A Box-Behnken experimental design for microwave assisted extraction optimization of pectin from citron peel

Document Type: Original research


Bioprocessing and Biodetection Laboratory, Department of Food Science and Engineering, University of Tehran, Karaj - Iran - Postal Code 31587-77871


Microwave assisted extraction technique was employed to extract pectin from citron peel. Box-Behnken design was applied to investigate the influence of irradiation time, microwave power and pH on the yield and DE of pectin. The finding indicated that the optimal conditions for the maximum yield of pectin (30.71%) were achieved at irradiation time of 3 min, microwave power of 700 W and pH of 1.5. DE of pectin ranged from 45.08 to 72.19% demonstrating that extracted pectin was classified as high methoxyl pectin. The emulsifier activity of extracted pectin under optimal conditions was 40.7%. Besides, the emulsions were 86.7 and 86.6% stable at 4°C and, 79.5 and 79.2 at 23°C after 1 and 30 days, respectively. The flow behavior of pectin solutions indicated that solutions in low (0.1, 0.5 and 1.0 %w/v) and high (2.0 %w/v) concentrations had Newtonian and pseudoplastic behaviors, respectively.


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.

Chaharbaghi, E., Khodaiyan, F., & Hosseini, S. S. (2017). Optimization of pectin extraction from pistachio green hull as a new source. Carbohydrate Polymers, 173, 107–113.

Chen, Y., Xie, M. Y., & Gong, X. F. (2007). Microwave-assisted extraction used for the isolation of total triterpenoid saponins from Ganoderma atrum. Journal of Food Engineering, 81(1), 162–170.

Chen, Y., Zhang, J. G., Sun, H. J., & Wei, Z. J. (2014). Pectin from Abelmoschus esculentus: Optimization of extraction and rheological properties. International Journal of Biological Macromolecules, 70, 498–505.

Cui, S. W., & Chang, Y. H. (2014). Emulsifying and structural properties of pectin enzymatically extracted from pumpkin. LWT - Food Science and Technology, 58(2), 396–403.

Dalev, P. G., & Simeonova, L. S. (1995). Emulsifying properties of protein–pectin complexes and their use in oil-containing foodstuffs. Journal of the Science of Food and Agriculture, 68(2), 203–206.

De Roeck, A., Sila, D. N., Duvetter, T., Van Loey, A., & Hendrickx, M. (2008). Effect of high pressure/high temperature processing on cell wall pectic substances in relation to firmness of carrot tissue. Food Chemistry, 107(3), 1225–1235.

Diaz-Rojas, E. I., Pacheco-Aguilar, R., Lizardi, J., Arguelles-Monal, W., Valdez, M. A., Rinaudo, M., et al. (2004). Linseed pectin: Gelling properties and performance as an encapsulation matrix for shark liver oil. Food Hydrocolloids, 18(2), 293–304.

Edashige, Y., Murakami, N., & Tsujita, T. (2008). Inhibitory effect of pectin from the segment membrane of citrus fruits on lipase activity. Journal of Nutritional Science and Vitaminology, 54(5), 409–415.

El-Nawawi, S. A., & Shehata, F. R. (1988). Effect of the extraction temperature on the quality characteristics of pectin extracted from Egyptian Orange Peel. Biological Wastes, 24(4), 307–311.

Espinal-Ruiz, M., Restrepo-Sánchez, L.-P., Narváez-Cuenca, C.-E., & McClements, D. J. (2016). Impact of pectin properties on lipid digestion under simulated gastrointestinal conditions: Comparison of citrus and banana passion fruit (Passiflora tripartita var. mollissima) pectins. Food Hydrocolloids, 52, 329–342.

Essien, E. P., Essien, J. P., Ita, B. N., & Ebong, G. A. (2008). Physicochemical properties and fungitoxicity of the essential oil of Citrus medica L. against groundnut storage fungi. Turkish Journal of Botany, 32(2), 161–164.

Faravash, R. S., & Ashtiani, F. Z. (2007). The effect of pH, ethanol volume and acid washing time on the yield of pectin extraction from peach pomace. International Journal of Food Science and Technology, 42(10), 1177–1187.

Gabriele, B., Fazio, A., Dugo, P., Costa, R., & Mondello, L. (2009). Essential oil composition of Citrus medica L. Cv. diamante (diamante citron) determined after using different extraction methods. Journal of Separation Science, 32(1), 99–108.

