Herbal medicines have a long therapeutic history and are still serving many of the health needs of a large population of the world. However, the quality control and quality assurance still remains a challenge because of the high variability of chemical components involved. Herbal drugs, singularly and in combinations, contain numerous compounds in complex matrices in which no single active constituent is responsible for the overall efficacy. This creates a challenge in establishing quality control standards and standardization of finished herbal drugs. Many preparations have been mentioned in Ayurvedic text books for the treatment of Urdhwaga Amlapitta (non-ulcer dyspepsia). Dashanga Kwatha is one such known formulation. In this study, Dashanga Kwatha was converted into tablet form to increase the shelf life, make it easy to dispense, for dose fixation, etc. The Dashanga Kwatha Ghana tablet was subjected to organoleptic analysis, phytochemical analysis, and qualitative analysis to detect the presence of various functional groups, and to high performance thin layer chromatography (HPTLC) examination by optimizing the solvent systems. The investigation revealed the presence of tannins, mucilage, ascorbic acid, alkaloids, saponins, glycosides, flavonoids and carbohydrates mainly.
Dashanga Kwatha; decoction; quality control; standardization; tablet
Tulasi Amla Yashti Ghrita is an Ayurvedic formulation, which is beneficial in the management of the side effects of Head and Neck Malignancies induced by Radiotherapy and Chemotherapy. A pharmacognostical study involving both the macroscopic and powder microscopy of raw drugs of Tulasi Amla Yashti Ghrita and a physicochemical analysis of the finished product were carried out, to evaluate the quality of the formulation. The specific gravity of the formulation was 0.9130 and pH was 3.5. Thin layer chromatography (TLC) and high performance thin layer chromatography (HPTLC) were carried out after organizing the appropriate solvent system, in which five spots were distinguished in TLC and nine spots in HPTLC. Most of the Rf values for the spots observed were identical. The observations could be considered to be the reference standards in future studies.
Chemotherapy; high performance thin layer chromatography; radiotherapy; Tulasi Amla Yashti Ghrita
Triphaladi Kwatha, a polyherbal Ayurvedic formulation, is recommended by Chakradatta and Yogaratnakara in the management of Prameha which has resemblance with type 2 diabetes mellitus. The present study deals with development of pharmacognostical and preliminary pharmaceutical profile of Triphaladi granules. The pH (5% aqueous extract) was 6.0, water-soluble extract 48.66% w/w, alcohol-soluble extract 33.91% w/w, ash value 5.97% w/w, and loss on drying at 105°C was 6.53% w/w. High performance thin layer chromatography were carried out after organizing appropriate solvent system in which maximum nine spots were distinguished and few of the Rf values were identical in the alcoholic extract.
High performance thin layer chromatography; Pharmacognosy; Prameha; Triphaladi granules
Brahmi Ghrita, a polyherbal Ayurvedic formulation is recommended in the management of various psychological disorders like Unmada, Apasmara and Graharogas. The present study deals with the pharmacognostical identification of ingredients of Brahmi Ghrita and its physico-chemical analysis. Pharmacognostical study containing both macroscopic and powder microscopy of raw drug revealed the quality and genuineness of all the constituents of Brahmi Ghrita. Organoleptic features of coarse powder made out of the crude drugs were within the standards prescribed. Acid value was 0.16075, saponification value 184.17, Refractive Index value 1.467 at room temperature, Iodine value 26.715, Specific gravity at room temperature was 0.9133. HPTLC was carried out after organizing appropriate solvent system in which maximum 9 spots were distinguished and most of the Rf values were identical in alcoholic extract which shows the presence of certain definite constituents in Brahmi Ghrita.
