Vinayak Upadhya

Evaluation of wound healing property of Caesalpinia mimosoides Lam.

ETHNOPHARMACOLOGICAL RELEVANCEnCaesalpinia mimosoides Lam. is one of the important traditional folk medicinal plants in the treatment of skin diseases and wounds used by healers of Uttara Kannada district of Karnataka state (India). However scientific validation of documented traditional knowledge related to medicinal plants is an important path in current scenario to fulfill the increasing demand of herbal medicine.nnnAIM OF THE STUDYnThe study was carried out to evaluate the claimed uses of Caesalpinia mimosoides using antimicrobial, wound healing and antioxidant activities followed by detection of possible active bio-constituents.nnnMATERIALS AND METHODSnExtracts prepared by hot percolation method were subjected to preliminary phytochemical analysis followed by antimicrobial activity using MIC assay. In vivo wound healing activity was evaluated by circular excision and linear incision wound models. The extract with significant antimicrobial and wound healing activity was investigated for antioxidant capacity using DPPH, nitric oxide, antilipid peroxidation and total antioxidant activity methods. Total phenolic and flavonoid contents were also determined by Folin-Ciocalteu, Swain and Hillis methods. Possible bio-active constituents were identified by GC-MS technique. RP-UFLC-DAD analysis was carried out to quantify ethyl gallate and gallic acid in the plant extract.nnnRESULTSnPreliminary phytochemical analysis showed positive results for ethanol and aqueous extracts for all the chemical constituents. The ethanol extract proved potent antimicrobial activity against both bacterial and fungal skin pathogens compared to other extracts. The efficacy of topical application of potent ethanol extract and traditionally used aqueous extracts was evidenced by the complete re-epithelization of the epidermal layer with increased percentage of wound contraction in a shorter period. However, aqueous extract failed to perform a consistent effect in the histopathological assessment. Ethanol extract showed effective scavenging activity against DPPH and nitric oxide free radicals with an expressive amount of phenolic and moderate concentration of flavonoid contents. Ethyl gallate and gallic acid were found to be the probable bio-active compounds evidenced by GCMS and RP-UFLC-DAD analysis.nnnCONCLUSIONnThe study revealed the significant antimicrobial, wound healing and antioxidant activities of tender parts of C. mimosoides and proved the traditional folklore knowledge.

Non-codified traditional medicine practices from Belgaum Region in Southern India: present scenario

BackgroundTraditional medicine in India can be classified into codified (Ayurveda, Unani, Siddha, Homeopathy) and non-codified (folk medicine) systems. Both the systems contributing equally to the primary healthcare in India. The present study is aimed to understand the current scenario of medicinal practices of non-codified system of traditional medicine in Belgaum region, India.MethodsThe study has been conducted as a basic survey of identified non-codified traditional practitioners by convenience sampling with semi structured, open ended interviews and discussions. The learning process, disease diagnosis, treatment, remuneration, sharing of knowledge and socio-demographic data was collected, analysed and discussed.ResultsOne hundred and forty traditional practitioners were identified and interviewed for the present study. These practitioners are locally known as “Vaidya”. The study revealed that the non-codified healthcare tradition is practiced mainly by elderly persons in the age group of 61 years and above (40%). 73% of the practitioners learnt the tradition from their forefathers, and 19% of practitioners developed their own practices through experimentation, reading and learning. 20% of the practitioners follow distinctive “Nadi Pariksha” (pulse examination) for disease diagnosis, while others follow bodily symptoms and complaints. 29% of the traditional practitioners do not charge anything, while 59% practitioners receive money as remuneration.Plant and animal materials are used as sources of medicines, with a variety of preparation methods. The preference ranking test revealed higher education and migration from villages are the main reasons for decreasing interest amongst the younger generation, while deforestation emerged as the main cause of medicinal plants depletion.ConclusionPatrilineal transfer of the knowledge to younger generation was observed in Belgaum region. The observed resemblance in disease diagnosis, plant collection and processing between non-codified traditional system of medicine and Ayurveda require further methodical studies to establish the relationship between the two on a more objective basis. However, the practice appears to be at crossroads with threat of extinction, because of non-inheritance of the knowledge and non-availability of medicinal plants. Hence conservation strategies for both knowledge and resources at societal, scientific and legislative levels are urgently required to preserve the traditional wisdom.

