In vitro Evaluation of Anti-Inflammatory Activity of Symplocos racemosa Using Protein Denaturation Assay

Introduction: Recently there is considerable awareness and interest in the field of herbal medicine due to its natural origin and lesser side effects compared to Allopathy. Selected herbal plants like Symplocos racemosa, commonly known as lodhra, are found mainly in plains and lower hills of Bengal. The word ‘Lodhra’ means ‘Propitious’. Symplocos racemosa is an important Indian traditional drug used in many Ayurvedic and herbal formulations for treatment of liver as well as uterine disorders and leucorrhea. Ethnobotanical Literature indicates use of Symplocos racemosa in treatment of eye disease, skin disease, ear disorders, liver and bowel complaints, tumours,uterine disorders, spongy and bleeding gums, asthma, fever, snakebite, gonorrhoea and arthritis. Aim: To analyse the anti-inflammatory activity of Symplocos racemosa using protein denaturation assay. Materials and Methods: 2 g of Lodhra bark powder is mixed with 100 ml distilled water & boiled for 20 min at 50°C. The extract is filtered using whatman filter paper & concentrated to 10 ml.1 ml each of Bovine serum albumin is added to various fixations of plant extract (10μL - 50 μL) and the anti - inflammatory activity was evaluated by analysing the percentage inhibition. Results: From this study, it is evident that Lodhra has significant anti-inflammatory activity. At 50μℓ concentration, the plant extract shower higher anti- inflammatory activity of 76%. Conclusion: Symplocos racemosa extract has proved to exhibit effective anti- inflammatory activity. Further studies have to be carried to analyse the other properties of this herb, which can be incorporated successfully in the pharmaceutical industry.

Gas chromatographic methods coupled to mass spectrometry detection of leaf extracts of Symplocos racemosa Roxb.

Symplocos racemosa Roxb., belonging to the family Symplocaceae, is an important medicinal plant. Chemometric profile of chloroform, methanol and ethanol leaf extracts of S. racemosa were performed by hyphenated gas chromatography mass spectrometry (GC/MS) technique. Chemometric profile of leaf extracts revealed the presence of 57 phyto-chemotypes, among them, Erucylamide (15.94%), Vitamin E (8.26%), 10-Heneicosene (8.97%), n-Hexadecanoic acid (5.21%), 1-Heneicosyl formate (5.11%), Tetracosanol (5.36%), 2,4-Hexanedione, 5,5-dimethyl- (11.94%), Linoleic acid (19.30%), Dibutyl phthalate (8.8%) and (Z)-9-Octadecenamide (19.33%) were found to be present in major amount. Eventually, in the present study we have found phenol, alcohol, ketone, alkane and ester as the major group of phytochemotypes in the different leaf extracts of S. racemosa. All these compounds identified by GC/MS analysis were further investigated for their biological activities and it was found that they possess a diverse range of positive pharmacological actions. In future, isolation of individual phyto-chemotypes and subjecting them to biological activity will definitely prove fruitful results in designing a novel drug.

DYEING OF WOOLEN FIBERS WITH RHODODENDRON ARBOREUM FLOWERS USING SYMPLOCOS RACEMOSA LEAVES AS MORDANT UNDER OPTIMUM CONDITIONS

Out of the multiple applications of the diverse natural products of Uttarakhand dyeing of fabrics with the help of various plant parts has always stood out. In the present investigation, an attempt has been made to extract natural dye from Rhododendron arboretum flowers and to optimize conditions of concentration and time for dyeing woolen sample from the extracted dye. The results showed that the colour strength (% Absorbance) increases with the increase in concentration of dye up to 0.03 g/ml concentration. Also, dyeing procedure has been optimized by using Symplocos racemosa leaves as mordant. The best results were obtained using simultaneous mordanting method with Symplocos racemosa leaves at 0.03 g/ml concentration.

Bioprospecting the antimicrobial, antibiofilm and antiproliferative activity of Symplocos racemosa Roxb. Bark phytoconstituents along with their biosafety evaluation and detection of antimicrobial components by GC-MS