Brahmi Ghrita; HPTLC; pharmacognosy; physico-chemical analysis
Amaranthus spinosus Linn. (Amaranthaceae) is found throughout India. This tree species has been of interest to researchers because it is a medicinal plant employed in the Indian traditional system of medicine. Pharmacognostic standardization; physico-and phytochemical evaluation of the roots of Amaranthus spinosus was carried out, to determine its macro-and microscopical characters, and also some of its quantitative standards. Microscopical studies were done by using the trinocular microscope. Total ash, water-soluble ash, acid-insoluble ash, sulfated ash values, and alcohol-and water-soluble extractive values were determined for physico-chemical evaluations. A preliminary phytochemical screening was also done to detect different phytoconstituents. Microscopically, the root showed cork, cortex, stellar region, and calcium oxalate crystals. Powder microscopy showed anamalous secondary growth in between the xylem vessels and Calcium Oxalate crystals in the cortex region. Total ash was approximately three times more than acid insoluble and water soluble ash. The ethanol soluble extractive was approximately the same as the water soluble extractive. Thin Layer Chromatography (TLC) of the Petroleum-ether extract using Benzene : Ethyl acetate (6 : 1), showed six spots. In the chloroform extract, using Benzene : Ethyl acetate (4 : 1) nine spots were seen, and in the ethanol extract, using Chloroform: Methanol (93 : 7), only four spots were observed, using Iodine vapor as a viewing medium. Phytochemically, the root exhibited terpenes, alkaloids, glycosides, and sugars. These findings might be useful to supplement information with regard to its identification parameters, which are assumed significant in the way of acceptability of herbal drugs, in the present scenario, which lacks regulatory laws to control the quality of herbal drugs.
Amaranthus spinosus Linn.; pharmacognostic standardization; physicochemical evaluations
Curcuma neilgherrensis Wight is a folk medicinal plant used in the management of diabetes mellitus. The leaves of this herb are said to be successful in managing high blood glucose levels. This study is aimed at assessing the scientific appraisal of C. neilgherrensis in the course of pharmacognostical characters and phytochemical parameters, as these are not yet been done. Pharmacognostic study mainly covered the macroscopic and microscopic features of the leaves including powder microscopy, and revealed the presence of trichomes, spiral vessels etc. Phytochemical parameters such as pH, total ash value, water-soluble extract and MeOH extract values were assessed in the preliminary physicochemical screening. Qualitative analysis revealed the existence of certain chemical constituents such as flavonoids, tannins, organic acids and saponin glycosides. The crude extract of leaves was subjected to TLC and HPTLC for the separation of components.
Curcuma neilgherrensis; HPTLC; pharmacognosy; phytochemistry; TLC
To perform aqueous ethanol soluble fraction (AESF) and dichloromethane extract of aerial parts of Maytenus obscura (A. Rich.) Cuf. using high performance thin layer chromatography (HPTLC) and to test anti-inflammatory activity of these extracts.
HPTLC studies were carried out using CAMAG HPTLC system equipped with Linomat IV applicator, TLC scanner 3, Reprostar 3, CAMAG ADC 2 and WIN CATS-4 software were used. The anti-inflammatory activity was tested by injecting different groups of rats (6 each) with formalin in hind paw and measuring the edema volume before and 1 h later formalin injection. Control group received saline i.p. The extracts treatment was injected i.p. in doses of 100 and 200 mg/kg 1 h before formalin administration. Indomethacin (30 mg/kg) was used as standard.
The results of preliminary phytochemical studies confirmed the presence of protein, lipid, carbohydrate, phenol, flavonoid, saponin, triterpenoid, alkaloid and anthraquinone in both extracts. Chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates with the green solvents toluene: ethyacetate: glacial acetic acid (5:3:0.2, v/v/v) as mobile phase. HPTLC finger printing of AESF revealed major eight peaks with Rf values in the range of 0.28 to 0.80 and the dichloromethane revealed major 11 peaks with Rf values in the range of 0.12 to 0.76. The purity of sample was confirmed by comparing the absorption spectra at start, middle and end position of the band. Treatment of rats (i.p.) with AESF and dichloromethane in doses of 100 and 200 mg/kg inhibited singnificantly (P<0.05, n=6) formalin-induced inflammation by 50%, 55.9%, 45.5%, and 51.4%, respectively.
HPTLC finger printing of AESF and dichloromethane of Maytenus obscura revealed eight major spots for alcoholic extracts and nine major spots for dichloromethane extracts. These HPTLC profiles may be of great usefulness in the quality control of herbal products containing these extracts. The anti-inflammatory activity of both extracts also revealed the medicinal importance of these extracts. The plant can be further explored for the isolation of phytoconstituents having anti-inflammatory activity.
Maytenus obscura; Phytochemical screening; Finger print; Anti-inflammatory; HPTLC
Cardiospermum halicacabum Linn (Sapindaceae) is an important medicinal plant in the traditional system of medicine, known as karṇasphoṭa. The root of it is officially included in Ayurvedic Pharmacopoeia for its therapeutic uses such as jvara, kuṣṭha, pāṇḍu, kṣaya and sandhivāta etc. As no detailed analysis of macroscopy, microscopy characters of the plant, except root, have been carried out till date, it was thought worth to carry out the detailed macroscopic and microscopic study of leaves and stem, following standard pharmacognostical procedures.
