Antioxidant Stability Research Articles

Identification and Functional Validation of Two Novel Antioxidant Peptides in Saffron.

Saffron (Crocus sativus L.) is one of the most expensive spices in the world, boasting rich medicinal and edible value. However, the effective development of active natural substances in saffron is still limited. Currently, there is a lack of comprehensive studies on the saffron stigma protein, and the main effect peptides have not been identified. In this study, the total protein composition of saffron stigmas was analyzed, and two main antioxidant peptides (DGGSDYLGK and VDPYFNK) were identified, which showed high antioxidant activity. Then, the stability of two peptides was further evaluated. Furthermore, our results suggested that these two peptides may protect HepG2 cells from H2O2-induced oxidative damage by significantly improving the activity of endogenous antioxidant enzymes and reducing the malondialdehyde (MDA) content. Collectively, we identified two peptides screened from the saffron protein possessing good antioxidant activity and stability, making them promising candidates for use as functional foods, etc., for health promotion. Our findings indicated that proteomic analysis together with peptide identification is a good method for exploitation and utilization of spice plants.

Breathable and washable MXene@gallic acid cross-linked textile assisted by borax for personal multi-protection wearables

Achieving multifunctional integration with comfortability and durability comparable to conventional textiles is one of the ultimate pursuits of personal protective wearables. Herein, a multi-protective textile with breathable, moisture-permeable and washable properties is reported, which is realized by cross-linking of electroactive MXene@gallic acid (MG) with the cationized cotton. Cationic modification reduces the charge repulsion between MG and cotton fibers, and promotes the deposition of conductive media on textile surface. After borax (STB) cross-linking, the obtained textile exhibits superior service stability and can be subjected to conventional washing, exhibiting antioxidant stability during natural storage. Importantly, the cross-linked textile inherits the properties of good breathability, softness and bendability as the original fabric. With the outstanding conductivity (260 mS/cm) and high absorptivity of the solar spectrum (0.920), the textile was used for Joule heating and solar heating, demonstrating their potential in the field of thermal management. Meanwhile, the textile demonstrates excellent antibacterial activity against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus, and maintained satisfactory antibacterial effect (>90 %) even after ten washing cycles. This facile strategy for fabricating personal protective textiles, based on cross-linking cotton fiber with electroactive mediators offers favorable durability, comfortability and multifunctionality, providing a scalable and sustainable approach for manufacturing multi-protective wearables.

Fabrication of EGCG‐loaded Spirulina platensis microcarrier with sustained release properties and their application in extending the shelf‐life of minced lamb meat

SummaryIn the present study, epigallocatechin‐3‐gallate (EGCG)‐loaded Spirulina platensis (SP) microcarriers (SP@EGCG) were constructed and used to extend the shelf‐life of minced lamb meat as an antioxidant with good sustained release properties. Results showed that the encapsulation efficiency and encapsulation yield of SP@EGCG microcarriers were 91.19 ± 1.18% and 133.19 ± 1.34 μg mg−1, respectively. Results of fluorescence microscopy, Fourier transform infrared spectroscopy, X‐ray diffraction, scanning electron microscopy and differential scanning calorimetry confirmed that the EGCG was successfully loaded in SP through hydrogen bonding interactions, and existing in an amorphous state. SP@EGCG displayed excellent antioxidant activity and stability. The SP@EGCG presented cumulative release in four food models (water, 3% acetic acid, 10% ethanol and 50% ethanol), and the cumulative release of SP@EGCG in 50% ethanol was the highest. These results indicated that SP can be used as a novel carrier of EGCG, and the prepared SP@EGCG microcarriers had potential applications in extending the shelf‐life of lamb meat.

Potential of Organic Acids, Essential Oils and Their Blends in Pig Diets as Alternatives to Antibiotic Growth Promoters.

Over the years, the use of management and feeding strategies to enhance pig productivity while minimizing the use of antibiotic growth promoters has grown. Antibiotic growth promoters have been widely used as feed additives to reduce diet-related stress and improve pig performance. However, increasing concern about the consequences of long-term and increased use of antibiotic growth promoters in animal production has led to a paradigm shift towards the use of natural organic alternatives such as plant essential oils and organic acids in pig nutrition to enhance growth. Antibiotic growth promoters endanger human health by allowing multidrug-resistant genes to be transferred horizontally from non-pathogenic to pathogenic bacteria, as well as directly between animals and humans. Scientific research shows that alternative growth promoters such as essential oils and organic acids appear to improve pigs' ability to prevent pathogenic bacteria from colonizing the intestinal system, stabilizing the gut microflora and promoting eubiosis, as well as improving immunity and antioxidant stability. The purpose of this review was to provide an in-depth review of organic acids and essential oils as growth promoters in pig production, as well as their effects on productivity and meat quality. Organic acids and essential oils in pig diets are a safe way to improve pig performance and welfare while producing antibiotic-free pork.

