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1.  Anti-arthritic activity of root bark of Oroxylum indicum (L.) vent against adjuvant-induced arthritis 
Pharmacognosy Research  2013;5(2):121-128.
Background:
Oroxylum indicum (Bignoniaceae) also known as Sonapatha is an indigenous medicinal plant widely used in Ayurvedic medicine for over thousands of years. It is an active ingredient of well-known Ayurvedic formulations such as Chyawanprash and Dasamula. Root bark of this plant has tonic and astringent properties and it is also used in rheumatism.
Objective:
The present investigation was carried out to evaluate the anti-arthritic activity of different extracts of root bark of Oroxylum indicum against adjuvant - induced arthritis in rats.
Materials and Methods:
Male Wistar rats were used in this study. Arthritis was induced by injecting 0.1 ml Freund's complete adjuvant intra-dermally into the left hind paw of the rats. The paw volume, hematological, biochemical, radiographic and histopathological aspects were evaluated.
Results:
The relative percentage inhibition potential of paw volume in rats treated with various extracts of Oroxylum indicum was found to be ethyl acetate extract (67.69%) >chloroform extract (64.61%) >n-butanol extract (58.46%) respectively. The hematological parameters like RBC count, hemoglobin content showed significant increase while there was a significant decrease in total WBC count and ESR in all the groups of animals pretreated with root bark extracts. The biochemical parameters such as catalase, glutathione contents showed a significant increase while the lipid peroxide and Cathepsin-D content decreased significantly only in case of ethyl acetate pretreated rats when compared to others.
Conclusion:
The present study suggests that the chloroform, ethyl acetate and n-butanol extracts of root bark of Oroxylum indicum exhibit anti-arthritic activity. The order of activity of extracts was found to be ethyl acetate >chloroform >n-butanol respectively.
doi:10.4103/0974-8490.110543
PMCID: PMC3685761  PMID: 23798888
Anti-arthritic activity; biochemical parameters; Freund's complete adjuvant; histopathology; Oroxylum indicum
2.  Anti-inflammatory activity of root bark and stem bark of Shyonaka 
Background:
Shyonaka (Oroxylum indicum Vent.; Bignoniaceae) root bark is one of the ingredients of dashamoola (a group of 10 roots), and is used for its anti-inflammatory and analgesic action in a number of compound formulations in Ayurveda.
Aim:
Ayurvedic Pharmacopoeia of India (API) recommends using the stem bark instead of root bark.
Material and Methods:
An attempt has been made to study the anti-inflammatory activity of both root bark and stem bark kashaya (decoction) experimentally.
Conclusion
Results showed significant anti-inflammatory activity of root bark and stem bark decoction.
doi:10.4103/0975-9476.104434
PMCID: PMC3545239  PMID: 23326090
Anti-inflammatory; Ayurveda; dashamoola; Oroxylum indicum; shyonaka
3.  PA02.19. Prospects of commercial utilization of water extracted Curcuma longa 
Ancient Science of Life  2013;32(Suppl 2):S64.
Purpose:
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.
Method:
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.
Result:
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.
Conclusion:
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.
doi:10.4103/0257-7941.123885
PMCID: PMC4147537
4.  Anti-Bacterial Activities and Phytochemical Screening of Extracts of Different Parts of Thalictrum Rhynchocarpum 
Parts of the plant Thalictrum rhyncocarpum are used in herbal medicine in Kenya to treat various infections. The aim of this study was to evaluate in-vitro anti-bacteria activities and phytochemical profiles of solvent extracts of the leaves, stem bark and root of Thalictrum rhyncocarpum against Bacillus subtilis-6633, Staphylococcus aures-SG 511, Escherichia coli SG 458, Pseudomonus aeruginosa-K799/61 and Mycobacterium vaccae-10670. Anti-bacterial activity tests were carried out using disc diffusion assay and tube dilution technique, and phytochemical screening was carried out through Thin Layer Chromatography. The crude extracts showed antibacterial effects on M. vaccae, P. aeruginosa and B. subtilis. M. vaccae was most sensitive, particularly to the methanol root extract. Phytochemical screening of the extracts suggested the presence of glycosides and alkaloids in the stem bark and root extracts, and flavonoids and triterpenes in the leaf extracts. The study showed interesting levels of activities of solvent extracts of different parts of T. rhyncocarpum against some of the bacteria tested (M. vaccae, P. aeruginosa and B. subtilis). The results provide some scientific rationale for the traditional use of the plant in Kenya to treat different microbial infections.
PMCID: PMC3847427  PMID: 24311847
Thalictrum rhyncocarpum; Ethno-medicinal plant; Anti-microbial activities; Phytochemical profile
5.  Pharmacognostic Standardization, Physico- and Phytochemical Evaluation of Amaranthus Spinosus Linn. Root 
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.
doi:10.4103/0975-1483.83770
PMCID: PMC3159276  PMID: 21897662
Amaranthus spinosus Linn.; pharmacognostic standardization; physicochemical evaluations
6.  Production of camptothecin in cultures of Chonemorpha grandiflora 
Pharmacognosy Research  2010;2(5):296-299.
Background:
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.
Methods:
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).
Results:
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.
