High Performance Thin Layer Chromatography Studies Research Articles

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

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

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.

Pharmacognostical and physicochemical evaluation of gandhakadi yoga vati: An ayurveda herbo-mineral formulation for thalassemia major

Background: Ayurveda is age-old medical system, practiced in India. Ayurveda medicines are being prepared from natural sources like herbs, animal products, and minerals. Ancient Ayurveda scholars have mentioned various pharmaceutical procedures to make it bioavailable. They also mentioned about various parameters to control the quality and standardize those formulations. But due to globalization and industrialization, it is need of the time to produce medicines in larger scale and to evaluate its quality with currently available tools for the global acceptance of Ayurveda. Gandhakadi Yoga Vati is an herbo-mineral formulation used as an adjuvant in the management of Thalassemia Major. Till date quality data of this formulation is not been evaluated. Aims and Objectives: To analyze the Gandhakadi Yoga Vati through pharmacognostically and physicochemical parameters. Materials and Methods: In this analytical study, preauthenticated raw drugs were procured from pharmacy, ITRA, Jamnagar. Drug was prepared at RSBK department and pharmacy, ITRA, Jamnagar. Organoleptic parameters and microscopic analysis of Vati were done at pharmacognosy department, ITRA. Physicochemical analysis and high-performance thin layer chromatography (HPTLC) were carried out at pharmaceutical laboratory, ITRA. Results: Microscopic characteristics of Gandhakadi Yoga Vati showed simple trichome, starch grain, epidermal cells, fibers, parenchymal cell of Agastya Patra (Sesbenia grandiflora Linn.), pitted stone cells, lignified stone cell, trichome, tanin content, prismatic crystal, brown content, spool cells of Vidanga (Embelia ribes Burm.f.), and black debris of Gandhaka. In physicochemical analysis, Ash value was 8.64%w/w, loss on drying was 0.85%w/w, pH was 6.4, acid-insoluble ash value was 2.49% w/w alcohol-soluble extractive was 14.9%w/w, and water-soluble extractive was 5.09%w/w. HPTLC study showed 06 and 07 peaks at 254 and 366 nm wave lengths, respectively. Conclusion: This study generated preliminary data on pharmacognostical, physicochemical parameters, and HPTLC of Gandhakadi Yoga Vati. These fingerprinting can be useful for future researchers to reproduce this formulation.

Pharmaceutico-Analytical standardization of amavatari rasa – A herbo-mineral formulation

Introduction: Amavatari Rasa (AR) (mercury-containing herbo-mineral formulation), widely used in the management of Amavata (rheumatoid arthritis) has been being manufactured by many pharmaceutical companies in India for long. However, till today, the standard manufacturing process (SMP) for this formulation is not available, and hence, the present study was carried out to set the SMP of AR along with its quality control parameters. Methods: Pharmaceutical procedures such as Shodhana (pre-processing) of Parada (mercury), Gandhaka (sulfur), Chitraka (Plambago zeylanica Linn.), and Guggulu (Commiphora wightii Arn.); preparation of lime water, Triphla Kwatha (decoction) and AR were carried out at Rasa shastra and Bhaishajya Kalpana Department of ITRA, Jamnagar. Physicochemical parameters, high-performance thin-layer chromatography (HPTLC), and inductively coupled plasma atomic emission spectroscopy (ICP-AES) were carried out at the pharmaceutical chemistry Lab, ITRA, Jamnagar and Sophisticated Analytical Instrument Facility, IIT Powai, Mumbai respectively. Results: Average of 54.33 g of castor oil was required for 354.33g of AR. Mean ± standard deviation (SD) of pH, loss on drying, ash value, acid insoluble ash, water soluble extractive, and alcohol soluble extractive of AR was 3.77 ± 0.20, 6.29 ± 0.41%w/w, 5.46 ± 0.72%w/w, 0.40 ± 0.15%w/w, 23.2 ± 5.15%w/w and 30.13 ± 7.19%w/w, respectively. HPTLC study showed 4 peaks in 254 nm and 5 peaks in 366 nm ultraviolet lights. AR contains 7.38% mercury, 0.0031% arsenic, and 0.0032% lead. Conclusion: The adopted method for preparation of AR was convenient to replicate and hence can be considered SMP. Results of physicochemical parameters, HPTLC, and ICP-AES of this study could be used as quality parameters for Amavatari Rasa prepared by this method.

