Optimum Extraction Condition of Clitorea ternatea Flower on Antioxidant Activities, Total Phenolic, Total Flavonoid and Total Anthocyanin Contents
Main Article Content
Abstract
Clitoria ternatea is a herbaceous plant with many health benefits. Extraction is crucial to obtain its bioactive components which contribute to its antioxidant properties. Therefore, this study was conducted to find an optimum extraction condition of C. ternatea flower on total phenolic content (TPC) and antioxidant activity (2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity) as well as to determine its total flavonoid content (TFC) and anthocyanin content based on the optimum extraction condition generated by Response Surface Methodology (RSM)-Design Expert 7.1.5. TPC, TFC and total anthocyanin of C. ternatea were conducted by Folin Ciocalteu (FC), calorimetric assay and pH differential method, respectively. The ranges of selected independent variables were ethanol concentration (30ºC–90% v/v), time (60–120 min) and temperature (30ºC–70ºC). The optimum extraction condition was obtained at 39.62% v/v ethanol concentration, 90 min and 44.24ºC. However, these values were slightly adjusted according to the convenience of equipment to operate in which ethanol concentration was adjusted to 37% v/v, time remain at 90 min and temperature at 45ºC. The predicted values of TPC and DPPH radical scavenging activity were 41.60 mg GAE/g dry samples and 68.12% inhibition and were experimentally verified to be 41.17 ± 0.5 mg GAE/g dry samples and 63.53 ± 0.95% inhibition of TPC and DPPH radical scavenging activity respectively. This result has showed RSM can optimise TPC and radical scavenging activity of C. ternatea. Upon the optimum condition, the TFC determined was 187.05 ± 3.18 mg quercetin/g dried sample which was higher than TPC and the total anthocyanin content was 28.60 ± 0.04 mg/L. Hence, the extractable phenolic, flavonoid and anthocyanin compounds indicated that C. ternatea is a good source of natural antioxidant.
Clitoria ternatea adalah tumbuhan herba yang banyak kebaikan kesihatan. Pengekstrakan adalah penting untuk mendapatkan komponen bioaktif yang menyumbang kepada sifat antioksidannya. Oleh itu, kajian ini dijalankan untuk mendapatkan keadaan pengekstrakan yang optimum ke atas bunga C. ternatea terhadap jumlah kandungan fenolik (TPC) dan aktiviti antioksidan (2,2-difenil-1-picrylhydrazyl (DPPH) aktiviti pemerangkapan radikal bebas) serta menentukan jumlah kandungan flavonoid (TFC) dan antosianin berdasarkan keadaan pengekstrakan optimum yang dihasilkan oleh Metodologi Respons Permukaan (RSM)-Design Expert 7.1.5. TPC, TFC dan jumlah antosianin C. ternatea masing-masing ditentukan oleh Folin Ciocalteu (FC), ujian kalorimetri dan kaedah pembezaan pH. Pembolehubah bebas yang dipilih adalah kepekatan ethanol (30%–90% v/v), masa (60–120 min) dan suhu (30ºC–70ºC). Kondisi pengekstrakan optimum diperolehi pada kepekatan ethanol 39.62% v/v, 90 min dan 44.24ºC. Walau bagaimanapun, nilai-nilai ini telah diselaraskan mengikut kemudahan peralatan untuk beroperasi iaitu ethanol diselaraskan kepada 37% v/v, masa kekal pada 90 min dan suhu pada 45ºC. Nilai ramalan kandungan fenolik dan aktiviti pemerangkapan radikal DPPH adalah 41.60 mg GAE/g sampel kering dan 68.12% perencatan radikal dan experimen pengesahan keadaan optimum mendapati keputusan TPC ialah 41.17 ± 0.5 mg GAE/g sampel kering dan keputusan pemerangkapan radikal DPPH ialah 63.53 ± 0.95%. Keputusan ini menunjukkan bahawa RSM dapat mengoptimumkan pengekstrakan TPC dan aktiviti antioksidan C. ternatea. Berdasarkan keadaan pengektrakan yang optimum, jumlah kandungan flavonoid yang ditentukan ialah 187.05 ± 3.18 mg quercetin/g sampel kering yang iaitu lebih tinggi daripada kandungan TPC dan jumlah kandungan antocianin adalah 28.60 ± 0.04 mg/L. Oleh itu, komponen fenolik, flavonoid dan antosianin yang boleh diekstrak menunjukkan bahawa C. adalah sumber antioksidan semula jadi yang baik.