Conclusion:
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.
doi:10.4103/0974-8490.72327
PMCID: PMC3093040  PMID: 21589755
Apocynaceae; callus; camptothecin; Chonemorpha grandiflora
7.  Pharmacognostical evaluation of Cardiospermum halicacabum Linn. leaf and stem 
Ancient Science of Life  2013;33(1):15-21.
Background:
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.
Results:
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.
Conclusion:
Various pharmacognostic characters observed in this study help in identification, quality, purity and standardization of C. halicacabum.
doi:10.4103/0257-7941.134561
PMCID: PMC4140016  PMID: 25161325
Cardiospermum halicacabum; fluorescence analysis; macroscopy; microscopy; physicochemical; phytochemical
8.  Anti-nephrotoxic activity of some medicinal plants from tribal rich pockets of Odisha 
Pharmacognosy Research  2014;6(3):210-217.
Background:
Gentamicin, a strong cationic drug accumulated at biological membranes causes net increase in oxidative stress and lipid peroxidation leading to necrotic changes in renal tubles and consequently precipitates acute nephrotoxicity. Several phytoconstituents and plants extracts demonstrated significant anti-oxidant and cyto-protective activities. Vitex negundo Linn. (VN), Oroxylum indicum Vent. (OI) and Barringtonia acutangula Linn. (BA) are widely found throughout the Asian sub-continent including India, used extensively in different forms of Indian traditional medicine like Ayurveda and Unani.
Objective:
Nephroprotective activity of extracts of VN roots, OI whole plant and BA leaves were investigated against experimentally induced acute nephrotoxicity [Gentamicin (i.p; 80mg/kg for 7 days)] in Wistar rats as test animals.
Materials and Methods:
The rats were treated with Cystone (5 mL/kg; p.o) taken as positive control and methanol-dichloromethane (1:1) extracts of VN, OI and BA (200 mg/kg; p.o) as test drugs for 7 days. Following the said treatments, biochemical parameters of urine (volume, creatinine and lactate dehydrogenase (LDH)) and serum (urea, creatinine, albumin and total protein) were estimated. Renal anti-oxidant markers viz., superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and lipid peroxidation (LPO) in renal tissue were assayed. Tissue sections of kidneys from different groups were made and histopathological features were observed.
Result:
The extracts of VN, OI and BA significantly attenuated the nephrotoxicity by elevation of body weight, CAT, GPx and SOD or lowering urine LDH and creatinine, serum urea; serum creatinine and LPO respectively. Histopathological score of VN, OI and BA treated groups were 1+, 2+ and 2+ respectively against 4+ of the toxic group.
Conclusion:
The findings suggested the significant nephroprotection of VN roots followed by OI whole plant and BA leaves.
doi:10.4103/0974-8490.132598
PMCID: PMC4080501  PMID: 25002801
Gentamicin; nephroprotection; Oroxylum indicum Vent. whole plants and Barringtonia acutangula Linn. leaves; Vitex negundo Linn. roots
9.  Antibacterial, anti-HIV-1 protease and cytotoxic activities of aqueous ethanolic extracts from Combretum adenogonium Steud. Ex A. Rich (Combretaceae) 
Background
Records have shown that Combretum adenogonium Steud. Ex A. Rich (Combretaceae) is used in traditional medicine systems of several tribes in Tanzania. This study focused on the investigation of antibacterial activity, anti-HIV-1 protease activity, toxicity properties and classes of phytochemicals in extracts from C. adenogonium Steud. Ex A. Rich (Combretaceae) to evaluate potential of these extracts for development as herbal remedies.
Methods
Dried plant material were ground to fine powder and extracted using 80% aqueous ethanol to afford root, leaf and stem bark extracts. The extracts were assayed for anti-HIV-1 protease activities, antibacterial activities using microdilution methods and cytotoxicity using brine shrimps lethality assay. Screening for major phytochemical classes was carried out using standard chemical tests.
Results
All extracts exhibited antibacterial activity to at least one of the test bacteria with MIC-values ranging from 0.31-5.0 mg/ml. Two extracts, namely, root and stem bark exhibited anti-HIV-1 PR activity with IC50 values of 24.7 and 26.5 μg/ml, respectively. Stem bark and leaf extracts showed mild toxicity with LC50 values of 65.768 μg/ml and 76.965 μg/ml, respectively, whereas roots were relatively non-toxic (LC50 = 110.042 μg/ml). Phytochemical screening of the extracts indicated presence of flavonoids, terpenoids, alkaloids, tannins, glycosides and saponins.
Conclusion
These results provide promising baseline information for the potential development of C. adenogonium extracts in treatment of bacterial and HIV/AIDS-related opportunistic infections.
