling of albedo, flavedo and juice samples. Profile differences were determined using a multi-analytical clustering approach based on high-performance thin-layer chromatography fingerprints and UHPLC-HRMS data. Finally, a validated HPLC method offering good precision (0.12–0.94%) and accuracy (95.25–103.93%) was proposed for the quantification of the major flavanones. A total of 37 secondary metabolites were characterized belonging to flavonoids, limonoids and coumarins. Their distribution was tissue-dependent and varied significantly from bergamots of other geographical locations. Surprisingly, neoeriocitrin was the major flavanone, reaching 1.69 ± 0.05 g/L in the juice and 5.24 ± 0.12 mg/g in albedo. This is the highest reported amount among Citrus species, rendering Ionian bergamots a promising candidate for novel functional products.

Adraka Maricha Khandavaleha: Preparation and Analysis

Therapeutic efficacy and safety of polyherbal ayurvedic preparations can only be assured by performing quality testing which is also called as Standardization. Adraka Maricha Khandavaleha is generally preferred in Allergic skin diseases, dermatitis, chronic bronchitis, asthma, emaciation, anorexia, bloating, edema and inflammation. This formulation is having spicy test. The aim of reported study was to Prepare the formulation at laboratory scale and to standardize it with respect to phytochemical screening, physicochemical analysis and High-performance thin layer chromatography (HPTLC) finger printing and stability studies. It is one of the common dosages from appreciated by all age group for their palatability and easy method of administration and feasibility. The formulation Adraka Maricha Khandavaleha was prepared by adopting reference of Bhaishajya Ratnavali. The results obtained from this study would help to set standards for preparation and analysis of the formulation. HPTLC fingerprint profile would serve as parameter to identity different phytoconstituents or marker compounds present in this poly herbal formulation. The results of other tests would be beneficial and can be taken as reference while developing standard formulation of great efficacy.

Phytochemical evaluation, HPTLC analysis and in-vitro antioxidant activity of hydroalcoholic leaf extract of Grewia hirsuta collected from Western Ghats forest

In-vitro antioxidant action of hydroalcoholic leaf extract of Grewia hirsuta (HAEGH) has been examined using one, “1-diphenyl-2-picryl-hydrazil (DPPH) free from radical scavenging” actions. The plant collected from the forest of the Western Ghats region of Karnataka province. The motive for plant collection from a specific location is the plant of forests exhibits the variation in growth, quantity, and quality of their active ingredients and secondary metabolites due to influence ecological factors like effect changes in location, soil, climate, etc. The work corresponds to preliminary phytochemical investigation for diverse phytoconstituents and quantitative phytochemical analysis of total phenolic, flavonoids &amp;amp; alkaloids content (TPC, TFC &amp;amp; TAC respectively) was evaluated with advanced methods. “HPTLC (High performance thin-layer” chromatography) fingerprint investigation was achieved for qualitative determination of the likely number of elements present in the hydroalcoholic extract. In-vitro antioxidant activity of HAEGH has been determined through hydroxyl radical scavenging assay that exhibited strong dose-dependent antioxidant activity as compared with standards compound, ascorbic acid. The IC50 value of HAEGH found, 25.90 % inhibition and for ascorbic acid, it was 17.68%. The Preliminary phytochemical estimation found presence of flavonoids, alkaloids, glycosides, phenol, proteins, diterpins and quantitative phytochemical analysis estimation of TPC, TFC &amp;amp; TAC found to be 3.627%, 4.059% &amp;amp; 5.671% respectively. HPTLC analysis of HAEGH at 354nm reveals the presence of a compound with Rf value 0.44 compare with Rf value 0.46 of quercetin. These outcomes indicated that the hydroalcoholic leaf extract of Grewia hirsuta plant contains phytoconstituents that exhibit antioxidant activity possible because of the existence of bioactive compounds.

Combining multivariate image analysis with high-performance thin-layer chromatography for development of a reliable tool for saffron authentication and adulteration detection.

In this work, high-performance thin-layer chromatography (HPTLC) coupled with multivariate image analysis (MIA) is proposed as a fast and reliable tool for authentication and adulteration detection of Iranian saffron samples based on their HPTLC fingerprints. At first, the secondary metabolites of saffron were extracted using ultrasonic-assisted solvent extraction (UASE) which was optimized using central composite design (CCD). Next, the RGB coordinates of HPTLC images were used for estimation of saffron origin based on principal component analysis (PCA). The PCA scores plot showed that saffron samples were clustered into two clear-cut groups which was 92% matched with the geographical origins of the samples. In the next step, common plant-derived adulterants of saffron including safflower, saffron style, calendula, and rubia were investigated with MIA analysis of HPTLC images using partial least squares-discriminant analysis (PLS-DA) at 5-35% (w/w) levels. The PLS-DA results showed proper classification of saffron and adulterants with sensitivity 99.14%, specificity 96.94%, error rate 1.96% and accuracy 98.04. Also, the effect of HPTLC injection volume on the performance of the proposed strategy was evaluated. The ability of the proposed method was then investigated by analyzing two additional sample sets including mixed samples of four plant-derived adulterants and adulterated commercial samples. Sensitivity and specificity of this model were 100% which confirmed its validity.

Geographical variation in total phenolics, flavonoids and antioxidant activities of Eucalyptus camaldulensis leaves in Burkina Faso

This study aimed to elucidate the effect of sampling location of Eucalyptus camaldulensis leaves on antioxidant activity, total phenolic content (TPC) and total flavonoid content (TFC). Leaves of E. camaldulensis collected at three different locations in Burkina Faso (north, central, south-west) were subjected to hydrodistillation to obtain a water extract and subsequent ethanol extraction of the solid residue. The extracts were analysed for antioxidant activity using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging, TPC, TFC and high-performance thin layer chromatography (HPTLC) fingerprint. TPC were higher in water extracts and presented considerable variation from 129±9 to 266±4 milligram Gallic Acid Equivalent per gram of extract (mg GAE/g) with the highest value 266±4 mg GAE/g at the south-west site. This aqueous extract also showed high antioxidant activity with EC50 8.3 µg/mL. Furthermore, HPTLC fingerprint of the extracts showed both band intensity and size differences. For TFC, we found a slight differences within both water and ethanolic extracts with the highest value of 38.3 ±4.2 mg QE/g for the south-west ethanolic extract. The noticeable impact of climate on chemical variability of E. camaldulensis leaves needs to be taken into consideration for any further applications based on TPC biological activity of the extracts. Key words: Eucalyptus camaldulensis, total phenolic content, total flavonoid content, antioxidant activity, sample location, chemical variation.