Phytochemical analysis of Clerodendron infortunatum Linn. leaves in various seasons by different solvent and extraction techniques

Clerodendron infortunatum Linn is widely used for the treatment of malaria and vermifuge, scorpion and snake bites, tumors, inflammation, bronchitis and in wound healing. The present study was carried out to estimate the variation in the amounts of Gallic acid, Rutin and Quercetin in Clerodendron infortunatum Linn.leaves extracts collected in different seasons using high-performance thin layer chromatography (HPTLC).In this study, leaves of C. infortunatum were subjected to methanol extractions using Soxhlet method and cold maceration. The leaves were collected in different seasons in the months of December (Sample A -Shishir rutu-Winter season) and July (Sample B -Varsha Rutu -Rainy season). The methanolic extracts were subjected to HPTLC study & using CAmag HPTLC system, employing a mixture of toluene:ethyl acetate:formic acid:methanol(3:6:1.6:0.4 ) as the mobile phase with densitometric scanning at 254nm. Quantity of Quercetin is found in both samples, extracted by Soxhlet method. Its quantity is higher in the winter season. Quantity of Gallic acidis found in both samples. Its quantity is the same in Sample B by Soxhlet extraction method & maceration method respectively. Quantity of Rutin is found in both the samples extracted by Soxhlet & maceration methods. It is higher in the sample-A. The present HPTLC method is used for the detection and quantization of Gallic acid, Quercetin and Rutin of C. infortunatum leaves. It needs to study the season wise and region wise variation of phytochemicals in useful parts of all other plants by different extraction methods.

Comparative study of High-performance thin-layer chromatography and Antioxidant potential of Hydrocotyle asiatica mother tincture used in Homoeopathy

Background: Hydrocotyle asiatica has a therapeutic significance in the Indian system of medicine due to its rich antioxidant activity. In Homoeopathy, Hydrocotyle asiatica is used for the treatment of jaundice, skin diseases, dropsy, elephantiasis, leprosy, gonorrhea, leucorrhea, and nervous debility. It contains the abundant triterpene glycoside Asiatic acid which shows cytotoxic activity on cancer cells. Its homoeopathic mother tincture is a major source of antioxidant compounds, which is responsible for its overall pharmacological activity. Objectives: This study was done to evaluate antioxidant activity and High-Performance Thin-Layer Chromatography study of Hydrocotyle asiatica in-house homoeopathic mother tincture and market samples. Materials and Methods: Antioxidant activity of in-house homoeopathic mother tincture (A) and three market samples (B, C, and D) were determined by 2, 2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free-radical scavenging activity, total phenol, and 2,2’-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assay methods. High-performance thin-layer chromatography (HPTLC) study performed on precoated silica gel 60 F254 TLC plate, mobile phase used was toluene: ethyl acetate: formic acid (5.5:4.5:1, v/v/v) and UV detection were performed at 254 and 366 nm. For derivatization, an anisaldehyde sulfuric acid reagent was used. Results: The homoeopathic mother tincture of Hydrocotyle asiatica had prominent antioxidant activity. HPTLC study indicated the presence of triterpene glycoside compound Asiatic acid in chloroform extract of Hydrocotyle asiatica. Conclusion: The mother tinctures prepared by authenticated plant samples showed maximum active constituents and prominent antioxidant activity as compared to the mother tinctures procured from the market. The present study justifies the homoeopathic usage of Hydrocotyle asiatica and highlights its healing properties.

