Evaluation of Antioxidant, Cytotoxic and Antidiabetic Effects of Single Bulb Black Garlic Extracts

Öz

Aim: It was aimed to determine the bioactivity of a single clove of black garlic, the fermented product obtained as a result of the treatment of Allium sativum L. (Garlic) plant with a certain degree of humidity and temperature.

Method: In the study, ethanol and water extract of black garlic bulb were prepared, and the separated husks were also extracted with ethanol, and their antioxidant activity was evaluated by scavenging DPPH● and ABTS●+ radicals. Its antidiabetic activity was determined by in vitro α-amylase inhibition test. The cytotoxic effect of the extracts on Colo-205 cells was determined using the MTT method.

Results: Among the black garlic extracts, the husk extract (A.S.K EtOH) was determined as the extract with the highest total phenol (61.40±0.48 mgGA/gextract) and total flavonoid (26.08±0.37 mgCA/gextract) content. In addition, at a concentration of 4mg/mL, A.S. SU had the highest DPPH● radical scavenging activity (% inhibition 64.66±1.94), while A.S.K EtOH extract had the highest ABTS radical scavenging activity (2.44±0.16mmol/L). /Trolox) was found. Among the extracts whose cytotoxic effect was evaluated in the human colorectal cancer cell line Colo-205, it was determined that A.S.K EtOH extract showed the most cytotoxic effect compared to other extracts in the concentration range of 1000-250 µg/mL. None of the extracts showed antidiabetic activity according to the α-amylase inhibition test, in which acarbose was used as a positive control.

Conclusion: The results are preliminary study on the biological activities of black garlic, which has been very popular in recent years. According to the data obtained, the active components of black garlic should be studied in more detail and the methods of obtaining and storing black garlic should be investigated in more detail. Thus, promising results can be obtained regarding the use and benefits of the fermented form of a medicinally important plant.

Anahtar Kelimeler

• Black garlic
• maillard reaction
• antioxidant
• cytotoxicity

Tek Diş Siyah Sarımsak Ekstrelerinin Antioksidan, Sitotoksik ve Antidiyabetik Etkilerinin Değerlendirilmesi

Öz

Amaç: Allium sativum L. (Sarımsak) bitkisinin belli bir derecede nem ve sıcaklık ile işlem görmesi sonucu, elde edilen fermente ürün tek diş siyah sarımsağın biyoaktivitesinin tespit edilmesi amaçlandı.

Yöntem: Çalışmada, siyah sarımsak dişlerinin etanol ve su ekstresi hazırlandı, yine ayrılan kabuk kısımları da etanol ile ekstre edilerek, DPPH● ve ABTS●+ radikalini süpürücü etki tayin yöntemi ile antioksidan aktivitesi değerlendirildi. In vitro α-amilaz inhibisyon testi ile antidiyabetik aktivitesi belirlendi. MTT yöntemi kullanılarak ekstrelerin Colo-205 hücrelerinde sitotoksik etkisi tayin edildi.

Bulgular: Siyah sarımsak ekstrelerinden kabuk ekstresi (A.S.K EtOH) en yüksek toplam fenol (61,40±0,48 mgGA/gekstre) ve toplam flavonoit (26,08±0,37 mgCA/gekstre) içeriğine sahip ekstre olarak tespit edildi. Ayrıca, 4mg/mL konsantrasyonda A.S. SU ekstresi DPPH● radikalini süpürücü aktivitesi (% inhibisyonu 64,66±1,94) en yüksek ekstre olarak tespit edilirken, A.S.K EtOH ekstresi 4mg/mL’de ABTS radikalini süpürücü aktivitesi en yüksek ekstre (2,44±0,16mmol/L/Trolox) olarak bulundu. İnsan kolorektal kanser hücre hattı Colo-205’te sitotoksik etkisi değerlendirilen ekstrelerden, S.K EtOH ekstresinin 1000-250 µg/mL konsantrasyon aralığında diğer ekstrelere oranla en fazla sitotoksik etki gösteren ekstre olduğu belirlendi. Ekstrelerden hiçbiri akarbozun pozitif kontrol olarak kullanıldığı α-amilaz inhibiyon testine göre antidiyabetik aktivite gösteremedi.

Sonuç: Sonuçlar son yıllarda oldukça popüler olan siyah sarımsağın biyolojik aktivitelerine dair ön çalışma verileridir. Elde edilen verilere göre siyah sarımsağın aktif bileşenlerinin daha detaylı bir şekilde çalışılması ve siyah sarımsak elde etme ve saklama yöntemlerinin daha detaylı araştırılması gerekmektedir. Böylelikle tıbbi açıdan oldukça büyük öneme sahip bir bitkinin fermente halinin kullanımı ve faydaları ile ilgili umut vaat edici sonuçlar elde edilebilir.

