EXTRACTION OF FUCOXANTHIN FROM SARGASSUM WIGHTII, FTIR SPECTRA AND IT’S ANTIOXIDANT AND ANTIBACTERIAL PROPERTIES

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EXTRACTION OF FUCOXANTHIN FROM SARGASSUM WIGHTII, FTIR SPECTRA AND IT’S ANTIOXIDANT AND ANTIBACTERIAL PROPERTIES

S. Doshi *, S. Mukadam, P. Aslot, M. Gohil and P. Patel

Department of Biochemistry and Biotechnology, St. Xavier’s College, Gujarat University, Ahmedabad, Gujarat, India.

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ABSTRACT: Sargassum wightii,a seaweed and a marine macroalgae, is known to offer numerous health benefits, and compounds isolated from it are observed as the alternative to conventional treatment as an affordable plant-derived drug. In the present study, the bioactivity of the two fractions from Sargassum wightiiw as derived using two different solvents, methanol and acetone: methanol. Acetone: Methanol fraction showed better yield. Both the fractions it was tested against both gram-positive Staphylococcus aureus and gram-negative Escherichia coli bacteria. The antioxidant potential was evaluated by the reducing power assay. Fucoxanthin extracted by maceration in methanol: acetone (9:1) showed more excellent antioxidant activity. The partially purified extract was also analysed through the FT-IR spectra. The extract prepared by methanol: Acetone extract performed better than streptomycin, but compared to streptomycin, none of the extracts showed significant antibacterial action at higher concentrations. The FT-IR spectra showed various secondary metabolites and their derivatives in the crude extracts.

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Keywords: Sargassum weightii, Antimicrobial Action, Antioxidant activity, FT-IR spectra

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INTRODUCTION: Seaweeds, also known as marine macroalgae, are aquatic plants that grow in both fresh and saltwater environments. They are categorised into three primary categories include: green, red, and brown seaweeds ¹. Marine macroalgae are significant due to their roles in aquatic ecosystems, their potential for producing food, oxygen, and bioactive compounds, and their participation in the global carbon cycle ². The Sargassum genus is a diversified collection of brown seaweeds usually found in tropical and subtropical waters worldwide. It is a member of the Sargassaceae family ³.

One of the members of the genus Sargassaceae, Sargassum wightii, also known as Wight's Sargassum, is found in the Arabian and Indian oceans ⁴. The seaweed Sargassum wightii is abundant in bioactive substances compressing of, polysaccharides, fucoidan, fucoxanthin, Xanthophyll, terpenoids, etc., ⁵.  Additionally, it has anti-obesity, anti-cancer, antioxidant, anti-bacterial and anti-inflammatory properties ⁶.

The Sargassum genus is also known to have skin-healing properties. The presence of these properties is attribute of the carotenoid along with Xanthophyll pigments, which give S. wightii a brown colour ⁵. One of the essential carotenoids is Fucoxanthin, which is promoted as a nutraceutical ⁷. Fucoxanthin is the marine carotenoid responsible for the brown colour of the seaweed; There are various types of seaweed species, comprising Undaria, Fucus, Sargassum and Myagropsis, in addition to microalgae such as Phaeodactylum, Cyclotella, Isochrysis,, Nitzschia and Prymnesium, contain, conjugated carbonyl groups and  an unusual allenic bond, a 5,6-monoepoxide group in their polyene chain. These compounds possess antioxidant, anti-obesity, anti-inflammatory, and anticancer properties ^{5, 8}. Many of these attributes are because fucoxanthin can protect against oxidative stress by inhibiting DNA-damaging agents. Further, it showed no toxicity or mutagenicity in mice under the experimental conditions ⁹. The extraction yield of Fucoxanthin from various seaweeds depends upon the species chosen, the growth conditions, and the extraction method.

The antibacterial actions of the fucoxanthin have not been explored much except by a few groups, as mentioned. Using the disc-diffusion method, Fucoxanthin isolated from the Himanthalia elongata showed antimicrobial activity against Listeria monocytogenes. The fucoxanthin isolated from the Undaria pinnatifida demonstrated significant antimicrobial properties targeting gram-positive bacteria. The crude leaf extract from the sargassum weight was shown to be effective against the periodontal pathogen ¹⁰.

