Phytochemical Characterisation, Antioxidant and Antidiabetic activity of extracts of Neptunia prostrata Linn.
Raja Chakraverty1*, Chowdhury Mobaswar Hossain2, Anjan Adhikari3,
Pranabesh Chakraborty4
1Doctoral Scholar (Pharmaceutical Technology), Maulana Abul Kalam Azad University of Technology,
Nadia - 741249, West Bengal.
2 Head, Department of Pharmaceutical Technology, Maulana Abul Kalam Azad University of Technology, Haringhata, Nadia - 741249, West Bengal.
4Director, School of Food, Pharmaceutical, Medical Science and Technology,
Makaut, Haringhata, Nadia - 741249, West Bengal.
3Head, Department of Pharmacology, MJN Medical College and Hospital, Coochbehar, West Bengal.
*Corresponding Author E-mail: rchakraborty20@yahoo.com
ABSTRACT:
The herb Neptunia prostrata Linn. Is belonging to the family Mimosaceae have been used in folkloric medicine in the North-eastern states of Indiaof Assam and Tripura by indigenous herbal healers since time immemorial but there is a scarcity of any background study documenting its use as an antimicrobial herb. For the same, plants were collected and authenticated. Following identification of these herbs methanolic, ethanolic, pet ether and chloroform extracts were prepared using soxhletion. Acute toxicity study as per OECD guidelines 420 was assessed in wistar albino rats and in swiss albino mice (n=5) of both sexes at doses of 2000mg/kg body weight and did not reveal any morbidity or mortality in the animals within the stipulated period.Phytochemical screening was performed on all four extracts of Neptunia prostrata. Phytochemical constituents depicted presence of glycoside, flavonoids in only ethanolic, methanolic and chloroform extracts. Alkaloids were present in the chloroform extract. The antimicrobial activity was performed by disc agar diffusion method with respect to amoxicillin at standard doses against ATCC strains of Gram positive Staphylococcus aureus, Bacillus subtilis and Gram negative Salmonella typhi. The herbs showed antioxidant activity comparable to standard antioxidants in-vitro such as Ascorbic acid (Vitamin C) with comparable IC 50 values. Results of the antibacterial assay on the basis of zone of inhibition (mm) and MIC values of extracts of the extract (NPHE) under study to suggest these two indigenous herb have conspicuous and potent putative role in the therapeutics of a vast plethora bacterial infections that need to be corroborated for the expansion of future prospective in-vivo studies with larger sample size.
KEYWORDS: Neptunia prostrata, Antioxidant, Antimicrobial activity, MIC.
INTRODUCTION:
The practice of indigenous medicine has time and again focused on herbs for their innate antimicrobial activity against a plethora of bacteria and moulds since ages1,2. Neptunia Prostrata, is one such herb growing in the states of Assam, Tripura of India and is regarded as a variant of the“Touch-me not plant”Mimosa pudica known locally by the synonym “Water- Mimosa”.
The leaves of this herb have found use in folkloric medicine and ancient vedic texts 3,4 since time immemorial.
The herb under study Neptunia prostrata Linn. (Synonym: Neptunia oleracea belonging to Family: Mimosaceae) is macroscopically a miniature aquatic herb that floats by its white spongy structure and has been reported to be a source of antioxidants5-8. Since traditionally antioxidants in aerial parts of many herbs have reported a close link to antimicrobial or antibacterial properties this led us to the objective of the study to elucidate any such properties associated with leaves this herb.
For the evaluation of the antimicrobial activity of the leaf extract of Neptunia prostrata literature review revealed that the indigenous ethnic groups of Tripura cultivate this plant both as vegetable as well as medicinal plant and prepare various tasty delicacy dishes with this vegetable. The plant has been used for different types of remedies like gastritis, acidity, constipation, dysentery, and reported to possess hepatoprotective, analgesic and antimicrobial activity 9-11.
MATERIAL AND METHOD:
Acute toxicity study as per OECD guidelines 420 11,12
The standard method of oral acute toxicity study was contacted on rodents as per OECD Guideline no 420. Wistar albino rats of both sexes (n=5) were administered 2000 mg/ kg dose of the Neptunia prostrata extract (NP-1) used for the study and observed for 14 days for mortality of any behavioural abnormalities.
Identification of plant materials:
The herbarium containing the dried plant materials were processed and identified as Neptunia prostrata respectively by the office of the Botanical Survey of India, Shibpur and a voucher was accordedthe number BST/Herb/2016/003 respectively. A copy of the same was maintained for further use.
Antioxidant assay:
Antioxidants play an important role as health protecting factor. Scientific evidence suggests that antioxidants reduce the risk for chronic diseases including cancer and heart disease. Primary sources of naturally occurring antioxidants are whole grains, fruits and vegetables. Plant sourced antioxidants like vitamin C, vitamin E, carotenes, phenolic acids etc. have been recognized as having the potential to reduce disease risk. Most of the antioxidant compounds in a typical diet are derived from plant sources and belong to various classes of compounds with a wide variety of physical and chemical properties16,17.
The free radical scavenging activity was found to be high in methanolic extract for DPPH, hydroxyl, nitric oxide, superoxide, hydrogen peroxide (H2O2) radical. The content of total phenols, flavonoids and tannins was also found to be high in methanolic extract which may be correlated to its antioxidant activity18- 20.