Physicochemical, Phytochemical, Qualitative HPTLC and Antioxidant Study of Medicinal Plant Leucas aspera

Introduction: Leucas aspera (family-Lamiaceae) has great therapeutic significance in the Indian system of medicine due to its rich antioxidant activity. The main objective of the present work was to evaluate physicochemical, phytochemical, qualitative high-performance thin layer chromatography (HPTLC), and antioxidant study of L. aspera in-house mother tincture (alcoholic extract) and 2 market samples of reputed brands. Materials and methods: Three samples of mother tincture were used for the study, in-house-mother tincture (A) of L. aspera and market samples (B and C). For qualitative HPTLC studies, precoated silica gel 60 F254plate was used, mobile phase used was toluene: ethyl acetate: formic acid (7:3:0.3, v/v/v), and UV detection was performed at 254 and 366 nm. For derivatization,anisaldehyde-sulphuric acid reagent was used. In the antioxidant study, total phenolic content (TPC), DPPH, and ABTS assay were studied. Results and discussion: Result shows that the mother tinctures prepared by authenticated plant sample showed maximum active constituents and prominent antioxidant activity as compared to the mother tinctures procured from the market. Conclusions: The present physicochemical, phytochemical, HPTLC, and antioxidant study justify L. aspera medicinal usage in Homeopathy. This is the reason for the cure and healing property of L. aspera whole plant extract.
 HIGHLIGHTS
 
 Physicochemical and phytochemical investigation of Leucas asperaplant extract justifies its medicinal usage, its healing properties, and usage in Homeopathy
 aspera mother tincture prepared from authenticated sample showed maximum active constituents and prominent antioxidant activity as compared to the mother tinctures procured from the market
 The physicochemical, HPTLC, and the antioxidant study revealed the presence of major phytochemical compounds including triterpenes ursolic acid and oleanolic acid responsible for asperaoverall antioxidant activity
 
 GRAPHICAL ABSTRACT

DNA bar-coding, pharmacognostical, and phytochemical analysis of Atalantia monophylla DC. leaves

BACKGROUND: Leaves of Atalantia monophylla DC. (Syn. Limonia monophylla L.), family Rutaceae, are traditionally used to treat rheumatoid pain, hemiplegia, itch and cutaneous complaints, cough, and as an antidote to snake bite, etc. The present study reports the detailed morphology, anatomy, genetic barcoding, physiochemical, and phytochemical aspects of A. monophylla leaves. METHODS: The sample of leaves of the plant were collected from one of its natural habitat, Athagarh forest, Odisha, India. The detailed macroscopic observations were made with naked eyes, and microscopy characters of leaf, stomatal index, and powder microscopy were done as per standard protocol. Physicochemical and qualitative preliminary phytochemical analysis, high-performance thin layer chromatography (HPTLC), and DNA bar-coding studies were carried out as per Ayurvedic Pharmacopoeia of India methods and other standard guidelines. RESULTS: Leaves were opposite, unifoliate, ovate-elliptic, entire, emarginate, and glabrous. Transverse section (TS) through petiole showed epidermis, cortex, pericyclic fibers, and vascular bundles along with trichome. TS of lamina passing through midrib shows lamina with upper and lower epidermis, lignified elements, interrupted by stomata and unicellular trichome. Microscopic characteristics of the leaves powder show the presence of oil globule, rhomboidal crystal, prismatic crystal, and fragment of paracytic stomata. The result of loss on drying, total ash, acid-insoluble ash, water, methanol-soluble extractive, and the obtained pH values were about 10.47 ± 0.40%, 6.94 ± 0.95%, 1.00 ± 0.00%, 30.41 ± 2.26%, 7.64 ± 0.56%, and 6.6, respectively. Qualitative phytochemical analysis shows the presence of carbohydrates, glycosides, saponins, alkaloids, tannins, and flavonoids and the HPTLC study revealed 29 spots at both short (254 nm) and long (366 nm) ultraviolet rays. CONCLUSION: Epidermal cells, oil globule, rhomboidal crystal, prismatic crystal, paracytic stomata, and simple unicellular trichome are key characters for identification of its leaves. The results obtained from pharmacognostical, phytochemical, physicochemical, and DNA bar-coding studies may serve as the diagnostic tool of A. monophylla toward its authentication and identification and also in maintaining of quality, strength, purity, and efficacy of the leaves.