This study investigates the antimicrobial efficacy of Fucoxanthin, obtained through ultrasonication from Sargassum weight, effective against both of gram-positive (GPB) and gram-negative (GNB) bacteria. The assays had been conducted using the microdilution method in a 96-well plate. In addition, the fractions had been characterized using Fourier Transform Infrared Spectroscopy (FTIR), and their antioxidant capacity was assessed. FTIR, and their antioxidant potential was also evaluated.

MATERIALS AND METHODS:

Preparation of Sargassum wightii Extracts: The extraction was performed using the method developed by Oliyaei N et al. ¹¹. With minor alterations. Two different solvent systems were tried to extract the fucoxanthin; in one of the methods, only acetone was used, while in another, methanol-acetone in the ratio of 9:1 was taken. The desiccated sample was immersed (at a ratio of 1:30 weight/volume) in each of these two solvent systems and placed on a rotary shaker operating at 100 revolutions per minute at room temperature for a duration of 24 hours, while being placed in dark. The extraction process was performed continuously until the solvent became colourless, and the extracts were pooled together and dried.

Fourier Transform Infrared Spectroscopy (FTIR) Analysis: Dried samples had been combined with powdered KBr and compressed into a fine pellet. FTIR spectra of the extracted compounds were measured within at the wave numbers range of 4000 cm^-1 and 500 cm^{-1 12}.

Antioxidant Activity Analysis: The reducing capacity of fucoxanthin obtained from both methods was assayed in accordance with the process of Husni et al., ¹³.

The brief, sample of 1 ml solutions with varying concentrations were combined with potassium ferricyanide aqueous solution (2.5 mL, 1%) and potassium phosphate buffer (2.5 mL, 0.2mol/L, pH 6.6). Mixture was then subjected incubated at temperature 50^oC for 20 minutes, subsequent addition of aqueous ferric chloride solution (0.5 mL, 1%) and trichloroacetic acid solution (2.5 mL, 1%) The optical density of the reaction mixture quantified at a wavelength of 700 nm. The reducing power exhibited a direct correlation with the optical density of the reaction mixture.

Antibacterial Activity Determination: The assay used two test strains: Staphylococcus aureus and Escherichia coli. Streptomycin antibiotic was employed as a positive control. The antibacterial activity of the compounds was assessed using the protocol developed by Sultanbawa et al., with the specified modifications ¹⁴.

Each 96-well plate contained single row of wells containing 200µl sterile medium (sterility control). One row contained 50µl bacterial solution with 150µl medium and no extracts (negative control). One row consisted of bacterial cultures with streptomycin dilutions ranging from 10µg/ml to 100mg/ml. The remaining rows contains bacterial cultures with extract dilutions ranging from 10µg/ml to 100 µg/ml. Each 300µl well contained 50µl of inoculum, 2µl of extract, and 148µl of nutrient broth. The plates placed in an incubator set at a temperature of 37⁰C for 22 hours. The measurement of optical density was conducted using a spectrophotometer at a wavelength of 595 nm.

RESULTS:

Extraction Yield: The efficiency of the extraction procedures followed was evaluated by calculating the percentage yield of extracts obtained for each method.

Fucoxanthin extracted by maceration using a 9:1 mixture of methanol-acetone showed the highest yield (13.787%), while fucoxanthin extracted using acetone as solvent showed comparatively less yield (0.537%) Fig. 1. Thus, the mixture of solvents (9:1 methanol-acetone) proved to be better for the extraction of fucoxanthin.

FIG. 1: YIELD OF EXTRACTION OF FUCOXANTHIN FROM SARGASSUM WIGHTII BY TWO DIFFERENT SOLVENTS. Methanol: Acetone (9:1) gave a significantly higher yield. P-value was computed using GraphPad Prism software (Version 8) using one-way ANOVA. (Data represents the mean value ± S.D., n=3). (****denote the statistical significant difference (p < 0.0001) between the yield of extraction).

Reducing Power Assay: An antioxidant capacity assessment was conducted using a reducing power assay to evaluate the effectiveness of fucoxanthin extracted from S. wightii. Ascorbic acid, a standard antioxidant, was employed as positive control. The reducing power of fucoxanthin amplified in a concentration dependent manner as the concentration of fucoxanthin increased from 5 µg/ml to 30 µg/ml. However, the reducing capacity exhibited a lower value compared to that of ascorbic acid Fig. 2. Fucoxanthin extracted by maceration in methanol: acetone (9:1) showed more excellent antioxidant activity.