Materials and Methods:
Pharmacognostic studies of C. halicacabum were carried out, and in this, the macroscopic, microscopic, physicochemical, fluorescence and phytochemical analyses were done. Physicochemical parameters such as total ash, moisture content, extractive values were determined by World Health Organization guidelines. The microscopic features of leaf and stem components were observed.
Macroscopically the leaves are bi-ternate, ovate-lanceolate in shape with dentate margin. Microscopically, leaf shows prominent midrib and thin dorsiventral lamina. The midrib shows the presence of epidermal layers, angular collenchyma, palisade cells and vascular strands comprised of thin walled xylem and thick walled phloem elements. The lamina shows prominent, narrow and cylindrical upper epidermis. The upper epidermal cells are large and contain mucilage, whereas lower epidermis possesses thin, small and elliptical epidermal cells. The mesophyll was differentiated into two zones upper and lower. The upper zones show narrow cylindrical palisade cells and lower zone shows 2-3 layers of loosely arranged spongy parenchyma cells. In the Paradermal section of the lamina we observe anomocytic stomata. The transverse section of stem shows a pentagonal appearance with five short blunt ridges and prominent cuticle. Parenchymatous cells, cortical sclerenchyma, lignified xylem fibers, phloem and pit were also found. In the powder microscopy of whole plant, glandular trichomes, non-glandular trichomes, fragments of lamina, xylem elements, parenchyma cells and fibers are observed. Phytochemical screening reveals that the C. halicacabum extract contains glycosides, carbohydrates, flavonoids, phytosterols, phenolic compounds and saponin.
Various pharmacognostic characters observed in this study help in identification, quality, purity and standardization of C. halicacabum.
Cardiospermum halicacabum; fluorescence analysis; macroscopy; microscopy; physicochemical; phytochemical
Combretum albidum Don belonging to family Combretaceae is an unexplored medicinal plant in the Indian medicinal system. According to ethnobotanical information, the leaves are used in the treatment of peptic ulcer and its fruits are used in diarrhoea and dysentery. Stem bark is used in the treatment of jaundice and skin diseases. The problem encountered in standardisation of this medicinal plant is its identification by source.
Materials and Methods:
The pharmacognostical studies were carried out in terms of organoleptic, macroscopic, microscopic, physicochemical, florescence and phytochemical analysis. Physicochemical parameters such as total ash, moisture content and extractive values are determined by World Health Organization guidelines. The microscopic features of leaf components are observed with Nikon lab photo device with microscopic units.
Macroscopically, the leaves are simple, obovate in shape, acuminate apex, entire margin and smooth surface. Microscopically, the leaves showed a large vascular strand that consists of thick walled xylem elements mixed with xylem fibres and phloem which is present in a thin layer along inner and outer portions of xylem. External to the xylem occur a thin line of sclerenchyma. Powder microscopy revealed glandular trichomes in the adaxial epidermal peelings also shows the non-glandular trichomes fairly common in powder and epidermis with anisocytic stomata. Vessels elements are narrow, long, cylindrical and dense multi-seriate bordered pits. Xylem fibres are thin and long, with thick walls, which are lignified. Preliminary phytochemical screening showed the presence of carbohydrate, glycoside, saponin, flavonoid, phytosterols and phenolic compounds.
The results of the study can serve as a valuable source of pharmacognostic information as suitable standards for identification of this plant material in future investigations and applications.
Combretum albidum; fluorescence analysis; macroscopy; microscopy; physicochemical; phytochemical
To establish the standardization parameters for complete pharmacognostic evaluation of stems of Thespesia lampas (T. lampas) (Cav.) Dalz & Gibs (Malvaceae), an important plant in the Indian system of medicine.
Morphological, microscopical, physico-chemical evaluations, florescence analysis of T. lampas stems were investigated and preliminary phytochemical analysis, GC-MS analysis and HPTLC fingerprinting were carried out for qualitative phytochemical evaluation of various extracts of stems of T. lampas.