Gfrerer, M., & Lankmayr, E. (2005). Screening, optimization and validation of microwave-assisted extraction for the determination of persistent organochlorine pesticides. Analytica Chimica Acta, 533(2), 203–211.

Gnanasambandam, R., & Proctor, A. (2000). Determination of pectin degree of esterication by diffuse reflectance Fourier transform infrared spectroscopy. Food Chemistry, 68, 327–332.

Happi Emaga, T., Garna, H., Paquot, M., & Deleu, M. (2012). Purification of pectin from apple pomace juice by using sodium caseinate and characterisation of their binding by isothermal titration calorimetry. Food Hydrocolloids, 29(1), 211–218.

Hokputsa, S., Gerddit, W., Pongsamart, S., Inngjerdingen, K., Heinze, T., Koschella, A., et al. (2004). Water-soluble polysaccharides with pharmaceutical importance from Durian rinds (Durio zibethinus Murr.): Isolation, fractionation, characterisation and bioactivity. Carbohydrate Polymers, 56(4), 471–481.

Hosseini, S. S., Khodaiyan, F., & Yarmand, M. S. (2016). Aqueous extraction of pectin from sour orange peel and its preliminary physicochemical properties. International Journal of Biological Macromolecules, 82, 920–926.

Inngjerdingen, K. T., Patel, T. R., Chen, X., Kenne, L., Allen, S., Morris, G. A., et al. (2007). Immunological and structural properties of a pectic polymer from Glinus oppositifolius. Glycobiology, 17(12), 1299–1310.

Jackson, C. L., Dreaden, T. M., Theobald, L. K., Tran, N. M., Beal, T. L., Eid, M.,  et al. (2007). Pectin induces apoptosis in human prostate cancer cells: correlation of apoptotic function with pectin structure. Glycobiology, 17(8), 805–819.

Jackson, J. C., Bourne, M. C., & Barnard, J. (1996). Optimization of blanching for crispness of banana chips using response surface methodology. Journal of Food Science, 61(1), 165–166.

Jafari, F., Khodaiyan, F., Kiani, H., & Hosseini, S. S. (2017). Pectin from carrot pomace: Optimization of extraction and physicochemical properties. Carbohydrate Polymers, 157, 1315–1322.

Joye, D. D., & Luzio, G. A. (2000). Process for selective extraction of pectins from plant material by differential pH. Carbohydrate Polymers, 43(4), 337–342.

Kim, K. N., Ko, Y. J., Yang, H. M., Ham, Y. M., Roh, S. W., Jeon, Y. J., et al. (2013). Anti-inflammatory effect of essential oil and its constituents from fingered citron (Citrus medica L. var. sarcodactylis) through blocking JNK, ERK and NF-κB signaling pathways in LPS-activated RAW 264.7 cells. Food and Chemical Toxicology, 57, 126–131.

Kozarski, M., Klaus, A., Niksic, M., Jakovljevic, D., Helsper, J. P. F. G., & Van Griensven, L. J. L. D. (2011). Antioxidative and immunomodulating activities of polysaccharide extracts of the medicinal mushrooms Agaricus bisporus, Agaricus brasiliensis, Ganoderma lucidum and Phellinus linteus. Food Chemistry, 129(4), 1667–1675.

Kratchanova, M., Pavlova, E., & Panchev, I. (2004). The effect of microwave heating of fresh orange peels on the fruit tissue and quality of extracted pectin. Carbohydrate Polymers, 56(2), 181–185.

Kumar, A., & Chauhan, G. S. (2010). Extraction and characterization of pectin from apple pomace and its evaluation as lipase (steapsin) inhibitor. Carbohydrate Polymers, 82(2), 454–459.

Kumbár, V., Nedomová, S., Pytel, R., Kilián, L., & Buchar, J. (2017). Study of rheology and friction factor of natural food hydrocolloid gels. Potravinarstvo Slovak Journal of Food Sciences, 11(1), 203–209.

Kurita, O., Fujiwara, T., & Yamazaki, E. (2008). Characterization of the pectin extracted from citrus peel in the presence of citric acid. Carbohydrate Polymers, 74(3), 725–730.

Lee, W. C., Yusof, S., Hamid, N. S. A., & Baharin, B. S. (2006). Optimizing conditions for enzymatic clarification of banana juice using response surface methodology (RSM). Journal of Food Engineering, 73(1), 55–63.

