# Development of nanoemulsion formulation loaded with Enteromorpha intestinalis extract: Characterization and evaluation for topical use

**Authors:** Shumaila Qadir, Shahlla Imam, Syed Abid Ali, Fatima Ramzan Ali, Wajiha Iffat, Rabia Ismail Yousuf, Rafia Usman, Shahana Wahid, Iqbal Azhar

PMC · DOI: 10.1371/journal.pone.0343626 · 2026-03-06

## TL;DR

This paper develops a nanoemulsion using green seaweed extract for topical use, showing it is stable, effective, and safe for skin application.

## Contribution

The study introduces a novel nanoemulsion formulation incorporating Enteromorpha intestinalis extract for the first time.

## Key findings

- The optimal nanoemulsion (F6) had a droplet size of 183.27 ± 20.04 nm and good skin compatibility.
- The nanoemulsion showed greater antioxidant efficacy than the seaweed extract alone with an IC50 of 163.19 μg/mL.
- The formulation significantly inhibited paw volume in an anti-inflammatory test and was non-irritating.

## Abstract

Enteromorpha intestinalis is a green seaweed enriched with diverse bioactive compounds that possess substantial pharmacological and biotechnological properties. Despite the focus on their therapeutic applications, research on the development of nano-formulations is limited. Therefore, this study aimed to develop an E. intestinalis (EI) extract-based nanoemulsion for topical use. Olive oil, Tween-80 (surfactant), and PEG-400 (co-surfactant) were selected for the formulation of the nanoemulsion. The Smix ratio was set to 1:1 using a pseudo-ternary phase diagram. Moreover, a design experiment ascertained the composition of the formulation, followed by physicochemical characterization. The optimal formulation, based on droplet size, was selected for further analysis. Stability studies, antioxidant and anti-inflammatory properties of selected nanoemulsion formulation were determined and acute dermal toxicity assay was also performed. The best formulation (F6) displayed a droplet size of 183.27 ± 20.04 nm, PDI of 0.6, and viscosity of 290 ± 5.77 m-Pa.S. The developed nanoemulsion exhibited good skin compatibility and a slightly acidic pH. Both the extract and nanoemulsion formulation exhibited concentration-dependent antioxidant activity. The nanoemulsion had a lower IC50 value of 163.19 μg/mL, showing greater efficacy than the seaweed extract alone. Formulation (F6) also significantly (p < 0.05) inhibited paw volume (8–31%) compared to the control, while diclofenac sodium achieved a maximum inhibition of 41%. The designed formulation was stable, effective, and non-irritating demonstrating its potential topical application. This study presents, for the first time, a nanoemulsion formulation that incorporates E. intestinalis extract. This advancement paves the way for further in vivo studies to assess the efficacy and safety of this formulation for clinical applications.

## Linked entities

- **Chemicals:** Tween-80 (PubChem CID 443315), diclofenac sodium (PubChem CID 5018304)

## Full-text entities

- **Diseases:** irritability (MESH:D001523), cancer (MESH:D009369), Paw edema (MESH:D004487), acute inflammation (MESH:D007249), Enteromorpha intestinalis (MESH:D005873), Skin irritation (MESH:D012871), rash (MESH:D005076), toxicity (MESH:D064420), erythema (MESH:D004890), dermal irritation (MESH:D016136), inflammatory drugs (MESH:D000081015)
- **Chemicals:** polysaccharides (MESH:D011134), quercetin (MESH:D011794), ferric chloride (MESH:C024555), hexadecane (MESH:C007932), 2, 2-diphenyl-1-picrylhydrazyl (MESH:C004931), rutin (MESH:D012431), polymer (MESH:D011108), Loliolide (MESH:C030425), diosmin (MESH:D004145), phosphate (MESH:D010710), Prussian blue (MESH:C000170), phytol (MESH:D010836), O (MESH:D010100), Carrageenan (MESH:D002351), PEG 400 (MESH:C000595213), alkane (MESH:D000473), Ascorbic acid (MESH:D001205), quinic acid (MESH:D011801), ethanol (MESH:D000431), heneicosane (MESH:C554094), Water (MESH:D014867), phenol (MESH:D019800), 2, 4- di-tert-butylphenol (MESH:C056559), carotenoids (MESH:D002338), Diclofenac Sodium (MESH:D004008), free radical (MESH:D005609), ethyl ester (MESH:C465446), stearic acid (MESH:C031183), Fe++ (MESH:D007501), peptides (MESH:D010455), cinnamic acid (MESH:C029010), F4 (MESH:C006011), apigenin (MESH:D047310), terpenes (MESH:D013729), coconut oil (MESH:D000074263), amino acids (MESH:D000596), eicosane (MESH:C050821), phenols (MESH:D010636), pentadecanoic acid (MESH:C117025), trichloroacetic acid (MESH:D014238), n-heptadecane (MESH:C016486), hydrocarbons (MESH:D006838), Oil (MESH:D009821), fatty acid (MESH:D005227), TMS (MESH:D013932), monounsaturated fatty acids (MESH:D005229), K3Fe(CN)6 (MESH:C028033), Olive oil (MESH:D000069463), Smix (-), DS (MESH:D003903), palmitoleic acid (MESH:C008757), elaidic acid (MESH:C011459), polyunsaturated fatty acids (MESH:D005231), hexahydrofarnesyl acetone (MESH:C014584), Tween 80 (MESH:D011136), oleic acid (MESH:D019301), phenanthrene (MESH:C031181), alcohols (MESH:D000438), ellagic acid (MESH:D004610), flavonoid (MESH:D005419)
- **Species:** Ulva intestinalis (hollow green seaweed, species) [taxon 3116], Rattus norvegicus (brown rat, species) [taxon 10116], Homo sapiens (human, species) [taxon 9606], Ulva prolifera (species) [taxon 3117], Gracilaria (genus) [taxon 2774], Ulva compressa (species) [taxon 63659]
- **Mutations:** E 30H

## Figures

22 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12965610/full.md

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Source: https://tomesphere.com/paper/PMC12965610