Antimicrobial screening of ethnobotanically important stem bark of medicinal plants.

BACKGROUND: The stem barks are the rich sources of tannins and other phenolic compounds. Tannins inhibited the growth of various fungi, yeast, bacteria and virus. Hence, ten stem barks of ethnomedicinally important plants were screened for antibacterial and antifungal activities against human pathogenic strains. METHODS: Air-dried and powdered stem bark of each plant was extracted with 50% aqueous ethanol, lyophilized and the dried crude extracts were used for the screening against 11 bacteria and 8 fungi. Antibacterial and antifungal activities were performed according to microdilution methods by NCCLS. RESULTS: The plants Prosopis chilensis, Pithecellobium dulce, Mangifera indica showed significant antibacterial and antifungal activities against Streptococcus pneumonia, Enterobacter aerogenes, Klebsiella pneumonia and Candida albicans with MIC of 0.08mg/ml. Pithecellobium dulce bark also showed significant antibacterial activity against Bacillus cereus. CONCLUSION: The bark of Pithecellobium dulce has more or less similar activity against the known antibiotic and may be considered as potent antimicrobial agent for various infectious diseases.

Chromatographic estimation of p-coumaric acid and triacontanol in an Ayurvedic root drug patala (Stereospermum suaveolens Roxb.).

A high-performance thin layer chromatographic method has been established for the identification and standardization of Stereospermum suaveolens extracts using two active biomarkers viz. triacontanol and p-coumaric acid in root samples collected from different geographical locations of India. These marker components have different therapeutic activities: p-coumaric acid is a good antioxidant and also has anti-diabetic properties; triacontanol has anti-inflammatory and anti-ulcer properties. Studies on root as well as stem samples showed characteristic bands of triacontanol and p-coumaric acid at R(f) 0.51 and 0.42, respectively. The amount of triacontanol varied from 0.035-8.472 mg/g in the root samples while the amount of p-coumaric acid was in the range 0.081-0.203 mg/g. The limit of detection for triacontanol was 100 ng/spot whereas that for p-coumaric acid was 10 ng/spot.

Molecular characterization of Desmodium species--an important ingredient of 'Dashmoola' by RAPD analysis.

Identification of medicinal plants by their molecular signature is a fast growing tool. The identification of Desmodium gangeticum (L.) DC. (Shalparni, a constituent of Ayurvedic formulation "Dashmoolarishtha") was carried out using genomic approach. Authentic samples of D. gangeticum(L.) DC., D. velutinum (Willd.) DC. and D. triflorum (L.) DC. were analyzed and compared to commercial samples of various origin. Within twenty primers used, eleven gave 223 RAPD fragments. RAPD profiles of three species showed very low similarity index (0.21-0.39), whereas market samples showed high similarity of 0.82-0.89 with authenticated D. gangeticum.

Chromium-induced changes in ultramorphology and secondary metabolites of Phyllanthus amarus Schum & Thonn. - an hepatoprotective plant.

Chromium (Cr) concentration in the environment is increasing day by day due to its excessive use in leather processing, refractory steel, and chemical manufacturing industries. Excess Cr in the environment causes hazardous effects on all living beings including plants. In this context attempts have been made to see the effect of Cr on a very important medicinal plant Phyllanthus amarus Schum and Thonn. It has focused as a hepatoprotective plant curing hepatitis 'B' in modern research. The P. amarus seedlings of approximately same height and weight were grown in similar conditions in experimental garden. After 30 days plants were harvested and chlorophyll, protein, sugar, starch and secondary metabolites concentration were estimated in these plants as per standard methods. Besides ultramorphological variations in leaves and Cr accumulation in roots and aerial parts were also checked with the help of scanning electron microscope and atomic absorption spectrophotometer respectively. The study revealed that Cr causes significant decrease in fresh and dry weight, length of root and shoot, protein, sugar, chlorophyll and carotenoids in P. amarus. On the other hand it is interesting to note that the concentration of therapeutically active secondary metabolites - phyllanthin and hypophyllanthin increased up to certain level of Cr. It may be due to stress caused by the accumulation of Cr. Besides, ultramorphological changes viz. wide stomatal opening and less wax deposition on both the surfaces of leaves were also observed.

Hepatoprotective potential of Fumaria indica Pugsley whole plant extracts, fractions and an isolated alkaloid protopine.