Chemo-microscopy revealed the presence of lignin, starch grains and calcium oxalate crystals. Physico-chemical evaluation used to determine numerical standards showed a result with total ash (9.03 ± 0.05) % w/w, acid insoluble ash (1.50 ± 0.01) % w/w, water soluble ash (2.51 ± 0.02) % w/w, sulphated ash (7.50 ± 0.01) % w/w, ethanol soluble extractive (0.24 ± 0.02) % w/w, water soluble extractive (0.08 ± 0.01) % w/w, moisture content (6.03 ± 0.05) % w/w and total crude fibre content of stem powder (47.36 ± 0.32) % w/w. Behavior characteristics of the stem powder showed presence of steroids, starch, alkaloid, flavonoids and proteins. Preliminary phytochemical analysis revealed presence of glycosides, phenolic compounds, tannins, steroids, saponins, flavonoids, carbohydrates and proteins. GC-MS analysis showed the presence of fatty acids such as dodecanoic acid, tetradecanoic acid, n-hexadecanoic acid, 9-tetradecenal and HPTLC fingerprinting revealed the presence of β-sitosterol and quercetin in stems of T. lampas.
The pharmacognostic standardization of T. lampas is useful towards establishing standards for quality, purity and sample identification.
Thespesia lampas; Stems; Pharmacognosy; Malvaceae; Physicochemical analysis; Preliminary phytochemical testing; GC-MS analysis; Phelloderm; Periderm; Xylem
A simple, sensitive, and specific thin layer chromatography (TLC) densitometry method has been developed for the simultaneous quantification of strychnine and brucine in the seeds of Strychnos nux-vomica.
Materials and Methods:
The method involved simultaneous estimation of strychnine and brucine after resolving it by high performance TLC (HPTLC) on silica gel plate with chloroform–methanol–formic acid (8.5:1.5:0.4 v/v/v) as the mobile phase.
The method was validated as per the ICH guidelines for precision (interday, intraday, intersystem), robustness, accuracy, limit of detection, and limit of quantitation. The relationship between the concentration of standard solutions and the peak response was linear within the concentration range of 50–1000 ng/spot for strychnine and 100–1000 ng/spot for brucine. The method precision was found to be 0.58–2.47 (% relative standard deviation [RSD]) and 0.36–2.22 (% RSD) for strychnine and brucine, respectively. Accuracy of the method was checked by recovery studies conducted at three different concentration levels and the average percentage recovery was found to be 100.75% for strychnine and 100.52% for brucine, respectively.
The HPTLC method for the simultaneous quantification of strychnine and brucine was found to be simple, precise, specific, sensitive, and accurate and can be used for routine analysis and quality control of raw material of S. nux-vomica and several unani and ayurvedic formulations containing this as an ingredient.
HPTLC; method development; strychnine; brucine; validation
Liquid-liquid extraction-thin layer chromatography (LLE-TLC) has been a common and routine combined method for detection of drugs in biological materials. Solid-phase extraction (SPE) is gradually replacing the traditional LLE method. High performance thin layer chromatography (HPTLC) has several advantages over TLC. The present work studied the higher efficiency of a new SPE-HPTLC method over that of a routine LLE-TLC method, in extraction and detection of urinary morphine. Fifty-eight urine samples, primarily identified as morphine-positive samples by a strip test, were re-screened by LLE-TLC and SPE-HPTLC. The results of LLE-TLC and SPE-HPTLC were then compared with each other. The results showed that the SPE-HPTLC detected 74% of total samples as morphine-positive samples whereas the LLE-TLC detected 48% of the same samples. We further discussed the effect of codeine abuse on TLC analysis of urinary morphine. Regarding the importance of morphine detection in urine, the present combined SPE-HPTLC method is suggested as a replacement method for detection of urinary morphine by many reference laboratories.
morphine detection; liquid-liquid extraction; thin-layer chromatography; solid-phase extraction; high-performance thin layer chromatography
A decoction (hot-water extract) comprised of Nigella sativa (seeds), Hemidesmus indicus (roots), and Smilax glabra (rhizome) has been reported to prevent chemically-induced hepatocarcinogenic changes in rats and to exert significant cytotoxic effects on human hepatoma (HepG2) cells. However, the decoction used in previous studies to determine cytotoxicity was not standardized. Further, during preparation of pharmaceuticals for clinical use, it is more convenient to use an ethanolic extract. Therefore this study was carried out to (a) develop standardized aqueous and ethanolic extracts of the plant mixture (N. sativa, H. indicus, and S. glabra) used in the preparation of the original decoction, and (b) compare the cytotoxic effects of these two extracts by evaluating cytotoxicity to the human hepatoma (HepG2) cell line.
