Phytochemical evaluation and in vitro antioxidant and photo-protective capacity of Calendula officinalis L. leaves

Abstract

ABSTRACT:The plant Calendula officinalis L. is widely applied due to its medicinal properties, which are mainly dermatological and ornamental. The goal of this study is to assess the phytochemical components in a hydroethanolic extract (HECO) from the leaves of Calendula officinalis L. using UV-VIS spectrophotometry and thin layer chromatography (TLC), as well as to identify and quantify the components related to its antioxidant capacity employing high performance liquid chromatographic (HPLC). The antioxidant capacity evaluation was performed using the DPPH method for superoxide and hydroxyl radicals. The photo-protective capacity was evaluated by UVspectrophotometry in order to determine the in vitro Sun Protection Factor(SPF). The results show the plant's strong antioxidant activity (DPPH and hydroxyl methods), which we believe to be related to the presence of flavonoids (24.67 mg/g), polyphenols (33.90 mg/g), condensed tannins (27.30 mg/g), and the amount of rutin (37.25 mg/g), and quercetin (6.09 mg/g) found during the study. The HECO presented a good antioxidant capacity, most likely due to the polyphenols, flavonoids, and tannins in its contents. However, the obtained SPF of 1.89 ± 0.05 does not allow the plant to be classified as a stand-alone sunscreen, and more studies are needed in order to test its ability to enhance sunscreens in existing cosmetic formulations.Keywords: Calendula officinalis. Marigold. Antioxidant. Phenolic Compounds. Photo-protection. RESUMO:Avaliação fitoquímica e capacidade antioxidante e fotoprotetora in vitro das folhas de Calendula officinalis L. A Calendula officinalis L. é uma planta amplamente utilizada por suas propriedades medicinais, principalmente dermatológica e ornamental. O objetivo deste estudo é avaliar constituintes fitoquímicos do extrato hidroetanólico das folhas de Calêndula officinalis L. (HECO) por espectrofotometria UV-visível e cromatografia em camada delgada (CCD), bem como detectar e quantificar os componentes relacionados com a capacidade antioxidante por Cromatografia Líquida de Alta Eficiência (CLAE). Posteriormente, foi avaliada a capacidade antioxidante pelo método do DPPH, radicais superóxido e hidroxila. Além disso, a capacidade fotoprotetora foi avaliada através de espectrofotometria UV para determinação do Fator de Proteção Solar in vitro (FPS). Os resultados evidenciaram que a planta apresentou uma excelente atividade antioxidante para o método do DPPH e do radical hidroxila, o que pode estar relacionada com a presença de flavonoides (24,67 mg/g), polifenóis (33,90 mg/g) e taninos condensados (27,30 mg/g) e da quantidade de rutina (37,25 mg/g) e quercetina (6,09 mg/g) encontrados neste estudo. O HECO apresentou uma notável capacidade antioxidante, provavelmente devido à presença de polifenóis, flavonoides e taninos. O valor do FPS encontrado (1,89 ± 0,05), não classifica a planta como um protetor solar isolado. No entanto, mais estudos são necessários para testar a capacidade de potencializar filtros solares em formulações cosméticas.

Plant polyphenols as green sunscreen ingredients: A systematic review

“…Among the 9 plants in a study conducted by Nunes et al (2018), ethanolic extracts of leaves of L. microphylla showed the highest SPF value of 26.82, and S. galheirensis has the lowest SPF value of 5.40 28 . The aqueous ethanolic extract of C. officinalis L. showed low SPF values of 1.89 29 . Yasmeen and Gupta (2016) reported the methanolic barks, leaves, and fruit extracts of D .…”

“…Deuschle (2015) found alkaloids, terpenoids, saponins, and volatile oils using thin‐layer chromatography, subsequently, they identified 25 mg/g of rutin and 6.09 mg/g of quercetin in the leaves of C. officinalis L. 29 . Raquel et al (2017) identify apigenin, catchehin, epicatechin, and gallic acid in the barks of S. brasiliensis by comparing the retention time of peaks analytical standards using HPLC–DAD analysis 32 .…”

“…Deuschle (2015) found alkaloids, terpenoids, saponins, and volatile oils using thin-layer chromatography, subsequently, they identified 25 mg/g of rutin and 6.09 mg/g of quercetin in the leaves of C. officinalis L.. 29 Raquel et al (2017) identify apigenin, catchehin, epicatechin, and gallic acid in the barks of S.brasiliensis by comparing the retention time of peaks analytical standards using HPLC-DAD analysis. 32 Gasparrini (2015) found myricetin-3-glucoside and myricetin-3-rhamnoside, as well as rutin and cyanidin-3-rutinoside, using HPLC-DAD-IT coupled with a MS. 33 Choi and co-researchers (2019), in a UPHLC study, compounds such as (i) phenolics (pcoumaric acid, caffeic acid, and chlorogenic acid), (ii) polyols (quinic acid and citric acid), and (iii) flavonoids and flavonoid derivatives (patuletin, catechin, epicatechin, eriodictyol, naringenin, quercetin, and quercetin derivatives) were detected in the leaves of P.villosa.…”

Marigold Metabolites: Diversity and Separation Methods of Calendula Genus Phytochemicals from 1891 to 2022

“…Evaluation of phenolic compounds in Calendula plants is an important task, as indicated by the known HPLC protocols found in the scientific literature. To separate target compounds, only RP C18 columns with varying lengths were used, such as 75 mm ProntoSIL-120-5-C18 [ 90 , 114 , 144 ]; 100 mm Phenomenex C18 [ 142 ], Zorbax SB-C18 [ 143 ], and LiChrosorb RP18 [ 100 , 153 ]; 150 mm Luna C18 [ 131 ], SiliaChrom C-18 [ 140 ], Eclipse XDB-C18 [ 146 ], Spherisorb S3 ODS-2 C18 [ 149 ], Zorbax Eclipse Plus C18 [ 151 ], and Aquity UPLC HSS T3 [ 154 ]; 220 mm Aquapore RP-300 [ 152 ]; 250 mm Shim-pack C-18 [ 96 ], Hypersil C18 [ 141 , 147 ], X-Bridge C18 [ 145 ], and Phenomenex C18 [ 148 ]; 300 mm Bondclone C18 [ 133 ]; and 1000 mm Hypersil Gold [ 75 ]. The presence of various eluents requires the frequent use of formic acid [ 75 , 96 , 142 , 150 , 151 ], acetic acid [ 133 , 141 , 148 ], phosphoric acid [ 140 , 146 ] as the polar eluent and methanol [ 133 , 140 ] and acetonitrile [ 96 , 114 , 141 , 142 ] as the non-polar eluent.…”