PHYSICO - CHEMICAL AND HPTLC STUDIES OF LIMONIA CRENULATA (ROOT)

Herbal medicines have gained popularity as potential therapeutic agents for the prevention and treatment of many diseases due to their high efficacy and low side effects. With increasing demand for safer drugs, attention has been drawn to the identity, purity and strength of the herbal medicines. Hence, there is a need for standardization and development of reliable quality protocols using modern techniques of analysis for the herbal drugs. In the present paper an attempt was made to lay down the pharmacopoeial standards of Limonia crenulata Roxb. (Root) based on its physico-chemical and HPTLC analysis. Determination of physico-chemical parameters such as total ash, acid insoluble ash, water and alcohol soluble extractive values and High-Performance Thin Layer Chromatography (HPTLC) studies of the chloroform extract of the plant material were carried out. The solvent system, Toluene: Ethyl acetate: Formic acid (5: 2: 0.1) efficiently resolved the components present in the crude extract. The physico-chemical and HPTLC studies on root of Limonia crenulata yielded a set of qualitative and quantitative parameters or standards that can serve as an important source of information to ascertain the identity and to determine the quality and purity of the plant material. These studies enable the identification of the plant material for future investigation and form an important aspect of drug studies.

Preliminary Phytochemical Screening and HPTLC Fingerprinting of Leaf Methanolic Extract of Spermacoce hispida

The main objective of the present work to established the fingerprint profile of S.hispida using high performance thin layer chromatography (HPTLC) technique. Preliminary phytochemical screening was done and HPTLC studies were carried out. The scanning was performed using 'CAMAG' densitometer scanner 3V equipped with CATS3 software, at a wavelength of 400 and 254 nm using deuterium lamp.Preliminary phytochemical screening of the methanolic extract S.hispida showed the presence of alkaloids, carbohydrates, phenols, tannins, flavanoids, gums and mucilage. HPTLC finger printing of S.hispida methanolic extract showed best results in Toluene: Ethyl Acetate: Formic acid: 5:4:1 solvent system. After scanning and visualizing the plates in absorbance mode at 254nm, 366 nm and visible light range (400-600nm after spraying with anisaldehyde sulphuric acid reagent) best results were shown at 400nm. The Rf values ranged from 0.06 to 0.99. The result showed that out of 15 components, the component with Rf values 0.95, 0.82 were found to be more predominant as the percentage area is more with 28.85% and 12.91% respectively.

Development and Evaluation of Talisapatradi and Vyoshadi Choorna Lozenges

In Ayurveda, Talisapatradi choorna (TPC) and Vyoshadi choorna (VSC) are commonly used medicines for cough, cold, asthma, and rhinitis. These symptoms are due to upper respiratory infections of predominantly of viral origin. Currently, there are no effective medicines except indiscriminate uses of antibiotics, local anesthetics, and pain killers. The conventional formulation of TPC and VSC is difficult to administer so an easy manufacturing lozenge formulation was developed. The phytochemical analysis was done by preliminary thin layer chromatography (TLC) derivatization studies. High-performance thin layer chromatography (HPTLC) analysis confirmed the presence of herbal actives in the lozenge formulations. The TLC analysis results showed that TPC and VSC contain phytochemicals of flavonoids, steroids and phytosterols, and alkaloids family. The herbal actives were found to be stable in the final formulation without any interference with the excipients used in the formulation. The lozenges formulated from TPC and VSC are found to be promising alternatives to traditional form for the traditional Ayurvedic preparation. Compatibility study was done using Fourier transform infrared spectroscopy and HPTLC study.