The present investigation demonstrates the hepatoprotective potential of 50% ethanolic water extract of whole plant of Fumaria indica and its three fractions viz., hexane, chloroform and butanol against d-galactosamine induced hepatotoxicity in rats. The hepatoprotection was assessed in terms reduction in histological damage, changes in serum enzymes (SGOT, SGPT, ALP) and metabolites bilirubin, reduced glutathione (GSH) and lipid peroxidation (MDA content). Among fractions more than 90% protection was found with butanol fraction in which alkaloid protopine was quantified as highest i.e. about 0.2mg/g by HPTLC. The isolated protopine in doses of 10-20mg p.o. also proved equally effective hepatoprotectants as standard drug silymarine (single dose 25mg p.o.). In general all treatments excluding hexane fraction proved hepatoprotective at par with silymarine (p

Toxic metals and organochlorine pesticides residue in single herbal drugs used in important ayurvedic formulation - 'Dashmoola'.

Herbal formulations are getting popularity throughout the world and commercialized extensively for various medicinal properties. WHO has emphasized the need for quality assurance of herbal products, including testing of heavy metals and pesticides residues. 'Dashmoola', a popular herbal formulation, with immunomodulator and febrifugal properties, consists of ten single root drugs. In view of WHO guidelines, single herbal drugs used in 'Dashmoola', were collected from different places of India for testing heavy metals and persistent pesticides residue. Although use of roots in 'Dashmoola' is prescribed in original ayurvedic literature but now many pharmacies use stem in place of roots. Therefore, in the present study both roots and stems were selected for estimation of six heavy metals namely arsenic (As), mercury (Hg), lead (Pb), cadmium (Cd), chromium (Cr) and nickel (Ni). Apart from these, the organochlorine pesticides residue viz. different metabolites of DDT, DDE, isomers of HCH and alpha-endosulfan were checked in total 40 samples of single crude drugs. Heavy metals except Hg, were present in most of the samples. In few samples Pb and Cd concentration were beyond the WHO permissible limits. Although alpha-HCH and gamma-HCH were present in almost all the samples, but other pesticides were not detected in these samples. DDT and DDE were found only in two samples.

Fatty acid composition of some medicinally useful seeds.

Fatty acid composition of seeds from the fruits of Butea monosperma, Jatropha glandulifera and Portulaca oleracea of three different families, namely Papilionaceae, Euphorbiaceae and Portulaceae were determined through Gas Liquid Chromatography to find novel natural sources of essential fatty acids for human health and of economic interest. Comparative studies were also performed to ascertain the utilization of each species for domestic as well as industrial purpose and the quality parameters developed can be utilized as marker characters for the aforesaid seeds used by the pharmaceutical industries.

Action of (2-->1)Fructo-oligopolysaccharide fraction of Chlorophytum borivilianum against Streptozotocin-Induced oxidative stress.

A fructo-oligosaccharide was isolated from Chlorophytum borivilianum and identified as O-beta-D-fructofuranosyl-(2-->1)-(beta-D-fructofuranosyl) (n)-(2-->1)-alpha-D-glucopyranoside (n = 5 - 30) using high-pressure anion exchange chromatography, MALDI-MS, NMR, GC, HPTLC and chemical analysis. The extract and the fructo-oligosaccharide were found to have significant antidiabetic activity with the blood sugar levels being 118.32 +/- 3.56 and 110.21 +/- 4.22, respectively, as compared to the control value of 231.25 +/- 3.03 along with moderate antioxidant activity in streptozotocin-induced diabetic animals.

Estimation of heavy metals in different berberis species and its market samples.

Popularity of herbal drugs is increasing all over the world because of lesser side effects as compared to synthetic drugs. Besides, it costs less and easily available to poor people particularly in developing countries. But quality assurance of herbal drugs is very necessary prior to its use. Because in today's polluted environment, even herbal drugs are not safe. Berberis spp. is very important medicinal plant, having various medicinal properties. It is also included in Indian and British pharmacopoeias. Its demand is quite high in herbal drug market. So, to check the quality of market samples of this drug, ten different samples were procured from different drug markets of India for heavy metal estimation. Besides, genuine samples of four species of Berberis viz. B. aristata, B. chitria, B. lycium, B. asiatica were also collected from natural habitats to compare heavy metal concentration in both market and genuine samples. It was found that market samples were much more contaminated than genuine samples. Lead (Pb) concentration is far beyond from WHO permissible limit (10 ppm) for herbal drugs, reaching to maximum 49.75 ppm in Amritsar market sample. Likewise, concentration of all other metals like Cd, Cr and Ni were also very high in market samples as compared to genuine samples.