doi:10.1186/1472-6882-12-163
PMCID: PMC3517472  PMID: 23013240
Combretum adenogonium; Combretaceae; Anti-HIV-1 protease; Antibacterial; Cytotoxicity
10.  Antimicrobial Activity and Phytochemical Study of Vernonia Glabra (Steetz) Oliv. & Hiern. in Kenya 
Infectious diseases are prevalent and life threatening in Kenya. Majority of the sick are seeking herbal remedies in search of effective, safe, and affordable cure. This project aims to investigate the antimicrobial activity and presence of active phytochemical compounds in different parts of Vernonia glabra; a plant used by herbalists in various regions of Kenya, for the treatment of gastrointestinal problems. The plant sample was collected in January 2010 in Machakos, and different parts dried at room temperature under shade, ground into powder and extracted in Dichloromethane: Methanol in the ratio 1:1, and water. These crude extracts were tested against Staphylococcus aureus, Escherichia coli, Candida albicans, and Aspergillus niger for antimicrobial activity using disc diffusion technique. Minimum inhibitory concentrations (MICs) for active crude extracts were done using disc diffusion technique after the failure of agar and broth dilution methods. It was observed that the organic crude extracts of flower, leaf, stem, root, and/or entire plant, showed activity against at least one of the four micro-organisms screened, and at concentrations lower than the aqueous crude extracts. Organic crude extract of the leaf showed the highest activity against Staphylococcus aureus (mean inhibition zone of 1.85), recording higher activity than the commercially used standard antibiotic (Streptomycin mean inhibition zone of 1.30). The organic crude extract of flower showed significant activity only against S. aureus, with the lowest MIC of 1.5625 mg/100µl, compared to streptomycin with M.I.C of 6.25 mg/100µl. Thin Layer Chromatography-Bioautography Agar-Overlay showed that, flower alkaloids (50% active), root sapogenins (43.8% active), and root terpenoids (38.5% active) were identified as the potential antibacterial compounds against S. aureus. These results suggest that, V. glabra contains phytochemicals of medicinal properties and justify the use of V. glabra in traditional herbal medicine for the treatment of microbial based diseases. However, research on toxicity which is missing in this study is recommended for V. glabra in order to verify, validate and document the safety of this medicinal plant to the society.
PMCID: PMC3746369  PMID: 24082337
Vernonia glabra; Antimicrobial activity; Phytochemicals
11.  Studies on the antibacterial activity of Khaya senegalensis [(Desr.) A. Juss)] stem bark extract on Salmonella enterica subsp. enterica serovar Typhi [(ex Kauffmann and Edwards) Le Minor and Popoff] 
Objective
To study the phytochemical screening and antibacterial activity of the stem bark extracts of Khaya senegalensis (K. senegalensis) against Salmonella enterica subsp. enterica serovar Typhi.
Methods
The plant components were extracted using methanol, ethanol and water. The phytochemical screening of the stem bark extracts were carried out using a standard method. The antibacterial assay of the stem bark extracts against Salmonella Typhi (S. Typhi) using the agar well diffusion method with different concentrations of 50, 100, 200, 400 and 500 mg/mL and the corresponding concentrations of the control was carried out and the result compared with a standard antibiotic, amoxicillin as the control.
Results
The results obtained from the phytochemical screening of the three plant bark extracts of K. senegalensis showed 10 plant secondary metabolites including saponins, tannins, reducing sugars, aldehyde, phlobatannins, flavonoids, terpenoids, alkaloids, cardiac glycoside and anthroquinones. The ethanol and aqueous extracts showed antibacterial activities against S. Typhi at concentration of 50 mg/mL with the zone diameter of inhibition (ZDI) of 14 mm and 15 mm respectively. The ethanol and aqueous extracts also showed zone diameter of inhibition of 23 mm and 25 mm respectively at 250 mg/mL and 27 mm each at 500 mg/mL. The ethanol and aqueous stem bark extracts gave the highest ZDI at 500 mg/mL while 100 mg/mL gave the least ZDI for ethanol extract and 50 mg/mL for the aqueous extract. This was followed by 400 mg/mL that gave 24 mm ZDI of the aqueous extract and 27 mm of the ethanol extract. The methanol extract showed intermediate susceptibility evidenced by ZDI of 10 mm at 100 mg/mL concentration. The methanol extract also showed antibacterial activity of 24 mm ZDI against the test organism at a higher concentration of 250 mg/mL and 26 mm at 500 mg/mL concentration. The methanol, ethanol and aqueous extracts displayed antibacterial activities against S. Typhi with a statistical significant difference at (P≤0.05). The extracts compared favourably with the standard antibiotic, the control. The minimum inhibitory concentration of the extracts was 250, 200, 200 and 100 mg/mL for methanol, ethanol, aqueous extracts and amoxicillin (control) respectively. The minimum lethal concentration of the extracts was 250, 250, 400 and 200 mg/mL for methanol, ethanol, aqueous extracts and control respectively.
Conclusions
The antibacterial properties of K. senegalensis stem bark extract can be harnessed for the production of new antibiotics or the enhancement of already existing antibiotics.
doi:10.12980/APJTB.4.2014C636
PMCID: PMC4025353  PMID: 25183098
Khaya senegalensis; Antibacterial; Zone diameter of inhibition; Minimal inhibitory concentration
12.  In vitro Antioxidant Potential of Different Parts of Oroxylum indicum: A Comparative Study 
The present study evaluated the in vitro antioxidant potential of different parts of Oroxylum indicum. 2,2-diphelyl 1-picrylhydrazyl (DPPH), nitric oxide, superoxide anion and hydroxyl radical scavenging potential and reductive ability assay of methanol extract of different parts i.e. root, root bark, stem, stem bark, leaves and fruits were performed. Leaves and bark extracts exhibits highest free radical scavenging activity than bark, stem and fruit extract. Leaves extract showed maximum reductive ability and found to contain maximum amount of polyphenolic compounds. The highest free radical activity may be due to presence of polyphenolic compounds.
doi:10.4103/0250-474X.65013
PMCID: PMC2929795  PMID: 20838540
Free radical scavenging; Oroxylum indicum; different parts; polyphenolic compounds
13.  Phytochemical investigation and pharmacognostic standardization of Cissampelos pareira root 
Ancient Science of Life  2012;31(4):181-184.