Antioxidant and Emulsion Stability of Citrus Peel Dressing

Antioxidant and Emulsion Stability of Citrus Peel Dressing

Encapsulation and Characterization of Proanthocyanidin Microcapsules by Sodium Alginate and Carboxymethyl Cellulose.

Proanthocyanidins are important compounds known for their antioxidant and radical scavenging properties, but they are highly sensitive to light, heat, oxygen, and pH. In our study, proanthocyanidin was encapsulated using sodium alginate and carboxymethyl cellulose to enhance controlled release, pH stability, metal ion tolerance, temperature resistance, time release, the microencapsulation of food additives stability, antioxidant capacity analysis, and the storage period tolerance of proanthocyanidin. Fourier transforms infrared (FTIR) analysis and full-wavelength UV scanning indicated the successful immobilization of proanthocyanidins into the polymeric microcapsules. The flowability and mechanical properties of the microcapsules were enhanced. Moreover, proanthocyanidin microcapsules exhibited higher thermal, pH, metal ion, time, and microencapsulation food additive stability. In addition, due to their high antioxidant properties, the proanthocyanidin microcapsules retained a greater amount of proanthocyanidin content during the gastric phase, and the proanthocyanidin was subsequently released in the intestinal phase for absorption. Thus, the study provided a systematic understanding of the antioxidant capabilities and stability of proanthocyanidin microcapsules, which is beneficial for developing preservation methods for food additives.

Multi-spectroscopic and molecular docking studies for the pH-dependent interaction of β-lactoglobulin with (−)-epicatechin gallate and/or piceatannol: Influence on antioxidant activity and stability

(−)-Epicatechin gallate (ECG) and piceatannol (PIC) are commonly polyphenols with excellent biological activities. β-Lactoglobulin (BLG) is a food-grade globule protein and its morphologies are sensitive to pH. This study used experimental and computational methods to determine the interaction of single or combined ECG and PIC with BLG at different pHs. The static quenching process was determined through fluorescence and ultraviolet–visible spectroscopy. Compared with ECG, PIC could significantly bind to BLG with higher affinity. Their binding affinity for BLG with different morphologies followed the tendency of monomer > dimer > tetramer. The negative contribution of van der Waals forces, electrostatic interactions, and hydrogen bonds to ΔHo exceeded the positive contribution of hydrophobic interactions in the spontaneous and exothermic process. The reduced binding affinity in the ternary systems demonstrated the competitive binding between ECG and PIC on BLG, and the hinder effect of ECG or PIC was enhanced with increasing pH. Molecular docking studies revealed the same binding sites of ECG and PIC on various conformations of BLG and identical driven forces as thermodynamic results. Tryptophan and tyrosine were the main participators in the BLG + ECG and BLG + PIC systems, respectively. The conformational changes in the binary and ternary systems could be ascertained through synchronous fluorescence, circular dichroism, and dynamic light scattering. Furthermore, the effects of pH and BLG encapsulation on the antioxidant capacity and stability of ECG or PIC were also implemented. ECG or PIC was the most stable in the (BLG + PIC) + ECG system at pH 6.0. This study could clarify the interaction mechanism between ECG/PIC and BLG and elucidate the pH effect on their binding information. The results will provide basic support for their usage in food processing and applications.