Background Plants provide a ray of hope to combat the ever increasing antibiotic resistance and Symplocos racemosa is a valuable medicinal plant. The study focused on highlighting the importance of this plant’s phytoconstituents as potential source of novel antimicrobials against planktonic as well as biofilm forming microorganisms, along with their antiproliferative activity. The biosafety of the phytoconstituents was also established, followed by detection of probable antimicrobial components. Methods The best organic extractant and major groups of phytoconstituents were tested for their antimicrobial activity against reference microbial strains and drug-resistant clinical isolates. The anti-proliferative potential of the most active group of phytoconstituents was evaluated against cancerous cell lines. The in vitro biosafety of phytoconstituents was evaluated by Ames and MTT assay, while in vivo biosafety of the most active phytoconstituents, i.e., flavonoids was determined by acute oral toxicity. Further, the probable antimicrobial components in the flavonoids were detected by TLC and GC-MS. Results Ethyl acetate extract was the most effective among various organic extracts, whereas phytoconstituents such as flavonoids, cardiac glycosides, saponins, tannins, triterpenes and phytosterols were the major groups present, with flavonoids being the most potent antimicrobials. The phytoconstituents displayed a significant antibiofilm potential, as exhibited by inhibition of initial cell attachment, disruption of the pre-formed biofilms and reduced metabolic activity of biofilms. The phytoconstituents were significantly active against the drug-resistant strains of E.coli, MRSA and Salmonella spp. Further, flavonoids showed significant cytotoxic effect against the cancerous cell lines but were non-cytotoxic against Vero (normal) cell line. All the test preparations were biosafe, as depicted by the Ames test and MTT assay. Also, flavonoids did not induce any abnormality in body weight, clinical signs, biochemical parameters and organs’ histopathology of the Swiss albino mice during in vivo acute oral toxicity studies. The flavonoids were resolved into 4 bands (S1-S4), where S3 was the most active and its GC-MS analysis revealed the presence of a number of compounds, where Bicyclo [2.2.1]heptan-2-one,1,7,7-trimethyl-, (1S)- was the most abundant. Conclusions These findings suggest that the phytoconstituents from Symplocos racemosa bark could act as potential source of antimicrobial as well as antiproliferative metabolites.

Enhanced Antibacterial Activity of Silver Nanoparticles Synthesised Using Symplocos Racemosa

The green synthesis methods for the nanoparticles are economically feasible. It does not require any adverse effect causing agents such as usage of temperatures, toxic chemicals, energy, high pressure in the scientific and medical applications. Oral pathogens such as Streptococcus mutans and Lactobacillus sp is the major causes for many oral diseases. The different types of antibiotics are used for the controlling of many oral diseases. In this tudy we enhance the activity of nanoparticles by adding antibiotics to control the oral pathogen growth in invitro. The silver nanoparticles are prepared from Symplocos racemosa plant extract. To study and compare about the enhanced antibacterial activity of silver nanoparticles synthesised using Symplocoa racemosa. The plant extract was prepared by using double distilled water. The microorganisms such as Streptococcus mutans, and Lactobacillus sp were chosen and cultured in respective culture media. The silver nanoparticles synthesised is introduced into cultured petri plates to observe the changes. The zone of inhibition of the growth of microorganisms was studied. The effect of the prepared silver nanoparticles is compared using effect with the normal antibiotics. As a result, it is evident that there is an increase in zone of inhibition comparing the normal antibiotic effect. Thus it can used as drug of the choice in future.

Bioprospecting the antimicrobial, antibiofilm and antiproliferative activity of Symplocos racemosa Roxb. Bark phytoconstituents along with their biosafety evaluation and detection of antimicrobial components by GC-MS

Background: The increased occurrence of the antibiotic resistance has added up to the misery of mankind. There is an immediate need to search alternatives and Symplocos racemosa is one such potent medicinal plant . The aim of the study was to explore various activities of its phytoconstituents, confirm their biosafety and identify the active components.Methods: The ethylacetate extract and the major group of phytoconstituents were tested for minimal inhibitory concentration, viable cell count studies, Post antibiotic effect, activity against 34 multi-drug resistant microorganisms, anti-biofilm and anti proliferative potential. The biosafety was evaluated by Ames and MTT assay. In vivo safety of flavonoids was determined by acute oral toxicity test using Swiss albino mice. The antimicrobial components in flavonoids were also identified by using TLC and GC-MS analysis.Results: Ethyl acetate was the best organic extractant. The phytochemical analyses revealed the presence of flavonoids, cardiac glycosides, saponins, tannins, triterpenes and Phytosterols. Flavonoids exhibited the highest antimicrobial potential whereas tannins, triterpenes and phytosterols were completely inactive. Their Minimum Inhibitory Concentration ranged from 0.5-10 mg/ml; exhibited a bactericidal mode of action and a prolonged post antibiotic effect. The phytoconstituents displayed a significant antibiofilm potential by inhibiting the initial cell attachment, disrupting the pre-formed biofilms and reducing the metabolic activity of biofilms. The phytoconstituents were significantly active against the drug-resistant strains of E.coli , MRSA and Salmonella spp. Flavonoids showed significant cytotoxic effect against the RD, L20B and Hep2 cell lines (IC 50 361-494 µg/ml). All the test extracts were biosafe as depicted by the Ames test, MTT assay. Also, flavonoids did not induce any abnormality in biochemical parameters and Organs’ histopathology of the Swiss albino mice during the in vivo acute oral toxicity studies. The flavonoids were resolve into 4 bands (S1-S4) using TLC, where S3 was the most active and its GC-MS analysis revealed the presence of Bicyclo [2.2.1]heptan-2-one,1,7,7-trimethyl-, (1S)- at an RT of 11.14 as the major compound.Conclusions: These findings suggest that Symplocos racemosa bark could be a potential source of antimicrobial metabolites, which could substantially contribute to the arsenal of existing natural drugs for combating the antimicrobial resistance.