Total polyphenolic contents and in vitro antioxidant properties of eight Sida species from Western Ghats, India

Background: Sida L., is a medicinally important genus, the species of which are widely used in traditional systems of medicine in India. Pharmacologically, roots are known for anti-tumor, anti-HIV, hepatoprotective, and many other properties. Phenolic antioxidants help in reducing oxidative stress occurring during treatment of such diseases. Objective: The study aimed to evaluate and compare polyphenol contents and antioxidant properties of eight selected species of Sida from Western Ghats, India. Materials and Methods: Methanolic root extracts (10% w/v) of Sida species, viz., S. acuta, S. cordata, S. cordifolia, S. indica, S. mysorensis, S. retusa, S. rhombifolia, and S. spinosa were analyzed. Results: Sida cordifolia possessed highest total phenolic content (TPC: 1.92 ± 0.10 mg Caffeic Acid Equivalent/g and 2.13 ± 0.11 mg Tannic Acid Equivalant/g), total flavonoid content (TF: 2.60 ± 0.13 mg Quercetin Equivalent/g) and also possessed highest antioxidant activities in 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging (51.31 ± 2.57% Radical Scavenging Activity, (RSA); Trolox Equivalent Antioxidant Capacity: 566.25 ± 28.31μM; Ascorbic acid Equivalent Antioxidant Capacity: 477.80 ± 23.89 μM) and Ferric Reducing Antioxidant Power assays (TEAC: 590.67 ± 29.53 μM; AEAC: 600.67 ± 30.03 μM). Unlike DPPH and Ferric Reducing Antioxidant Power (FRAP) activity, 2, 2Ͳ-Azinobis (3-ethyl Benzo Thiazoline-6-Sulfonic acid) ABTS + antioxidant activity was highest in S. indica (TEAC: 878.44 ± 43.92 μM; AEAC 968.44 ± 48.42 μM). It was significant to note that values of AEAC (μM) for all the antioxidant activities analyzed were higher than that of TEAC. Conclusion: The high contents of phenolic compounds in the root extracts of selected Sida species have direct correlation with their antioxidant properties. Conclusively, roots of S. cordifolia can be considered as the potential source of polyphenols and antioxidants.

Accumulation and trends in distribution of three triterpenoids in various parts of Achyranthes coynei determined using RP-UFLC analysis

Background: Achyranthes coynei Sant. (Family: Amaranthaceae) is a rare endemic medicinal plant used by local traditional practitioners to treat various diseases. The plant has been reported for promising antibacterial and antioxidant activities. However, the plant is not explored for its phytocompounds, especially triterpenoids. Objective: To study the accumulation and trends in distribution of triterpenoids: betulinic, oleanolic and ursolic acids (BA, OA and UA) in leaf, stem, root and inflorescence of A. coynei. Materials and Methods: Extraction was achieved using refluxing and reversed phase-ultra flow liquid chromatographic (RP-UFLC) technique was employed for determination. Separation of triterpenoids was achieved on a Hibar 250-4.6 mm, 5 μ, Lichrospher 100, C18e column using methanol and water (90:10) as mobile phase (pH adjusted to 5.0 using GAA) in an isocratic mode. Results: Oleanolic acid was higher in leaf (0.172 ± 0.009%) followed by stem (0.035 ± 0.002%) and root (0.028 ± 0.001%). Ursolic acid was accounted to be highest in the inflorescence (0.099 ± 0.005%). The contents of BA and UA were lower than OA in leaf and stem while it was remained undetected in roots of A. coynei. Conclusion: The triterpenoids: BA, OA and UA were detected, quantified and reported for the first time from A. coynei. In the present study leaves were found to be the major source of BA and OA, whereas inflorescence was for UA.