Improvement of antioxidant, emulsification properties and thermal stability of egg white protein by covalent binding to gallic acid/tannic acid and xanthan gum

This study aimed to improve the functional properties of egg white protein (EWP) by covalent modification of EWP with xanthan gum (XG) and gallic acid (GA)/tannic acid (TA) to form covalent complexes. The structure and functional properties of EWP before and after modification were explored. The results showed that the grafting degree of EWP-GA-XG and EWP-TA-XG reached 37.23% and 44.01% respectively measured by the OPA method. The secondary structure of EWP became loose after reaction with GA/TA and XG, with a decrease in the content of α-helix and an increase in the β-sheet content, thus contributing to the improvement of its functional properties. The DPPH and ABTS radical scavenging rate and iron reduction ability of EWP-GA-XG (44.94%, 56.38%, 18.29 mg VC/μmol) and EWP-TA-XG (85.45%, 88.39%, 39.26 mg VC/μmol) complexes were significantly improved compared with EWP (7.90%, 12.94%, 0.12 mg VC/μmol). The emulsion activity index and emulsion stability index of EWP-GA-XG (10.05 m2/g, 96.43%) and EWP-TA-XG (11.04 m2/g, 96.53%) complexes were also improved as compared to those of EWP (5.94 m2/g, 51.45%). Moreover, the peak transition temperatures of EWP-GA-XG (138.09 °C) and EWP-TA-XG (143.96 °C) complexes were higher than that of EWP (125.00 °C). This study may provide the theoretical basis for improving the functional properties of EWP and expanding its application in the field of food.

Anti-fatigue effect of Lateolabrax japonicus peptides in mice and the underlying action mechanism via in vitro and in vivo assays

Fatigue is a common health issue in today's society worldwide. Finding anti-fatigue functional substances seems warranted to alleviate fatigue-related issues. The Lateolabrax japonicus peptides (LPH) obtained via enzymatic hydrolysis were identified being rich in Glu, Gly, and hydrophobic amino acids, which exhibited antioxidant properties. However, other biological functions of LPH have yet to be reported. This study main goal was to investigate the anti-fatigue effects of LPH via a fatigue mice model, and further to elucidate its underlying mechanism using in vivo assay. The results indicated that LPH exhibited strong antioxidant activity and stability in vitro. Meanwhile, LPH significantly extended the swimming time of mice, while also decreasing fatigue marker levels (LA, BUN, CK, and MDA) in serum concurrently by increased levels of antioxidant enzymes, blood glucose, and Na+-K+-ATPase. Histopathological examination revealed that LPH effectively improved tissue damage caused by fatigue. Mechanistically, LPH upregulated the gene and protein expression of Nrf2 and AMPK, indicating its regulatory role in Keap1/Nrf2 and AMPK/SIRT1/PGC-1α/NRF1 signaling pathways. Furthermore, the study demonstrated that LPH could modulate the abundance and balance of fatigue-related gut microbiota (Butyricimonas, Helicobacter, Monoglobus, and Turicibacter). In summary, these findings indicated that LPH exerted its anti-fatigue effect by improving energy metabolism and reducing oxidative stress, suggesting its potential as a promising functional anti-fatigue nutraceutical in food industry.

Effect of three different insect larvae on growth performance and antioxidant activity of thigh, breast, and liver tissues of chickens reared under mild heat stress

This study investigated the potential of insect-based diets to mitigate heat stress impact on broiler chickens, focusing on growth performance and antioxidant stability. Four dietary groups were examined, including a control and three treated groups with Tenebrio molitor (TM), Hermetia illucens (HI), and Zophobas morio (ZM) larvae, respectively, at a 5% replacement ratio. Temperature and relative humidity of the poultry house were monitored. Under heat stress conditions, the HI-fed group consistently exhibited the highest body weight, demonstrating their remarkable growth-promoting potential. TM-fed broilers also displayed commendable growth compared to the control. Insect larvae inclusion in the diet improved feed intake during early growth stages, indicating their positive influence on nutrient utilization. Regarding antioxidant stability, malondialdehyde (MDA) levels in the liver, an oxidative stress and lipid peroxidation marker, were significantly lower in the TM-fed group, suggesting reduced oxidative stress. While the specific insect-based diet did not significantly affect MDA levels in thigh and breast tissues, variations in the total phenolic content (TPC) were observed across tissues, with HI larvae significantly increasing it in the breast. However, the total antioxidant capacity (TAC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) levels did not differ significantly among dietary groups in the examined tissues. Results suggest that insect-based diets enhance broiler growth and potentially reduce oxidative stress, particularly in the liver. Dietary presence of bioactive compounds may contribute to these benefits. Further research is required to fully elucidate the mechanisms underlying these findings. Insect-based diets seem to offer promise as feed additives in addressing the multifaceted challenges of oxidative stress and enhancing broiler health and resilience under heat stress conditions.

