UVB-Protective Fermented Plant Extracts

Production of Fermented Plant Extracts with UVB Protective Activity for Use in Cosmetic Products  

Abstract

In this study, fermented plant extracts from carrot (Daucus carota), ginger (Zingiber officinale), ginkgo (Ginkgo biloba), butternut squash (Cucurbita moschata), apple (Malus domestica), banana (Musa sapientum) and strawberry (Fragaria ananassa) were prepared using Saccharomyces ceravisea fermentation. Changes in phenolic compounds between fermented plant extracts and control group extracts were determined. Additionally, anti-UVB activities of fermented plant extracts were determined through SPF measurement and calculations. In our study, fermented plant extracts were obtained as a result of fermentation of plant parts with S. ceravisea under controlled conditions. Following fermentation, a significant increase in phenolic compounds was determined in some plant extracts. It was determined that fermented plant extracts exhibit anti-UVB activity, with SPF values in the range of 0.43 ± 0.04 and 48.17 ± 3.04.  

Introduction

Cosmetics are stable mixtures containing one or more active ingredients, additives and certain preservatives that help improve the physical appearance of the user. Today, cosmetic use and the production and product diversity of the cosmetic industry are increasing. The cosmetic industry aims to develop new products with innovative active ingredients or improve existing products. Recently, active ingredients derived from natural sources and their effects on skin have come to the forefront [1]. Our skin has a protective function that extends throughout the human body, protecting internal tissues and organs. Human skin consists of three main layers; epidermis, dermis and hypodermis [2]. Human skin is frequently exposed to sunlight and other environmental factors. Sunscreens are topical products used to protect skin against damage caused by ultraviolet (UV) radiation. UV radiation can cause irreversible skin aging, sun spots, sunburn and skin cancer in skin tissue. The harmful effects of sun rays generally originate from the ultraviolet (UV) region of the electromagnetic spectrum. This region is divided into three parts: UVA (320-400 nm), UVB (290-320 nm) and UVC (200-290 nm). While UVC radiation is filtered by the ozone layer before reaching Earth, UVA and UVB radiation cannot be completely filtered by the ozone layer [3]. UVB radiation penetrates the epidermis of the skin, causing oxidative stress, accumulation of DNA damage and skin aging [4,5]. Today, the destruction of stratospheric ozone and increased UVB radiation pose a major threat to skin. The generation of reactive oxygen species (ROS) causing oxidative damage can be triggered by UV radiation. Repeated exposure to UVA and UVB radiation triggers the generation of ROS species, leading to problems such as premature aging, sunburn, skin cancer and immune system suppression [6]. Additionally, UV radiation prevents collagen production, thereby causing the skin to lose elasticity, weakening of subcutaneous fat tissue and various biochemical and physiological changes [7]. Traditionally, plant extracts and other natural compounds in cream form have been used to prevent UVB damage. Plant extracts contain phenolic compounds with anti-UVB and anti-aging activity [8]. Phenolic compounds are phytochemicals found in all plants. These compounds are synthesized in plants partly in response to ecological and physiological stresses such as pathogen and insect attacks, UV radiation and injury. Over the past 25 years, significant advances have been made in research focused on the extraction, identification and quantification of phenolic compounds. The main method used to extract phenolic compounds is organic solvent extraction [9, 10]. According to research, fermented plant extracts contain more phenolic compounds than traditional plant extracts. Fermentation helps break down complex compounds into simpler compounds, allowing extraction of bioactive components at higher yields. The high phenolic compounds in fermented plant extracts are relevant here [11]. Due to the intense phenolic compounds they contain, they may show strong activity against UVB and aging. Today there is a continuously increasing interest in cosmetics in anti-UVB and anti-aging products based on natural ingredients [12]. In our study, we aimed to compare the phenolic compounds and anti-UVB activities of seven fermented plant extracts—carrot, ginger, ginkgo, butternut squash, apple, banana and strawberry—using Saccharomyces ceravisea and discuss potential cosmetic applications.  

Materials and Methods

1. Preparation of Fermented Plant Extracts The drug parts of plants to be fermented were prepared by shredding to increase the surface area. The fermentation medium consisted of sterile distilled water, plant drug and S. ceravisea. Fermentation was carried out in the dark at 180 rpm and 37°C with shaking for 2 days. Following fermentation, extraction was performed with dry matter, deionized water and 95% ethyl alcohol (1:1:3). The prepared extract mixture was left for incubation at room temperature for 2 days. The liquid portion was then collected from the solid portion using filter paper. Ethyl alcohol was evaporated from the aqueous phase using a cooled rotary evaporator. The obtained aqueous extract was stored at 4°C in a sealed container for subsequent analyses [13].  

