Table No.2 Pharmacological activity of various parts of the plant

Sl. No	Plant Part used	Method	Activity	Reference
1.	Stem Bark	STZ Induced Diabetic Rat.	Cardioprotective activity of Pet Ether Extract of P. Pinnata Stem Bark in diabetic rats.	41
2.	Flower	α-amylase inhibitory method	Different isolated active constituents from the flowers of P. pinnata were screened for antidiabetic activity	42
3.	Leaf	Alloxan-induced diabetic model and OGTT.	Antidiabetic effect of P. pinnata leaves in alloxan-induced diabetic rats.	43
4.	Leaf	Alloxan β- cytotoxin induced diabetes model.	Antidiabetic potential of the polyherbal formulation which is a mixture of ethanolic extract of Caesalpinia bonducella, Mucona puriens, Pongamia pinnata	44
5.	Leaf	Agar well diffusion method	Antifungal activity of Pongamia pinnata L. extracts on saprolegnia isolated from diseased fishes.	45
6.	Leaf	Agar well diffusion method	Antibacterial properties of leaf extracts of P. pinnata against some human pathogens,	47
7.	Leaf	Disc diffusion method	Antibacterial properties of leaves of P. pinnata, against enteric bacterial pathogens.	48
8.	Leaf	Agar well diffusion and Disc diffusion method	Antibacterial activity of leaf extract of P. pinnata, C. long. and M. arvensis. against S. aureus bactria	51
9.	Leaf	In vitro anti-inflammatory activity by protein denaturation technique	Anti-inflammatory activities of methanolic extracts of P. pinnata and Diclofenac Sodium, using heat-induced protein denaturation technique.	60
10.	Leaf	Agar diffusion method	The antibacterial potential of Pongamia pinnata leaf extracts was tested against selected food-borne pathogens, highlighting its potential application as a natural antimicrobial agent in the pharmaceutical or food industry.	52
11.	Seed	Agar well diffusion and Disc diffusion method	Antibacterial activity of P. pinnata seed extracts using organic and aqueous solvent, against multidrug human pathogens.	50
12.	Seed	Inhibition of protein denaturation model & HRBC membrane stabilization model	Anti-arthritic activity of ethanolic extract of seeds of P. pinnata and methanolic extract of Punica granatum linn by in-vitro technique.	54
13.	Seed	Paw oedema method	Anti-inflammatory potential of different solvent fractionated seed extracts of P. pinnata, using chemically induced paw inflammation in rats.	61
14.	Seed	Nitric oxide and superoxide quenching properties, ABTS, iron chelating activities.	The seed extracts of Pongamia pinnata were assessed for their antioxidant and free radical scavenging properties, using a standard spectrophotometric protocol for estimation..	62
15.	Seed	Agar Well Diffusion Method	Antimicrobial efficacy of seed extracts of P. pinnata against P. aeruginosa, S. aureus, S. epidermidis, M. furfur, and C. albicans.	58
16.	Seed, Bark, Leaf, Root	Well diffusion method	Fungitoxic properties of P. pinnata against two human pathogenic fungi and two plant pathogenic fungal strains.	46
17.	Leaf and Seed	Cup diffusion method	Evaluation of antibacterial potential of P. pinnata against Xanthomonas axonopodis punicae, phytopathovar of Bacterial blight of Pomegranate.	49
18.	Leaf, Bark, Flower, and Root.	BSL and THP-1 Cell Line study/ Invitro Antimalarial activity.	Different extracts of P. pinnata were screened for In vitro antimalarial activity against P. falciparum 3D7 strain.	53
19.	Bark	Acetic acid-induced abdominal writhing & ELISA in carrageenan and cotton pellet granuloma test	Alcoholic extract of stem bark of P. pinnata was studied for its analgesic and anti-inflammatory activity in rats, Serum TNF-alpha and IL-1beta were measured by ELISA.	59
20.	Bark	Agar Well Diffusion Method	The hydroalcoholic bark extract of Pongamia pinnata was evaluated for its antimicrobial potential against various pathogens, including Bacillus subtilis, Escherichia coli, and Aspergillus niger.	55
21.	Leaf, Stem, Bark, Flower.	Agar Well Diffusion Method	Different parts of P. pinnata were screened for its antimicrobial activity against bacterial pathogens.	56
22.	Leaf, Bark, Seed	Disc diffusion method	Various extracts of P. pinnata were examined for antimicrobial potential against Gram + and – bacteria and fungal species.	57
23.	Leaf, Bark, Seed	DPPH radical scavenging activity	Different parts of P. pinnata were studied for its antioxidant properties and maximum extraction yield of antioxidant components from bark, leaves, seeds were estimated.	63

