INTRODUCTION:

Inflammation is the body's normal response to injury, infection, or harmful stimuli, and it is characterised by heat, redness, pain, and swelling¹. It is the reaction of the body to inactivate or kill the invading organisms by releasing chemical mediators such as cytokines, interleukins, and prostaglandins etc., from the site of tissue injury.

These released chemicals increase blood flow to the affected area causing vasodilation and permeability of capillaries. Inflammation can be acute or chronic. Acute inflammation is the body's first reaction to foreign particles, and it is triggered by a continuous influx of plasma and leucocyte-like components from the blood to the injured area². Acute inflammation may last for few minutes only. Prolonged inflammation or chronic inflammation is condition in which there is gradual movement of cells present in the inflammation site followed by tissue destruction and healing³. Chronic inflammation causes serious damage to the host tissue and results in various chronic disorders including cancer, neurogenerative disorders, metabolic and cardiovascular diseases¹.

Chronic inflammation in the damaged tissue is marked by dominant involvement of macrophages. Although these cells are protective agents in the body the toxins they release (including reactive oxygen and nitric oxide species) are injurious to the organisms and invading agents.

One of the characteristic features of inflammation is the increased oxygenation of eicosanoic acid, which is incorporated by two enzyme pathways; 5- lipooxygenase and cyclooxygenase (COX 1 and COX2)) that leads to generation of inflammatory mediators like prostaglandins and leukotrienes. COX 2 which helps in the oxidative metabolism of arachidonic acid, play a crucial role in inflammatory responses⁴. Therefore, numerous non-steroidal anti-inflammatory drugs (NSAID’s) are developed which inhibit the release of these inflammatory mediators. Although these drugs are effective but their use is limited because of side effects like gastric irritation, ulcer and cardiovascular disorders. Therefore, in recent years search for natural bioactive compounds from medicinal plants having pharmacologic activities has increased considerably. Screening of these phytochemical compounds from medicinal plant lead to development of new herbal drugs that shows effective treatment against various diseases.

Flacourtia indica (syn. Flacourtia ramontchi), sometimes known as the Batako plum or Indian plum, belongs to the Flacourtiaceae family. It is an important plant used in dietary supplements and is widely distributed in Bangladesh and India, from Punjab to Bihar, the Deccan, and the Southern Peninsula⁵. It is a little bushy shrub or tree that can reach a height of 900 meters and produces greenish-yellow blooms that are either unisexual or bisexual. The blooming season lasts from December to April. The fruits are fleshy and round, about 8-10 mm in diameter⁶. Unripe fruits are green, whereas ripe fruits are red. Fruit ripens during the months of March and July. Fruits can be eaten fresh or made into jams, jellies, and preservatives. This plant is regarded as a significant herb in Ayurvedic medicine, where plant infusions are used to cure a wide range of ailments⁷. Fresh leaf juice is used to treat fevers. The dried leaves are used to treat bronchitis, asthma, and bladder catarrh. Urinary calculi are treated with a root decoction⁸. Although research on other plant parts have indicated anti-inflammatory, antimicrobial⁹, antioxidant¹⁰, hepatoprotective¹¹, antimalarial¹², anti-diabetic⁶, anti-asthmatic¹³, and antibacterial activity¹⁴, the therapeutic efficacy of fruit extract remains unknown. As a result, the purpose of this study is to evaluate the anti-inflammatory potential of Flacourtia indica fruit extracts.

MATERIALS AND METHODS:

Materials:

Collection of plant material:

Flacourtia indica fruits were collected from the Mangalore area in Karnataka from March to August. The Chairman of the Department of Applied Botany Dr. Raju Krishna Chalannavar, at Mangalore University, authenticated the plant. Fruits were cleaned and washed in deionized water, then dried for two weeks in the shade. The shade-dried Flacourtia indica fruits were mechanically crushed and kept in an airtight container.

