INTRODUCTION:

Costus igneus commonly known as Insulin plant has been recently introduced in India as herbal cure for diabetes. The plant belongs to the family Costaceae and is widely grown in the southern parts of India as an ornamental plant.¹ Its leaf is consumed for keeping a check on the sugar levels in serum.^2,3 Studies have revealed the presence of steroids, triterpenoids, alkaloids, tannins, flavonoids, glycosides, saponins, carbohydrates, and proteins.⁴It has been reported that the leaves of the plant are rich in ascorbic acid, α‑tocopherol, β‑carotene, terpinoids, steroids, and flavonoids.⁵ Isolation of bis (2’‑ethylhexyl)‑1,2‑benzenedicarboxylate, ergastanol, quercetin and diosgenin has also been reported from the leaves.^6,7Several scientific studies have been carried out the leaves of C. igneus for its antidiabetic potential.^8-10 Other pharmacological actions like hypolipidemic¹¹, diuretic¹², antioxidant¹³, antimicrobial¹⁴, anticancer¹⁵ and anti-urolithiatic¹⁶ have also been reported.

Inflammation includes a long chain of molecular reactions and cellular activity, which are designed to restore a tissue. Free radicals are known to cause inflammation in human by cellular damages. In living system, among the radical species, oxygen derived radicals are most important and it is called reactive oxygen species (ROS).¹⁷ ROS can be produced from several endogenous sources, such as xanthine oxidase, cytochrome oxidase, cyclooxygenase, mediated unsaturated fatty acid oxidation, oxidation of catecholamines, mitochondrial oxidation, inflammation, phagocytosis, ischemic reperfusion injury, exercise activation of leukocyte nicotinamide adenine dinucleotide phosphate oxidase, iron release, and reduction-oxidation reaction cycling.¹⁸ Antioxidants are compounds that combat the free radicals like ROS and help the body restore the normal physiological functioning.

In the present work, our aim was to study and ability of the aqueous extract from the leaves of C. igneus in inhibiting the inflammatory response caused by lipoxygenase enzyme.

MATERIAL AND METHODS:

Collection of plant material:

The plant material was collected from the locality of Satna and was authenticated in the botany department of the institute. The collected leaves were dried, powdered and used for extraction.

Extraction and phytochemical screening:

The leaf powder was extracted using purified water as the extraction solvent with the aid of soxhlet apparatus. Briefly, 15g of the leaf powder was packed in a thimble and the extractor of the soxhlet apparatus and extracted with 75mL purified water. The extraction process was carried out for about 2h and the hot extract was filtered through Whatman filter paper. The volume of solvent was reduced by evaporation on water bath to obtain the extract.¹⁹The extract was qualitatively tested for alkaloids, glyocosides, saponins, phenolics, tannins, flavonoids and sterols.²⁰

Total Phenolic Content:

The total phenolic content in the leaf extract was determined quantitatively using Folin-Ciocalteu reagent method, using gallic acid as the reference standard. Briefly, 200μL of sample was mixed with 1.4mL purified water and 100μL of Folin-Ciocalteu reagent. After incubating at room temperature for 15min, 300μL of 20% Na₂CO₃ aqueous solution was added and the mixture was allowed to incubate at room temperature for 2h.²¹The absorbance of the solution was measured at 760nm with a UV-Vis spectrophotometer. Standard solutions of gallic acid (10-100ppm) were similarly treated to plot the analytical curve. The control solution contained 200μL of methanol and suitable reagents, and it was prepared and incubated under the same conditions as the rest of the samples. Results were expressed as milligrams of gallic acid equivalent (GAE) per 100g of the dry sample.

Anti-inflammatory evaluation:

Preparation of extract sample:

The ethanolic extract (10mg) of the leaf was dissolved in ethanol (10mL). From this solution, 1mL was withdrawn and diluted to 10mL with ethanol. Aliquots of 2.5, 5.0, 7.5mL were taken and diluted up to 10mL with ethanol to obtain the desired concentrations of 25, 50 and 75 µg/mL whereas aliquot of 10mL was pipetted out directly for 100µg/mL solution.