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
References
Abu Bakar M F, Mohamed M, Rahmat A and Fry J. (2009). Phytochemicals and antioxidant activity of different part of bambangan (Mangifera pajang) and tarap (Artocarpus odorotassimus). Food Chemistry 113: 479–483. https://doi.org/10.1016/j.foodchem.2008.07.081
Association of Official Analytical Chemists (AOAC). (2000). Official methods of analysis. USA: AOAC.
Benmeziane F, Djamai R, Cadot Y and Seridi R. (2014). Optimization of extraction parameters of phenolic compounds from Algerian fresh table grapes (Vitis Vinifera). International Food Research Journal 21(3): 1061–1065.
Bezerra M A, Santelli R E, Oliveira E P, Villar L S and Escaleira L A. (2008). Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta 76(5): 965–977. https://doi.org/10.1016/j.talanta.2008.05.019
Borrás-Linares I, Fernández-Arroyo S, Arráez-Roman D, Palmeros-Suárez P, Val-Díaz R D, Andrade-Gonzáles I, Gonzáles A and Segura-Carretero A. (2015). Characterization of phenolic compounds, anthocyanidin, antioxidant and antimicrobial activity of 25 varieties of Mexican Roselle (Hibiscus sabdariffa). Industrial Crops and Product 69: 385–394. https://doi.org/10.1016/j.indcrop.2015.02.053
Bravo L. (2009). Polyphenols: Chemistry, dietary sources, metabolism, and nutritional significance. Nutrition Reviews 56(11): 317–333. https://doi.org/10.1111/j.1753-4887.1998.tb01670.x
Braca A, Sortino C, Politi M, Morelli I and Mendez J. (2002). Antioxidant activity of flavonoids from Licania linaniaeflora. Journal of Ethnopharmacology 79: 379–381. https://doi.org/10.1016/S0378-8741(01)00413-5
Buci?-Koji? A, Planini? M, Tomas S, Joki? S, Muji? I, Bili? M and Veli?, D. (2011). Effect of extraction conditions on the extractability of phenolic compounds from lyophilised fig fruits (Ficus carica L.). Polish Journal of Food and Nutrition Sciences 61(3): 195–199. https://doi.org/10.2478/v10222-011-0021-9
Carlo G D, Mascolo N, Izzo A A and Capasso F. (1999). Flavonoids: Old and new aspects of a class of natural therapeutic drugs. Life Sciences 65(4): 337–353. https://doi.org/10.1016/S0024-3205(99)00120-4
Chayaratanasin P, Barbieri M A, Suanpairintr N and Adisakwattana S. (2015). Inhibitory effect of Clitoria ternatea flower petal extract on fructose-induced protein glycation and oxidation-dependent damages to albumin in vitro. BMC Complementary and Alternative Medicine 18(15): 27. https://doi.org/10.1186/s12906-015-0546-2
Chen X and Gu Z. (2013). Absorption-type optical pH sensitive film based on immobilized purple cabbage pigment. Sensors and Actuators B: Chemical 178: 207–211. https://doi.org/10.1016/j.snb.2012.12.094
Chew K K, Khoo M Z, Ng S Y, Thoo Y Y, Wan Aida M and Ho C W. (2011). Effect of ethanol concentration, extraction time and extraction temperature on the recovery of phenolic compounds and antioxidant capacity of Orthosiphon stamineus extracts. International Food Research Journal 18(4): 1427–1435.