Preparation of extracts:

Ethanolic extract: A total of 400g of finely powdered powder was extracted over 7 days using a cold maceration process using ethanol as the solvent. Following the extraction technique, the solvent was removed via a simple distillation method. The recovered solvent was employed for the subsequent extraction process. The finely obtained ethanolic extract has been concentrated under vacuum and stored within a desiccator. ¹⁵.

Aqueous extract: 400g of powder were subjected to a 24-hour cold maceration process utilizing distilled water as the solvent. The concentrated, finely recovered semisolid residue was maintained in a desiccator¹⁵.

Gas Chromatography-Mass Spectroscopy Analysis (GC-MS) of Extracts:

GC-MS analysis of ethanolic and aqueous extracts of fruits of Flacourtia indica was performed by using the equipment Clarus 680 GC. A fused silica column filled with Elite-5MS (5% biphenyl, 95% dimethylpolysiloxane, 30 m, 0.25 mm ID, 250 m df) is a feature of the instrument. Helium is a common carrier gas used with a flow rate of 1 ml/min. The injector was operated at a temperature of 260 °C during the chromatographic run. The oven temperature was programmed initially at 60 °C for 2 min and gradually increased to 300°C for 6 min¹⁶.

The mass-spectrum data was interpreted using the National Institute of Standards and Technology (NIST) database. The spectra of the unknown component were compared to the spectra of recognized components in the NIST collection.

In vitro-anti-inflammatory activity:

Protein denaturation inhibition method:

When a protein is subjected to an external stressor or substance, such as a potent acid or base, a concentrated inorganic salt, an organic solvent, or heat, the protein loses its tertiary and secondary structure. A protein loses its biological activity when it is denatured. Protein denaturation is a well-known cause of inflammation. Nonsteroidal anti-inflammatory drugs (NSAIDs), which have been shown to be beneficial in avoiding protein denaturation, are frequently prescribed interventions in the world. However, these drugs cause serious side effects, plant extracts have been used to treat inflammatory diseases without side effects ¹⁷.

a) Protein denaturation by Bovine serum albumin method:

Stock solutions of fruit extracts were prepared in ethanol and distilled water at a concentration of 1000 µg/ml. Aliquots of 1ml of fruit extracts approximating stock solution concentrations (50-300 g/ml) were added to test tubes containing 0.45ml of bovine serum albumin solution. The pH of above solution was maintained at 6.3 by adding 1N HCl. Incubated the mixture at 37⁰C for 20 minutes and heated at 57 ⁰C for 5 min. cooled the solution and added 2.5ml of phosphate buffer. The absorbance of above sample mixture was recorded at 660nm. Diclofenac taken as the standard drug. The percentage inhibition of denaturation was calculated by the formula:

Abs Control – Abs Extract

% Inhibition = --------------------------------- x 100

Abs Control

Where Abs control and Abs extract gives absorbance values of control and extract respectively.

b) Protein denaturation by Egg albumin method:

2.8 ml of phosphate buffer and 2ml of fruit extracts of different concentrations (50-300 (µg/ml) were mixed together. To the above solution mixture added 0.2ml of egg albumin and incubate for 15min at 37⁰C. The mixture was further heated at 70⁰C for 5min. After cooling the mixture, absorbance was measured at 660 nm ¹⁷.

Abs Control – Abs Extract

% Inhibition = --------------------------------- x 100

Abs Control

Where Abs Control is the absorbance of control and Abs Extract is the absorbance of test extract.