Aqueous and ethanolic extracts have been standardized by evaluating organoleptic characters, physicochemical properties, qualitative and quantitative analysis of chemical constituents, and analysis of High Performance Liquid Chromatography (HPLC) and Thin Layer Chromatography (TLC) profiles. Cytotoxic potentials of the above standardized extracts were compared by evaluating their effects on the survival and overall cell activity of HepG2 cells by use of the 3-(4, 5-dimethylthiazol-2yl) -2, 5 – biphenyl tetrazolium bromide (MTT) and Sulphorhodamine B (SRB) assays.
Results from MTT and SRB assays demonstrated that both extracts exerted strong dose-dependent in vitro cytotoxicity to HepG2 cells. The standardized aqueous extract showed a marginally (though significantly, P<0.05) higher cyotoxic potential than the ethanolic extract. Thymoquinone, an already known cytotoxic compound isolated from N. sativa seeds was only observed in the standardized ethanolic extract. Thus, compounds other than thymoquinone appear to mediate the cytotoxicity of the standardized aqueous extract of this poly-herbal preparation.
It may be concluded that results obtained in the present study could be used as a diagnostic tool for the correct identification of these aqueous or ethanolic extracts and would be useful for the preparation of a standardized pharmaceutical product that may be used in the future for clinical therapy of hepatocellular carcinoma.
Cytotoxicity; Hemidesmus indicus; MTT and SRB assays; Nigella sativa; Smilax glabra; Standardization
Pharmacodynamics, in Ayurveda has been described in terms of Rasadipanchaka. Rasa, on one side indicates the Bhautika composition of the drug and on the other side predicts the action. Different analytical techniques, pharmaceutical processes are being used in Ayurveda for the purpose of standardization of raw drugs.
In this study an attempt has been made to apply chromatographic technique in determination of Kashaya (astringent) Rasa (taste).
Materials and Methods:
Two important Kashaya dominant drugs Kulattha (Dolichos biflorus Linn.) and Kanchanara (Bauhinia variegata Linn.), falling under Vichitra and Samana Pratyayarabdha category respectively, were subjected to physicochemical parameters and qualitative tests followed by High-Performance Thin-Layer Chromatography (HPTLC). In light of chromatographic fingerprinting; sample preparation protocol is modified to incorporate taste threshold in correlation. Column chromatography is used for first-level discrimination technique followed by HPTLC. Kashaya Rasa Dominant Zone (KsRDZ) was separated and subjected to TLC fingerprinting. The KsRDZ fraction was designated as Botanical Reference Material (BRM) in further analysis.
Ash value, Alcohol and water soluble extract value were more in B variegata as compared to D biflorus. Presence of tannin in both the samples was confirmed through qualitative test. The KsRDZ fraction separated at Rf 0.46 and 0.48 for Kulattha and Kanchanara respectively.
The results showed that the planner chromatography technique seems very useful when BRM hypothesis was adjunct to method that explains the categorization according to traditional Rasa domain classification method.
Column chromatography; high-performance thin-layer chromatography; Rasa; spectral comparison; taste threshold
The present study deals with antimicrobial activity of ayurvedic drugs containing single herb (Amalaki Choorna and Yastimadhu Choorna) and combination of herbs (DN-90 and Asanadi Kwatha Choorna). Disc diffusion method was used to assess antibacterial activity and antifungal activity was tested using Poison food technique. Absence of bacterial growth around the discs impregnated with the aqueous extracts of drugs and reduction of fungal growth in poisoned plates indicated antimicrobial activity. Further, the results of antibacterial activity of Amalaki choorna were comparable with standard drug Streptomycin. Asanadi Kwatha Choorna inhibited bacteria to more extent than Yastimadhu choorna and DN-90. Among fungi tested, more antifungal activity was observed against Mucor sp. The antimicrobial activity of drugs tested could be due to active principles present in them.
Ayurvedic drugs; antimicrobial activity; inhibition zone; Disc diffusion method; Poison food technique
Background of the study: Water extraction is a major activity in the processing of formulations such as Kashayam, Arishtam, Medicated Oil and Lehyam. The herbal residues produced after the water extraction are mostly discarded. This study is on one such residue of a potent herb namely Curcuma longa. Curcuma longa generally known as turmeric is an important and potential drug widely used in different formulations of Ayurveda. The officinal part used is rhizome. It has been Phytochemicaly explored by different researchers. The prominent active principle in it is considered to be the phenolic compound Curcuminoids. Curcumin is the major compound in Curcuminoids. It is responsible for the yellow colour of turmeric and is practically insoluble in water. An assessment of phytochemicals in Curcuma longa rhizomes, before and after water extraction, and the fate of water insoluble compounds and Curcumin is explored in this paper. Aim: To explore bioactive principles retained with the remnants of curcuma longa rhizomes, after taking water decoctions and the scope of commercial utilization of the remnants or the active principles.