Antimicrobial, wound healing and antioxidant activity of Plagiochasma appendiculatum Lehm. et Lind.

Plagiochasma appendiculatum (Aytoniaceae) of the order Marchantiales is widely used in the form of paste ethnomedicinally by Gaddi tribe in Kangra valley for treating skin diseases. In this context, antimicrobical potential of Plagiochasma appendiculatum against a wide range of microorganisms was studied. To validate the ethnotherapeutic claims of the plant in skin diseases, wound healing activity was studied, besides antioxidant activity to understand the mechanism of wound healing activity. The plant (alchoholic and aqueous extract) showed significant antibacterial and antifungal activity against almost all the organisms: Micrococcus luteus, Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Streptococcus pneumoniae, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhimurium, and eight fungi Candida albicans and Cryptococcus albidus-dimorphic fungi, Trichophyton rubrum-dermatophyte fungi, Aspergillus niger, Aspergillus flavus, Aspergillus spinulosus, Aspergillus terreus and Aspergillus nidulans-systemic fungi, with especially good activity against the dermatophyte (Trichophyton rubrum) and some infectious bacteria (Escherichia coli, Proteus mirabilis and Salmonella typhimurium) with an MIC of 2.5 microg/disc. The results show that Plagiochasma appendiculatum extract has potent wound healing capacity as evident from the wound contraction and increased tensile strength. The results also indicated that Plagiochasma appendiculatum extract possesses potent antioxidant activity by inhibiting lipid peroxidation and increase in the superoxide dismutase (SOD) and Catalase activity.

Comparative pharmacognostic studies of three Phyllanthus species.

Different species of Phyllanthus are considered to be very effective hepatoprotective agents in the Indian indigenous systems of medicine and are considered bitter, astringent, stomachic, diuretic, febrifuge, deobstruant and antiseptic. Still ayurvedic practitioners prescribed fresh juice of 'Bhuiamlki' for jaundice. Various species of Phyllanthus are being sold in India under the trade name 'Bhuiamlki'. During market surveillance of herbal drug, it was observed that almost all the commercial samples, either comprise of Phyllanthus amarus Schum & Thonn. or Phyllanthus maderaspatensis Linn. or mixture of Phyllanthus amarus, Phyllanthus fraternus Webster. and Phyllanthus maderaspatensis. Therefore, in this context the detailed pharmacognostical evaluation of all the three species has been carried out with the aim to establish the identification markers of this important hepatoprotective agent (effective in hepatitis B too). The study conclude that all the three species can be differentiated on the basis of macro and microscopic characters, physico-chemical values, HPTLC fingerprint profile, and detection of phyllanthin and hypophyllanthin as marker components. Besides, an interesting conclusion can also be drawn that phyllanthin and hypophyllanthin said to protect hepatocytes against carbon tetrachloride and galactosamine induced toxicity, may not be exclusively responsible for hepatoprotective activity as these are present only in Phyllanthus amarus while Phyllanthus fraternus and Phyllanthus maderaspatensis also possess significant hepatoprotective activity.

Free radical scavenging potential of Chlorophytum tuberosum Baker.

Chlorophytum tuberosum Baker commonly referred as 'Musli' has been widely used as a potent 'Rasayana' drug in 'Ayurveda' as a rejuvenator and tonic. Antioxidant potential of Chlorophytum tuberosum has been investigated for their ability to scavenge 1,1,diphenyl picryl hydrazyl (DPPH), nitric oxide radical along with their capacity to reduce lipid peroxidation in rat liver homogenate, chelation of ferrous ion, radical scavenging potential using chemiluminescence and their total antioxidant capacity. Sugar, starch, protein, and Vitamin C content were estimated spectrophotometrically along with the percentages of the individual amino acids by HPLC and individual sugars by using HPTLC as standardization tool. The extract has been found to possess antioxidant activity in all the models tested as evident by IC50 values being 225.31, 888.44, 809.22 and 422.97 microg/ml for scavenging of DPPH, nitric oxide, lipid peroxidation and ferry bi-pyridyl complex, respectively, along with a integral anitoxidant activity of 2.986 nmol ascorbic acid/g equivalents in photochemiluminescence assay.