Effect of drying and storage on antioxidant activity parameters of celery, parsnip and root parsley chips

Vegetable chips are extremely popular due to high content of antioxidants, valuable biological activity and long-term storage. During the investigation optimal methods of drying and chips storage from celery (Egor and Dobrynya cvs), parsnip (Krugly, Bely aist and Zhemchug) and parsley (Zolushka cv) roots were developed using the criteria parameters of taste, aroma, color and antioxidant content. The highest organoleptic properties were recorded for celery chips of Dobrynya cv, parsnip chips of Krugly cv and root parsleyThe highest differences in chips organoleptic parameters between convectional and freeze-drying methods reached 1.5 times for parsnip Bely aist cv. while the lowest differences were recorded for root parsley (1.05 times). Decrease in antioxidant activity during freeze-drying was the lowest in root parsley and celery Dobrynya cv. Convectional drying did not affect chips total antioxidant activity and demonstrated the highest preservation effect after 8 months storage. On the contrary, vitamin C losses were the highest during convectional drying though during storage vitamin C content in such chips decreased much less than in freezedried products. Aluminum foil provided 1.2-1.8 times higher content of antioxidants than paper and polyethylene package. Significant inter species and varietal differences in antioxidant stability during various methods of drying and storage were revealed.

Nanocomplex of (−)-epigallocatechin-3-gallate and glycated ovalbumin based on natural deep eutectic solvents: Binding analysis and functional improvement

Natural deep eutectic solvents (NADES) are considered an efficient reaction medium for the production of glycated proteins. By optimizing the Maillard reaction conditions through response surface analysis, a glycation product (GOVA) of ovalbumin (OVA) with high grafting degree (92.2%) was prepared in choline chloride/glucose NADES. The binding information of (−)-epigallocatechin-3-gallate (EGCG) to OVA/GOVA was acquired by fluorescence spectroscopy and molecular docking. The interaction strength of GOVA + EGCG was higher than that of OVA + EGCG. Hydrogen bonding and van der Waals forces were the major driving forces in the binding process. Glycation and EGCG binding reduced the surface hydrophobicity, α-helix content and digestibility of OVA. Compared with OVA, the particle sizes of GOVA and GOVA + EGCG were increased by 54.8% and 87.7%, respectively. In addition, glycation increased the emulsifying activity index and foaming capacity of OVA by 32.6% and 20.0%, respectively, which was further increased in the presence of EGCG. The encapsulation of OVA, especially GOVA, improved the antioxidant activity, stability and bioaccessibility of EGCG. The results suggest that the glycated proteins based on NADES may provide an effective alternative for the protection and transport of bioactive ingredients in the food industries.

Emulsified sausages with yeast protein as an animal fat replacer: Effects on nutritional composition, spatial structure, gel performance, and sensory quality

This paper investigated the effect of yeast protein (YP)-fat replacement on the nutritional composition, spatial structure, gel performance, and sensory quality of emulsified sausages. YP is enriched with essential amino acids (36.49 g/100 g), which improved the nutritional quality of sausages whereas reducing its fat content. Moreover, YP could absorb water and fat, thus the YP-added sausages exhibiting an amount-dependent increase in emulsion stability and water migration. The microstructure illustrated that YP acted as a filler to improve structural homogeneity and compactness of the pork gel network. And YP-fat replacement could significantly enhance the hardness, gel strength and elasticity of sausages whereas decreasing the viscosity. Additionally, at partial or full YP-fat replacement (25–100%), the YP-added sausages scored higher in odor and texture, as well as better antioxidant stability than controls. Overall, YP can be employed as a new fat substitute for the preparation of healthy and nutritional sausages, while maintaining the sensory quality.

Thermal processing of pomegranate seed oils underscores their antioxidant stability and nutritional value: Comparison of pomegranate seed oil with sesame seed oil.