Background:
In ethno-medicinal practices, the roots of Cissampelos pareira(Patha) are used in the treatment of various ailments related to urinary problems and skin infections, and in tumorinhibitor activity, antibacterial, antimalarial, diuretic activity,anticonvulsant activity etc., The main problem encountered in standardization of Ayurvedic drugs is proper identification of the source plant.
Materials and Methods:
The macroscopic features of each anatomical component have been observed by a high-resolution camera. The moisture content, total ash, acid-insoluble ash, and water—alcohol and ether extractive values of the powdered sampleswere determined by the method as per WHO guidelines. The phytochemical investigation of the methanolic extract of Cissampelos pareira root was performed by the standard chemical ltests and by TLC on silica gel G using solvent systems proposed by Harborne.
Result:
The microscopic characteristics showed the wavy epidermis with unicellular trichomes. Lignified xylem vessels, radial medullary rays, and prismatic calcium oxalate crystals had also been found. Phytochemical screening revealed that the Cissampelos pareira root extract contains terpenoids, alkaloids, tannins, amino acid proteins, and carbohydrates. Alkaloids and essential oil were detected in TLC of the Cissampelos pareira root extract developed using blends of methanol:concentrated ammonia (200:3) and n-butanol:acetone:water (3:1:1) and benzene:ethyl acetate:formic acid(9:7:4) as solvent systems for alkaloid whereas chloroform(100%), benzene (100%), chloroform:benzene(1:1),and ether:benzene(1:1) as solvent systems for essential oil.
Conclusion:
These findings will be useful toward establishing pharmacognostic standards on identification, purity,quality, and classification of the plant, which is gaining relevance in plant drug research.
doi:10.4103/0257-7941.107352
PMCID: PMC3644755  PMID: 23661865
Cissampelos pareira; pharmacognostic standards; pharmacognostical and phytochemical investigation
14.  Pharmacognostical and physicochemical analysis of Tamarindus indica Linn. stem 
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.
doi:10.4103/0975-9476.93939
PMCID: PMC3326798  PMID: 22529673
Ayurveda; Chincha; powder microscopy; tamarind; thin layer chromatography
15.  Differential effects of Oroxylum indicum bark extracts: antioxidant, antimicrobial, cytotoxic and apoptotic study 
Cytotechnology  2012;65(1):83-95.
Stem bark of Oroxylum indicum (L) (SBOI) is used by ethnic communities of North East India as health tonic and in treating diseases of humans and animals. The objective of this research was to carry out a detailed investigation including total phenolic and flavonoid content, antioxidant, antimicrobial, cytotoxic and apoptotic activities of different solvent extracts of SBOI and to establish correlation between some parameters. Among petroleum ether (PE), dichloromethane and methanol (MeOH) extract of SBOI, MeOH extract contained the highest amount of total phenolic (320.7 ± 34.6 mg Gallic acid equivalent/g extract) and flavonoid (346.6 ± 15.2 mg Quercetin equivalent/g extract) content. In vitro antioxidant activity (IC50 22.7 μg/ml) was highest in MeOH extract (p > 0.05) and also a significant inverse correlation was observed between phenolic (r = 0.886)/flavonoid (r = 0.764) content and corresponding DPPH IC50. Only MeOH extract inhibited both bacteria and fungi. Although, individual extract showed cytotoxicity on HeLa cells with characteristic features of apoptosis, PE extract caused maximum cytotoxicity (IC50 of 112.3 μg/ml, p < 0.05) and apoptotic activity (33.2 % sub-G0/G1 population) on HeLa cells. But, there was a significant non-inverse correlation of the MTT IC50 with total phenolic (r = 0.812, p < 0.05)/flavonoid (r = 0.998, p < 0.05) content in the three solvent extracts. TLC analysis showed three unique compounds in PE extract which may have a role in apoptosis mediated cytotoxicity. These results called for futher chemical characterisation of MeOH and PE extract of SBOI for specific bioactivity.
doi:10.1007/s10616-012-9463-0
PMCID: PMC3536868  PMID: 22821054
Oroxylum indicum; Antioxidant; Antimicrobial; Cytotoxicity; Apoptosis
16.  Detailed pharmacognostical studies on Berberis aristata DC plant 
Ancient Science of Life  2013;32(4):234-240.
Background:
Berberis aristata DC (Berberidaceae) commonly known in Hindi as “Dāruhaldi” and “Citra,” is an important medicinal herb native to Northern Himalaya region. The plant is used traditionally in Indian system of medicine as an antibacterial, antiperiodic, antidiarrheal and anticancer and it is also used in the treatment of ophthalmic infections. Its root, stem and leaves also find their use in treatment of various ailments and hence is used extensively in Ayurveda.