Antioxidant activity of Desmodium gangeticum and its phenolics in arthritic rats.

Total alcoholic extract of Desmodium gangeticum, which exhibited significant anti-inflammatory activity, was evaluated for the possible mode of action by studying its antioxidant potential in adjuvant-induced arthritic rats. Activity guided fractionation and isolation were carried out. The phenolics fraction showed maximum potency. Solid phase extraction followed by preparative HPLC of the active phenolic fraction yielded for the first time two potent antioxidant compounds, caffeic acid and chlorogenic acid, from this plant. The biological antioxidant defense system, involving superoxide dismutase, glutathione and catalase, showed a significant increase with their levels close to the normal control with a decrease in the lipid peroxide content upon administration of D. gangeticum extract (100 mg kg(-1)) and its phenolics (50 mg kg(-1)) in arthritic rats, thereby indicating the extracts antioxidant property under arthritic conditions.

Effect of cadmium on growth, ultramorphology of leaf and secondary metabolites of Phyllanthus amarus Schum. and Thonn.

The pollution is increasing in the environment by different kinds of human activities, which results in the accumulation of heavy metals including cadmium in the soil and water and it causes different types of problems to living beings. As the plants are utilized by human being as food and medicine, therefore, it is mandatory to see the effect of metals on plants. In this context, efforts have been made to observe the effect of different concentration of Cadmium (Cd) on Phyllanthus amarus Schum. and Thonn., because Cd is the widespread metal and the plants response to low and high level of exposure is a complex phenomenon. P. amarus is mostly grown as weed in agricultural and waste lands. It is a reputed plant used in Indian indigenous systems of medicine with hepatoprotective, diuretic, stomachic properties and is recently being used for the treatment of hepatitis B. The study revealed that Cd causes significant decrease in fresh and dry weight, length of root and shoot, protein, chlorophyll, carotenoids and sugar and increase in starch content. It is interesting to note that the therapeutically active compounds-phyllanthin and hypophyllanthin, enhanced at certain levels of Cd due to abiotic stress. Besides, the ultramorpholical changes were also observed in stomatal opening and wax deposition on both the surfaces of leaves.

Antioxidant potential of Anogeissus latifolia.

Anogeissus latifolia is widely used in the Indian indigenous system of medicine and is reported to contain leucocyanidins and tannoid principles like ellagic acid and its derivatives. In view of its wide use and its chemical composition, this study was aimed at examining the antioxidant activity of the extract of A. latifolia. The extract was studied for total antioxidant capacity, hydrogen-donating ability, nitric oxide, superoxide scavenging activity, hydrogen peroxide decomposition activity along with lipid peroxidation. Integral antioxidative capacity was determined by chemiluminescence assay. The extract was also studied for lipid peroxidation assay by thiobarbituric acid-reactive substances (TBARS) method using rat liver homogenate. The results indicate that A. latifolia extract has potent antioxidant activity. Also to ascertain the possible reason for the potent activity, percentage of gallic acid was estimated and was found to be 0.95%, which could be one of the reasons for potent antioxidant activity exhibited by the plant.

Healing potential of Anogeissus latifolia for dermal wounds in rats.

Wound healing potential of ethanolic extract of Anogeissus latifolia bark (ALE) for treatment of dermal wounds in rats was studied on excision and incision wound models. HPTLC of the total extract was recorded for the purpose of standardization. Various parameters of incision wound, viz. epithelization period, scar area, tensile strength and hydroxyproline measurements along with wound contraction, were used to evaluate the effect of A. latifolia on wound healing. The results obtained indicate that A. latifolia accelerates the wound healing process by decreasing the surface area of the wound and increasing the tensile strength. Nitrofurazone ointment was used as a positive control. Complete epithelization was observed within 15 days with ALE. Measurements of the healed area and the hydroxyproline level were in agreement. Antibacterial activity of ALE was studied against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae) compared to erythromycin and tetracycline. Moderate activity was observed against all organisms. The present study provides a scientific rationale for the traditional use of Anogeissus latifolia in the management of skin diseases such as sores, boils and itching.

