Antioxidant Activity and Hemolytic Effect of Hydro-methanolic Extract and its Phenolic Enriched Fractions From Leaves and Stems of Salvia officinalis L. from Algeria

Abstract

This study aimed to assess an in vitro antioxidant and hemolytic activities of crude hydromethanolic extract, ethyl acetate and n-butanol fractions from leaves and stems of Salvia officinalis L. Extract and fractions prepared by maceration from S. officinalis were determined for their phytochemical composition and their contents in total phenolic and flavonoid. Antioxidant activity was evaluated by the methods of DPPH radical scavenging, ferric reducing antioxidant power and total antioxidant capacity. Hemolysis assay was carried out to evaluate the toxicity of the studied samples. Crude extract and fractions from S. officinalis contain different secondary metabolites and considerable contents of phenolic and flavonoids. They exhibit high antioxidant activity and low hemolytic effect. However, ethyl acetate fraction is characterized by the highest total phenolic (362.75 ± 0.07 µg GAE/mg DE) and flavonoid (263.27 ± 0.1 µg CE/mg DE) contents. Furthermore, this fraction shows the best antioxidant activity tested by DPPH, FRAP and TAC assays. The antiradical activity of ethyl acetate fraction against DPPH (IC50 = 208.51 ± 5.77 μg/mL) is close to that of ascorbic acid (IC50 = 206.43 ± 4.16 μg/mL). It exhibits a powerful activity to reduce iron (EC50 = 250 ± 0.001 μg/mL) and the best total antioxidant capacity (817.33 ± 0.06 μg/mL). It reveals the lowest hemolysis rate of 3.77 ± 0.02% with 200 mg/mL of fraction, after one hour of incubation. The obtained results suggest that crude extract, ethyl acetate and n-butanol fractions from Salvia officinalis L. are considered efficient natural antioxidants and safe for human erythrocytes.

REFERENCES:

Andrei, M., Cadmiel, M., Simone, C., Marcello, L., Marina, S., & Gianina, C. (2018).
Determination of phenolic compounds in different species of salvia from Romania
(Salvia transsylvanica, Salvia glutinosa, Salvia officinalis) and their biological
activities. Acta Medica Marisiensis, 64, 14.
Bruneton, J. (1999). Pharmacognosie-Phytochimie-Plantes médicinales, 3ème édition, Tec &
Doc. Lavoisier: Paris.
Burci, L.M., da Silva, C.B., Rondon, J.N., da Silva, L.M., de Andrade, S.F., Miguel, O.G., de
Fátima Gaspari Dias, J., & Miguel, M.D. (2019). Acute and subacute (28 days) toxicity,
hemolytic and cytotoxic effect of Artocarpus heterophyllus seed extracts. Toxicology
Reports, 6, 1304–1308. https://doi.org/10.1016/j.toxrep.2018.02.006
Cheng, K., Dong, W., Long, Y., Zhao, J., Hu, R., Zhang, Y., & Zhu, K. (2019). Evaluation of
the impact of different drying methods on the phenolic compounds, antioxidant activity,
and in vitro digestion of green coffee beans. Food science and nutrition, 7(3), 1084–
1095. https://doi.org/10.1002/fsn3.948
Dandlen, A.S., Sofia Lima, A., Marta Mendes, D., Graça Miguel, M., Leonor Faleiro, M.,
João Sousa, M., Pedro Luis, G., Barroso José, G., & Cristina Figueiredo, A. (2010).
Antioxidant activity of six Portuguese thyme species essential oils. Flavour and
fragrance journal, 25(3), 150–155. https://doi.org/ 10.1002/ffj .1972.
Duletić-Laušević, S., Alimpić Aradski, A., Ţivković, J., Gligorijević, N., Šavikin, K.,
Radulović, S., Ćoćić, D., & Marin, P.D. (2019). Evaluation of bioactivities and
phenolic composition of extracts of Salvia officinalis L. (Lamiaceae) collected in
Montenegro. Botanica Serbica, 43(1), 47–58.
https://doi.org/10.2298/BOTSERB1901047D.
El khoudri, N., Baali, A., & Amor, H. (2016). Maternal morbidity and the use of medicinal
herbs in the city of Marrakech, Morrocco. Indian journal of traditional knowledge,
15(1), 79–85.
Farzaneh, V., & Carvalho, I.S. (2015). A review of the health benefits potentials of herbal
plant infusions and their mechanism of actions. Industrial Crops and Products, 65,
247–258. https://doi.org/ 10.1016/j.indcrop.2014.10.057
Garcia, C.S.C., Menti, C., Lambert, A.P.F., Barcellos, T., Moura, S., Calloni, C., Branco,
C.S., Salvador, M., Roesch-Ely, M., & Henriques, J.A.P. (2016). Pharmacological
perspectives from Brazilian Salvia officinalis (Lamiaceae): antioxidant, and antitumor
in mammalian cells. Anais da Academia Brasileira de Ciências, 88(1), 281–292.
https://doi.org/10.1590/0001-3765201520150344.

