HPTLC Fingerprint Analysis Research Articles

Preparation, Quality control and Standardization of a Destructive Distillate <i>Swasakasa Nei</i> (SKN), A Traditional Siddha Herbal Pharmaceutic for <i>Swasakasam</i> (Bronchial Asthma)

Standardization is essential to ensure the quality and reliability of traditional medicines. This study aimed to collect, authenticate and purification of raw materials, preparation of <i>Swasakasa Nei</i> (SKN) according to SOP, and Quality evaluation and Standardization procedures following Pharmacopoeial Laboratory of Indian Medicine (PLIM) guidelines. SKN, prepared as <i>Kuzhithylam</i>, is used to treat <i>Swasakasam</i> (Bronchial Asthma) in the Siddha system of medicine. The organoleptic characters of <i>Swasakasa Nei</i> reveal that SKN was a black, thick semisolid with a smoky odour, greasy consistency, and free-flowing nature drug. Physicochemical analysis showed values for Saponification Value (202.19), Iodine Value (30), Acid Value (7.25%), Peroxide Value (0.58%), Refractive Index (1.462), Specific Gravity (0.9131 g/ml), Loss on Drying (34.17%), Total Fatty Matter (60.18%), Free Fatty Acids (1.06%), and Unsaponifiable Matter (0.98%). The congealing point was 19°C, with no mineral oil and definite oxidation on rancidity. HPTLC fingerprint analysis revealed multiple prominent peaks at various wavelengths. Microbial contamination tests showed no specific microbes, and aflatoxin assays confirmed SKN was free from Aflatoxins B1, B2, G1, and G2. Pesticide residue analysis detected no organochlorine, organophosphorus, or pyrethroids, and heavy metal analysis showed no trace elements or heavy metals. These standards affirm the authenticity, genuineness, and safety of SKN, contributing to its scientific validation.

Identification of the Secondary Metabolites of Sargassum Tenerrimum and their Molecular Docking Analysis against the Targets of Anxiety, Depression and Cognitive Disorder

Objective: This article aimed to identify the bioactive compounds present in the brown algae Sargassum tenerrimum using TLC and HPTLC fingerprinting analysis and followed in silico molecular docking against a potential target of anxiety, depression, and cognitive disorder with identified compounds. Methods: Bioactive compounds were identified from the methanolic extract of Sargassum tenerrimum through TLC and HPTLC fingerprinting analysis. In silico molecular docking against a potential target of anxiety, depression, and cognitive disorder was performed on the latest version of AutoDock Vina v.1.2.0 software. The pharmacokinetic profile and possible bioactivities of the compounds were predicted using SwissADME. Results: Fucoxanthin, β-Cryptoxanthin, and Canthaxanthin were identified from the brown algae Sargassum tenerrimum through TLC and HPTLC fingerprinting analysis. Fucoxanthin showed the highest fitness score of -9.7 kcal/mol, -9.6 kcal/mol, and -9.7 kcal/mol against the target protein GABA-A, 5ht2c, and AchE, respectively. β-Cryptoxanthin showed the highest fitness score of -9.4 kcal/mol against target SERT compared with Fucoxanthin and Canthaxanthin. Canthaxanthin exhibited the highest fitness score- 7.5 kcal/mol, -9.0 kcal/mol, -9.7 kcal/mol, -9.1 kcal/mol, -9.1 kcal/mol, -7.4 kcal/mol, -7.9 kcal/mol and - 7.6 kcal/mol against the target receptor trkB, 5ht1A, D2, DAT, MOA-A, COMT, NMDA and 7nAchR respectively on the comparing with Fucoxanthin and β-Cryptoxanthin. Conclusion: In silico docking and ADME analysis concluded that the canthaxanthin acted through various targets and was safer than the fucoxanthin and β-Cryptoxanthin. Hence, canthaxanthin can be the best potential compound in the therapy of neuropsychological disorders.