Pharmacognostic Study:

Phytochemically leaf powder of P. pinnata was reported to have alkaloids, flavonoids, steroids, triterpenes, cardiac glycosides and saponins^33,34.Ethanolic extracts of leaves have been studied for various phytocompounds and report disclosed the presence of Flavonoids, alkaloids, carbohydrates, proteins, terpenoids, steroids and glycosides³⁵. Seed extract shows the presence of phenols, flavonoids, tannins, saponins, alkaloids, fats and fixed oils. The total flavonoids content of hydro alcoholic extract of plant was 1.398 mg/100mg³⁶. Acetone and chloroform extracts of pinnata contains flavonoids, tannins, and cardiac glycosides in acetone extract and alkaloids and saponins were present in chloroform extract³⁷_.Pharmacognostic studies were carried out to evaluate organoleptic, microscopic, physicochemical parameters of stem bark of Pongamia pinnata. Results revealed various microscopical characters like Phellogen, Parenchymatous cells, Phloem fibres, Wavy medullary rays and Physicochemical investigation of bark revealed total ash value 10.94%w/w, water soluble ash 1.96%w/w, acid insoluble ash 1.47%w/w and sulphated ash 15.8% w/w respectively. The alcohol and water-soluble extractive value were 9.6 & 18.4%w/w.³⁸. P. pinnata stalk was used for extracting cellulosic natural fibres which were characterised and through compositional analysis it revealed fibres have 54% cellulose, 12% hemi cellulose, 15% ligands and 11% ash. The tensile strength of fibre was 310 MPa which is equivalent to cotton and linen. The tensile strength of P. pinnata fibre reinforced polypropylene composite was 17.96 MPa which is equivalent to other natural fibre based composite³⁹_.Karanjin was extracted from seeds using Ultrasound assisted extraction technique which was the most efficient mode of extraction using methanol as solvent. Extraction temperature of 57.85⁰C, extraction time of 25.45 minute and solvent to drug ratio of 86.4709 W/V was the optimum condition for extraction. The non thermal method of extraction, (UAE) is most efficient mode of extraction and polar solvents are most promising solvents for extraction of Karanjin⁴⁰_.

Biological activities:

Antidiabetic activity:

Petroleum ether extract of stem bark of plant was studied for its hypoglycaemic activity. Findings suggested that blood glucose level was reduced with improved electro cardiography, hemo-dynamic parameters and enhanced oxidative stress in STZ induced diabetic rats⁴¹_.Three isomeric furano flavonoids from hexane extract of flowers of P. pinnata were studied for antidiabetic screening using in-vitro model which showed higher anti diabetic activity as that of standard Acarbose⁴²_. Ethanolic and aqueous extracts of leaves were compared with standard Glibenclamide in alloxan induced diabetic rats, the extracts revealed their potency and proved their anti-diabetic effects in rats and reduced mortality rate⁴³_.P. pinnata in combination with Caesalpinia bonducella, Mucona puriens were subjected to investigate the antihyperglycemic activity in alloxan induced diabetic rats and revealed better results due to restoration or regeneration of beta cells of pancreas⁴⁴.

Antifungal activity:

P. pinnata extracts were used for evaluation of antifungal activity on Saprolegnia parasitica isolated from cultured and wild fishes. The extracts showed highest growth inhibition. It proved to be a potent antifungal remedy⁴⁵_.Extracts of different plant parts of (seeds, bark, leaves, roots) were subjected to investigate fungi toxic properties against two human pathogens- Epidermophyton floccosum and Candida albicans and 2 plant pathogens- Alternaria solani and Helminthosporiumturcicun. Amongst all extracts the seed extract showed higher antifungal activities followed by roots, barks while the leaves extract showed the least activity. E. Floccosum was more susceptible to all extracts followed by C. albicans, whereas H. turcicun was more resistant to all extracts followed by A. Solani⁴⁶.