Materials and Methods:
Samples of the whole plants of B. aristata were collected and identified. Hand and microtome sections were taken, stained and mounted and the cell content and cell wall structure were studied according to the method described by Kay and Johansen. Representative sketches were made with the help of camera Lucida. Methods for determining the quantitative values were the same as described elsewhere. For fluorescence analysis, the powder of the root, stem and leaf were examined under ultraviolet light. Total ash, acid insoluble ash and water-soluble ash values and water- and alcohol-soluble extractives were determined.
Results:
The detailed investigations carried on the pharmacognosy of the root; stem and leaf of B. aristata have brought out some salient diagnostic features, which allow one to differentiate it from other substitutes and or adulterants. The determination of quantitative values, fluorescence analysis and the use of lycopodium spore analysis has specifically contributed to this differentiation.
Conclusion:
From the foregoing observation on the pharmacognosy of root, stem and leaf of B. aristata DC, the salient diagnostic characters of three parts have been presented, which can allow one to differentiate it from other substitutes and or adulterants.
doi:10.4103/0257-7941.131981
PMCID: PMC4078475  PMID: 24991073
Berberis aristata; fluorescence analysis; macroscopic characters; microscopic characters; physico-chemical properties
17.  Marker based standardization of polyherbal formulation (SJT-DI-02) by high performance thin layer chromatography method 
Background:
Preparation of highly standardized herbal products with respect to chemical composition and biological activity is considered to be a valuable approach in this field. SJT-DI-02 polyherbal formulation was successfully developed at our institute and filed for patent at Mumbai patent office.
Objective:
The present work was marker based standardization of patented, novel and efficacious polyherbal formulation namely SJT-DI-02 for the treatment of diabetes. The SJT-DI-02 was comprised of dried extracts of rhizomes of Acorus calamus, leaves of Aegle marmelose, fruits of Benincasa hispida, roots of Chlorophytum arendinaceum, seeds of Eugenia jambolana, leaves of Ocimum sanctum, pericarp of Punica granatum, seeds of Tamarindus indica. Selected plants were collected, dried and extracted with suitable solvents. The formulation was prepared by mixing different fractions of extracts.
Materials and Methods:
For successful and best standardization, first of all selection and procurement was carried out. Selection is done on the basis of therapeutic efficacy and amount of the marker present in the particular plant part. At the time of procurement side by side phytochemical screening and estimation of phytoconstituents was carried out. After completion of preliminary screening using characterized markers, we tried to develop best TLC systems using selected solvent composition. Finally well-developed TLC systems were applied in HPTLC. In the present study polyherbal formulation was standardized by using different four markers. TLC Densitometric methods were developed using HPTLC for the quantification of these marker compounds. Solvent systems were optimized to achieve best resolution of the marker compounds from other components of the sample extract. The identity of the bands in the sample extracts were confirmed by comparing the Rf and the absorption spectra by overlaying their UV absorption spectra with those of their respective standards. The purity of the bands due to marker compounds in the sample extracts were confirmed by overlaying the absorption spectra recorded at start, middle and end position of the band in the sample tracks. After conforming all these things fingerprints were developed for all three formulations which will be act as authentification and quality control tool.
Results:
% w/w of asarones is 3.61, % w/w of marmelosin is 4.60, % w/w of gallic acid is 10.80 and % w/w of lupeol is 4.13. The method was validated in terms of linearity, precision, repeatability, limit of detection, limit of quantification and accuracy. In well-developed mobile phase system linearity was found to be in the range of 0.983-0.995, % recovery was found to be in the range of 97.48-99.63, % RSD for intraday and interday was found to be 0.13- 0.70 and 0.32 -1.41 and LOD and LOQ was found to be in the range of 0.15- 0.61 and 0.45 -1.83 microgram per ml.
Conclusion:
Thus High performance thin layer chromatography (HPTLC) methods were developed and validated in terms of linearity, precision, repeatability, limit of detection, limit of quantification and accuracy. The methods were rapid, sensitive, reproducible and economical. It does not suffer any positive or negative interference due to common other component present in the formulation and would also serve as a tool for authentication of herbal products containing marmelosin, gallic acid, lupeol and asarones. Thus this work provides standardized and therapeutically active polyherbal formulations for the different ailments.
doi:10.4103/0975-7406.135249
PMCID: PMC4097936  PMID: 25035642
Asarones; gallic acid; HPTLC; lupeol; marmelosin; SJT-DI-02
18.  Antioxidant activities of ethanol extracts and fractions of Crescentia cujete leaves and stem bark and the involvement of phenolic compounds 
Background
Antioxidant compounds like phenols and flavonoids scavenge free radicals and thus inhibit the oxidative mechanisms that lead to control degenerative and other diseases. The aim of this study was to investigate the antioxidant activity in vitro, total phenolic and flavonoid contents in ethanol extracts and fractions of Crescentia cujete leaves and stem bark.
Methods
Crescentia cujete leaves and bark crude ethanol extract (CEE) and their partitionates petroleum ether (PEF), chloroform (CHF), ethyl acetate (EAF) and aqueous (AQF) were firstly prepared. Different established testing methods, such as 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radical, ferric reducing power (FRP), and total antioxidant capacity (TAC) assays were used to detect the antioxidant activity. Further, the total yield, total phenolic (TPC) and total flavonoid contents (TFC) of CEE and all the fractions were determined. Ethanol extracts of both leaves and stem bark were also subjected to preliminary phytochemical screening to detect the presence of secondary metabolites, using standard phytochemical methods (Thin layer chromatography and spray reagents).