Determination of DPPH radical scavenging activity:
DPPH radical scavenging assay is a widely used method to evaluate the free radical scavenging ability of natural compounds. This assay is based on the measurement of the scavenging ability of antioxidant substances towards the stable radical. The free radical scavenging activity of the extracts was examined in vitro using DPPH radical 21, 22.
The free radical DPPH (1,1-Diphenyl 1-2-pieryl-hydrazil) 0.1 mM solution of DPPH in ethanol was prepared and 1ml of this solution was added to 3ml of various concentrations of extracts of Neptunia prostrata (25, 50, 75, 100, 125, 150 μgm/ml) of ethanol extracts. After 30mints put in the incubators. Then absorbance was measured at 517nm. The percentage of inhibition was calculated by comparing the absorbance of controls and test samples 23.
The IC 50 value of the sample, which is the concentration of sample required to inhibit 50% of the DPPH free radical, was calculated using Log dose inhibition curve. Lower absorbance of the reaction mixture indicated higher free radical activity 24.
Where a control is the absorbed of the control reaction A and A control and A testis the absorbance in the presence of the samples of the extracts. The antioxidant activity of the extract was expressed as IC 50. The IC 50 values are defined as the concentrations in (micro gram/ml) of extracts that inhibits the formation of DPPH radicals by 50% 25.
RESULT:
Diphenyl pycrylhydrazyl (DPPH) assay of the extracts (Summarised in Figure 1)
The IC50value of the extract was 11.2 µg/mlwhereas the same for Ascorbic acid was 18.02 µg/ml
Figure 1: IC 50 value of NP-1 extracts is 11.02µg/ml (IC 50Ascorbic acid=18.02µg/ml)
Figure 2: IC50 value of extract of Neptunia prostrata (NP-2) = 101.03 µg/ml, IC50value of Ascorbic acid (Vitamin C) = 93 µg/ml.
Extracts used for Phytochemical Screening (summarised in Table 1)
|
Phytoconstituents |
NP1 extract |
NP2 extract |
|
Tannins |
- |
- |
|
Alkaloids |
++ |
++ |
|
Flavonoids |
+++ |
+ |
|
Glycosides |
+++ |
++ |
Indicated the presence of alkaloids, glycosides and flavonoids in NP 1 and NP 2 leaf extracts respectively:
The antimicrobial study using the extracts revealed that the plant possesses chemical constituents in the extract and has definite antimicrobial activity against the three strains of bacteria and their laboratory cultures as evident from the Nutrient agar method. IC 50 values of 15, 18 and 19 mcg/ml was comparable to amoxicillin and give a clearer picture bout the potency and strong antibacterial property of the extracts.
DISCUSSIONS:27,42-45
There is indeed a great dearth of credible literature related to reporting about the use of Neptunia prostrata from North eastern states of India. Only a handful number of studies focussed on the antioxidant nature of the leaves and other aerial parts but the real novelty of the study lies in linking the antibacterial effect on select strains of bacteria (Both Gram positive and negative) to its which may justify the robust antioxidant profile of the herb.
The study was aimed at assessing the plausible antibacterial and antimicrobial role of extracts of leaves of Neptunia prostrata Linn. (NP).Gross phytochemical screening tests revealed the presence of flavonoids and tannins predominantly in the leaves of the plant. However, as a limitation of the study may be not isolating and characterising any lead moiety that is responsible for the bioactivity of NP and study of any isolated medicinal lead moiety could have been facilitated and was a ground for connecting the potential antioxidant nature and antibacterial property of the herb extract on the basis of its structure and chemistry.
The study stands out to report the safety of the herb through oral acute toxicity study as per OECD guidelines-420 and secondly with respect to the strong antioxidant and practically comparable IC50 values with standard antioxidants (such as ascorbic acid) and antibacterial property comparable to the beta lactam antibiotic amoxicillin) through the disc diffusion assay methodology in selected strains of bacterium.
CONCLUSION:
From the above mentioned study we came to know about the preliminary phytochemical screening of the extracts of the plant and also the antimicrobial effects of the plant against bacterial strains.The study results revealed that the plant has some antimicrobial activity as evident from the study. The IC50 value of the extract was 11.2µg/ml whereas the same for Ascorbic acid was 18.02µg/ml for the DPPH assay. With regards to the Hydrogen peroxide antioxidant assay IC50 value of extract of Neptunia prostrata (NP-2) was 101.03µg/ml while IC50value of Ascorbic acid (Vitamin C) was found to be about 93µg/ml.
Acute toxicity study as per OECD guidelines 420 was assessed in wistar albino rats and in swiss albino mice (n=5) of both sexes at doses of 2000 mg/kg body weight and did not reveal any morbidity or mortality in the animals within the stipulated period pointing out towards its possible safety profile. Antimicrobial studies on other strains of bacteria may be also performed to understand the spectrum of antimicrobial activity of the plant extracts under study.The pharmacological in-vivo activities of the plant and its toxicity studies as per OECD guidelines need to be ascertained using a large sample size in animal models of disease to corroborate the present findings and reaffirm its putative role in therapeutics.
ACKNOWLEDGEMENT:
The authors would like to gratefully acknowledge all cooperation from their respective institutions.
ETHICS CLEARANCE:
BST/IAEC/Feb-2018/02.
CONFLICT OF INTEREST:
None.
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Received on 27.01.2022 Modified on 14.06.2022
Accepted on 12.11.2022 © RJPT All right reserved
Research J. Pharm. and Tech 2023; 16(5):2411-2414.
DOI: 10.52711/0974-360X.2023.00397