The samples were collected from The Arya Vaidya Pharmacy (Coimbatore) Limited. The preliminary phytochemical studies were done using the methods by Harborne and the antioxidant properties were estimated by DPPH radical scavenging activity.
Preliminary phytochemical studies on Curcuma longa rhizome samples before and after extraction show the presence of bioactive principles like phenols, alkaloids, flavonoids and terpenoids in both samples. The physicochemical parameters such as Alcohol extractive values, Total Ash, and Acid insoluble Ash were also compared. Thin layer chromatography was carried out for phytochemical comparison of methanolic extract of Curcuma longa before and after extraction and the profile shows comparable spots at same Rf values. The isolated Curcumin was compared with the standard Curcumin by UV–Visible Spectroscopy and the ?max was obtained at 421nm. The yield of Curcumin obtained was 3.91%. The antioxidant study gave IC50 values for the fresh sample, extracted herb and Curcumin are obtained at concentrations 49μg/ml, 85μg/ml and 12.1μg/ml respectively.
The present study shows that considerable amount of secondary plant metabolites are retained in the herbal residue. The presence of phenols and other secondary metabolites in the herb even after extraction suggest that they can be used either as such for the isolation of Curcumin, a natural colouring compound. Both turmeric and Curcumin has wide application in Pharmaceutical, Food and Textile industries. Usually a large amount of such extracted curcuma longa is available from Ayurvedic Industry. Scope of the present study thus can be extended further to the possible utilization of the extracted Curcuma longa.
Tamarindus indica Linn. fruits (Chincha) are extensively used in culinary preparations in Indian civilization. Its vast medicinal uses are documented in Ayurvedic classics and it can be used singly or as a component of various formulations. Besides fruit, the Kasta (wood) of T. indica L. is also important and used to prepare Kshara (alkaline extract) an Ayurvedic dosage form. Pharmacognostical and physicochemical details of Chincha Kasta are not available in authentic literature including API (Ayurvedic Pharmacopoeia of India). The study is an attempt in this direction. T. indica L. stem with heartwood was selected and morphological, microscopic and physicochemical standardization characters along with TLC finger print, and fluorescence analysis were documented. Transverse section of stem showed important characters such as phelloderm, stone cells layer, fiber groups, calcium oxalate, crystal fibers, and tylosis in heartwood region. Four characteristic spots were observed under UV long wave, in thin layer chromatography with the solvent combination of toluene: ethyl acetate (8:2). The study can help correct identification and standardization of this plant material.
Ayurveda; Chincha; powder microscopy; tamarind; thin layer chromatography
Caesalpinia bonduc (L.) Roxb. (Kuberaksha) is an Ayurvedic herb used in the management of malaria, liver disorders, worms, edematous conditions, etc. Based on classical Ayurvedic textual indications and recent pharmacological studies, its leaf powder was selected for studying its effect clinically on filaria. Before conducting the clinical trails, this leaf powder was subjected to certain chemical studies to find the pH, ash value, extractive values, High Performance Thin Layer Chromatography (HPTLC), etc. for standardization of the drug.
Acid insoluble ash; analytical study; Caesalpinia bonduc; Kuberaksa leaf powder; loss on drying; pH value; total ash
Sesbania grandiflora (L.) Pers., commonly known as Agasti, is widely used in Ayurveda for the treatment of diseases and for processing of various formulations in Rasashastra. It is used for its astringent, antihistaminic, anxiolytic, anticonvulsive and febrifugal activities. Moreover, because of its edible nature, the leaves and pods are used as flavoring items in the cuisine of South India. A detailed investigation of fresh and powder of leaves of Agasti was carried out. The diagnostic characters of this plant include stomatal characters, presence of resins, oil globules, appressed epidermal hair and mucilage cells. Physicochemical studies revealed loss on drying (0.6%), total ash (10.75%), acid insoluble ash (0.045%), alcohol-soluble extractive (21.7%), and water-soluble extractive (30.72%). Preliminary analysis for the presence of various functional groups revealed the presence of alkaloids, saponins, phenols and proteins. Thin layer chromatographic study of the alcoholic extract showed the presence of five, six and seven spots in short UV, long UV and after spraying developing reagent, respectively. The information generated by this particular study will provide relevant pharmacognostical and physicochemical data needed for proper identification and authentication of leaves of this particular species.