Crozier A, Clifford M and Ashihara H. (2006). Plant secondary metabolites: Occurrence, structure and role in the human diet. Blackwell Publishing. https://doi.org/10.1002/9780470988558
Dent M, Dragovi-Uzelac V, Peni M, Brn~i M, Bosiljkov T and Levaj B. (2013). Polyphenols from Dalmatian Wild Sage. Food Technology and Biotechnology 51(1): 84–91.
Do Q D, Angkawijaya A E, Tran-Nguyen P L, Huynh L H, Soetaredjo F E, Ismadji S and Ju Y. (2014). Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatica. Journal of Food and Drug Analysis 22(3): 296–302. https://doi.org/10.1016/j.jfda.2013.11.001
Gomez S M and Kalamani A. (2003). Butterfly pea (Clitoria ternatea): A nutritive multipurpose forage legume for the tropics–an overview. Pakistan Journal of Nutrition 2(6): 374–379. https://doi.org/10.3923/pjn.2003.374.379
Holiman P C, Hertog M G and Katan M B. (1996). Analysis and health effects of flavonoids. Food Chemistry 57(1): 43–46. https://doi.org/10.1016/0308-8146(96)00065-9
International Legume Database and Information Service (ILDS). (1994). Plants and their constituents. In: Bisby F A (ed.), Phytochemical dictionary of the Leguminosae. New York: Chapman and Hall, 1–748.
Kaisoon O, Siriamornpun S, Weerapreeyakul N and Meeso N. (2011). Phenolic compounds and antioxidant activities of edible flowers from Thailand. Journal of Functional Foods 3(2): 88–99. https://doi.org/10.1016/j.jff.2011.03.002
Kamkaen N and Wilkinson J M. (2009). The antioxidant activity of Clitoria ternatea flower petal extracts and eye gel. Phytotherapy Research 23(11): 1624–1625. https://doi.org/10.1002/ptr.2832
Kazuma K, Noda N and Suzuki M. (2003). Malonylated flavonol glycosides from the petals of Clitoria ternatea. Phytochemistry 62(2): 229–237. https://doi.org/10.1016/S0031-9422(02)00486-7
Katsube T, Tabata H, Ohta Y, Yamasaki Y, Anuurad E and Shiwaku K. (2004). Screening for antioxidant activity in edible plant products: Comparison of low-density lipoprotein oxidation assay, DPPH radical scavenging assay, and Folin–Ciocalteu assay. Journal of the Agricultural and Food Chemistry 52: 2391–2396. https://doi.org/10.1021/jf035372g
Khuri A I and Mukhopadhyay S. (2010). Response surface methodology. Wiley Interdisciplinary Reviews: Computational Statistics 2(2): 128–149. https://doi.org/10.1002/wics.73
Kim S, Hwang G, Lee S, Zhu J-Y, Paik Nguyen T T, Kim J and Oh E. (2017). High ambient temperature represses anthocyanin biosynthesis through degradation of HY5. Front Plant SCI 8: 1787. https://doi.org/10.3389/fpls.2017.01787
Kungsuwan K, Singh K, Phetkao S, and Utama-ang N. (2014). Effects of pH and anthocyanin concentration on color and antioxidant activity of Clitoria ternatea extract. Food and Applied Bioscience Journal 2(1): 31–46.
Liyana-Pathirana C and Shahidi F. (2005). Optimization of extraction of phenolic compounds from wheat using response surface methodology. Food Chemistry 93(1): 47–56. https://doi.org/10.1016/j.foodchem.2004.08.050
Li J, Li X.-d, Zhang Y, Zheng Z.-d, Qu Z.-y, Liu M, Zhu S.-h, Liu S, Wang M and Qu L. (2013). Identification and thermal stability of purple-fleshed sweet potato anthocyanins in aqueous solutions with various pH values and fruit juices. Food Chemistry 136(3–4): 1429–1434. https://doi.org/10.1016/j.foodchem.2012.09.054
Mazza G and Miniati E. (1993). Anthocyanins in fruits, vegetables, and grains. Legumes. Boca Raton, FL: CRC Press, 1–28.