Physicochemical characterization, Phytochemical analysis and Anti-microbial activity of Elathy mathirai

Elathy Mathirai is a Siddha herbomineral formulation indicated to cure sori(itching), sirangu(Scabies), suram(fever), vaayu, maandham(gastro intestinal disorder) . The aim of the study is to standardize Elathy Mathirai to ensure its quality, purity and safety through its physicochemical, chromatographic, phytochemical, heavy metal, pesticide residue and aflatoxins analysis and the secondary objective is to evaluate its antimicrobial property. This study determines the physicochemical paramaters that are loss on drying, total ash, acid insoluble ash, water soluble extractive, alcohol soluble extractive and pH as 5.667% , 5%, 0.014 %, 14.93 %, 8.3% and 5.6 respectively denotes purity and quality of the drug. From the study results it is ensured that the test drug Elathy mathirai is free from pesticide toxicity and aflatoxins. Qualitative preliminary phytochemical analysis shows the presence of alkaloids, Flavanoids,steroids, triterpenoids, saponins, coumarins, phenol, tannin, sugar and betacyanin. HPTLC finger printing analysis reveals the presence of seven prominent peaks Rf value ranges from 0.04 to 0.39 corresponds to the presence of seven components present within it. It was observed from the results of In vitro anti microbial assay that the formulation EM reveals significant level of activity against bacteria Klebsiella pneumonia and observed moderate activity against Staphylococcal aureus, Pseudomonas aeruginosa and posess consistent anti fungal activity against Candida albicans and there is no significant effect on Aspergillus niger. The obtained results shows that the EM complies with the regulatory standard and also possess significant anti microbial property against the tested microbes.

Exploring Morpho-Anatomical Attributes, Phytochemical, and HPTLC Profile of Enicostema axillare (Poir. ex Lam.) A.Raynal

Enicostema axillare (Lam.) A. Raynal, known as the Indian whitehead, is member of Gentiaceae family and has been used medicinally for a very long time both in India and around the world. The objective of the present work comprises collection, identification, macroscopical, microscopical, and phytochemical evaluation and HPTLC fingerprint analysis on Leaves of E. axillare. The surface studies of the leaf are amphistomatic, with anisocytic stomata and trichome complex absent in both surfaces. The T.S of Leaf is slightly flat adaxially and ribbed on abaxially at midvein, presence of one layered epidermis. Mesophyll is heterogenous, differentiated into palisade and spongy tissues, and interspersed with sphaerocrystaliferous idioblasts. The ground tissue of the mid vein consists of palisade 1-2 layered and parenchyma 1-3 layered. The midvein consists of a single oval-shaped vascular bundle, at the center. The microscopic and organoleptic characters of the powder are provided. The phytochemical analysis following hot continuous and successive extraction by Soxhlet apparatus on various extracts indicates the presence of alkaloids, phenolic glycosides, tannins, steroids and terpenoids flavonoids, saponins, phytosterols, fixed oils, gums and mucilage’s, etc., It is discovered that the number of spots, and Rf values of HPTLC are distinct diagnostic features of the plant for identification. The information accumulated through the combination of chemical and botanical screening could potentially be utilized for establishing pharmacopeial standards as well as creating a standard for the identification and authentication of E. axillare.

In Vitro Antioxidant Activity, Pharmacognostical Evaluation, HPTLC and FTIR Fingerprinting of Phyllanthus Acidus L. Stem Bark Extract for Better Application in Phytotherapy

Background: For safe and efficacious use of natural products, quality control of the crude drug is of paramount importance. Due to lack of scientific investigations pertaining to quality control phytochemical and pharmacological properties of Phyllanthus acidus (L.) Skeels, the present study was undertaken. Objective: The aim of the study is to establish various pharmacognostical and phytochemical parameters, including both macro and microscopic studies, along with HPTLC and FTIR fingerprinting analysis, total phenolic and flavonoid content of stem bark of Phyllanthus acidus, followed by exploring its anti-oxidant potential. Method: The determination of total phenolic and flavonoid content was done by various colorimetric assays. DPPH assay was used to establish antioxidant activity. Physiochemical analysis was carried out and presence of various functional groups was determined using various techniques like HPTLC, FTIR analysis and colorimetric assays. Result: The phytochemical screening showed the presence of various phytoconstituents like phenols, carbohydrates, flavonoid, tannins and terpenoids. in various solvent systems. Total phenolic was established as 189.74±0.52mg GAE/g and flavonoid content was found to be38.92±0.47 mg QE/g. Concentration of heavy metal was within acceptable limits. The stem bark showed comparable antioxidant activity in methanolic and aqueous extract showed IC50 values of 26.92 and 26.52 respectively compared to ascorbic acid having IC50 value of 31.82. HPTLC fingerprinting envisaged the presence of several phytoconstituents in Phyllanthus acidus. The FTIR analysis established the presence of phenolic functional groups. Conclusion: The referential information provided by this study will be useful to determine and manage adulterations in raw material. The study also provides insight into antioxidant property of this plant validating its ethno pharmacological use as a natural antioxidant.