In the present study, the oxidative stability and antioxidant activity of seed oils were investigated in three Iranian pomegranate cultivars, Shirin Khafr, Torsh Sabz, and Rabab, along with the sesame (Sesamum indicume L. cv Dezful) seed oil. Punicic acid was the primary fatty acid in the pomegranate seed oils, with contents ranging from 75.5 to 80.9% (w/w). The tocopherol levels in pomegranate seed oils ranged from 1439 to 2053 mg/kg, whereas the phenolics ranged from 130 to 199.3 mg/kg, respectively. Comparatively, in the seed oil of sesame "Dezful," these substances' contents were 1053 and 79 mg/kg, respectively. Contrary to common perception, the seed oil of the three pomegranate cultivars cultivated in Iran had high oxidative stability and antioxidative activity during the 32 h of thermal processing at 170°C. The oxidation stability assayed by peroxide value, p-anisidine value, and TOTOX index revealed that the pomegranate seed oils had a much higher resistance to the oxidation process than the sesame oil. The content of tocopherols increased during thermal processing due to the regeneration phenomenon. Tocopherols are not always free and may form a matrix with themselves or other compounds. Changes in the antioxidant activity during the thermal processing assessed by DPPH free radical scavenging power and by the FRAP test were consistent with those for the antioxidants. Therefore, these oils can be added to other edible oils as a natural antioxidant to improve their oxidative stability.

Easy preparation of porous boron nitride with excellent cycle stability for water purification

The structure of hexagonal boron nitride (h-BN) is similar to that of graphite, which has strong reducibility and adsorbability. In this study, boron nitride precursor was made using melamine as a nitrogen source and boric acid as a source of boron under hydrothermal conditions, and the precursor was subsequently burned to create porous BN at a certain temperature. Effect of NH4HCO3 content on the porosity and structure of the materials was investigated to improve the adsorption performance of h-BN. The morphology, composition and structure of the materials were analyzed by scanning electron microscope, energy spectrum analyzer and X-ray diffraction. According to the findings, BN has a greater specific surface area of 169.29 m2/g under optimal preparation circumstances, which increases its capacity for adsorption. When the concentration of methylene blue was 200 mg/L, the adsorption capacity of the material reached to 377 mg/g in 120 min. In addition, the remarkable anti-oxidation and structural stability of the employed BN materials make it simple to regenerate them through burning or ethanol elution. In five cycles, the adsorption performance did not decrease.

Potential of White Jack Bean (Canavalia ensiformis L. DC) Kefir as a Microencapsulated Antioxidant.

Oxidative stress plays a major role in the pathogenesis and progression of noncommunicable diseases. Kefir is a fermented food that has been reported to repress oxidative stress. This study aimed to assess the antioxidant activity, bioactive composition, and encapsulation efficiency of white jack bean (WJB) kefir. The following procedures were conducted: WJB was prepared and converted into juice using water solvent. The sterilized WJB juice was then fermented with kefir grain (10%) for 24∼72 h. Every 24 h, the kefir was evaluated for antioxidant activity, and the dominant bioactive component suspected to be the source of the antioxidant activity was identified. The final stage was the encapsulation process. WJB kefir showed high antioxidant activity, inhibiting DPPH radicals by 90.51±4.73% and ABTS radicals by 86.63±2.34% after 72 h of fermentation. WJB kefir contained 0.35±0.01 mg GAE/g total phenolics and 0.08 mg/g total flavonoids. The LC/MS identification suggested that the bioactive antioxidant components of the WJB kefir were from the alkaloid, saponin, phenolic, and flavonoid groups. The encapsulation with maltodextrin using freeze drying resulted in microencapsulation of WJB kefir with a particle size of 6.42±0.13 μm. The encapsulation efficiency was 79.61%, and the IC50 value was 32.62 ppm. The encapsulation method was able to maintain the antioxidant stability of the kefir and extend its shelf life. WJB kefir, a nondairy, lactose-free kefir, can be used as an antioxidant functional food.

Enzymatic Acylation of Black Rice Anthocyanins and Evaluation of Antioxidant Capacity and Stability of Their Derivatives.

In this study, the structure of the anthocyanin fractions isolated from black rice (Oryza sativa L.) was modified by the enzyme catalysis method using caffeic acid as an acyl donor. At the same time, the effects of the acylation on the lipophilicity, antioxidant activity, and stability of black rice anthocyanins were comprehensively evaluated. The structural analyses of acylated derivatives based on ultraviolet-visible spectroscopy, Fourier-transform infrared spectroscopy, ultra-high-performance liquid chromatography-high-resolution mass spectrometry, and thermogravimetric analysis revealed that caffeic acid was efficiently grafted onto the anthocyanins of black rice through an acylated reaction, while the acylation binding site was on glucoside. When the mass ratios of anthocyanins to caffeic acid were 1:1, the A319/AVis-max value of acylated anthocyanins reached 6.37. Meanwhile, the lipophilicity of acylated derivatives was enhanced. The antioxidant capacity (DPPH and FRAP) and stability (thermal, pH, and light stability) were significantly increased. Overall, the study results provide deeper insights into controlling anthocyanin homeostasis in food processing, broadening the application of colored grain products.