Total Polyphenols, Antioxidant and Antimicrobial Activity of Leaves and Stem Bark Extracts of Allophylus cobbe (L.) Raeusch

Allophylus cobbe is a small tree and an ethnomedicinally important plant species belonging to the family Sapindaceae, traditionally used to treat various ailments. Previous work on the species led to show ethanolic extracts to be active against Ranikhet disease virus and have showed effects on central nervous system and hypothermia. However, literature on its polyphenolic content, antioxidant and antimicrobial activity is inadequate. Because of its wide use in traditional medicine, it was decided to evaluate total polyphenols, antioxidant activity and to screen the antimicrobial activity of leaves and stem bark of A. cobbe. Leaves yielded 63.20xa0mg tannic acid equivalent per gram total phenolics and 17.39xa0mg quercetin equivalent per gram total flavonoids. A 0.01xa0% extract was sufficient to produce 32.78xa0% radical scavenging activity (RSA). 2,2′-Azinobis (3-ethyl benzo thiazoline-6-sulfonic acid) activity using ascorbic acid and Trolox® as standards were higher than ferric reducing antioxidant power activity. Both the extracts were more effective on gram positive organism, than gram negative organisms and were ineffective on fungal strains. The results herein report methanolic leaf and stem bark extracts of A. cobbe to possess good antibacterial and antioxidant activity.Allophylus cobbe is a small tree and an ethnomedicinally important plant species belonging to the family Sapindaceae, traditionally used to treat various ailments. Previous work on the species led to show ethanolic extracts to be active against Ranikhet disease virus and have showed effects on central nervous system and hypothermia. However, literature on its polyphenolic content, antioxidant and antimicrobial activity is inadequate. Because of its wide use in traditional medicine, it was decided to evaluate total polyphenols, antioxidant activity and to screen the antimicrobial activity of leaves and stem bark of A. cobbe. Leaves yielded 63.20xa0mg tannic acid equivalent per gram total phenolics and 17.39xa0mg quercetin equivalent per gram total flavonoids. A 0.01xa0% extract was sufficient to produce 32.78xa0% radical scavenging activity (RSA). 2,2′-Azinobis (3-ethyl benzo thiazoline-6-sulfonic acid) activity using ascorbic acid and Trolox® as standards were higher than ferric reducing antioxidant power activity. Both the extracts were more effective on gram positive organism, than gram negative organisms and were ineffective on fungal strains. The results herein report methanolic leaf and stem bark extracts of A. cobbe to possess good antibacterial and antioxidant activity.

Evaluating Nothapodytes nimmoniana population from three localities of Western Ghats using camptothecin as phytochemical marker and selection of elites using a new-content range chart method

Background: Nothapodytes nimmoniana (Grah.) Mabb. is a high valued medicinal plant endemic to Western Ghats of India, distributed in fragmented populations. The plant is valued for potent anticancer drug camptothecin (CPT). Objective: The study compares and expounds variation in CPT content from leaves and stems of N. nimmoniana obtained from three populations of Western Ghats, India. The study also describes a method for categorizing these populations using content range chart (CRC) method for percent yield of CPT. Materials and Methods: A total of 60 samples were investigated including ten each of leaves and stems from three localities. Micro-extraction method was implemented to extract CPT. reversed phase ultra-performance liquid chromatography photo diode array technique was used to quantify CPT. Results: Leaf samples of an individual collected from Joida, yielded lowest CPT content (0.002 ± 0.000 g/100 g), whereas a stem sample from Amgaon, yielded highest CPT content (0.123 ± 0.006 g/100 g). The findings suggest great variation in individuals producing and accumulating CPT. Using this data along with earlier published work, five categories of CPT yielding plants were made viz. I: Very low: <0.020, II: Low: 0.021-0.039, III: Moderate: 0.040-0.059, IV: High: 0.060-0.079 and V: Very high: >0.080. Based on CPT content in leaves, majority of individuals were under very low category (Ist) and on the other hand stem samples were in ‘II’ category. Besides, very few individuals were observed in category ‘V’. Conclusion: The study expounds use of CRC method for identifying elite population and suggests the need for its conservation.

Pharmacognostic Screening of Piper trichostachyon Fruits and its Comparative Analysis with Piper nigrum Using Chromatographic Techniques.