Li, D. Q., Jia, X., Wei, Z., & Liu, Z. Y. (2012). Box-Behnken experimental design for investigation of microwave-assisted extracted sugar beet pulp pectin. Carbohydrate Polymers, 88(1), 342–346.

Lota, M.-L., de Rocca Serra, D., Tomi, F., Bessiere, J.-M., & Casanova, J. (1999). Chemical composition of peel and leaf essential oils of Citrus medica L. and C. limonimedica Lush. Flavour and Fragrance Journal, 14(3), 161–166.

Ma, S., Yu, S. J., Zheng, X. L., Wang, X. X., Bao, Q. D., & Guo, X. M. (2013). Extraction, characterization and spontaneous emulsifying properties of pectin from sugar beet pulp. Carbohydrate Polymers, 98(1), 750–753.

Maran, J. P., Manikandan, S., Priya, B., & Gurumoorthi, P. (2015). Box-Behnken design based multi-response analysis and optimization of supercritical carbon dioxide extraction of bioactive flavonoid compounds from tea (Camellia sinensis L.) leaves. Journal of Food Science and Technology, 52(1), 92–104.

Maran, J. P., & Priya, B. (2014). Ultrasound-assisted extraction of polysaccharide from Nephelium lappaceum L. Fruit peel. International Journal of Biological Macromolecules, 70, 530–536.

Maran, J. P., Priya, B., & Nivetha, C. V. (2015). Optimization of ultrasound-assisted extraction of natural pigments from Bougainvillea glabra flowers. Industrial Crops and Products, 63, 182–189.

Maran, J. P., Swathi, K., Jeevitha, P., Jayalakshmi, J., & Ashvini, G. (2015). Microwave-assisted extraction of pectic polysaccharide from waste mango peel. Carbohydrate Polymers, 123, 67–71.

Mastrocola, D., Dalla Rosa, M., & Massini, R. (1997). Freeze-dried strawberries rehydrated in sugar solutions: Mass transfers and characteristics of final products. Food Research International, 30(5), 359–364.

Matia-Merino, L., Lau, K., & Dickinson, E. (2004). Effects of low-methoxyl amidated pectin and ionic calcium on rheology and microstructure of acid-induced sodium caseinate gels. Food Hydrocolloids, 18(2), 271–281.

Menichini, F., Tundis, R., Bonesi, M., de Cindio, B., Loizzo, M. R., Conforti, F., et al. (2011). Chemical composition and bioactivity of Citrus medica L. cv. Diamante essential oil obtained by hydrodistillation, cold-pressing and supercritical carbon dioxide extraction. Natural Product Research, 25(8), 789–799.

Mesbahi, G., Jamalian, J., & Farahnaky, A. (2005). A comparative study on functional properties of beet and citrus pectins in food systems. Food Hydrocolloids, 19(4), 731–738.

Migliori, M., Gabriele, D., Checchetti, A., & Battipede, B. (2010). Compatibility analysis of pectin at different esterification degree from intrinsic viscosity data of diluted ternary solutions. Reactive and Functional Polymers, 70(10), 863–867.

Minkov, S., Minchev, A., & Paev, K. (1996). Modelling of the hydrolysis and extraction of apple pectin. Journal of Food Engineering, 29(1), 107–113.

Mohnen, D. (2008). Pectin structure and biosynthesis. Current Opinion in Plant Biology, 11(3), 266–277.

Moorthy, I. G., Maran, J. P., Surya, S. M., Naganyashree, S., & Shivamathi, C. S. (2015). Response surface optimization of ultrasound assisted extraction of pectin from pomegranate peel. International Journal of Biological Macromolecules, 72, 1323–1328.

Morris, G. A., Ralet, M. C., Bonnin, E., Thibault, J. F., & Harding, S. E. (2010). Physical characterisation of the rhamnogalacturonan and homogalacturonan fractions of sugar beet (Beta vulgaris) pectin. Carbohydrate Polymers, 82(4), 1161–1167.

Nangia-Makker, P., Hogan, V., Honjo, Y., Baccarini, S., Tait, L., Bresalier, R., et al. (2002). Inhibition of human cancer cell growth and metastasis in nude mice by oral intake of modified citrus pectin. JNCI: Journal of the National Cancer Institute, 94(24), 1854–1862.

Nicolosi, E., La Malfa, S., El-Otmani, M., Negbi, M., & Goldschmidt, E. E. (2005). The search for the authentic citron (Citrus medica L.): Historic and genetic analysis. HortScience, 40(7), 1963–1968.