Results
Phytochemical screening of crude ethanol extract of both leaves and stem bark revealed the presence of steroids, flavonoids, saponins, tannins, glycosides and terpenoids. All the fractions and CEE of leaves and bark exhibited antioxidant activities, however, EAF of leaves showing the highest antioxidant activity based on the results of DPPH, FRP and TAC assay tests. The above fraction has shown the significant DPPH scavenging activity (IC50 = 8.78 μg/ml) when compared with standard ascorbic acid (IC50 =7.68 μg/ml). The TAC and FRP activities increased with increasing crude extract/fractions content. The TPC (371.23 ± 15.77 mg GAE/g extract) and TFC (144.64 ± 5.82 mg QE/g extract) of EAF of leaves were found significantly higher as compared to other solvent fractions for both leaves and bark. TPC were highly correlated with the antioxidant activity (R2 = 0.9268 and 0.8515 in DPPH test for leaves and bark, respectively).
Conclusion
The results of the study show that leaves of C. cujete possesses significant free radical scavenging properties compared with stem bark and a clear correlation exists between the antioxidant activity and phenolic content.
doi:10.1186/1472-6882-14-45
PMCID: PMC3937116  PMID: 24495381
Calabash tree; Oxidative stress; Crude extracts; Free radicals; Anti-aging
19.  Phytochemical analysis & Antibacterial activity of Nerium oleander 
Ancient Science of Life  2007;26(4):24-28.
Nerium indicum [Family:Apocynaceae] is commonly known as Arali [Tam] found throughout India, and has been used in the treatment of cancer, cardiotonic, leprosy and skin diseases. Plant parts such as root, bark and leaves are used. The present study is therefore undertaken to analyse its phyto chemical constituents in solvents like Benene, Chloroform and Alcohol and to screen its antibacterial activity. The dried leaf sample is extracted with solvents by cold maceration. The phytochemical analysis showed the presence of Alkaloids, Terpenoids, Cardiac glycosides, Saponins, Tannins & Carbohydrates in all the solvents. All the extracts were screened for antibacterial activity by Disc Diffusion Method. Out of the cultures used Staphylococccus aureus, Pseudomonas aeruginosa and Salmonella typhimurium showed better zone of inhibition which is 10mm, 9mm & 7 mm respectively.
PMCID: PMC3330883  PMID: 22557246
20.  C-Glucoside xanthone from the stem bark extract of Bersama engleriana 
Pharmacognosy Research  2010;2(4):229-232.
Background:
The genus Bersama belongs to the Melianthaceae family and comprises of four species (B. swinnyi, B. yangambiensis, B. abyssinica, and B. engleriana) all of which are very high trees; the latter two detected species are found in Cameroon. Previous phytochemical investigation on B. yangambiensis, B. swinnyi, and B. abyssinica led to the isolation of triterpenes, saponins, flavonoids, and xanthones.
Method:
The stem bark of B. engleriana were collected in the village, Baham near Bafoussam city, Cameroon in August 2003 and identifi ed by Dr. Onana National Herbaruim, Yaoundι, Cameroon. The air dried and powdered stem bark of B. engleriana (1 kg) was extracted at room temperature with CH2Cl2-MeOH (1:1) 5 L for 48 hours. The mixture of the solvent was removed by evaporation to yield 200 g of crude extract. The latter was then dissolved in CH2Cl2 to give the CH2Cl2 soluble fraction of 5 g and a remaining gum of 195 g. Part of the remaining gum (22 g) was dissolved in water and extracted four times with butanol to give 12 g of red oil; which was then separated by paper chromatography, with butanol-acetic acid-water (4:1:5), to give 3 g of orange gum; purification was carried out on HPLC with MeOH (100%) to yield 2 g of mangiferin (1) as red oil. The CH2Cl2 soluble extract was eluted on silica gel n-hexane-CH2Cl2 gradient ratio and Sephadex LH-20 (n-hexane -CH2Cl2 -MeOH, (7:4:0.5) to afford compounds swinniol (2), Δ4-stigmaster-3β-ol (3), 4-methylstigmaster-5,23-dien-3β-ol(4).
Results:
Herein, we carried out a phytochemical study of the stem bark of B. engleriana, and we report herein the isolation and structural elucidation of mangiferin, in addition to three triterpenes, previously reported from other species of the genus.[35] The assignment of the signals of mangiferin was determined using 1H, 13C-NMR, and 2D-NMR spectral data (HMQC, COSY, HMBC). The terpenoids were identifi ed by comparison of their 1H and 13C-NMR spectra with the literature data. Fractionation of the CH2Cl2-MeOH (1:1) extract of the stem bark of B. engleriana Guike gave mangiferin (1), in addition to three previously reported triterpenes, swinniol (2), Δ4-stigmaster-3β-ol (3), and 4-methylstigmaster-5,23-dien-3-β-ol (4).