Gessner, D.K., Ringseis, R., Eder, K. (2017). Potential of plant polyphenols to combat
oxidative stress and inflammatory processes in farm animals. Journal of Animal
Physiology and Animal Nutrition, 101(4), 605–628. https://doi.org/10.1111/jpn.12579.
Giacometti, J., Bursa´c Kovaˇcevi´c, D., Putnik, P., Gabri´c, D., Biluši´c, T., & Kreši´c, G.
(2018). Extraction of bioactive compounds and essential oils from mediterranean herbs
by conventional and green innovative techniques. Food Research International, 113,
245–262.
Harborne, J. (1998). Phytochemical Methods. In: A Guide to Modern Techniques of Plant
Analysis, 3rd ed, 203–234. Chapman and Hall Thomson Science, UK.
Henneberg, R., Otuki, M.F., Furman, A.E.F., Hermann, P., Nascimento, A.J., & Leonart,
M.S.S. (2013). Protective effect of flavonoid against reactive oxygen species
production in sickle cell anemia patients treated with hydroxy urea. Revista Brasileira
de Hematologia e Hemoterapia, 35(1), 52–55. https://doi.org/10.5581/1516-
8484.20130015.
Jasicka-Mislak, I., Poliwoda, A., Petecka, M., Buslovych, O., Shlyapnikov, V.A., &
Wieczorek, P.P. (2018). Antioxidant phenolic compounds in Salvia officinalis L. and
Salvia sclarea L. Ecological Chemistry Engineering S., 25(1), 133–142.
https://doi.org/10.1515/eces-2018-0009
Kaliora, A.C., Kogiannou, D.A.A., Kefalas, P., Papassideri, I.S., & Kalogeropoulos, N.
(2014). Phenolic profiles and antioxidant and anticarcinogenic activities of Greek
herbal infusions; Balancing delight and chemoprevention? Food Chemestry, 142, 233–
241. https://doi.org/10.1016/j.foodchem.2013.07.056.
Karagözler, A.A., Erdag, B., Emec, Ç.Y., & Uygunet, D.A. (2008). Antioxidant activity and
proline content of leaf extracts from Dorystoechas hastata. Food chemestry, 111, 400–
407.
Korkmaz, M., Karakuş, S., Özçelik, H., & Selvi, S. (2016). An ethnobotanical study on
medicinal plants in Erzincan, Turkey. Indian journal of traditional knowledge, 15(2),
192–202. http://hdl.handle.net/123456789/33967.
Kumar, S., Yadav, A., Yadav, M., & Yadav, J.P. (2017). Effect of climate change on
phytochemical diversity, total phenolic content and in vitro antioxidant activity of Aloe
vera (L.). Burm.f. BMC Research Notes, 10(1), 60. https://doi.org/10.1186/s13104-017-
2385-3
Li, H.B., Cheng, K.W., Wong, C.C., Fan, K.W., Chen, F., & Jiang, Y. (2007). Evaluation of
antioxidant capacity and total phenolic content of different fraction of selected
microalgae. Food Chemistry, 102(3), 771–776.
Martins, T.F., Palomino, O.M., Álvarez-Cilleros, D., Martín, M.A., Ramos, S., & Goya, L.
(2020). Cocoa Flavanols Protect Human Endothelial Cells from Oxidative Stress. Plant
Foods for Human Nutrition, 75(2), 161–168. https://doi.org/10.1007/s11130-020-
00807-1.

Phuse, S.S., & Khan, Z.H. (2018). Assessment of hemolytic effect of Cassia flower extracts
on human RBCs. Journal of Drug Delivery & Therapeutics, 8(6-s), 18–20.
https://doi.org/10.22270/JDDT.V8I6-S.2169.
Prieto, P., Pineda, M., & Aguilar, M. (1999). Spectrophotometric quantitation of antioxidant
capacity through the formation of a phosphomolybdenum complex: specific application
to the determination of vitamin E. Analytical Biochemestry, 269, 337–341.
https://doi.org/10.1006/ABIO.1999.4019.
Rguez, S., Msaada, K., Hammami, M., Daami-Remadi, M., Chayeb, I., Ksouri, R., &
Hamrouni‐Sellami, I. (2019). Diurnal variation of the chemical composition and its
repercussion on the biological activity polyphenolics of Salvia officinalis aerial parts.
International Journal of Plant Science and Horticulture, 1(1), 07–21.
https://doi.org/10.36811/ijpsh.2019.110002.
Saba, A., Suhayla, M.K., & Raheem, Z.H. (2019). Study the Antioxidant Activity of Sage
(Salvia Officinalis) Leaves Extract. Indian Journal of Public Health Research &
Development, 10(3), 556. https://doi.org/10.5958/0976-5506.2019.00557.6.
Venkatesan, T., Choi, Y. W., & Kim, Y. K. (2019). Impact of Different Extraction Solvents
on Phenolic Content and Antioxidant Potential of Pinus densiflora Bark Extract.
BioMed Research International, 2019, 1–14. https://doi.org/10.1155/2019/3520675.
Yashaswini, S., Fagan, J., & Schaefer, J. (2019). Ethnobotany, phytochemistry, cultivation
and medicinal properties of Garden sage (Salvia officinalis L.). Journal of
Pharmacognosy and Phytochemistry, 8(3), 3139–3148.
Zheng, S., Wang, Y., Liu, H., Chang, W., Xu, Y., & Lin, F. (2019). Prediction of Hemolytic
Toxicity for Saponins by Machine-Learning Methods. Chemical Research in
Toxicology, 32, 1014–1026. https://doi.org/10.1021/acs.chemrestox.8b00347.
Zhishen, J., Mengcheng, T., & Jianming, W. (1999). The determination of flavonoid contents
in mulberry and their scavenging effects on superoxide radicals. Food Chemistry,
64(4), 555–559. https://doi.org/10.1016/S0308-8146 (98)00102-2.