Sains Malaysiana 52(11)(2023): 3147-3162

http://doi.org/10.17576/jsm-2023-5211-10

Optimization of Roselle (Hibiscus sabdariffa Linn.) Anthocyanin Extraction Parameter by Response Surface Modeling and Potential of Roselle Agro-Waste as Alternative Sources of Anthocyanin

(Pengoptimuman Parameter Pengekstrakan Antosianin Rosel (Hibiscus sabdariffa Linn.) Mengikut Pemodelan Permukaan Tindak Balas dan Potensi Sisa Agro Rosel sebagai Punca Alternatif Antosianin)

NURUL NADIAH ABU BAKAR¹, JALIFAH LATIP¹, SYAIFUZAH SAPIAN² & ROZIDA MOHD KHALID^1,*

¹Department of Chemical Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

²Centre for Diagnostic, Therapeutic and Investigative Studies, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Malaysia

Diserahkan: 8 Jun 2023/Diterima: 24 Oktober 2023

Abstract

Roselle or Hibiscus sabdariffa Linn. is a perennial plant with a high concentration of anthocyanin in the calyx. This study optimized the extraction conditions for maximum anthocyanin yield in the calyx using response surface methodology (RSM) and face-centered central composite design (FCCCD), which showed that ultrasound-assisted extraction (UAE) method with 70% ethanol concentration (v/v) at 35 minutes had the highest total anthocyanin content (TAC) which at 5.277 mg Cya3G/g DW. Using this optimised extraction method, roselle agro-waste which are calyx residue, leaf and stem were extracted and tested for TAC. The highest TAC was found in the calyx residue (3.011 mg Cya3G/g DW), followed by stems (0.077 mg Cya3G/g DW), and leaves (0.073 mg Cya3G/g DW). This study also quantified the concentration of two major anthocyanins using high-performance liquid chromatography (HPLC) and compared the total phenolic content (TPC) and total flavonoid content (TFC) of calyx and calyx residue extracts. Our findings showed that the TAC of calyx residue extracts was significantly low compared to calyx extracts. However, there were no significant differences in the TPC and TFC values. These results suggest that calyx residue, which is typically discarded after processing of calyx products, is a good source of phenolics and flavonoids and compatible with fresh calyx. In addition, calyx residues are still a good source of anthocyanins, making it an added-value for the roselle industry.

Keyword: Anthocyanins; face-centered central composite design; HPLC; Response Surface Methodology; roselle

Abstrak

Rosel atau Hibiscus sabdariffa Linn. ialah sejenis tumbuhan berumur panjang yang mengandungi kepekatan tinggi antosianin di kaliksnya. Kajian ini mengoptimumkan keadaan pengekstrakan bagi mendapatkan hasil antosianin maksimum di kaliks menggunakan metodologi rangsangan permukaan (RSM) dan reka bentuk komposit berpusatkan muka (FCCCD) yang menunjukkan bahawa kaedah pengekstrakan dengan bantuan gelombang bunyi (UAE) menggunakan kepekatan etanol 70% (v/v) selama 35 minit mempunyai kandungan antosianin keseluruhan tertinggi (TAC) iaitu pada nilai 5.277 mg Cya3G/g BK. Dengan menggunakan kaedah pengekstrakan yang telah dioptimumkan ini, sisa pertanian roselle seperti sisa kaliks, daun dan batang diekstrak dan diuji untuk TAC. TAC tertinggi ditemui dalam sisa kaliks (3.011 mg Cya3G/g BK), diikuti oleh batang (0.077 mg Cya3G/g BK) dan daun (0.073 mg Cya3G/g BK). Kajian ini juga mengkuantifikasi kepekatan dua jenis antosianin utama menggunakan kromatografi cecair prestasi tinggi (HPLC) serta membandingkan kandungan fenol keseluruhan (TPC) dan kandungan flavonoid keseluruhan (TFC) antara ekstrak kaliks dan sisa kaliks. Hasil kajian kami menunjukkan bahawa TAC ekstrak sisa kaliks adalah jauh lebih rendah berbanding ekstrak kaliks. Namun begitu, tidak terdapat perbezaan yang signifikan dalam nilai TPC dan TFC. Hasil ini mencadangkan bahawa sisa kaliks yang biasanya dibuang selepas proses penghasilan produk kaliks adalah sumber yang baik untuk fenol dan flavonoid, sejajar dengan kaliks segar. Selain itu, sisa kaliks masih merupakan sumber yang baik untuk antosianin dan menjadikannya tambahan nilai bagi industri rosel.