Membrane stabilization test by Heat Induced Hemolytic Method:

HRBC membrane stabilization has been used to assess in vitro anti-inflammatory effects, since the erythrocyte membrane has similarity with lysosomal membrane. Lysosomal stabilization dampens the inflammatory response by inhibiting the release of activated neutrophil lysosomal components, which produces further tissue inflammation. Various disorders are caused by the lysosomal enzymes produced during inflammation. The extracellular activity of these enzymes has been linked to acute and chronic inflammation. Nonsteroidal medicines function by blocking or stabilizing lysosomal enzymes or the lysosomal membrane ¹⁸.

a) Preparation of suspension of red blood cells (RBCs) :

The fresh human blood (10ml) was drawn from a healthy volunteer and transferred to a centrifuge tube for the experiment. The tubes were centrifuged at 3000 rpm for 10 minutes and then rinsed three times with normal saline. Blood density was measured and then reconstituted with a 10% v/v normal saline suspension.

b) Heat induced haemolysis:

1ml of test extract of various concentration (50-300µg/ml) and 1ml of 10% RBC suspensions were mixed in a test tube. Incubate above mixture in water bath for 30min at 56⁰C. Saline was taken as control and diclofenac sodium was used as standard drug. All the tubes were cooled under running tap water and again centrifuged for 5 min at 2500rpm. The supernatant was collected, and the absorbance at 560nm was measured. The percentage of lysis was calculated by

Abs Control – Abs Extract

% Inhibition = ----------------------------------- x 100

Abs Control

Where Abs Control is the absorbance of control and Abs Extract is the absorbance of test extract

Statistical Analysis:

The results were expressed as Mean±SEM, statistical analysis was performed by one-way ANOVA followed by Dunnet Multiple comparison tests using graph pad software, P values <0.05 were considered as significant.

RESULTS:

The Practical yield of Flacourtia indica fruit extracts:

About 400g of dried fruits of Flacourtia indica were sequentially extracted by using ethanol and water as solvents by maceration method. The color, Consistency and Practical yield of these crude extracts were shown in Table 1

Table 1. Color, Consistency and Practical yield (w/w) of different solvent fruit extracts of Flacourtia Indica

Solvent	Color	Consistency	Practical yield(g)
Ethanol	Dark brown	Non-sticky	25
Water	Brown	Non-Sticky	18.4

GC-MS Analysis of Extracts:

Gas chromatography mass spectrometry (GCMS) is a hybrid technology that combines the properties of gas chromatography with mass spectrometry to identify distinct substances in a test sample. The concentrated extract is injected into the GC/MS apparatus. The sample is volatilized at the injection port and eluted through a capillary column as temperature rises. As the sample passes through the column, individual components are separated according to their attraction for the stationary phase of the column, and they can be recognized by retention time¹⁹.

From the GC-MS analysis, a total of 24 compounds were identified in ethanol extract shown in Table-2 and a total of 6 Phytocompounds were detected in aqueous extracts shown in Table-3. The number of compounds found in the ethanol extracts were larger than that of aqueous extract since the GC-MS analysis is limited to only volatile compounds. The compounds peak area, molecular weight, and molecular formula were used to identify them. Two compounds namely N-Hexadecanoic acid and Oleic acid were found to be major in ethanol extract with 18.032% and 35.247% peak area respectively. Other minor compounds found in the ethanol and aqueous extracts are Heptanoic Acid, 6-Oxo-(1.238%), 2-Pentanone, 5-(Acetyloxy)-(1.238%), Tetrahydro-4h-Pyran-4-Ol(1.238%), Carbamic Acid, (3,4,4-Trimethyl-1,2-Dioxetan-3-Yl) Methyl Ester(2.493%), Ethanethioic Acid, S-(Dihydro-2,5-Dioxo-3-Furanyl) Ester (2.493%), Acetamide, N-Aminocarbonyl (2.493%), 1-Heptanamine, N-Heptyl-N-Nitro-(18.032%), 1,4-Butanediol, Diacetate(2.303%), 2,3-Anhydro-D-Galactosan(18.032%), 2-Pentadecanol Acetate(2.120%), Hexadecanoic Acid, Methyl Ester(5.733%), Pentadecanoic Acid, Ethyl Ester(5.733%), Nonadecanoic Acid, Ethyl Ester(6.862%), 1-Hexyl-2-Nitrocyclohexane(16.796%), Eicosanoic acid(16.796%), Z-8-Methyl-9-Tetradecenoic Acid (35.247%), Heneicosanoic Acid, Methyl Ester(1.096%), Pseduosarsasapogenin-5,20-Dien(8.080%),Beta.-sitosterol(8.080%), 2(3h)-Benzofuranone, Hexahydro-4,4,7a-Trimethyl-(1.096%), Oxirane, (Fluoromethyl)-( 9.564%), Silane, Methyl-(9.564%), Methane, Dichloronitro(27.132%), etc shown in Table 2 and Table 3. The chromatogram of ethanolic and aqueous extracts were shown in Figure -1 and Figure-2.