Swertia chirayita mediated modulation of interleukin-1beta, interleukin-6, interleukin-10, interferon-gamma, and tumor necrosis factor-alpha in arthritic mice.

The effect of aqueous extract of Swertia chirayita stem on the pro- and anti-inflammatory cytokines balance in primary joint synovium of adjuvant-induced arthritic mice has been studied. The level of pro-inflammatory cytokines was found elevated in the joint synovium of arthritic mice in comparison to normal joints. Administration of S. chirayita extract in varying doses through the oral route did not modulate the proinflammatory cytokines on day 2. In contrast, by day 12, a dose dependent (0, 11.86 and 23.72 mg/kg body weight) reduction of tumor necrosis factor-alpha (INF-alpha) interleukin-1beta, (IL-beta) and interferon-gamma, (IFN-gamma) and elevation of Interleukin-10 (IL-10) was observed in the joint homogenates of arthritic mice. Interleukin-6 (IL-6) was not down regulated in joint homogenate of arthritic mice at the dose 11.86 mg/kg but at higher doses (23.72 and 35.58 mg/kg) significant reduction was observed. The aqueous extract was found to possess two polar compounds, amerogentin and mangiferin but was devoid of swerchirin, chiratol, methyl swetianin, and swertanone. Mangiferin has been reported to possess potent anti-inflammatory property and we presume its presence in the aqueous extract of S. chirayita is responsible for reducing TNF-alpha, IL-1beta, IL-6, and IFN-gamma and/or elevating IL-10 in the joint homogenates of arthritic mice on day 12. This study will help in our understanding of the mechanism of anti-inflammatory action of S. chirayita in the light of proinflammatory and anti-inflammatory cytokine balance.

Studies on the antioxidant activities of Desmodium gangeticum.

Desmodium gangeticum is herbal species which is widely used in the indigenous system of medicine and is reported to contain flavone and isoflavanoid glycosides. In view of its wide use and it's chemical composition, this study was aimed at examining the antioxidant activity of the extract of D. gangeticum. The extract was studied for diphenyl picryl hydrazyl (DPPH), nitric oxide, ferryl-bipyridyl and hypochlorous acid scavenging activity along with lipid peroxidation. Nitric oxide was generated using sodium nitroprusside and was studied using Griess reagent. In order to study the iron chelating capacity of the extract, the percentage ferryl-bipyridyl inhibition was studied. Hypochlorous acid scavenging activity was tested by measuring the inhibition of 5-thio-2-nitrobenzoic acid oxidation. The extract was also studied for lipid peroxidation assay by thiobarbituric acid-reactive substances (TBARS) method using rat brain homogenate. The results indicate that D. gangeticum extract has potent antioxidant activity.

Spectrophotometric method for estimation of alkaloids precipitable with Dragendorff's reagent in plant materials.

A rapid, easy, and simple spectrophotometric method was developed for the estimation of total alkaloids precipitated by Dragendorff's reagent (DR) in plant materials. It is based on the formation of yellow bismuth complex in nitric acid medium with thiourea. The yellow-colored complex formed obeys Lambert-Beer's law in the concentration range of 0.06-50 microg/mL with lambdamax at 435 nm. Using this method, the alkaloidal percentage of certain alkaloids (ajamalicine, papaverine, cinchonine, piperine, berberine) and some plant materials containing alkaloids (Berberis aristata, Solanum nigrum, and Piper longum) were determined. The method was compared with other methods. It can be used for routine analysis of commercial samples by industries dealing with herbal drugs for standardization of plant materials containing alkaloids and for alkaloid-containing pharmaceutical products.

Free radical scavenging potential of Picrorhiza kurrooa Royle ex Benth.

For assessing free radical scavenging potential of P. kurrooa, the antioxidant activity of P. kurrooa extract was studied by lipid peroxidation assay using rat liver homogenate. The extract (1 mg/ml) showed marked protection (up to 66.68%) against peroxidation of liver phospholipids. Besides, reduced glutathione showed very encouraging activity. The extract also exhibited significant scavenging activity. Thus augmenting the wide use of plant in the indigenous system of medicine, which may partly be due to antioxidant and free radical scavening activity of the extract.