Biopigments Produced by Monascus purpureus Through Solid State Fermentation: HPTLC Finger Printing Analysis

Biopigments Produced by Monascus purpureus Through Solid State Fermentation: HPTLC Finger Printing Analysis

Optimization of Extraction Conditions using Response Surface Methodology and HPTLC Fingerprinting Analysis of Indian Propolis

The potential of Indian propolis as a suitable therapeutic agent is still to be explored to the fullest. As variation in the chemical composition across different geographical regions has been a major concern, it is, therefore, worthwhile to systematically investigate, and assess the quality of Indian propolis and set nationalized standards. The propolis collected from three different regions were authenticated for the major plant source, followed by the evaluation of physicochemical properties. Different extraction methods were compared, with their influence on balsam content, polyphenol, and flavonoid content. The sample coded HAR using ultrasonication gave the highest extraction yield of 71.87%. Further, the process variables in the ultrasonication method (i.e. Amplitude, extraction time, the concentration of ethanol) at three levels were optimized using the Box-Behnken response design. The model was found significant with the concentration of ethanol having a maximum influence on the responses. The model was successfully verified by setting the amplitude at 48%, extraction time to 5.6 mins, and ethanol concentration to 87% v/v. The extraction efficiency of 83.86±1.17% was achieved with a substantial diminution of extraction time. The total phenolic, total flavonol, and total flavanone content of the optimized batch were 28.74±0.13 mgGAE/g, 4.66±0.78 mgQE/g, and 3.16±0.14 mgPE/g of propolis respectively. Further HPTLC fingerprint image of the optimized extract revealed the presence of quercetin, p-coumaric acid, kaempferol, chrysin, pinocembrin, and galangin.

A comparative study on bioactive extracts of endophytic fungi of the leaves and stems of Tithonia diversofolia

Natural products as potential sources for novel medications are one of the key goals of phytochemistry. Phytochemicals offer useful lead compounds that can be changed appropriately for subsequent medication development. The Mexican sunflower plant, Tithonia diversifolia, is being investigated in the current study. Sesquiterpenoids, diterpenoids, and flavonoids, among other secondary metabolites, have been identified from Tithonia sp. Endophytic fungi are an important source of phytomolecules, which are bioactive chemicals derived from plants. These substances, which are unique from those found in their host plants, are crucial for improving both the adaptation of endophytic fungus and the plants they live on. In the present study, leaves and stems were examined and endophytic fungi, Colletotrichum gloeosporioides and Cladosporium sphaerospermum were isolated. Hexane and EtOAc extracts of these fungi were analyzed for their phytoconstituents. Flavonoids, tannins, terpenoids and alkaloids are the major class of compounds identified by using the HPTLC finger printing analysis. The extracts showed significantly high phenolic content. All the endophytic extracts showed cytotoxicity against human leukemic monocyte THP-1 cells and indicated possible anti-cancer activity potential.