FIG. 2: THE GRAPH SHOWS THE ANTIOXIDANT ACTIVITY OF SARGASSUM WIGHTII IN TERMS OF ASCORBIC ACID EQUIVALENT. The methanol and acetone extracts showed more potential at all concentrations.

Antibacterial Activity: The antibacterial activity of fucoxanthin extracted from S. wightii was determined with respect to streptomycin on 2 bacterial strains, E. coli, gram negative and S. aureus, gram positive Fucoxanthin significantly inhibits growth of E. coli at higher concentrations. Fucoxanthin extracted from S. wightii by maceration using two different solvent systems showed increasing antimicrobial activity with increasing concentration. Fucoxanthin was more effective than the positive control at lower concentrations, but a higher concentration of streptomycin was more effective than fucoxanthin extracts Fig. 3.

FIG. 3: THE FIGURE DEPICTS THE ANTIBACTERIAL ACTION OF THE TWO EXTRACTS PREPARED THROUGH DIFFERENT SOLVENT. The data represents the mean ± S.D. The data was examined in triplicate. The P value was computed using GraphPad Prism software through a two-way ANOVA analysis, resulting in a value of 0.001 for the comparison between S. aureus and E. coli.

FTIR Analysis: The FT-IR analysis of extracts from S. wightii showed peaks similar to the standard fucoxanthin. The distinctive wave frequencies of characteristic functional groups identified in both purified samples were as follows: C–H stretch (2918 - 2929 cm⁻¹), allene (1929cm⁻¹), OH group (3374 - 3421cm⁻¹,C=O acetate and conjugated C=O (1634 – 1639cm⁻¹) and trans-distributed –C=C– (1201 – 958cm⁻¹) ¹⁵.

However, the peak value obtained at 1929cm⁻¹ corresponding to the allenic group of fucoxanthin ¹⁵ and is only obtained in the extract obtained by maceration using a mixture of methanol-acetone. The allene stretch is present but not significant in the other extracts Table 1. In this study, the extracts prepared by both solvents showed different degrees of antioxidant activities.

TABLE 1: FTIR PEAK VALUES AND FUNCTIONAL GROUPS OF SARGASSUM WIGHTII EXTRACTS

Extracts	Peak Value	Functional Group	Functional Group Name	Vibration
Fucoxanthin Maceration (Acetone)	719.96	Unidentified	Benzene derivative	Bending
	1161.37	S=O	Sulfonic acid	Stretching
	1462.78	C-H	Alkane	Bending
	2849.85	C-H	Alkane	Stretching
	2918.42	C-H	Alkane	Stretching
	3411.25	N-H	Amine	Stretching
Fucoxanthin Maceration (Methanol: Acetone; 9:1)	1074.23	S=O	Sulfoxide	Stretching
	1639.91	C=C	Alkene	Stretching
	2358.45	O=C=O	Carbon Dioxide	Stretching
	2919.85	C-H	Alkane	Stretching
	3395.53	N-H	Amine	Stretching

DISCUSSION: In this present study, fucoxanthin was partially purified using maceration as the extraction technique and acetone and methanol acetone as the solvents. The extract obtained using two different solvents showed differences in their FT-IR spectra and antibacterial activities. Several different solvent systems have been employed to extract fucoxanthin from the seaweed previous findings show that fucoxanthin extracts were more effective than other antibiotics like clotrimazole and chloramphenicol ^{16, 17}.

Fucoxanthin exhibited greater efficacy against Gram Positive bacteria (S. aureus) compared to Gram negative bacteria (E. coli), aligning with obtained results ^{16, 18, 19}. Acetone and methanol were considered tobe the best solvents to extract antimicrobial components from the different types of seaweed, as described by Cox et al., and hence, the utilization of acetone and methanol is justified in this study, with particular emphasis on the use of acetone ²⁰.

In another report, Moubayed NM ET. Alcon ducted the screening study of antimicrobial potency of sea weeds encapsulated from the coast of Saudi Arabia. Seaweed extracts have been found to be more effective against GPB, according to report.  This aligns with our result in which it was also noted that Sargassum weightii is less efficient against E. coli than S. Aureus ²¹. These might be because of the variance in the composition of the cell walls of GNB and GPB. FTIR chemical analysis reveals the presence of various bioactive components, which are known to have antimicrobial and antioxidant activities. These findings is in agreement with the one reported by.