Table – 2. Phytochemicals reported in GC-MS analysis of Ethanolic fruit extract of Flacourtia indica

Si No	IUPAC name of the compound	Retention time	Area %	Molecular formula	Molecular weight
1	Heptanoic Acid, 6-Oxo-	15.018	1.238	C₇H₁₂O₃	144
2	2-Pentanone, 5-(Acetyloxy)-	15.018	1.238	C₇H₁₂O₃	144
3	Tetrahydro-4h-Pyran-4-Ol	15.018	1.238	C₅H₁₀O₂	102
4	Carbamic Acid, (3,4,4-Trimethyl-1,2-Dioxetan-3-Yl)Methyl Ester	15.544	2.493	C₇H₁₃O₄N	175
5	Ethanethioic Acid, S-(Dihydro-2,5-Dioxo-3-Furanyl) Ester	15.544	2.493	C₆H₆O₄S	174
6	Acetamide, N-(Aminocarbonyl)	15.544	2.493	C₃H₆O₂N₂	102
7	1,4-Butanediol, Diacetate	15.939	2.303	C₈H₁₄O₄	174
8	2,3-Anhydro-D-Galactosan	18.700	18.032	C₆H₈O₄	144
9	1-Heptanamine, N-Heptyl-N-Nitro-	18.700	18.032	C₁₄H₃₀O₂N₂	258
10	N-Hexadecanoic Acid	18.700	18.032	C₁₆H₃₂O₂	256
11	2-Pentadecanol Acetate	19.420	2.120	C₁₇H₃₄O₂	270
12	Hexadecanoic Acid, Methyl Ester	19.610	5.733	C₁₇H₃₄O₂	270
13	Pentadecanoic Acid, Ethyl Ester	19.610	5.733	C₁₇H₃₄O₂	270
14	Octadecanoic acid	19.610	5.733	C₁₈H₃₆O₂	284
15	Nonadecanoic Acid, Ethyl Ester	20.350	6.862	C₂₁H₄₂O₂	326
16	1-Hexyl-2-Nitrocyclohexane	20.661	16.796	C₁₂H₂₃O₂N	213
17	Eicosanoic acid	20.661	16.796	C₂₀H₄₀O₂	312
18	Oleic Acid	21.746	35.247	C₁₈H₃₄O₂	282
19	Z-8-Methyl-9-Tetradecenoic Acid	21.746	35.247	C₁₅H₂₈O₂	240
20	2(3h)-Benzofuranone, Hexahydro-4,4,7a-Trimethyl-	24.777	1.096	C₁₁H₁₈O₂	182
21	Heneicosanoic Acid, Methyl Ester	24.777	1.096	C₂₂H₄₄O₂	341
22	2,6-Pyrazinediamine	24.777	1.096	C₄H₆N₄	110
23	Pseduosarsasapogenin-5,20-Dien	27.008	8.080	C₂₇H₄₂O₃	414
24	Beta.-sitosterol	27.008	8.080	C₂₉H₅₀O	414

Figure 1. Chromatogram of ethanolic fruit extract of Flacourtia Indica

Table- 3. Phytochemicals reported in GC-MS analysis of aqueous fruit extracts of Flacourtia indica