Kata kunci: Antosianin; HPLC; Metodologi Rangsangan Permukaan; reka bentuk komposit berpusatkan muka; rosel

RUJUKAN

Ali, S.A.M., Latip, J. & Zain, C.R.C.M. 2019. Growth and phenolic constituents production in roselle (Hibiscus sabdariffa var. UKMR-2) as influenced by irrigation treatment. AIP Conference Proceedings 2111(1): 050013.

Ali, S.A.M., Mohd, C.R.C. & Latip, J. 2019. Comparison of phenolic constituent in Hibiscus sabdariffa cv. UKMR-2 calyx at different harvesting times. Sains Malaysiana 48(7): 1417-1424.

Ali, S.A.M., Zainalabidin, S. & Latip, J. 2019. Quantitative analysis of phenolics content in two roselle varieties (Hibiscus sabdariffa) by high performance liquid chromatography. Malaysiana J. Anal. Sci. 23: 715-724.

Amor, B.B. & Allaf, K. 2009. Impact of texturing using instant pressure drop treatment prior to solvent extraction of anthocyanins from Malaysian Roselle (Hibiscus sabdariffa). Food Chemistry 115(3): 820-825.

Bettiol, V.R., Bilibio, D., Priamo, W.L., Bergmeier, D., Berres, P.H.D. & Filippi, D. 2014. Extraction of total polyphenols from hibiscus (Hibiscus sabdariffa L.) and waxweed/‘sete-sangrias’(Cuphea carthagenensis) and evaluation of their antioxidant potential. Acta Scientiarum Technology 36(3): 545-551.

Bezerra, M.A., Santelli, R.E., Oliveira, E.P., Villar, L.S. & Escaleira, L.A. 2008. Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta 76(5): 965-977.

Celli, G.B., Ghanem, A. & Brooks, M.S.L. 2015. Optimization of ultrasound-assisted extraction of anthocyanins from haskap berries (Lonicera caerulea L.) using Response Surface Methodology. Ultrasonics Sonochemistry 27: 449-455.

Chen, S., Zeng, Z., Hu, N.A., Bai, B., Wang, H. & Suo, Y. 2018. Simultaneous optimization of the ultrasound-assisted extraction for phenolic compounds content and antioxidant activity of Lycium ruthenicum Murr. fruit using response surface methodology. Food Chemistry 242: 1-8.

Chumsri, P., Sirichote, A. & Itharat, A. 2008. Studies on the optimum conditions for the extraction and concentration of roselle (Hibiscus sabdariffa Linn.) extract. Songklanakarin Journal of Science & Technology 30: 133-139.

Duy, N.Q., Binh, M.L.T., Thuan, M., Van, N.T.T., Lam, T.D., Tran, T.H. & Nhan, P.N.T. 2019. Effects of extraction conditions on total phenolic content and total flavonoid content of roselle (Hibiscus sabdariffa L.) extracts. Key Engineering Materials 814: 469-474.

Enaru, B., Drețcanu, G., Pop, T.D., Stǎnilǎ, A. & Diaconeasa, Z. 2021. Anthocyanins: Factors affecting their stability and degradation. Antioxidants 10(12): 1967.

Galgano, F., Tolve, R., Scarpa, T., Caruso, M.C., Lucini, L., Senizza, B. & Condelli, N. 2021. Extraction kinetics of total polyphenols, flavonoids, and condensed tannins of lentil seed coat: Comparison of solvent and extraction methods. Foods 10(8): 1810.

Giusti, M.M. & Wrolstad, R.E. 2001. Characterization and measurement of anthocyanins by UV-visible spectroscopy. Current Protocols in Food Analytical Chemistry (2001). New York: John Wiley and Sons Inc.

Halim, N.A.A., Abidin, Z.Z., Siajam, S.I., Hean, C.G. & Harun, M.R. 2021. Optimization studies and compositional analysis of subcritical water extraction of essential oil from Citrus hystrix DC. leaves. The Journal of Supercritical Fluids 178: 105384.

Hapsari, B.W. & Setyaningsih, W. 2021. Methodologies in the analysis of phenolic compounds in Roselle (Hibiscus sabdariffa L.): Composition, biological activity, and beneficial effects on human health. Horticulturae 7(2): 35.