Anahtar Kelimeler

• Siyah sarımsak
• maillard reaksiyonu
• antioksidan
• sitotoksisite

Kaynakça

1. Yang P, Song H, Wang L, Jing H. Characterization of key aroma-active compounds in black garlic by sensory-directed flavor analysis. J. Agric. Food Chem. 2019;67(28):7926-7934. doi.org/10.1021/acs.jafc.9b03269.
2. İbret B. Türkiye’deki sarımsak tarımı ve taşköprü sarımsağı üzerine coğrafi açıdan bir inceleme. Marmara Coğrafya Dergisi. 2013;12:17-50.
3. Çiçek İS. Siyah sarımsağın kardiyovasküler hastalıklar üzerine etkisi. Fenerbahçe Üniversitesi Sağlık Bilimleri Dergisi. 2022;2(1):299-311.
4. Ryu JH, Kang D. Physicochemical properties, biological activity, health benefits, and general limitations of aged black garlic: A review. Molecules. 2017;22(6):919. doi:10.3390/molecules22060919.
5. Halimah LS, Nawangsih EN, Hasan K. Analysis of antibacterial and antioxidant activities of a single bulb of garlic fermented into black garlic. In: Proceedings of the 12th Annual Scientific Meeting International Symposium on" Emergency Preparedness and Disaster Response during COVID 19 Pandemic"; 2021; Universitas Jenderal Achmad Yani, Indonesia. Volume 37.
6. Wu J, Jin Y, Zhang M. Evaluation on the physicochemical and digestive properties of melanoidin from black garlic and their antioxidant activities in vitro. Food Chem. 2021;340:127934. doi:10.1016/j.foodchem.2020.127934.
7. Tran GB, Pham TV, Trinh NN. Black garlic and its therapeutic benefits. In: Hassan B eds. Medicinal Plants-Use in Prevention and Treatment of Diseases. IntechOpen. 2019. doi: 10.5772/intechopen.85042.
8. Riwanti P, Ma’arif B, Jannah MZ. Potential Effect of Black Garlic (Allium sativum L.) as Antioxidant. In: 2nd International Conference on Education and Technology; Sep 6, 2021; Atlantis Press.

9. Singleton Vernon L, Rudolf O, Rosa ML. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Meth Enzymol. 1999;14:152-178. doi.org/10.1016/S0076-6879(99)99017-1.
10. Zhıshen J, Mengcheng T, Jıanmıng W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem. 1999;64(4): 555-559.
11. Gyamfi MA, Yonamine M, Aniya Y. Free-radical scavenging action of medicinal herbs from Ghana: Thonningia sanguinea on experimentally-induced liver injuries. Vasc. Pharmacol. 1999;32:661-67.
12. Re R, Nicoletta P, Anna P, Ananth P, Min Y, Catherine R. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999;26:1231-37.
13. Ali H, Houghton P, Soumyanath A. α-Amylase inhibitory activity of some Malaysian plants used to treat diabetes; with particular reference to Phyllanthus amarus. J Ethnopharmacol. 2006;107(3):449–455. doi:10.1016/j.jep.2006.04.004.
14. Büyüktuncel E. Toplam fenolik içerik ve antioksidan kapasite tayininde kullanılan başlıca spektrofotometrik yöntemler. Marmara Pharm J. 2013;17(2):93-103. doi:10.12991/201317377.
15. Liu C, Lu L, Yang C, et al. Effects of thermal treatment on alliin and its related sulfides during black garlic processing. LWT. 2022;159.
16. Kahyaoğlu DT. Comparison of the antioxidant activity of garlic cloves with garlic husk and stem: Determination of utilization potential of garlic agricultural wastes. Türk Tarım ve Doğa Bilimleri Dergisi. 2021;8(2):463-469.
17. Noda Y, Asada C, Sasaki C, Nakamura Y. Effects of hydrothermal methods such as steam explosion and microwave irradiation on extraction of water soluble antioxidant materials from garlic husk. Waste and Biomass Valorization. 2019;10(11):3397-3402. doi:10.1007/s12649-018-0353-3.
18. Kim J, H Nam SH, Rico CW, Kang MY. A comparative study on the antioxidative and anti‐allergic activities of fresh and aged black garlic extracts. Int J Food Sci. 2012;47(6):1176-1182. doi:10.1111/j.1365-2621.2012.02957.x.
19. Marie AMA, Wijayanti N. The antioxidant properties of black garlic aqueous extract in Vero Cell line. In: Proceeding International Conference on Science and Engineering. April 4, 2020; Vol.3. https://doi.org/10.14421/icse.v3.474.
20. Barido FH, Jang A, Pak JI, Kim YJ, Lee SK. Combined effects of processing method and black garlic extract on quality characteristics, antioxidative and fatty acid profile of chicken breast: Scientific section: Processing and products. Poultry Science. 2022:101723. doi:10.1016/j.psj.2022.101723.
21. Mondal A, Banerjee S, Bose S, et al. Garlic constituents for cancer prevention and therapy: From phytochemistry to novel formulations. Pharmacol Res. 2022;175:105837. doi:10.1016/j.phrs.2021.105837.
22. Dong M, Yang G, Liu H, et al. Aged black garlic extract inhibits HT29 colon cancer cell growth via the PI3K/Akt signaling pathway. Biomedical Reports. 2014;2(2):250-254. doi:10.3892/br.2014.226.
23. Permatasari E, Farida, Widiyanto S. Cytotoxic effects and apoptosis of solo black garlic (Allium sativum L.) extract on T47D breast cancer cell line. In: Proceedings AIP Conference; 2020. https://doi.org/10.1063/5.0015736.
24. Ahmed MU, Ibrahim A, Dahiru NJ, Mohammed HUS. Alpha amylase inhibitory potential and mode of inhibition of oils from Allium sativum (garlic) and Allium cepa (onion). Clin Med Insights Endocrinol Diabetes. 2020;7(13):1179551420963106. doi:10.1177/1179551420963106. eCollection 2020.