Antibacterial activity:

P. pinnata leaf ethanolic extract has probable antibacterial effect against some human pathogens- C. violaceum, C. frendii, S. aureus and M. luteus. The activity was measured by Agar well diffusion method. Zone of inhibition was studied and highest zone of inhibition of ethanolic extract was observed against gram positive bacteria M. luteus (38mm) and smallest was observed against C. freundii (17.6mm)⁴⁷. Various extracts of leaves (petroleum ether, chloroform, ethyl acetate, methanol) were screened by disc diffusion method against some enteric bacterial pathogens such as E. coli, S.aureus, E. aerogenes, P. aeruginosa, S. typhimurium, S. typhi, S. epidermidis and P. vulgaris. The methanolic extracts exhibits wide range of antibacterial activity on these bacterial pathogens than pet ether extracts, whereas the ethyl acetate extracts show slightly higher than chloroform extract⁴⁸.The aqueous and other solvent extracts of P. pinnata leaves and seeds were tested against Xanthomonas axonopodispv. punicae, phytopathovar of bacterial blight of Pomegranate using agar cup diffusion method. The extracts were prepared in 6 solvents namely; water, ethanol, methanol, diethyl ether, benzene and chloroform. Aqueous, ethanol, methanol, extracts of leaves and seeds showed significant activity against X. axonopodispunicae. Minimum inhibitory concentration of these extracts ranged between 400- 500µg/ml. The Methanol extract of leaves and aqueous extract of seeds shows maximum antibacterial activity against the pathogen⁴⁹_.The aqueous, chloroform, ethanol, methanol, petroleum ether extracts of plant were tested against multi drug human pathogens. The ethanolic extract showed highest anti- S. typhi activity and was effective against all bacterial strains. Methanol, chloroform, aqueous extracts showed moderate activity and petroleum ether showed minimum inhibitory activity⁵⁰_.Anti-bacterial activity of P. pinnata, Curcuma longa and Mentha arvensisa comparative studies were performed against Staphylococcus aureus. Aqueous extract of M. arvensis is more effective than methanolic and ethanolic extracts; ethanolic extract of P.pinnatais more effective than the aqueous and methanolic extract; ethanolic extract of Curcuma longa leaf extract is more effective than methanolic and aqueous extract⁵¹_.The organic extracts of leaves were tested against some food born pathogenic bacteria. The organic extracts of chloroform, ethylacetate and methanol at concentration of 2500µg/ ml showed good effect against B. subtilis, S. aureus, L. monocytogenes, P. aeruginosa and S. typhimurium. The zones of inhibition were in range of 8.1 to 18mm and MIC value from 125 to 1000µg/ml. The chloroform, ethyl acetate and methanol extracts show greater activity as compared to streptomycin⁵².

Antimalarial activity:

To assess the in vitro anti-malarial efficacy against the Plasmodium falciparum 3D7 strain, natural extracts from methanol, chloroform, hexane, ethyl acetate, and aqueous extracts of dried leaves, bark, and roots of Pongamia pinnata were screened. The methanolic bark extract demonstrated significant in vitro anti-malarial activity, with an IC50 value of 11.67µg/ml.⁵³

Antiarthritic activity:

Ethanolic extract of P. pinnata and methanolic extract of Punica granatum in comparison were examined for antiarthritic activity by in vitro techniques i.e., inhibition of protein denaturation and HRBC membrane stabilization, Diclofenac sodium was used as standard. Protein denaturation was inhibited and stabilization toward the HRBC membrane was observed by ethanolic extract of P.pinnata and Methanolic extract of Punica granatum. Study revealed that MEPG has more potent activity than EEPP⁵⁴_.