2. Phenolic Compound Determination

Phenolic compound determination of fermented plant extracts and control group extracts was performed using the Folin-Ciocalteu method and calculated as gallic acid equivalents. Gallic acid standards were prepared in distilled water at 25 μg/ml, 50 μg/ml, 75 μg/ml, 100 μg/ml, 125 μg/ml and 150 μg/ml to determine the amount of phenolic compounds. 50 μl of extract or standard was mixed with 50 μl Folin-Ciocalteu Solution (1X). The mixture was incubated at room temperature for 5 minutes. 50 μl of 5% sodium carbonate solution was added to the mixture and vortexed for 10 seconds. The mixture was incubated at room temperature in a dark environment for 90 minutes. After the period, 200 μl of the mixture was taken and loaded onto a 96-well plate and the absorbance value was measured at 734 nm. Distilled water was used as blank in absorbance readings [14].  

3. Anti-UVB Activity Determination

Anti-UV activity of fermented fruit peel extracts was measured by in vitro determination of sun protection factor (SPF), which is considered one of the most frequently used indicators for classifying the level of protection provided by sun protection products, mainly against sunburn caused by harmful UVB radiation. Photoprotective activity against UVB was measured using spectrophotometric method at wavelengths between 290 and 320 nm at 5 nm intervals, and SPF values were calculated using the Mansur equation. EE(λ) represents erythema spectrum, I(λ) solar intensity spectrum, A(λ) absorbance, CF correction factor (10). EE(λ) × I(λ) values are constant and were determined by Sayre et al. (1979) [15].  

4. Statistical Analysis

Data obtained from the experiments were analyzed using GraphPad Prism v.8.4.3 software and presented as mean ± SD in triplicate. One-way analysis of variance (ANOVA) was performed to determine significant differences.  

Results and Discussion

In our study, fermented plant extracts were obtained as a result of fermentation of plant parts with S. ceravisea under controlled conditions. It was determined that fermentation caused an increase in phenolic compound amounts in some plants while causing no change in others. According to our statistical analysis, significant changes were determined between Ginger-Fermented Ginger, Apple-Fermented Apple, Ginkgo-Fermented Ginkgo and Strawberry-Fermented Strawberry. Particularly, compared to non-fermented Ginkgo extract, nearly a 2-fold increase in phenolic compounds was determined in Fermented Ginkgo extract (Figure 1).   [caption id="attachment_143686" align="aligncenter"] Figure 1: Phenolic compounds of fermented and control plant extracts were calculated as gallic acid equivalents. Changes in phenolic compounds following fermentation were compared with control plant extracts.[/caption]   When examining the anti-UVB activities of fermented plant extracts, results reaching 48.17 ± 3.04 SPF were obtained. Compared to phenolic compound amounts, high SPF values were achieved in some fermented plant extracts while low SPF values were encountered in others. For example, although fermented butternut squash had higher phenolic compound content than fermented banana, it was determined to have a lower SPF value than fermented banana. In this case, it can be considered that the type of phenolic compound contained in the extract determines anti-UVB activity rather than the total phenolic compound amount (Table 1).   [caption id="attachment_143687" align="aligncenter"] Table 1: Photoprotective activity of fermented plant extracts against UVB was measured at wavelengths between 290 and 320 nm at 5 nm intervals using spectrophotometric method and calculated using the Mansur equation [15].[/caption] Our results demonstrated that some fermented extracts exhibit higher phenolic compounds compared to non-fermented plant extracts. Additionally, an increase in UVB protective activity was detected following fermentation. A correlation is thought to exist between UVB protective activity and phenolic compound amount. At the same time, the type of phenolic compound is considered to play an important role in anti-UVB activity. Fermented plant extracts displaying UVB preventive activity and their mixtures were evaluated as potentially usable potential UVB filters in sun protection formulations. It is anticipated that evaluation of various potential bioactivities of fermented plant extracts will contribute to the development of functional products such as shampoos, soaps, creams and serums.     References [1] Ortega, P. A., Guzmán, M. E., Vera, L. R., Velázquez, J. A., and Abuín, E. B. (2000). Dihydroeuparin as sunscreen. Boletín de la Sociedad Chilena de Química, 45(4), 629-636. [2] BS Sivamaruthi, C Chaiyasut, P Kesika. (2018). Cosmeceutical importance of fermented plant extracts: A short review. Int. J. Appl. Pharm, 10, 31-34. [3] Park, J. J., An, J., Lee, J. D., Kim, H. Y., Im, J. E., Lee, E., Ha, J., Cho, C. H., Seo, D. and Kim, K. B. (2020). Effects of anti-wrinkle and skin-whitening fermented black ginseng on human subjects and underlying mechanism of action. 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Revista Brasileira de Ciências Farmacêuticas, 40(3), 381-385.     Haluk Çelik Research and Development Engineer Akten Cosmetics   Damla Gül Product Development and Application Specialist Akten Cosmetics   Sinem Asılı Research, Development and Quality Manager Akten Cosmetics