The Methanol: acetone extracts of Sargassum weightii showed more reduced power activity. The reducing power ensures that the antioxidant compounds serve as donor of electron, reducing the oxidized intermediate of the lipid peroxidation process. Additionally, they can act as primary 1⁰ and secondary 2⁰ antioxidants (Yen and Chen, 1995). The concentration-dependent antioxidant activity was studied about reducing power, providing a comprehensive studying of the antioxidants available in the sample. It had been observed that the reducing power increased with higher concentrations in all the samples ²².

Based on the present studies, Sargassum weight can be used as a beneficial antimicrobial agent; however, further studies are needed to purify its secondary metabolites further, and their efficacy can be tested on more microorganisms.

CONCLUSION: The presented study evaluates the antibacterial and antioxidant activity of acetone and 9:1 methanol: acetone extracts of S. wightii. Two methods, one conventional (maceration) and the other advanced (ultrasonication), were used to extract fucoxanthin and fucoidan using the stated solvent systems. The extraction yields were significantly low with acetone as solvent. FTIR spectra of the extracted compounds were inconclusive regarding the purity of extracts, indicating the presence of certain other pigments and polysaccharides alongside fucoidan and fucoxanthin. The antioxidant activity of fucoxanthin extracted by maceration was the highest, followed by fucoxanthin extracted by UAE.

ACKNOWLEDGEMENT: We want to express our sincere appreciation to the Department of Biochemistry and Biotechnology, St. Xavier’s College, Ahmedabad, and all its faculty, whose contributions and support have greatly enhanced the quality and rigour of this research.

CONFLICT OF INTEREST: All authors declare that they have no conflicts of interest.

REFERENCES:

1. El-Beltagi HS, Mohamed AA, Mohamed HI, Ramadan KMA, Barqawi AA and Mansour AT: Phytochemical and Potential Properties of Seaweeds and Their Recent Applications: A Review. Mar Drugs 2022; 20(6): 342. doi:10.3390/md20060342
2. Cotas J, Leandro A and Monteiro P: Seaweed Phenolics: From Extraction to Applications. Mar Drugs 2020; 18(8): 384. doi:10.3390/md18080384
3. Sanjeewa KKA, Kang N, Ahn G, Jee Y, Kim YT and Jeon YJ: Bioactive potentials of sulfated polysaccharides isolated from brown seaweed Sargassum spp in related to human health applications: A review. Food Hydrocoll 2018; 81: 200-208. doi:10.1016/j.foodhyd.2018.02.040
4. Ajith S, Rojith G and Zacharia PU: Production, Characterization and Observation of Higher Carbon in Sargassum wightii Biochar from Indian Coastal Waters. J Coast Res 2019; 86(1): 193-197. doi:10.2112/SI86-029.1
5. Afreen AB and Rasool F: Section Zoology, School of Sciences, Maulana Azad National Urdu University, Hyderabad-500 032, India,  Bioactive properties of brown seaweed, Sargassum wightii and its nutritional, therapeutic potential and health benefits: A review. J Environ Biol 2023; 44(2): 146-158. doi:10.22438/jeb/44/2/MRN-5081
6. Lee MK, Ryu H and Lee JY: Potential Beneficial Effects of Sargassum spp. in Skin Aging. Mar Drugs 2022; 20(8): 540. doi:10.3390/md20080540
7. Hong DD, Thom LT and Ha NC: Isolation of Fucoxanthin from Sargassum oligocystum Montagne, 1845 Seaweed in Vietnam and its Neuroprotective Activity. Biomedicines 2023; 11(8): 2310. doi:10.3390/biomedicines11082310
8. Oliyaei N, Moosavi-Nasab M, Tanideh N and Iraji A: Multiple roles of fucoxanthin and astaxanthin against Alzheimer’s disease: Their pharmacological potential and therapeutic insights. Brain Res Bull 2023; 193: 11-21. doi:10.1016/j.brainresbull.2022.11.018
9. Oliyaei N, Moosavi-Nasab M, Tamaddon AM and Tanideh N: Antidiabetic effect of fucoxanthin extracted from Sargassum angustifolium on streptozotocin-nicotinamide-induced type 2 diabetic mice. Food Sci Nutr 2021; 9(7): 3521-3529. doi:10.1002/fsn3.2301
10. Bk N, Ac K, Puladas H, KS, KP and SG: Antibacterial efficacies of brown sea weed Sargassum wightii on Periodontal Pathogens: An in-vitro Microbiological Analysis. J Ayurveda Integr Med Sci 2022; 7(11): 52-58. doi:10.21760/jaims.7.11.9
11. Oliyaei N and Moosavi‐Nasab M: Ultrasound‐assisted extraction of fucoxanthin from Sargassum angustifolium and Cystoseira indica brown algae. J Food Process Preserv 2021; 45(11). doi:10.1111/jfpp.15929
12. Wongsa P, Phatikulrungsun P and Prathumthong S: FT-IR characteristics, phenolic profiles and inhibitory potential against digestive enzymes of 25 herbal infusions. Sci Rep 2022; 12(1): 6631. doi:10.1038/s41598-022-10669-z
13. Husni A, Izmi N, Ayunani FZ, Kartini A, Husnayain N and Isnansetyo A: Characteristics and antioxidant activity of fucoidan from Sargassum hystrix: Effect of Extraction Method. Int J Food Sci 2022; 2022(1): 3689724. doi:10.1155/2022/3689724
14. Sultanbawa Y, Cusack A, Currie M and Davis C: An innovative microplate assay to facilitate the detection of antimicrobial activity in plant extracts. J Rapid Methods Autom Microbiol 2009; 17(4): 519-534. doi:10.1111/j.1745-4581.2009.00187.x
15. Selvaraj B and Ganapathy D: Exploration of Sargassum wightii: extraction, phytochemical analysis, and antioxidant potential of polyphenol. Cureus published online 2024. doi:10.7759/cureus.63706
16. Liu Z, Sun X, Sun X, Wang S and Xu Y: Fucoxanthin Isolated from Undaria pinnatifida Can Interact with Escherichia coli and lactobacilli in the Intestine and Inhibit the Growth of Pathogenic Bacteria. J Ocean Univ China 2019; 18(4): 926-932. doi:10.1007/s11802-019-4019-y
17. Ibrahim EA, El-Sayed SM, Murad SA, Abdallah WE and El-Sayed HS: Evaluation of the antioxidant and antimicrobial activities of fucoxanthin from Dilophys fasciola and as a food additive in stirred yoghurt. South Afr J Sci 2023; 119(3/4). doi:10.17159/sajs.2023/13722
18. Rajauria G and Abu-Ghannam N: Isolation and Partial Characterization of Bioactive Fucoxanthin from Himanthalia elongata Brown Seaweed: A TLC-Based Approach. Int J Anal Chem 2013; 2013: 1-6. doi:10.1155/2013/802573
19. Karpiński TM and Adamczak A: Fucoxanthin an antibacterial carotenoid. Antioxidants 2019; 8(8): 239. doi:10.3390/antiox8080239
20. Cox S, Abu-Ghannam N and Gupta S: An Assessment of the Antioxidant and Antimicrobial Activity of Six Species of Edible Irish Seaweeds. In: Dublin Institute of Technology 2010. doi:10.21427/D7HC92
21. Moubayed NMS, Al Houri HJ, Al Khulaifi MM and Al Farraj DA: Antimicrobial, antioxidant properties and chemical composition of seaweeds collected from Saudi Arabia (Red Sea and Arabian Gulf). Saudi J Biol Sci 2017; 24(1): 162-169. doi:10.1016/j.sjbs.2016.05.018
22. M J and A J: In-vitro evaluation of antioxidant potential and total phenolic content of Barleria longiflora leaf extracts. Asian J Pharm Clin Res. Published online 2020: 212-215. doi:10.22159/ajpcr.2020.v13i1.35468
    

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    How to cite this article:

    Doshi S, Mukadam S, Aslot P, Gohil M and Patel P: Extraction of Fucoxanthin from Sargassum wightii, FTIR spectra and it’s antioxidant and antibacterial properties. Int J Pharm Sci & Res 2025; 16(2): 445-50. doi: 10.13040/IJPSR.0975-8232.16(2).445-50.

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