Antimicrobial activity:

The hydro alcoholic bark extract was subjected for antimicrobial activity organisms included for activity were B. subtilis, E. coli and A. niger. The MIC of extract against all organism was 100µg/ml which is consistent. The extract shows synergistic effect with antibiotics and reduces the MIC of respective antibiotics towards pathogenic bacteria⁵⁵_. In another study the bioactive compounds from leaves, seed, bark, flowers were extracted by soxhlet among all parts the seeds and leaves extract exhibited maximum zone of inhibition in most bacterial pathogens at a concentration of 400µg/ml⁵⁶_.Different polarity solvents like pet. ether and ethyl acetate extracts were examined against gram positive and negative bacteria using the disc diffusion method. The extracts of leaves, bark, seeds from pet ether and ethyl acetate extracts showed an inhibition zone on B. subtilis and S. aureus. Extracts from seeds of pet ether (72.00±0.57) and ethyl acetate (72.00±0.00) showed maximum inhibition zone on B. subtilis, while leaf extract of Pet ether (17.66±0.66) and ethyl acetate (21.16±0.16) showed similar activity against S. aureus, E. coli, C. Albicans⁵⁷_. Ethyl acetate seed extracts evaluated against P. aeruginosa, S. aureus, S. epidermidis, M. furfur and C. albicans by Agar well diffusion essay, resulted that S. epidermidis was the most sensible strain among all the tested pathogens. Ethyl acetate extract of seeds show the inhibition zone of 41.05±0.7mm⁵⁸.

Anti-inflammatory activity:

Alcoholic extract of stem bark of P. pinnata was evaluated for its analgesic and anti-inflammatory activity in rats. Serum TNF alpha and IL 1 beta were measured by Eliza the extract exhibited notable analgesic and anti-inflammatory activity with decreased serum TNF alpha and IL 1 beta in acute as well as chronic anti-inflammatory studies⁵⁹. Methanolic extract of leaves was studied for in-vitroanti-inflammatory activity using heat induced protein denaturation technique where diclofenac sodium was used as reference standard. The concentration of leaf extract and diclofenac sodium was 50 to 250µg/ml. Diclofenac sodium and methanolic extract of leaves showed IC₅₀ value up to 18.36 and 82.24µg/ml respectively⁶⁰_. Different solvent fractionated seed extracts were tested in chemically induced paw inflammation in rats. The activity was best seen against Bradykinin and PGE1 induced inflammation and minimal effect was seen against histamine and 5 HT induced inflammation. The polar constituents in seeds appear to have anti-inflammatory principles⁶¹.

Antioxidant activity:

The antioxidant properties of Pongamia pinnata seed oil were evaluated using various scavenging assays. Karanjin, pongapin, and yellow oil were isolated from the hexane extract. Four different assays were conducted: superoxide quenching, nitric oxide scavenging, iron chelation, and total antioxidant status, all measured via spectrophotometry. The active compounds exhibited varying degrees of superoxide, nitric oxide, and iron chelating activity, contributing to a strong overall antioxidant capacity. Superoxide scavenging activity was tested at different concentrations, with the highest activity observed in pongapin (37.97%), followed by Karanjin (34.47%) and yellow oil (6.47%). In nitric oxide scavenging, ascorbic acid, used as a standard, showed maximum inhibition (89.14%). Among the seed oil components, Karanjin had the highest inhibition (88.47%), followed by pongapin (62.31%) and yellow oil (2.4%). Superoxide scavenging was also evaluated across different concentrations, where pongapin had the highest activity (0.61%), exceeding that of Karanjin (0.25%) and yellow oil (0.75%). In iron chelation, pongapin showed the greatest inhibition (87.12%), followed by yellow oil (81.27%) and Karanjin (30.17%)⁶². Antioxidant properties of different parts (bark, leaves, seeds) of P. pinnata was studied. The obtained bark extract from aqueous methanol showed higher DPPH radicals scavenging activity with IC 50 value of 3.21 µg/ml followed by leaves and seed extract. Bark has an enormous opportunity for antioxidants application in medications⁶³.

Neuro-protective activity:

Ethanolic extract of stem bark of P. Pinnata was studied for monosodium glutamate-induced neurotoxicity in rats. According to the study, ethanolic extract of stem bark has significant neuroprotective effects on albino rats⁶⁴.

Antiviral activity:

Isolated chemical 1, bis (2-methylheptyl) phthalate, from the P. pinnata extract effectively suppressed the White Spot Syndrome Virus (WSSV). After being exposed to WSSV for 15 days, the Penaeus monodon group that was fed the1, bis(2-methylheptyl) phthalate and experimentally challenged with the virus exhibited 60% and 50% survival rates, respectively, at 100 and 150 µg/gm per day. The results of this study demonstrated that 1, bis(2-methylheptyl) phthalate is the antiviral WSSV chemical found in P. pinnata leaves⁶⁵_.