SI No	IUPAC name of the compound	Retention time	Area %	Molecular formula	Molecular weight
1	Oxirane, (Fluoromethyl)-	2.744	9.564	C₃H₅OF	76
2	O-Methylisourea Hydrogen Sulfate	2.744	9.564	C₂H₆ON₂	74
3	Silane, Methyl-	2.744	9.564	CH₆Si	46
4	Methane, Dichloronitro-	3.409	27.132	CHO₂NCl₂	129
5	Methyl Hydrogen Disulfide	5.039	14.679	CH₄S₂	80
6	1-Propanethiol	6.095	48.625	C₃H₈S	76

Figure- 2. Chromatogram of aqueous fruit extract of Flacourtia Indica

In vitro-anti-inflammatory activity:

Protein denaturation method:

a) Bovine serum albumin method:

The in vitro anti-inflammatory efficacy of Flacourtia indica fruit extract was investigated using the Bovine serum albumin assay and compared to the standard drug sodium diclofenac. Table 4 shows the percentage of protein denaturation inhibition on increasing doses of each extract. Aqueous extract shows the highest percentage of inhibition at 20.72% -65.76% on varying concentrations of 50-300μg/ml. The Aqueous extract's IC₅₀ value (201.61 μg/ml) is comparable to sodium diclofenac, making it an effective protein denaturation agent. Standard sodium diclofenac inhibited protein denaturation (22.52% - 76.57%) at doses ranging from 50-300μg/ml, with an IC₅₀ of 177.10 μg/ml indicating higher potency and therapeutic efficacy.

Table 4. Effect of aqueous and ethanolic Flacourtia indica fruit extracts on Bovine serum albumin denaturation method

Tested material	Concentration (µg/ml)	% inhibition on BSA Denaturation ± SEM	IC₅₀ value
Diclofenac	50 100 150 200 250 300	22.52 ± 0.90 33.33 ± 0.90 49.54 ± 0.90 61.26 ± 0.90 68.46 ± 0.90 76.57 ± 0.90	177.10
Ethanolic extract of Flacourtia indica	50 100 150 200 250 300	18.91 ± 1.56 31.53 ± 0.90 45.94 ± 1.56 50.45 ± 0.90 53.15 ± 0.90 61.26 ± 0.90	214.79
Aqueous extract of Flacourtia indica	50 100 150 200 250 300	20.72 ± 1.80 30.63 ± 2.38 50.45 ± 1.80 53.15 ± 0.90 57.65 ± 0.90 65.76 ± 1.80	201.61

Each value represents the mean ± SEM. N= 3, experimental groups were compared with control P<0.05, considered extremely significant.

Figure- 3: BSA Denaturation Method

b) Egg albumin denaturation method

The anti-denaturation of egg albumin technique was used to assess anti-inflammatory efficacy. The aqueous extract exhibited maximum inhibition of 70.23±1.19% (300µg/ml) with an IC₅₀ value of 197.60, while ethanolic extract showed percentage inhibition of 66.66±1.37% with IC₅₀ value 215.16 as shown in Table 5. The observations were compared with standard drug diclofenac which exhibited 74.60±1.58% at concentration of 300µg/ml. With increasing concentration, the absorbance values decreased, indicating substantial anti-inflammatory effect.

Table- 5. Effect of aqueous and ethanolic Flacourtia indica fruit extracts on Egg albumin denaturation method

Tested material	Concentration (µg/ml)	% Inhibition on egg albumin denaturation ± SEM	IC₅₀ value
Diclofenac	50 100 150 200 250 300	16.66 ± 1.37 26.19 ± 1.37 44.44 ± 1.58 57.93 ± 1.58 63.49 ± 1.58 74.60 ± 1.58	191.25
Aqueous extract of Flacourtia indica	50 100 150 200 250 300	12.69 ± 0.79 24.60 ± 0.79 37.30 ± 0.79 43.65 ± 0.79 57.93 ± 0.79 66.66 ± 1.37	215.16
Ethanolic extract of Flacourtia indica	50 100 150 200 250 300	19.04 ± 1.19 26.19 ± 1.19 40.47 ± 1.19 55.95 ± 2.38 58.33 ± 1.19 70.23 ± 1.19	197.60

Each value represents the mean ± SEM. N= 3, experimental groups were compared with control P<0.05, considered extremely significant.