Ifie, I., Ifie, B.E., Ibitoye, D.O., Marshall, L.J. & Williamson, G. 2018. Seasonal variation in Hibiscus sabdariffa (Roselle) calyx phytochemical profile, soluble solids and α-glucosidase inhibition. Food Chemistry 261: 164-168.

Jiang, T., Mao, Y., Sui, L., Yang, N., Li, S., Zhu, Z., Wang, C., Yin, S., He, J. & He, Y. 2019. Degradation of anthocyanins and polymeric color formation during heat treatment of purple sweet potato extract at different pH. Food Chemistry 274: 460-470.

Khazaei, K.M., Jafari, S.M., Ghorbani, M., Kakhki, A.H. & Sarfarazi, M. 2016. Optimization of anthocyanin extraction from saffron petals with response surface methodology. Food Analytical Methods 9(7): 1993-2001.

Kouakou, T.H., Konkon, N.G., Ayolie, K., Obouayeba, A.P., Abeda, Z.H. & Kone, M. 2015. Anthocyanin production in calyx and callus of Roselle (Hibiscus sabdariffa L.) and its impact on antioxidant activity. Journal of Pharmacognosy and Phytochemistry 4(3): 09-15.

Lin, J.Y. & Tang, C.Y. 2007. Determination of total phenolic and flavonoid contents in selected fruits and vegetables, as well as their stimulatory effects on mouse splenocyte proliferation. Food Chemistry 101(1): 140-147.

Liu, X., Mu, T., Sun, H., Zhang, M. & Chen, J. 2013. Optimisation of aqueous two-phase extraction of anthocyanins from purple sweet potatoes by response surface methodology. Food Chemistry 141(3): 3034-3041.

Maciel, L.G., Carmo, M.A.V., Azevedo, L., Daguer, H., Molognoni, L., de Almeida, M.M., Granato, D. & Rosso, N.D. 2018. Hibiscus sabdariffa anthocyanins-rich extract: Chemical stability, in vitro antioxidant and antiproliferative activities. Food and Chemical Toxicology 113: 187-197.

Meziant, L., Benchikh, Y. & Louaileche, H. 2014. Deployment of response surface methodology to optimize recovery of dark fresh fig (Ficus carica L., var. Azenjar) total phenolic compounds and antioxidant activity. Food Chemistry 162: 277-282.

Mohd-Esa, N., Hern, F.S., Ismail, A. & Yee, C.L. 2010. Antioxidant activity in different parts of roselle (Hibiscus sabdariffa L.) extracts and potential exploitation of the seeds. Food Chemistry 122(4): 1055-1060.

Norhisamudin, N.A., Sabani, N., Shamimin, M.M., Juhari, N., Shaari, S., Ahmad, M.F. & Zakaria, N. 2021. The effect of different solvents in natural dyes from Roselle (Hibiscus sabdariffa) and green tea leaves (Camellia sinensis) for dye-sensitized solar cell. Journal of Physics: Conference Series 1755(1): 012024.

Ojulari, O.V., Lee, S.G. & Nam, J.O. 2019. Beneficial effects of natural bioactive compounds from Hibiscus sabdariffa L. on obesity. Molecules 24(1): 210.

Patras, A., Brunton, N.P., O'Donnell, C. & Tiwari, B.K. 2010. Effect of thermal processing on anthocyanin stability in foods; mechanisms and kinetics of degradation. Trends in Food Science & Technology 21(1): 3-11.

Peng, W.L., Mohd-Nasir, H., Setapar, S.H.M., Ahmad, A. & Lokhat, D. 2020. Optimization of process variables using response surface methodology for tocopherol extraction from Roselle seed oil by supercritical carbon dioxide. Industrial Crops and Products 143: 111886.

Peredo Pozos, G.I., Ruiz-López, M.A., Zamora Natera, J.F., Alvarez Moya, C., Barrientos Ramirez, L., Reynoso Silva, M., Rodriguez Macias, R., García-López, P.M., Gonzalez Cruz, R., Salcedo Perez, E. & Vargas Radillo, J.J. 2020. Antioxidant capacity and antigenotoxic effect of Hibiscus sabdariffa L. extracts obtained with ultrasound-assisted extraction process. Applied Sciences 10(2): 560.