Lipoxygenase inhibition assay:

Lipoxygenase inhibition activity of the extracts (25, 50, 75 and 100µg/mL) was assayed according to the method of Wu.²² In short, a mixture of a solution of sodium borate buffer (1mL, 0.1M, pH 8.8) and lipoxygenase (10 µL, ﬁnal concentration 8000U/mL) was incubated with 10µL leaf extract in a 1mL cuvette at room temperature (30±2^◦C) for 5min. The reaction was initiated by the addition of 10µL linoleic acid substrate (10mmol). The absorbance of the reaction solution was measured at 234 nm using a UV/VIS spectrophotometer. Phosphate buffer solution was used as the control, and the percentage inhibition of lipoxygenase was calculated using the following equation:

% Inhibition = 100 × (absorbance of the control − absorbance of the sample)/absorbance of the control

RESULTS AND DISCUSSION:

Extraction, phytochemical screening and total phenolic content:

The extract was pale-brown in color and obtained as sticky materialwith and extraction yield of17.2%. It revealed the presence of alkaloids, glycosides, phenolics, saponin, tannins, flavonoids and sterols (Table 1).

Table 1. Phytochemical analysis of Morinda citrifolia fruit aqueous extract

Test	Observation	Inference
Mayer test	-	Alkaloid absent
Dragendrof test	-	Alkaloid absent
Wagner test	+	Alkaloid present
Gelatin	+	Phenolic present
Vanillin hydrochloride test	+	Phenolic present
Alkaline reagent	+	Phenolic present
Shinoda	-	Flavonoid absent
Zinc hydrochloride	+	Flavonoid present
Alkaline reagent	+	Flavonoid present
Ninhydrin	-	Protein absent
Salkowski	+	Sterol present
Froth	+	Saponin present
Borntrager	+	Glycoside present

The total phenolic content was determined using Folin-Ciocalteau method and was found to be 27.72 GAE mg/g of the extract.

Anti-inflammatory activity:

The anti-inflammatory action was assessed using the ability of the abstract to inhibit lipoxygenase enzyme in in vitro test. The extract was tested at four concentrations and ibuprofen at 25 µg/mL concentration was used as standard. The percent inhibition of lipoxygenase enzyme ranged from 6.50 to 47.54 % for the extract solutions (Table 2).

Figure 1: Effect of extract on lipoxygenase inhibition

Table 2: % inhibition of lipoxygenase

Sample	Absorbance at 234nm	% inhibition of lipoxygenase
Control	1.251	-
25µg/mL extract	1.183	6.505±4.554
50µg/mL extract	0.987	21.827±2.368
75µg/mL extract	0.906	29.214±5.642
100µg/mL extract	0.681	47.536±6.708
Ibuprofen (25µg/mL)	0.447	64.299±0.454

Lipoxygenases are the key enzymes in the biosynthesis of leukotrienes. Leukotrienes play an important role in several inﬂammatory diseases, such as arthritis, asthma, cancer, and allergic diseases.²³ The mechanism of anti-inﬂammation may involve a series of events in which the metabolism of arachidonic acid plays an important role.²⁴ In this process, arachidonic acid is cleaved from the membrane phospholipids upon appropriate stimulation of neutrophils, and can be converted to leukotrienes and prostaglandins through lipoxygenase and cyclooxygenase pathways, respectively.²⁵Lipoxygenase catalyzes deoxygenation of polyunsaturated fatty acids to produce cis, trans-conjugated dienehydroperoxides, such as leukotrienes, which are essential mediators in a variety of inﬂammatory events.²⁵ Previous studies have shown that polyphenols may block or interfere with the cascade process of arachidonic acid metabolism by inhibiting lipoxygenase activity and, also, they may serve as scavengers of various reactive free radicals which are produced during arachidonic acid metabolism.²⁵

CONCLUSION:

The present investigation had thrown light on the remarkable potential of commonly available plant Costus igneus in terms of its pharmacological benefits it offers. The aqueous extract of the leaves of Costus igneus was found to be effective in the inhibition of inflammatory response. Further investigations need to be carried out for determining the active principle in extract responsible of the anti-inflammatory action.

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Received on 29.03.2024 Revised on 30.12.2025

Accepted on 02.02.2026 Published on 20.05.2026

Available online from May 25, 2026

Research J. Pharmacy and Technology. 2026;19(5):2273-2275.

DOI: 10.52711/0974-360X.2026.00327

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