Pharmacognostical study; physicochemical study; thin layer chromatography
Standardization of a compound Ayurvedic formulation is a critical and essential issue to be considered in assuring the therapeutic efficacy and safety and to rationalize their use in the health care. Sitopaladi churna is a reputed polyherbal formulation of Ayurveda. It is prescribed for the treatment of pleurodynia, intercostal neuralgia, cold, cough associated with bronchitis, pneumonia, tuberculosis, viral respiratory infection, and in pharyngeal and chest congestion.
The present study aimed at physico-chemical standardization of in-house and two marketed brands of Sitopaladi churna.
Materials and Methods:
In our investigation, in-house churna and two commercial brands of Sitopaladi churna were standardized based on powder microscopy, physico-chemical evaluations, thin layer chromatography (TLC) and high performance thin layer chromatography (HPTLC) finger printing as per standard procedures.
The set parameters were sufficient to evaluate the churna based on various physico-chemical parameters.
The data evolved can be adopted for laying down the standards for the manufacturing units of Sitopaladi churna.
High performance thin layer chromatography; physico-chemical; Sitopaladi churna; standardization
People in Indian region often apply Shankhpushpi and other Sanskrit-based common name to Evolvulus alsinoides, Convolvulus pluricaulis, Canscora decussata, and Clitorea ternatea. These are pre-European names that are applied to a medicinal plant. Before the establishment of British rule, like the other books, ayurvedic treatises were also hand written. This might be one of the reasons due to which ayurveda could not stand parallel to the western medicine and an ambiguity is reflected in the interpretation of names and description of drugs found in the books like Charaka Samhita and Sushruta Samhita. The most widespread application of Shankhpushpi is for mental problems, but they have been considered for an array of other human maladies. The present investigation deals with the comparative pharmacognostical evaluation of four ethanobotanicals of Shankhpushpi. A comparative morphoanatomy of the root, stem, and leaves has been studied with the aim to aid pharmacognostic and taxonomic species identification. Various physicochemical, morphological, histological parameters, comparative high-performance thin-layer chromatography (HPTLC), and comparative high-performance liquid chromatography (HPLC), chromatogram of methanolic extract presented in this communication may serve the purpose of standard parameters to establish the authenticity of commercialized varieties and can possibly help to differentiate the drug from the other species. All the parameters were studied according to the WHO and pharmacopoeial guidelines.
Ethanobotanicals; HPLC; HPTLC; physiochemical; Shankhpushpi
Malvastrum coromandelianum belongs to the family Malvaceae, commonly known as false mallow. Ethnobotanical survey revealed that it is used to treat various disorders. Pharmacological screening revealed that the plant possess antinoceceptive, anti-inflammatory, analgesic, and antibacterial activities. Lack of standardization parameters for herbal raw material is a great hindrance in ensuring the purity of M. coromandelianum. The present work was taken up to with a focus to set standardization parameters for M. coromandelianum.
Materials and Methods:
The plant was subjected to macroscopic and microscopic studies. Physicochemical parameters such as ash value and extractive value were determined by standard procedures. Different extracts were screened for the presence of secondary metabolites. Phenolic and flavonoid contents were estimated. Plant was subjected for high performance thin layer chromatography (HPTLC) analysis using standard chromatographic procedure.
The microscopic characteristics showed the dorsiventral nature of leaf. Two types of trichomes were observed: Covering, unicellular, uniseriate, and bi-cellular head sessile glandular. Vascular bundle was surrounded by spongy parenchyma. Phytochemical screening revealed the presence alkaloids, tannins, amino acid proteins, and carbohydrates. The phenolic and flavonoid content estimation revealed the presence of appreciable amount of these constituents, while HPTLC analysis showed the presence of β-sitosterol in petroleum ether extract.
These findings will be useful for the establishment of standardization parameters for M. coromandelianum.