Qualitative and quantitative estimation of gallic acid in flower galls of Crataeva religiosa through FT-IR and HPTLC techniques

In the present study, FT- IR (FourierTransform Infrared spectroscopy) analysis of galled flower was conducted to qualitatively identify gallic acid while comparative HPTLC (High-Performance ThinLayer Chromatography) studies of normal and galled flower extracts confirmed the gallic acid quantitatively. The quantitative estimation of gallic acid was carried out on silica gel 60 F24HPTLC plates (stationary phase). The plates were developed in a solvent system as prepared with Toluene, Acetone and Glacial acetic acid at a ratio of 3:1:2, in a CAMAG glass twin trough chamber of 20X10 cm, at a temperature of 2 20C. Detection was carried out densitometrically at 220 nm. The FTIR analysis was done with a spectrophotometer system, which was used to identify and analyze the characteristic peaks and their respective functional groups. HPTLC analysis revealed that gallic acid content increased almost two folds in gall tissues as compared to normal tissues. C. religiosa flower galls can be utilized medicinally due to enhanced gallic acid.This is the first report of the occurrence of gallic acid in flower galls of C.religiosa.

Impact of Modern Analytical Tools In The Standardization of Kharaliya Rasayana (Arogyavardhini Rasa)

Background: Arogyavardhini Rasa is a well known Ayurvedic herbo-mineral formulation widely used for a broad spectrum of therapeutic indications mainly in the treatment of Kamala (jaundice) and other Yakrit Vikara (liver disorders).
 Aim: To standardized Arogyavardhini Rasa prepared with and without Rasavarga Dravya (ingredients of Metallo-mineral origin) by using high-performance thin-layer chromatography (HPTLC) & computational by principal component analysis (PCA) as an analytical tool. 
 Materials & Method: Arogyavardhani Rasa (AVR) was prepared as per classical reference and further modified this preparation by omitting Rasavarga dravya (ingredients of Metallo-mineral origin) of Arogyavardhani Rasa (MAVR). Formulations were investigated for physicho-chemical parameters, chromatography and using the building block of chemometrics technique such as PCA was applied to discriminate the formulations AVR and MAVR.
 Result & Discussion: HPTLC study revealed a total of 11 and 8 bands at 254 nm and 366 nm in both AVR & MAVR, with only two similar Rf values in the sample. PCA revealed that both formulations were found similarities & dissimilarities within a batch of AVR and MAVR with its physicho-chemical constant.
 Conclusion: The analytical tools applied (HPTLC and PCA) for the study revealed that the classical processing like Bhavana (Lavigation ), Mardana (Trituration) etc. had a significant role in the preparation of Rasaaushadhis (mercurial medicine) and process validate the Indian system of medicinal pharmaceutics.

Development and Validation of HPTLC Method for Simultaneous Estimation of Resveratrol and Piperine in Its Pharmaceutical Dosage Form

Resveratrol is a plant compound that work as antioxidant and it also has anti-aging properties whereas Piperine is a alkaloid and it majorly used in spices. A HPTLC (High performance thin layer chromatography) study is conducted for estimation of Resveratrol and Piperine. The Mobile phase used was Chloroform: Ethyl Acetate (50:50 v/v). Rf Value of Resveratrol 0.59 and Piperine 0.79 was found. Stationary phase of Silica gel 60 F254 was used. Densitometric analysis was performed at 325 nm. The method was found to be linear. Recovery (ranging from 97% to 102.24 %), limit of detection (40.26 ng/spot, 1.64 ng/spot respectively for resveratrol & piperine), limit of quantification (122.02 ng/spot, 4.98 ng/spot respectively for resveratrol & piperine) and precision (≤ 2.00%) were found to be satisfactory. Validation performed with linearity, accuracy, precision, specificity, robustness, limit of detection and limit of quantification. Every parameter found within the range. The developed method allows to confirm that an accurate and reliable potency measurement of a pharmaceutical preparation can be performed.