Figure- 4. Egg Albumin Denaturation Method

HRBC membrane stabilization:

In the current investigation, the percentage inhibition with the HRBC membrane stabilization technique increased dose-dependently in both the ethanolic and aqueous fruit extracts of Flacourtia indica. The potent inhibitory activity was exhibited by aqueous extract with percentage inhibition of 64.23% with IC₅₀ value of 263.83, compared with standard drug showing a percentage inhibition of 70.66 ± 0.85% with IC₅₀ value 199.22 shown in Table 6.

Table- 6. Effect of aqueous and ethanolic Flacourtia indica fruit extracts on HRBC membrane stabilization method

Tested material

Concentration (µg/ml)

% membrane stabilization ± SEM

IC₅₀ value

Diclofenac

100

150

200

250

300

16.46 ± 0.49

28.23 ± 0.99

37.08 ±0.44

50.75 ±0.41

65.81± 0.27

70.66 ± 0.85

199.22

Aqueous extract of

Flacourtia indica

100

150

200

250

300

12.45 ± 1.68

18.96 ± 1.66

27.37 ± 1.49

31.67 ± 1.88

43.58 ± 1.63

59.46 ±1.98

241.77

Ethanolic extract of

Flacourtia indica

100

150

200

250

300

14.01 ±1.59

20.22 ± 0.34

31.87 ± 0.32

45.09 ± 2.86

50.87 ± 0.36

64.23 ± 2.05

263.83

Each value represents the mean ± SEM. N= 3, experimental groups were compared with control P<0.05, considered extremely significant

Figure- 5. HRBC membrane stabilization method

DISCUSSION:

Inflammation is a positive sign that the living tissue is raising an alarm about an irritation, infection, or injury that is occurring inside or outside the body²⁰. Although this tissue's usual response is to protect itself, when it becomes uncontrollable, repetitive, or chronic, it has been linked to disorders such as asthma, obesity, and rheumatoid arthritis. The most frequent treatments for inflammation are steroidal and non-steroidal drugs; however, their usage is limited due to their severe adverse effects; hence, interest is developing in the development of alternative therapies derived from natural sources that do not pose health hazards.

GC-MS is a highly accurate approach for identifying secondary metabolites in plant extracts, using the NIST library. The Identified compounds by GC-MS analysis possess various biological activities as shown in Table 7. Among the compounds identified N-Hexadecanoic acid was the most abundant phytocompound (18.032%) present in the ethanolic extract of Flacourtia indica fruit. N-Hexadecanoic Acid is a saturated fatty acid which are known to have anti-inflammatory and anti-fungal properties. Fatty acids have the ability to modulate immune responses by acting directly on T cells. According to several studies, Polyunsaturated fatty acids play a role in regulating the anti-inflammatory pathway²⁰. An enzyme kinetic study²¹reported that Saturated fatty acids block phospholipase A2, an enzyme that catalyzes the release of arachidonic acid, a precursor for the synthesis of inflammatory cytokines, at the sn-2 position of membrane phosphor.