β-sitosterol; high performance thin layer chromatography analysis; Malvastrum coromandelianum; phytochemical screening; total flavonoid; total phenolic
Barringtonia acutangula (L.) Gaertn. (Family: Lecythidaceae) is an evergreen tree with simple, alternate leaves, long pendulous racemes, dark scarlet flowers, and ellipsoid to ovoid berries containing one ovoid black seed. The present study deals with a detailed pharmacognostical study on the leaf of the crude drug, B. acutangula. Morphoanatomy of the leaf was studied using light and confocal microscopy and World Health Organization (WHO) guidelines on quality control methods for medicinal plant materials. Literature reveals that the phytoconstituents like tanginol, barrinic acid, and barringenic acid are present in the wood and fruits of this plant. Our preliminary phytochemical studies of the powdered leaves revealed the presence of terpenes, flavanoids, carbohydrates, tannins, steroids, and glycosides. The physico-chemical, morphological, histological parameters, and High Performance-Thin Layer Chromatographic (HPTLC) profile presented in this paper may be proposed as parameters to establish the authenticity of B. acutangula and can possibly help to differentiate the drug from its other species and the pharmacognostic profile of the leaves presented here will assist in standardization viz., quality, purity, and sample identification.
Barringtonia acutangula; high performance thin layer chromatographic profile; pharmacognosy
Chonemorpha grandiflora (Syn. Chonemorpha fragrans (Apocynaceae) is an endangered medicinal plant. It is used in different preparations, such as sudarsanasavam and kumaryasavam used in Kerala Ayurvedic system. C. grandiflora is used for the treatment of fever and stomach disorders. Phytochemical investigations have revealed the presence of steroidal alkaloids, such as chonemorphine and funtumafrine in C. grandiflora. Camptothecin, a well-known anticancer alkaloid has been detected in ethanolic extracts of stem with bark and callus cultures derived from C. grandiflora.
Callus cultures of C. grandiflora were raised on Murashige and Skoog’s medium supplemented with 2, 4-D. Stem with bark and callus were used for phytochemical analysis mainly the alkaloids. Detection and identification of camptothecin was carried out using thin-layer chromatography (TLC), high-performance thin-layer chromatography, (HPTLC) and high-performance liquid chromatography (HPLC).
An important anticancer alkaloid, camptothecin was detected in ethanolic extracts of stem with bark and callus cultures of C. grandiflora. camptothecin content was 0.013 mg/g in stem with bark and 0.003 mg/g in callus.
This is the first report on in vivo and in vitro production of camptothecin in C. grandiflora. Camptothecin is known to occur only in six plant sources so, alternative sources for camptothecin are needed. Thus of C. grandiflora could be a new promising alternative source of camptothecin.
Apocynaceae; callus; camptothecin; Chonemorpha grandiflora
To evaluate and compare the anti-inflammatory activity and to develop HPTLC fingerprint profile of ethanolic extract of Maytenus obscura (M. obscura) and Maytenus parviflora (M. parviflora).
Preliminary phytochemical screening was done and HPTLC studies were carried out using CAMAG HPTLC system equipped with Linomat IV applicator, TLC scanner 3, Reprostar 3, CAMAG ADC 2 and WIN CATS-4 software. The anti-inflammatory activity was tested by injecting different groups of rats (6 each) with formalin in hind paw and measuring the edema volume before and 1 h after formalin injection. Control group received saline i.p. The extract treatment was injected i.p with doses of 200 and 400 mg/kg 1 h before formalin administration. Indomethacin (30 mg/kg) was used as standard.
Treatment of rats (i.p.) with M. obscura and M. parviflora in doses of 200 and 400 mg/kg inhibited significantly (P<0.05) formalin-induced inflammation by 55.9%, 63.2% and 77.9%, 82.4%, respectively. Preliminary phytochemical studies were done which confirmed the presence of protein, lipid, carbohydrate, phenol, flavonoid, saponin, triterpenoid, alkaloid and anthraquinone. Chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates with the solvent system: Toluene: ethylacetate: glacial acetic acid (5:2:0.1, v/v/v) as mobile phase. HPTLC finger printing of M. obscura revealed major 8 peaks with Rf values in the range of 0.27 to 0.77 and the M. parviflora revealed maximum 9 peaks with Rf values in the range of 0.17 to 0.76. The purity of sample was confirmed by comparing the absorption spectra at start, middle and end position of the band.
HPTLC of M. parviflora revealed 8 major spots and 9 spots for M. obscura. HPTLC finger printing of ethanolic extract of M. obscura and M. parviflora may become potential tool for checking authenticity of these species. It may help in quality control against adulterant and act as a biochemical marker for these medicinally important plants in the pharmaceutical industry and plant systematic studies. The anti-inflammatory potential of these plants also reveals its medicinal importance. It might be further explored for the isolation of phytoconstituents having anti-inflammatory potential.
Maytenus obscura; Maytenus parviflora; Phytochemical screening; Finger print; Standardization; HPTLC