Colchicine (a high-priced alkaloid) accumulation and HPTLC quantification in different stages of in vitro developed tuber of Gloriosa superba L.

BackgroundGloriosa superba L. belongs to Colchicaceae, which is an important medicinal plant containing high-priced alkaloid colchicines and other potent phytochemicals. Due to its extensive importance at the industrial level, this plant is overexploited. Moreover, indiscriminate harvesting for raw material leads to a decline in the population of this plant in the natural environment. Thus, the present study deals with the optimization of colchicine accumulation from the different intervals of in vitro and in vivo tubers of Gloriosa.ResultTo obtain in vitro tuberization, shoot tip explants were inoculated on Murashige and Skoog medium prepared with 3 mg/l BA and 0.5 mg/l Kn + 1 mg/l 2, 4-D followed by 2 mg/l BA + 0.2 mg/l NAA. In the high-performance thin-layer chromatography study (HPTLC), the linearity range of colchicine was set at a concentration range of 100–1000 ng/spot with a regression value (r) of 0.99. Its Rf value (0.25) was recorded at 254 nm. The colchicine amount in the in vivo tuber was 7.75 ± 0.25% dry weight, while the nearby amount of 7.7 ± 0.40% dry weight of colchicine was produced from 2 weeks in vitro old tuber. This value was followed by the 4th-week old tuber with 6.35 ± 0.17% dry weight and then a gradual decrease in its accumulation.ConclusionThe significant results for the accumulation of colchicine at different stages were observed. Hence, this strategy of colchicine production creates a new possibility for improved production of colchicine under in vitro conditions which will be helpful to various pharmaceutical industries without damaging the plants from the natural environment.Graphical abstract

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.
 Objectives: Objectives of present research was to establish HPTLC fingerprinting of methanolic extract of Piper betle L. leaves.
 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.
 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.
 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

HPTLC study to determine the antioxidant activity of dried leaves of Portulaca oleracea L.

This present study involves the assessment of the anti-oxidant activity study of the sample which was obtained from the methanolic extracts of dried leaves of Portulaca Oleracea L.(common name Purslane). Purslane is a rich source of Vitamin A, Vitamin-C and some other B-complex vitamins like riboflavin, niacin, pyridoxine and carotenoids which are known powerful natural anti-oxidants. Anti-oxidants are compounds that inhibit oxidation. This methanolic extract of leaves was evaluated for the determination of its anti-oxidant efficiency by using 1,1–diphenyl-2-picryl-hydrazyl (DPPH) by using Silica TLC plates on Camag High-Performance Thin Layer Chromatography (HPTLC) system using visionCATS software. Densitograms and chromatographs obtained show the presence of anti-oxidant activity. It is a rapid, inexpensive and straightforward method to measure anti-oxidant properties of substances after separation by HPTLC. It involves the use of the free radical, 2, 2-Diphenyl-1- picrylhydrazyl (DPPH) which is widely used to test the ability of compounds to act as free radical scavengers or hydrogen donors and to evaluate anti-oxidant activity. When Anti-oxidants substances react with DPPH, which is a stable free radical becomes paired off in the presence of a hydrogen donor (e.g., a free radical scavenging anti-oxidant) and is reduced to the DPPHH. As a consequence, the absorbance's decreased from the DPPH.

Phytochemical and high-performance thin-layer chromatography analysis of Ashawattha (Ficus religiosa Linn.) Kaanda Twaka (outer portion of stem bark) Churna (powder)