Conclusions:
A chemical investigation of the CH2Cl2-MeOH extract of the stem bark of Bersama engleriana afforded a xanthone C-glucoside (mangiferin) and fi rst isolation of three terpenoids from this species: swinniol (2), Δ4-stigmaster-3β-ol (3), and 4-methylstigmaster-5,23-dien-3-β-ol (4). The complete 1H and 13C chemical shift assignments of mangiferin were determined using 1D and 2D NMR spectroscopic data (COSY, HMQC, HMBC, DEPT). The structures of the terpenoids were determined from their 1H and 13C NMR data and compared with the literature data.
doi:10.4103/0974-8490.69110
PMCID: PMC3141132  PMID: 21808572
Bersama engleriana; melianthaceae; terpenoids; xanthone; mangiferin
21.  A review on chemical and biological properties of Cayratia trifolia Linn. (Vitaceae) 
Pharmacognosy Reviews  2011;5(10):184-188.
Cayratia trifolia Linn. Domin Syn. Vitis trifolia (Family: Vitaceae) is commonly known as Fox grape in English; Amlabel, Ramchana in Hindi and Amlavetash in Sanskrit. It is native to India, Asia and Australia. It is a perennial climber having trifoliated leaves with 2-3 cm long petioles and ovate to oblong-ovate leaflets. Flowers are small greenish white and brown in color. Fruits are fleshy, juicy, dark purple or black, nearly spherical, about 1 cm in diameter. It is found throughout the hills in India. This perennial climber is also found in the hotter part of India from Jammu and Rajasthan to Assam extending into the peninusular India upto 600 m height. Whole plant of Cayratia trifolia has been reported to contain yellow waxy oil, steroids/terpenoids, flavonoids, tannins upon preliminary phytochemical screening. Leaves contain stilbenes (piceid, reveratrol, viniferin, ampelopsin). Stem, leaves, roots are reported to possess hydrocyanic acid, delphinidin and several flavonoids such as cyanidin is reported in the leaves. This plant also contains kaempferol, myricetin, quercetin, triterpenes and epifriedelanol. Infusion of seeds along with extract of tubers is traditionally given orally to diabetic patients to check sugar level of blood. Paste of tuberous is applied on the affected part in the treatment of snake bite. Whole plant is used as diuretic, in tumors, neuralgia and splenopathy. Its climbers wrapped around the neck of frantic bullock and poultice of leaves are used to yoke sores of bullock. The bark extract shows the antiviral, antibacterial, antiprotozoal, hypoglycemic, anticancer and diuretic activity. This article focuses on the upgraded review on chemical and biological properties of Cayratia trifolia Linn. and triggers further investigation on this plant.
doi:10.4103/0973-7847.91117
PMCID: PMC3263053  PMID: 22279376
Biological; Cayratia trifolia; chemical; review
22.  A study of antimicrobial activity, acute toxicity and cytoprotective effect of a polyherbal extract in a rat ethanol-HCl gastric ulcer model 
BMC Research Notes  2012;5:546.
Background
The decoction of the aerial parts of Rhynchosia recinosa (A.Rich.) Bak. [Fabaceae] is used in combination with the stem barks of Ozoroa insignis Del. (Anacardiaceae), Maytenus senegalensis (Lam.) Excell. [Celastraceae] Entada abyssinica Steud. ex A.Rich [Fabaceae] and Lannea schimperi (Hochst.)Engl. [Anacardiaceae] as a traditional remedy for managing peptic ulcers. However, the safety and efficacy of this polyherbal preparation has not been evaluated. This study reports on the phytochemical profile and some biological activities of the individual plant extracts and a combination of extracts of the five plants.
Methods
A mixture of 80% ethanol extracts of R. recinosa, O. insignis, M. senegalensis, E. abyssinica and L. schimperi at doses of 100, 200, 400 and 800 mg/kg body wt were evaluated for ability to protect Sprague Dawley rats from gastric ulceration by an ethanol-HCl mixture. Cytoprotective effect was assessed by comparison with a negative control group given 1% tween 80 in normal saline and a positive control group given 40 mg/kg body wt pantoprazole. The individual extracts and their combinations were also tested for antibacterial activity against four Gram negative bacteria; Escherichia coli (ATCC 25922), Salmonella typhi (NCTC 8385), Vibrio cholerae (clinical isolate), and Klebsiella pneumoniae (clinical isolate) using the microdilution method. In addition the extracts were evaluated for brine shrimp toxicity and acute toxicity in mice. Phytochemical tests were done using standard methods to determine the presence of tannins, saponins, steroids, cardiac glycosides, flavonoids, alkaloids and terpenoids in the individual plant extracts and in the mixed extract of the five plants.
Results
The combined ethanolic extracts of the 5 plants caused a dose-dependent protection against ethanol/HCl induced ulceration of rat gastric mucosa, reaching 81.7% mean protection as compared to 87.5% protection by 40 mg/kg body wt pantoprazole. Both the individual plant extracts and the mixed extracts of 5 plants exhibited weak to moderate antibacterial activity against four G-ve bacteria. Despite Ozoroa insignis being toxic to mice at doses above 1000 mg/kg body wt, the other plant extracts and the combined extract of the 5 plants were tolerated by mice up to 5000 mg/kg body wt. The brine shrimp test results showed the same pattern of toxicity with Ozoroa insignis being the most toxic (LC50 = 10.63 μg/ml). Phytochemical tests showed that the combined extract of the five plants contained tannins, saponins, steroids, cardiac glycosides, flavonoids and terpenoids. Flavonoids, tannins and terpenoids are known to have antioxidant activity.