Analytical study:

HPTLC:

The leaf extract of Pongamia pinnata was analyzed using the HPTLC fingerprinting technique at a wavelength of 254nm, revealing various phytochemicals with Rf values ranging from 0.01 to 0.97. Among these, the highest concentration of phytochemicals was 32.66%. Several secondary metabolites, including alkaloids, carbohydrates, steroids, tannins, flavonoids, terpenoids, coumarins, phenols, and quinones, were identified in the extract.⁶⁶ A sensitive, selective, and precise TLC method was developed and validated for the analysis of Karanjin. The separation was carried out using TLC with a mobile phase of ethanol and ethyl acetate in a 7:3 v/v ratio. The retention factor (Rf) was determined to be 0.69. The calibration curve showed linearity within the concentration range of 0.1 to 1.2 µg/spot. The developed method demonstrated linearity, precision, accuracy, and specificity⁶⁷. Phytochemical screening and HPTLC analysis of methanolic extracts from seeds and callus were conducted using a mobile phase of toluene and ethyl acetate in a 9:1 ratio. The HPTLC results revealed a common band with an Rf value of 0.36 in seed oil, methanolic seed extract, and callus, indicating the presence of a novel compound in both the methanolic extracts of seeds and callus.⁶⁸

HPLC:

A novel, stable HPLC method was developed for the estimation of Karanjin, utilizing an RP-C18 column with detection at 260nm using a PDA detector. The mobile phase consisted of methanol, acetonitrile (ACN), and water in a 70:15:15 v/v ratio. The developed method was precise, specific, and accurate⁶⁹. A reverse-phase HPLC method was developed to analyze Karanjin in Pongamia pinnata leaves. The analysis was performed on a C18 column using a mobile phase consisting of a mixture of methanol, water, and acetic acid (85:13.5:1.5 v/v) at a flow rate of 1ml per minute, with UV detection at 300nm. The method was validated and found to be specific, accurate, precise, reliable, and reproducible. It showed good linear correlation coefficients, with detection and quantification limits of 4.35µg and 16.56 µg, respectively. This method can be used for quantitative analysis and quality control of leaves, with Karanjin serving as a marker compound.⁷⁰

CONCLUSION:

A significant proportion of the population in developing countries, more than 80% as per the World Health Organization, receive their primary medical care through herbal remedies. The well-documented safety and effectiveness of natural compounds, particularly those sourced from plants, have sparked considerable interest in their utilization in traditional and ethnobotanical medicine. It is essential to adopt a conventional strategy in the exploration of novel substances to tackle a spectrum of diseases. Pongamia pinnata has been commonly used for treating a wide array of illnesses. Its therapeutic attributes can be found in every part of the plant, which are applied in the treatment of wounds, piles, ulcers, and skin conditions. Analysis of phytochemicals has indicated that the plant contains a variety of secondary metabolites. In traditional medicine, the plant is recognized for its anti-diabetic, anti-inflammatory, antifungal, antibacterial, anti-malarial, antimicrobial, antiviral, and antioxidant properties. The primary focus of this review was to provide an overview of the pharmacological uses of P. pinnata, the diverse chemical constituents within the plant, and the analytical methods used for standardization. The future perspective of this medicinal plant is to isolate desired phytocompounds of therapeutic interest and development of novel drug delivery thereof.

ACKNOWLEDGEMENT:

The authors thank Principal, KLE College of Pharmacy and Department of Pharmacognosy, KLE College of Pharmacy Belagavi, for all the support to complete this review.

CONFLICT OF INTEREST:

The authors declare that they have no conflict of interest.

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Received on 05.08.2024 Revised on 18.01.2025

Accepted on 23.03.2025 Published on 02.08.2025

Available online from August 08, 2025

Research J. Pharmacy and Technology. 2025;18(8):3953-3960.

DOI: 10.52711/0974-360X.2025.00568

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Kingdom	Plantae
Subkingdom	Tracheobionta
Super division	Spermatophyta
Division	Magnoliophyta
Class	Magnoliopsida
Subclass	Rosidae
Order	Fabales
Family	Fabaceae
Genus	Pongamia
Species	Pinnata