Table -7. Activity of Phytoconstituents identified in ethanol and aqueous fruit extracts of Flacourtia indica

SI NO	Compound name	Structure	Uses
1	Eicosanoic acid		Alpha-glucosidase inhibitory activity, Anti-inflammatory²²
2	Hexadecanoic acid, methyl ester		Anti-inflammatory²², Anti-fungal²³
3	N-Hexadecanoic acid		Anti-inflammatory²⁴, anti-oxidant, Hypo-cholestrolemic, Alpha-reductase inhibitor activities²⁵
4	Octadecanoic acid		Anti-inflammatory, Anti-oxidant²⁶, Anti-cancer²⁷
5	Oleic acid		Anti-Inflammatory²⁸, Cancer preventive, Anti-androgenic, Insectifuge
6	Beta- sitosterol		Anti-inflammatory²⁹, anti-diabetic, anti-oxidant, Prostatic cancer treatment³⁰
7	Pseduosarsapogenin-5,20-Dien		Anti-diabetic activity (alpha amylase inhibitory activity³¹
8	2,6-Pyrazinediamine		Anti-diabetic. Anti-tubercular, anti-cancer, Antiinflammatory³²
9	Z-8-Methyl-9-Tetradecenoic Acid		Anti-oxidant, anti-cancer, 5α- reductase inhibitor, haemolytic³³

The Inflammatory process begins with neutrophil activation and the release of bactericidal enzymes and proteases. These enzymes generate lipid peroxidation and produce lysosomal enzymes that break down macromolecules. Thus, lysosomal membrane stability is required to prevent inflammatory reactions³⁴. Thus, suppression of HRBC membrane lysis was employed to evaluate the anti-inflammatory activity of Flacourtia indica fruit extracts. Membrane stability prevents fluid and serum protein leaks into tissues during periods of increased permeability caused by inflammatory mediators.

Reducing protein denaturation is another technique to exhibit anti-inflammatory properties. According to certain research, protein denaturation is one of the causes of inflammation³⁵. Protein denaturation is hypothesized to be produced by changes in electrostatic, hydrogen, hydrophobic, and disulfide bonds. In addition, a complicated chain of events occurs, including enzyme activation, mediator release, cell migration, tissue disintegration, and repair, causing the protein to denature, or lose its molecular structure and activities. As a result, substances that may prevent these changes and decrease thermal or heat-induced protein denaturation could be employed as anti-inflammatory medications³⁶

The current investigation evaluated the anti-inflammatory properties of fruit extracts from Flacourtia indica at varying concentrations (50-300μg/ml) against HRBC membrane and for prevention of protein denaturation using invitro methods. The results are similar to those of the widely used medication diclofenac sodium. The highest percentage of suppression of main inflammatory mediator release and stability of the cell membrane was demonstrated by the Flacourtia indica ethanol extract. According to GC-MS study, it can be because of the presence of bioactive phytoconstituents such oleic acid and hexadecenoic acid. The goal of the research is to find natural anti-inflammatory drugs that are as effective as pharmaceutical treatments but have fewer side effects.

CONCLUSION:

The results of Present study, GC MS analysis of ethanolic and aqueous extracts of Flacourtia indica revealed the presence of various bioactive phytochemical compounds which possess significant biological activities. In-vitro anti-inflammatory activity of ethanolic and aqueous fruit extracts were screened against HRBC membrane and protein denaturation methods with egg albumin and bovine serum albumin. The ethanolic extracts showed significantly higher anti-inflammatory activity in a concentration dependent manner. It may due to the presence of active principles identified by GC-MS analysis. Thus, we conclude that Flacourtia indica extract can be an effective anti-inflammatory herbal product and it gives hopes that new drugs can be derived from Flacourtia indica extracts which may be useful for the treatment of various inflammatory conditions.

CONFLICT OF INTEREST:

The authors have no conflicts of interest in this study.

ACKNOWLEDGMENTS:

The authors are thankful to the NGSM Institute of Pharmaceutical Sciences and NITTE (Deemed to be University) for their motivation and lab facilities provided to carry out this research work successfully.

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Received on 16.02.2024 Revised on 06.11.2024

Accepted on 10.04.2025 Published on 02.05.2025

Available online from May 07, 2025

Research J. Pharmacy and Technology. 2025;18(5):2081-2089.

DOI: 10.52711/0974-360X.2025.00298

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