HPTLC Fingerprint of an ayurvedic combination- Panchavalkala

Background: Panchavalkala is one of the most versatile ayurvedic combination of plants known for being used to reduce infections, inflammations and in wound healing and having a wide spectrum of medicinal uses. It has been formulated into various formulations like- Panchavalkala choorna, Lepa, Kashaya, kwatha, ointment, gel hand wash, decoction etc. It is It includes a group of five drugs- Ficus benghalensis L, Ficus racemosa L., Ficus religiosa L., Ficus lacor Buch. Ham, Thespesia populnea. WHO guidelines emphasises the need for modern and sophisticated modern instrumental method like HPTLC to standardize herbal materials. Aim and objective: To establish fingerprint profile of Panchavalkala combination and its individual drugs using HPTLC technique. Method: TLC method was developed for aqueous extract of individual drugs of Panchavalkala and Panchavalkala combination using solvent system- Toulene: ethyl acetate: methanol: formic acid (4:4:1:0.1). HPTLC fingerprinting was developed by scanning the chromatogram at 254, 366, 280 nm and at 250nm post derivatization using vanillin sulphuric acid. Results: The chromatograms were analyzed for number of peaks and common peaks in aqueous extract of individual drugs and Panchavalkala combination. HPTLC fingerprinting of all the individual extracts was established to identify them. The presence of common peaks found in individual extracts was confirmed in Panchavalkala combination, indicating that the developed HPTLC method can be successfully used to identify and standardize the combination. Conclusion: It can be concluded that HPTLC fingerprint analysis of Panchavalkala combination and its individual drugs can be used for Standardisation.

Standardization And Analytical Evaluation Of Siddha Herbal Formulation Aavarai Kirutham In The Treatment Of Madhumega Avathai Vii: Diabetic Peripheral Neuropathy

Aavaarai kirutham is a Siddha herbal confection which literarily recommended for the treatment of diabetic peripheral neuropathy (Mathumega Avathaigal) in the authentic Siddha textbook " Sigicha rathina deepam". There are no standards for this antidiabetic herbal preparation. Standardization of the individual ingredients helps to prepare a quality test drug which responsible for the desired pharmacological effect. So, the present study has been performed to standardize unique ingredients and the herbal formulation 'Aavaarai kirutham' as a preliminary study by using three groups of test drug samples. The parameters used to standardize the preparation are organoleptic characters, Biochemical analysis, Physicochemical properties,HPTLC fingerprint analysis, Heavy metal analysis by ASS, Test for specific pathogens, Sterility test by pour plate method, Aflotoxin analysis, Pesticide residue analysis,and Thin layer chromatography. The organoleptic characteristics such as color, taste, odor, smell, and consistency of the ingredients and the test drug were evaluated. Physico-chemical parameters were determined as per WHO guidelines and reported as total ash, insoluble acid ash, water soluble extractive, ethanol soluble extractive, total sugar, reducing sugar content, fat content, and loss on drying. Thin layer chromatography developed for different solvent systems and detection of TLC fingerprint profiles were carried out under UV light at the wavelength of 366nm and also viewed after spraying with anisaldehyde sulfuric acid spray reagent. Colors and Rf Values of major spots were noted.

In vitro regeneration and chromatographic fingerprint analysis of Aphanamixis polystachya (Wall.) Parker by HPTLC technique

• In vitro regeneration on nodes of Aphanamixis polystachya tree showed breakage of recalcitrance. • Biomass content was higher on cytokinins 2-ip (N 6 -(2-Isopentyl) Adenine) supplemented Murashige and Skoog's medium. • HPTLC analysis of showed higher diversity of phytochemicals in the in vitro shoots without any supplementation of elicitors. • Comparison between the in vitro shoots and mature plants shown the presence of majority of phytochemicals in both groups. Aphanamixis polystachya is an evergreen tree belonging to the Meliaceae family used for centuries in traditional medicines. Various metabolites that were isolated from the barks showed high therapeutic value. Isolation of pharmaceutically active compounds imposes pressure on harvesting the plants from the wild, resulting in the endangerment of the tree. The development of an alternate method for the isolation of phytochemicals is the need of the hour. The current study evaluated the phytochemical screening of bioactive compounds from the tree parts and in vitro regenerated shoots of A. polystachya . Due to its recalcitrancy, tissue culture data remains inaccessible, and the present investigation was first attempt on developing a protocol for generating plantlets using in vitro culture. Murashige and Skoog's medium fortified with cytokinins like BAP (6-Benzylaminopurine), Kinetin, 2-ip (N 6 -(2-Isopentyl) Adenine), and TDZ (Thidiazuron) were used, and maximum biomass was produced from 3 mg/L of 2-ip. Meanwhile, HPTLC fingerprinting analysis on the tree parts (bark and leaves) and in vitro shoots showed similar diversity among metabolites. The secondary metabolites produced in the in vitro plants were the same with mature trees for standards like Rutin, Stigmasterol, β-Sitosterol, Piscidinol A, Bourjotinolone B, and Niloticin.