AIM: The aim of this study was to evaluate phytochemical constituent and high-performance thin-layer chromatography (HPTLC) analysis of Ashawattha Kandatwaka (the outer portion of stem bark) Churna (powder). INTRODUCTION: Ashawattha Kandatwaka Churna is a major constituent of numerous ayurvedic formulations. The plant has possessed their various pharmacological activities such as antifungal, anthelmintic, hypoglycemic, antibacterial, antioxidant, anticonvulsant, hypolipidemic, immunomodulatory, and wound healing activities. MATERIALS AND METHODS: Experiments were performed on authenticated plant materials following standard procedures. The methanol extract of stem bark of Ficus religiosa Linn. was used for the HPTLC study. Toluene (9 ml) and ethyl acetate (1 ml) were used for stem bark of F. religiosa Linn. as a mobile phase. RESULTS: Identification of alkaloids, steroids, and amino acids was confirmed through the phytochemical parameter. In the HPTLC study, the Rf values obtained at 254 nm were 0.02 and at 366 nm were 0.01, 0.17, and 0.53, respectively. CONCLUSION: Physiochemical analysis including HPTLC of Ashawattha Kaanda Twaka Churna will help in further standardization.

Comparative quantitative phytochemical and HPTLC analysis of two Euphorbiaceae family plants under the name Dugdhika

Euphorbia hirta and Euphorbia thymifolia are plants with the same regional name “Dudheli” in Gujarat region, India. The present study has been conducted with comparative analysis of two plants, E. hirta and E. thymifolia, for total flavonoid content, total phenolic content, and quantification of biomarkers (rutin, gallic acid, and quercetin) by using high-performance thin-layer chromatography (HPTLC). Total phenolic contents (TPC) and total flavonoid contents (TFC) were evaluated by Folin‒Ciocalteu method and aluminum chloride colorimetric method, respectively. An improved HPTLC method was developed and validated for the simultaneous estimation of rutin, quercetin, and gallic acid. TPC was found more in the methanolic extract of E. hirta, and TFC was found almost the same in ethyl acetate extracts of both plants. Both plants showed the presence of rutin, quercetin, and gallic acid. The results of the HPTLC study provided a valuable bioactive marker content comparison between two species of the Euphorbiaceae family.

Pinus Roxburghii Sarg. Ameliorates Alzheimer?s disease- Type Neurodegeneration and Cognitive Deficits Caused by Intracerebroventricular-Streptozotocin in Rats: An In Vitro and In Vivo Study

Pinus roxburghii Sarg. is traditionally used as a nerve tonic, hemostatic and anticonvulsant. The present study was aimed to investigate the neuroprotective activity of Pinus roxburghiiSarg. stem bark for memory and cognition using in vitro and in vivo methods. Plant drug was extracted with methanol using Soxhlet extraction. In vitro cell viability activity of Pinus roxburghiiSarg. was assessed using PC-12 cell lines. The in vivo neuroprotective activity of Pinus roxburghii Sarg. was tested on Wistar albino rats (either sex) and intracerebroventricular streptozotocin (3 mg/kg, bilateral) was given to cause memory deficits. High-performance thin layer chromatography fingerprinting was performed for the presence of quercetin and gallic acid in the plant extract. The phytochemical testing of extract revealed the presence of flavonoids, alkaloids, steroids, and tannins. Pinus roxburghii Sarg. showed significant cell viability at 10, 50 and 100 μg/ml in an in vitro test on PC-12 cell lines. In the in vivo activity, intracerebroventricular streptozotocin caused significant deterioration in memory and cognition, tissue oxidative stress and acetylcholinesterase activity. Pinus roxburghiiSarg. 100, 200 and 300 mg/kg, per os and donepezil (3 mg/kg, per os) significantly (p<0.05) reversed the behavioral changes in rats when tested on morris water maze and elevated plus maze. Increased levels of lipid peroxidation, acetylcholinesterase activity and decreased the level of glutathione were significantly (p<0.05) antagonized by Pinus roxburghii Sarg., similar to donepezil in rat brain. Quercetin and gallic acid were identified quantitatively in the plant extract in High-performance thin layer chromatography study. The methanolic extract of Pinus roxburghii Sarg. ameliorated intracerebroventricular streptozotocin induced cognitive deficits and memory impairment possibly due to the presence of quercetin and gallic acid.