Conclusion
The combined extract of the five plants exhibited a dose-dependent protective activity in the rat ethanol-HCl gastric ulcer model. The extracts also exhibited weak antibacterial activity against four Gram negative bacteria and low acute toxicity in mice and brine shrimps. Although the results support claims by traditional healers who use a decoction of the five plants for treatment of peptic ulcers, more models of gastric ulceration and proper animal toxicity studies are needed to validate possible clinical use of the polyherbal extract. It is also evident that the doses of the crude extracts showing protection of the gastric mucosa are too large for realistic translation to direct clinical application, but further studies using bioassay guided fractionation are important to either identify more practical fractions or active compound/s.
doi:10.1186/1756-0500-5-546
PMCID: PMC3532137  PMID: 23031266
Ozoroa insignis; Maytenus senegalensis; Entada abyssinica; Lannea schimperi; Gastroprotection; Toxicity
23.  Antibacterial activities and phytochemical analysis of Cassia fistula (Linn.) leaf 
Cassia fistula Linn. which belongs to family Leguminosae is a medium-sized tree and its different parts are used in ayurvedic medicine as well as home remedies for common ailments. Sequential extraction was carried out using solvents viz. petroleum ether, chloroform, ethanol, methanol and water from leaf of the plant were investigated for preliminary phytochemical and antibacterial property. Results of the study showed that all the extracts had good inhibitory activity against Gram-positive test organism. Although all five extracts showed promising antibacterial activity against test bacterial species, yet maximum activity was observed in ethanol extract. The minimum inhibitory concentration ranged in between 94 to 1 500 μg/ml. Evaluation of phytochemicals such as alkaloids, flavonoids, carbohydrates, glycosides, protein and amino acids, saponins, and triterpenoids revealed the presence of most of constituents in polar extracts (ethanol, methanol, and aqueous) compared with nonpolar extracts (petroleum ether and chloroform). Furthermore, the ethanol extract was subjected to TLC bioautography and time-kill study against Staphylococcus epidermidis. All the findings exhibit that the leaf extracts have broad-spectrum activity and suggest its possible use in treatment of infectious diseases.
doi:10.4103/2231-4040.79814
PMCID: PMC3217677  PMID: 22171295
Cassia fistula; human pathogenic bacteria; minimum inhibitory concentration; Similipal Biosphere Reserve; TLC bioautography
24.  A Review on the Taxonomy, Ethnobotany, Chemistry and Pharmacology of Oroxylum indicum Vent 
Oroxylum indicum Vent. (O. indicum) is a tree commonly called Indian trumpet tree found in tropical countries, such as India, Japan, China, Sri Lanka, Malaysia. The chemical constituents obtained from different parts of plant include baicalein-7-O-diglucoside (Oroxylin B), baicalein-7-O-glucoside, chrysin, apegenin, prunetin, sitosterol, oroxindin, biochanin-A, ellagic acid, baicalein and its 6- and 7-glucuronides, scutellarein, tetuin, antraquinone and aloe-emodin. Various parts of the plant are used in Ayurveda and folk medicine for the treatment of different ailments such as cancer, diarrhea, fever, ulcer and jaundice. Recent in vivo and in vitro studies have indicated its antiinflammatory, antiulcer, hepatoprotective, anticancer, antioxidant, photocytotoxic, antiproliferative, antiarthritic, antimicrobial, antimutagenic and immunostimulant properties. Exhaustive literature survey reveals that there are some activities which are still not proven scientifically. This article is an attempt to compile an up-to-date and comprehensive review on O. indicum covering its traditional and folk medicinal uses, phytochemistry and pharmacology.
doi:10.4103/0250-474X.98981
PMCID: PMC3425058  PMID: 22923859
Ethnomedicinal; ethnopharmacology; O. indicum; phytochemical
25.  In Vitro Phytochemical, Antibacterial, and Antifungal Activities of Leaf, Stem, and Root Extracts of Adiantum capillus veneris 
The Scientific World Journal  2014;2014:269793.
Adiantum capillus veneris is a medicinally essential plant used for the treatment of diverse infectious diseases. The study of phytochemical and antimicrobial activities of the plant extracts against multidrug-resistant (MDR) bacteria and medically important fungi is of immense significance. Extracts from the leaves, stems, and roots of Adiantum capillus veneris were extracted with water, methanol, ethanol, ethyl acetate, and hexane and screened for their antimicrobial activity against ten MDR bacterial strains and five fungal strains isolated from clinical and water samples. Ash, moisture, and extractive values were determined according to standard protocols. FTIR (Fourier transform infrared Spectroscopy) studies were performed on different phytochemicals isolated from the extracts of Adiantum capillus Veneris. Phytochemical analysis showed the presence of flavonoids, alkaloids, tannins, saponins, cardiac glycosides, terpenoids, steroids, and reducing sugars. Water, methanol, and ethanol extracts of leaves, stems, and roots showed significant antibacterial and antifungal activities against most of the MDR bacterial and fungal strains. This study concluded that extracts of Adiantum capillus veneris have valuable phytochemicals and significant activities against most of the MDR bacterial strains and medically important fungal strains.
doi:10.1155/2014/269793
PMCID: PMC3925560  PMID: 24592156

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