HPTLC Finger print Analysis and In vitro Cytotoxicity Study against A549 cell line for Methanol and Chloroform extract of bark of Erythrina stricta Roxb.

Plant provides various important phytoconstituents in the form of primary and secondary metabolites. Secondary metabolites obtained from the plants possesses significant biological activities. The plant phytochemicals are useful for scavenging free radicals and also in the treatment of cell injury. The proper identification and authentication of the plant secondary metabolites is important for quality control purpose. There are various chromatographic tools like HPTLC, HPLC and GC are interest of researcher for carrying out the authentication of the plant secondary metabolites. Among these, HPTLC is used widely for the plant authentication for its fingerprint ability for plant constituents. In this study, methanol (ESM) and chloroform (ESC) extracts of Bark of Erythrina stricta Roxb. were selected for its HPTLC fingerprint profile and In vitro cytotoxicity study for A549 cell lines (lung cancer). Plants were collected from in and around of Aizawl city and authenticated from BSI, Shillong. Bark of plants were prepared and extracted using soxhlet extractor with different solvents gradually increasing their polarities (Petroleum ether, chloroform and methanol). Solvent systems for chromatography were developed and HPTLC fingerprint was carried out. MTT assay for cytotoxicity were performed against standard doxorubicin and IC50 concentrations were calculated. The HPTLC fingerprint showed the presence of various phytochemicals in chloroform and methanol extract. Cytotoxicity study suggested that the plants extracts reduce viable cells by exerting cytotoxic effect. These studies can be used further for exploration of other pharmacological actions including anticancer activity.

PHARMACOGNOSTIC STUDIES ON LEAVES OF GYMNANTHEMUM AMYGDALINUM WITH SPECIAL REFERENCE TO A NEW ADDITION TO THE FLORA OF SOUTH INDIA

There has been a tremendous increase in the demand for plant-based drugs over the last several years. Due to this, there is a need for standardization of the plant raw materials. Although modern techniques are being widely used for the standardization process, the pharmacognostic approach is still reliable for identifying the raw material. The current study deals with a detailed investigation of macro-and micromorphological characters, powder characteristics, organoleptic characters, HPTLC fingerprinting and preliminary phytochemical analysis on the leaves of Gymnanthemum amygdalinum. It belongs to the family Compositae. Commonly known as a bitter leaf, in Telugu “sugarchettu” it is found throughout Bihar, Madhya Pradesh, Odisha, West Bengal and has been recently reported in Sriramagiri Village, Mahabubababad District, Telangana, South India. The leaves are used ethnomedicinally and in folk-lore medicine, as antidiabetic and in treatment of gastritis and hepatitis. The present work can serve as a useful tool in identifying, authentication and standardization of the plant material and quality control in the pharmaceutical industry.

Microscopic and HPTLC Fingerprint Analysis a Tool for Authentication and Quality Control of Nelumbo nucifera

Microscopic and HPTLC Fingerprint Analysis a Tool for Authentication and Quality Control of Nelumbo nucifera

Neuropharmacological evaluation and HPTLC fingerprint profile of phytocompound-enriched chloroform fraction of methanolic extract of Lagenaria siceraria (Molina) Standley fruits - a potent Asian ethno-medicinal vegetable plant.

Lagenariasiceraria (Molina) Standley (Cucurbitaceae) is a traditional vegetable plant, popularly known as bottle gourd (English) and lauki (Hindi). It is a climbing herb characterized with a number of therapeutic properties. Traditionally Lagenariasiceraria (LS) fruits were used for their cardioprotective, hepatoprotective, diuretic, and purgative effects, but there is very little scientific data available on its neuroprotective potential.

HPTLC Fingerprinting analysis of phytoconstituents from Bixa orellana and Beta vulgaris plant

HPTLC Fingerprinting analysis of phytoconstituents from Bixa orellana and Beta vulgaris plant

Pharmacognostic evaluation, physicochemical standardization and HPTLC fingerprint analysis of pomegranate (Punica granatum L.) leaf and seed

Pharmacognostic evaluation, physicochemical standardization and HPTLC fingerprint analysis of pomegranate (Punica granatum L.) leaf and seed

Application of HPTLC Multiwavelength Imaging and Color Scale Fingerprinting Approach Combined with Multivariate Chemometric Methods for Medicinal Plant Clustering According to Their Species.