McCann D, Barrett A, Cooper A, Crumpler D, Dalen L, and Grimshaw K, Kitchin E, Lok K, Porteous L, Prince E, Sonuga-Barke E, Warner J O and Stevenon J. (2007). Food additives and hyperactive behaviour in 3-year-old and 8/9-year-old children in the community: A randomised, double-blinded, placebo-controlled trial. Lancet 370(9598): 1560–1567. https://doi.org/10.1016/S0140-6736(07)61306-3
Morris J B. (2008). Characterization of butterfly pea (Clitoria ternatea L.) accessions for morphology, phenology, reproduction and potential nutraceutical, pharmaceutical trait utilization. Genetic Resources and Crop Evolution 56(3): 421–427. https://doi.org/10.1007/s10722-008-9376-0
Nickavar B, Alinaghi A and Kamalinejad M. (2008). Evaluation of the antioxidant properties of five mentha species. Iranian Journal of Pharmaceutical Research 7(3): 203–209.
Pandey A and Tripathi S. (2013). Concept of standardization, extraction and pre phytochemical screening strategies for herbal drug. Journal of Pharmacognosy and Phytochemistry 2(5): 115–119.
Prasad N K, Yang B, Dong X, Jiang G, Zhang H, Xie H and Jiang Y. (2009). Flavonoid contents and antioxidant activities from Cinnamomum species. Innovative Food Science and Emerging Technologies 10: 627–632. https://doi.org/10.1016/j.ifset.2009.05.009
Sies H, Stahl W and Sundquist A R. (1992). Antioxidant functions of vitamins. Annals of the New York Academy of Sciences 669(1): 7–20. https://doi.org/10.1111/j.1749-6632.1992.tb17085.x
Siti Azima A M, Noriham, A and Manshoor, N. (2014). Anthocyanin content in relation to the antioxidant activity and colour properties of Garcinia mangostana peel, Syzigium cumini and Clitoria ternatea extracts. International Food Research Journal 21(6): 2369–2375.
Spigno G, Tramelli L and Faveri D M. (2007). Effects of extraction time, temperature and solvent on concentration and antioxidant activity of grape marc phenolics. Journal of Food Engineering 81(1): 200–208. https://doi.org/10.1016/j.jfoodeng.2006.10.021
Stankovic M S. (2011). Total phenolic content, flavonoid concentration and antioxidant activity of Marrubium peregrinum L. extracts. Kragujevac Journal of Science 33: 63–72.
Suppadit T. (2011). Use of anthocyanin extracted from natural plant materials to develop a pH test kit for measuring effluent from animal farms. African Journal of Biotechnology 10(82): 19109–19118. https://doi.org/10.5897/AJB11.2263
Tanaka Y, Brugliera F and Chandler S. (2009). Recent progress of flower colour modification by biotechnology. International Journal of Molecular Sciences 10(12): 5350–5369. https://doi.org/10.3390/ijms10125350
Turkmen N, Sari F and Velioglu Y S. (2006). Effects of extraction solvents on concentration and antioxidant activity of black and black mate tea polyphenols determined by ferrous tartrate and Folin–Ciocalteu methods. Food Chemistry 99(4): 835–841. https://doi.org/10.1016/j.foodchem.2005.08.034
Wang J, Sun B, Cao Y, Tian Y and Li X. (2008). Optimisation of ultrasound-assisted extraction of phenolic compounds from wheat bran. Food Chemistry 106(2): 804–810. https://doi.org/10.1016/j.foodchem.2007.06.062
Williams R J, Spencer J P, and Rice-Evans, C. (2004). Flavonoids: Antioxidants or signalling molecules? Free Radical Biology and Medicine 36(7): 838–849. https://doi.org/10.1016/j.freeradbiomed.2004.01.001
Yang Z and Zhai, W. (2010). Identification and antioxidant activity of anthocyanins extracted from the seed and cob of purple corn (Zea mays L.). Innovative Food Science and Emerging 11(1): 169–176. https://doi.org/10.1016/j.ifset.2009.08.012