Biological stabilization of Arthrospira bioactive-peptides within biopolymers: Functional food formulation; bitterness-masking and nutritional aspects

Physicochemical/biological instability and bitterness are among the major challenges limiting the shelf life and direct use of bioactive-peptides in the production of functional foods. This study focused on the structural modification and improvement of biological, nutritional and techno-functional properties of blue-green algae protein by alcalase (30–180 min). Then, the optimal sample (H-120) was stabilized with maltodextrin (MD), gum-arabic (GA), WPC, alginate (A), and pectin (P) biopolymeric-carriers by the spray-drying process. The MD-WPC-encapsulated peptide presented the highest production yield (∼65%), retention of antioxidant activity, functional, flowability, and stability properties. Morphological (formation of wrinkled, irregular, and dented particles) and chemical (FTIR) evaluations respectively indicated the effect of feed composition on the structural properties and entrapment of peptides (through hydrogen bonds and hydrophobic reactions) in the polymer matrix. Finally, the optimal sample (SDP: MD-WPC) was used at different ratios (1, 2, and 4% w/w) for pan-bread fortification. The examination of physicochemical properties (moisture content, water activity, and specific volume), textural (porosity and compressive strength) indices, a color histogram (crust and crumb), antioxidant activity (DPPH• and ABTS• scavenging), and sensory analysis (color, texture, chewiness, taste, and overall acceptability) indicated biological stability, bitterness-masking, and the feasibility of functional bread production with 2%-SDP.

Effects of cluster dextrin encapsulation on the properties and antioxidant stability of fractionated Riceberry protein hydrolysate powder prepared by bromelain

In this work, the biological properties of fractionated Riceberry bran protein hydrolysate obtained by ultrafiltration (URBPH) were evaluated and the possibility of using cluster dextrin to produce hydrolysate powder by spray-drying was investigated. Fractionation into peptides < 3 kDa was observed to improve antioxidant activity. URBPH < 3 kDa was then freeze-dried (FD-URBPH) and spray-dried (SD-URBPH) at different inlet air temperatures of 100–160 °C. The water solubility and antioxidant activity of FD-URBPH were higher than those of SD-URBPH. Nevertheless, encapsulation of hydrolysate with 10% cluster dextrin and an inlet temperature of 120 °C was also successful in maintaining protein qualities, which showed high 2,2′-azino-bis 3-ethylbenzthiazoline-6-sulfonic (ABTS•+) scavenging activity (89.14%) and water solubility index (92.49%) and low water activity (aw = 0.53). Moreover, encapsulation preserved the antioxidant activity of peptides during gastrointestinal digestion better than the free form. URBPH and its spray-dried microcapsules could be used as bioactive ingredients in functional drinks or foods.

Evaluation of Antioxidant Activity, Tyrosinase Inhibition, and Stability of Face Mask Cream Formulation from Sweet Granadilla (Passiflora ligularis Juss) Seed Fraction

The face mask has been widely used by consumers as a skin care product. In addition, masks made from natural ingredients have become one of the consumers' choices. This study aimed to evaluate facial mask cream from sweet granadilla fraction related to antioxidant activity, tyrosinase inhibition, and physical stability of the preparation for 12w. The fraction used was the ethyl acetate fraction from sweet granadilla seeds (Passiflora ligularis Juss), which was formulated in a face mask cream preparation and tested for antioxidant activity (DPPH and FRAP), tyrosinase inhibition, and stability of the preparation during 12w of storage. Face mask cream showed very strong antioxidant activity at F2(2% seed ethyl acetate fraction) at a temperature of 30±2oC and 40± 2oC compared to F1(1% seed ethyl acetate fraction). Meanwhile, the best inhibition of tyrosinase was at F2 at a temperature of 40±2oC. The physical stability test still met the requirements. However, a slight decrease in antioxidant activity and tyrosinase inhibition occurred after 12 w of storage at temperatures 30±2oC and 40±2oC (F1 and F2). In conclusion, the 2% seed ethyl acetate fraction formulation met the standards and was relatively stable at 30±2oC during 12w of storage.