In the current study, multiwavelength detection combined with color scales HPTLC fingerprinting procedure and chemometric approach were applied for direct clustering of a set of medicinal plants with different geographical growing areas. The fingerprints profiles of the hydroalcoholic extracts obtained after single and double development and detection under 254 nm and 365 nm, before and after selective spraying with specific derivatization reagents were evaluated by chemometric approaches. Principal component analysis (PCA) with factor analysis (FA) methods were used to reveal the contribution of red (R), green (G), blue (B) and, respectively, gray (K) color scale fingerprints to HPTLC classification of the analyzed samples. Hierarchical cluster analysis (HCA) was used to classify the medicinal plants based on measure of similarity of color scale fingerprint patterns. The 1-Pearson distance measurement with Ward’s amalgamation procedure proved to be the most convenient approach for the correct clustering of samples. Data from color scale fingerprints obtained for double development procedure and multiple visualization modes combined with appropriate chemometric methods proved to detect the similar medicinal plant extracts even though they are from different geographical regions, have different storage conditions and no specific markers are individually extracted. This approach could be proposed as a promising tool for authentication and identification studies of plant materials based on HPTLC fingerprinting analysis.

Cytotoxic evaluation and phytochemical screening of an ethnomedicinal plant: Pterospermum rubiginosum from southern Western Ghats

Pterospermum rubiginosum is an evergreen tree belonging to the family Sterculiaceae, different solvents like hexane, chloroform, acetone, and carbinol were used for sequential soxhlet extraction of air dried powdered bark, the obtained extracts stored at desiccators for phytochemical screening. The phytochemical analysis revealed the presence of alkaloids, phenolics, flavonoids, saponins, terpenoids, and phenolic acids. The main objective of this article is to evaluate the different phytochemicals present in bark extracts for further detailed study and to enable proper documentation for conserving our traditional knowledge about endemic medicinal plants for the future generations. HPTLC fingerprint analysis of Pterospermum rubiginosum bark extract showed the presence of tannins, its derivatives and gallic acid. The cell viability of both methanolic and aqueous extracts was evaluated by MTT assay against lung fibroblast (L929) cell line and LC50 Value were found to be 150.32µg/mL and 135.17µg/mL for PRAQ and PRME respectively.

Detection of Adulteration in Rubia cordifolia – A Chromatographic Approach

According to traditional medicinal texts as well as modern research, Rubia cordifolia (Desi Manjistha) stem cure various diseases of blood, skin diseases, inflammations, kidney stones, fever and various Kapha disorders. However, stems of Rubia cordifolia (Desi Manjistha) are often adulterated with stems of Rubia tinctorum (Irani Manjistha) due to morphological similarity. This adulteration is extremely harmful as R. tinctorum has Lucidin as one of the chemical constituents, which has severe genotoxic effects. The present work focuses on developing a comparative HPTLC fingerprint and GC-MS analysis of R. cordifolia and R. tinctorum, so as to detect the adulteration by R. tinctorum in R. cordifolia raw material form and in formulations. Methanol extracts of the individually powdered stems were used to develop a suitable mobile phase, such that a clear spot was obtained in TLC of R. tinctorum but not in TLC of R. cordifolia using the same mobile phase. This mobile phase was further used to develop a comparative HPTLC fingerprint of the two species. Methanol extracts of R. cordifolia and R. tinctorum were used for investigation of phytoconstituents by GC-MS analysis. The mobile phase n-butanol: ethanol (6:4) was the one which gave a clear single spot at Rf 0.89 in TLC of methanol extract of R. tinctorum but not in TLC of methanol extract of R. cordifolia, at 254nm and 366nm. Further, HPTLC analysis gave results clearly differentiating the two species using the same mobile phase. Further, GC-MS analysis of R. cordioflia revealed the presence of 9 phytoconstituents while R. tinctorum revealed the presence of 6 phytoconstituents, which are different from each other. Thus, these simple yet sophisticated techniques will be very useful for herbal industry in standardization and detection of adulteration of R. tinctorum in R. cordifolia formulations, thereby benefitting the patients who consume Manjistha formulations.