Background: Piper trichostachyon is a wild, endemic Piper species from Western Ghats of India. The folklore healers of Belagavi region use this plant, similar to Piper nigrum. Aims: The present study investigates the comparison between P. nigrum and P. trichostachyon using pharmacognostic parameters. Materials and Methods: Pharmacognostic evaluation was carried out in terms of morphological, microscopic characters, and phytochemical analysis using standard methods. Comparative physicochemical analysis between P. trichostachyon and P. nigrum was also carried out through estimation of micro-macro nutrients, high-performance thin layer chromatography (HPTLC) investigation and using piperine as a marker compound for reversed phase-ultra flow liquid chromatographic (RP-UFLC) technique. Results: P. trichostachyon grows in the forests, and the fruits are morphologically similar to P. nigrum fruits, so the name in Kannada “Kaadu Kalu menasu” (wild/forest black pepper). The microscopy revealed the presence of stone cells, starch grains, oil cells and globules, beaker cells, and yellowish brown pigment layer, parenchymatous cells. The presence of alkaloids, oil, and tannins were observed in P. trichostachyon fruits. The HPTLC studies visibly indicated differences among two species with 12 peaks and varied banding pattern. RP-UFLC results showed less amount of piperine in P. trichostachyon (0.05 ± 0.002 mg/g) than in P. nigrum (16.14 ± 0.807 mg/g). Conclusion: The study reports on pharmacognostic parameters of P. trichostachyon for the 1st time and will be useful for the identification and authentication. The comparative HPTLC and RP-UFLC studies resolve the differentiation impasse among two species. However, further biological efficacy studies are required to establish its use in traditional medicine. SUMMARY Piper trichostachyon grows in the forests, and the fruits are morphologically similar to Piper nigrum fruits The microscopy of P. trichostachyon revealed the presence of stone cells, starch grains, oil cells and globules, beaker cells and yellowish brown pigment layer, parenchymatous cells The high-performance thin layer chromatography studies visibly indicated differences among two species with varied banding pattern Reversed phase-ultra flow liquid chromatographic results showed less amount of piperine in P. trichostachyon than in P. nigrum. Abbreviation used: HPTLC: High Performance Thin Layer Chromatography, RP-UFLC: Reversed phase-ultra fl ow liquid chromatographic analysis, DST: Length of line, Maj: Length of large half axis for ellipse RDS - radius for circle, Rf: Retention Factor, TS: Transverse Section, TLC: Thin Layer Chromatography.

Pharmacognostic evaluation of Achyranthes coynei: Leaf

Abstract Achyranthes coynei Sant., (Amaranthaceae) is a rare, medicinal shrub, endemic to India. The plant especially, leaves are used in treatment of various disorders by folk healers. It has been scientifically evaluated for its antimicrobial and antioxidant properties. Pharmacognostic studies have not been carried out so far in this plant. So, the present study was undertaken to evaluate pharmacognostic characters of leaf of A. coynei. The studies were carried out in terms of morphological, microscopic characters and physicochemical parameters using standard methods. Leaf size (15–23 × 7–10 cm) and petiole nature (pink above and green beneath) were the distinguishing morphological character observed in the present study. Transverse section of leaf and petiole showed presence of long, multi-cellular, warty trichomes with pointed apex and short, glandular trichomes with blunt apex on thick walled epidermis; conjoint, collateral closed, endarch vascular bundles; rosette crystals of calcium oxalate in ground tissue. Ash and extractive values, micro and macro elements and nutritive factors were estimated. Leaf powder showed fluorescence under the influence of different solvents. Preliminary phytochemical screening showed the presence of alkaloids, saponins and triterpenoids. High Performance Thin Layer Chromatorgraphic (HPTLC) analysis yielded eight bands in leaf extract. The study forms the first report on pharamcognostic characters for A. coynei, which could be useful for identification and authentication of the plant.

Preliminary pharmacognostic screening of Achyranthes coynei stem

Achyranthes coynei is a rare, endemic perennial shrub reported from Karnataka and Maharashtra states of India. The plant is used to treat various disorders by folk healers and was proven to have antimicrobial and antioxidant properties. The present study was undertaken to evaluate microscopic and macroscopic characters of A. coynei stem, along with its physicochemical parameters. ProgRes ® CapturePro and Microsoft Excel were used for statistical analysis. Perennial, shrubby nature and woody stem were the distinguishing morphological characters observed. Transverse section (TS) illustrated quadrangular outline of the stem and showed the presence of two types of trichomes on the thick-walled epidermis. TS also showed number of rosette calcium oxalates crystals; prismatic and microsphenoid crystals; conjoint, collateral open secondary vascular bundles; and two amphixylic medullary bundles in the pith. Ash and extractive values, micro and macro elements and nutritive factors were estimated in the present study. The presence of alkaloids, saponins and triterpenoids were observed in preliminary phytochemical screening. High-performance thin layer chromatographic analysis yielded different bands and also indicated the presence of oleanolic acid. The studied parameters for A. coynei stem will be useful for identification and authentication of the plant material.