Pham, T.N., Nguyen, T.N.P., Lam, T.D., Tran, T.H., Nguyen, D.C., Vo, D.V.N., Le, X.T., Do, S.T. & Bach, L.G. 2019. Effects of various solvent concentration, liquid-solid ratio, temperatures and time values on the extraction yield of anthocyanin from Vietnam Hibiscus sabdariffa L. (Roselle). IOP Conference Series: Materials Science and Engineering 542(1):012033.

Pinela, J., Prieto, M.A., Pereira, E., Jabeur, I., Barreiro, M.F., Barros, L. & Ferreira, I.C. 2019. Optimization of heat-and ultrasound-assisted extraction of anthocyanins from Hibiscus sabdariffa calyx for natural food colorants. Food Chemistry 275: 309-321.

Pragalyaashree, M.M., Tiroutchelvame, D. & Sashikumar, S. 2018. Degradation kinetics of anthocyanin extracted from roselle calyx (Hibiscus sabdariffa). Journal of Applied Pharmaceutical Science 8(11): 057-063.

Purwanto, U.R.E., Ariani, L.W. & Pramitaningastuti, A.S. 2019. Formulasi serum liposom antosianin dari kulit buah naga merah (Hylocereus polyrhizus) untuk antiaging. Cendekia Journal of Pharmacy 3(2): 96-105.

Rajha, H.N., El Darra, N., Hobaika, Z., Boussetta, N., Vorobiev, E., Maroun, R.G. & Louka, N. 2014. Extraction of total phenolic compounds, flavonoids, anthocyanins, and tannins from grape byproducts by response surface methodology. Influence of solid-liquid ratio, particle size, time, temperature, and solvent mixtures on the optimization process. Food and Nutrition Sciences 5(04): 397-409.

Ryu, D. & Koh, E. 2018. Application of response surface methodology to acidified water extraction of black soybeans for improving anthocyanin content, total phenols content and antioxidant activity. Food Chemistry 261: 260-266.

Samadi, S. & Fard, F.R. 2020. Phytochemical properties, antioxidant activity and mineral content (Fe, Zn and Cu) in Iranian produced black tea, green tea and roselle calyx. Biocatalysis and Agricultural Biotechnology 23: 101472.

Sapian, S., Taib, I.S., Katas, H., Latip, J., Zainalabidin, S., Hamid, Z.A., Anuar, N.N.M. & Budin, S.B. 2022. The role of anthocyanin in modulating diabetic cardiovascular disease and its potential to be developed as a nutraceutical. Pharmaceuticals 15(11): 1344.

Shalini, S., Kumar, T.S., Prasanna, S. & Balasundaraprabhu, R. 2020. Investigations on the effect of co-doping in enhancing the performance of nanostructured TiO₂ based DSSC sensitized using extracts of Hibiscus sabdariffa calyx. Optik 212: 164672.

Singh, P., Khan, M. & Hailemariam, H. 2017. Nutritional and health importance of Hibiscus sabdariffa: A review and indication for research needs. J. Nutr. Health Food Eng 6(5): 00212.

Sirichan, T., Kijpatanasilp, I., Asadatorn, N. & Assatarakul, K. 2022. Optimization of ultrasound extraction of functional compound from Makiang seed by response surface methodology and antimicrobial activity of optimized extract with its application in orange juice. Ultrasonics Sonochemistry 83: 105916.

Witek-Krowiak, A., Chojnacka, K., Podstawczyk, D., Dawiec, A. & Bubała, K. 2014. Application of response surface methodology and artificial neural network methods in modelling and optimization of biosorption process. Bioresource Technology 160: 150-160.

Wu, H.Y., Yang, K.M. & Chiang, P.Y. 2018. Roselle anthocyanins: Antioxidant properties and stability to heat and pH. Molecules 23(6): 1357.

Yang, P., Yuan, C., Wang, H., Han, F., Liu, Y., Wang, L. & Liu, Y. 2018. Stability of anthocyanins and their degradation products from cabernet sauvignon red wine under gastrointestinal pH and temperature conditions. Molecules 23(2): 354.

Zannou, O., Koca, I., Aldawoud, T.M. & Galanakis, C.M. 2020. Recovery and stabilization of anthocyanins and phenolic antioxidants of roselle (Hibiscus sabdariffa L.) with hydrophilic deep eutectic solvents. Molecules 25(16): 3715.

*Pengarang untuk surat-menyurat; email: rozidakhalid@ukm.edu.my