Pagán, J., Ibarz, A., Llorca, M., Pagán, A., & Barbosa-Cánovas, G. V. (2001). Extraction and characterization of pectin from stored peach pomace. Food Research International, 34(7), 605–612.

Pinheiro, E. R., Silva, I. M. D. A., Gonzaga, L. V., Amante, E. R., Teófilo, R. F., Ferreira, M. M. C., et al. (2008). Optimization of extraction of high-ester pectin from passion fruit peel (Passiflora edulis flavicarpa) with citric acid by using response surface methodology. Bioresource Technology, 99(13), 5561–5566.

Prakash Maran, J., & Manikandan, S. (2012). Response surface modeling and optimization of process parameters for aqueous extraction of pigments from prickly pear (Opuntia ficus-indica) fruit. Dyes and Pigments, 95(3), 465–472.

Prakash Maran, J., Mekala, V., & Manikandan, S. (2013). Modeling and optimization of ultrasound-assisted extraction of polysaccharide from Cucurbita moschata. Carbohydrate Polymers, 92(2), 2018–2026.

Prakash Maran, J., Sivakumar, V., Thirugnanasambandham, K., & Sridhar, R. (2013). Optimization of microwave assisted extraction of pectin from orange peel. Carbohydrate Polymers, 97(2), 703–709.

Prakash Maran, J., Sivakumar, V., Thirugnanasambandham, K., & Sridhar, R. (2014). Microwave assisted extraction of pectin from waste Citrullus lanatus fruit rinds. Carbohydrate Polymers, 101(1), 786–791.

Raji, Z., Khodaiyan, F., Rezaei, K., Kiani, H., & Hosseini, S. S. (2017). Extraction optimization and physicochemical properties of pectin from melon peel. International Journal of Biological Macromolecules, 98, 709–716.

Ridley, B. L., O’Neill, M. A., & Mohnen, D. (2001). Pectins: Structure, biosynthesis, and oligogalacturonide-related signaling. Phytochemistry, 57(6), 929–967.

Sáenz, C., Sepúlveda, E., & Matsuhiro, B. (2004). Opuntia spp mucilage’s: A functional component with industrial perspectives. Journal of Arid Environments, 57(3), 275–290.

Santos, J. D. G., Espeleta, A. F., Branco, A., de Assis, S. A., David, J., Santos, G.,  et al. (2013). Aqueous extraction of pectin from sisal waste. Carbohydrate Polymers, 92(2), 1997–2001.

Shi, X. Q., Chang, K. C., Schwarz, J. G., Wiesenborn, D. P., & Shih, M. C. (1996). Optimizing pectin extraction from sunflower heads by alkaline washing. Bioresource Technology, 58(3), 291–297.

Sparr Eskilsson, C., & Björklund, E. (2000). Analytical-scale microwave-assisted extraction. Journal of Chromatography A, 902(1), 227–250.

Thakur, B. R., Singh, R. K., Handa, A. K., & Rao, M. A. (1997). Chemistry and uses of pectin-A review. Critical Reviews in Food Science and Nutrition, 37(1), 47–73.

Torralbo, D. F., Batista, K. A., Di-Medeiros, M. C. B., & Fernandes, K. F. (2012). Extraction and partial characterization of Solanum lycocarpum pectin. Food Hydrocolloids, 27(2), 378–383.

Verzera, A., Trozzi, A., Zappala, M., Condurso, C., & Cotroneo, A. (2005). Essential oil composition of Citrus meyerii Y. Tan. and Citrus medica L. cv. Diamante and their lemon hybrids. Journal of Agricultural and Food Chemistry, 53(12), 4890–4894.

Vriesmann, L. C., & Petkowicz, C. L. O. (2013). Highly acetylated pectin from cacao pod husks (Theobroma cacao L.) forms gel. Food Hydrocolloids, 33(1), 58–65.

Wai, W. W., Alkarkhi, A. F. M., & Easa, A. M. (2010). Effect of extraction conditions on yield and degree of esterification of durian rind pectin: An experimental design. Food and Bioproducts Processing, 88(2–3), 209–214.

Wang, S., Chen, F., Wu, J., Wang, Z., Liao, X., & Hu, X. (2007). Optimization of pectin extraction assisted by microwave from apple pomace using response surface methodology. Journal of Food Engineering, 78(2), 693–700.

Yapo, B. M., Robert, C., Etienne, I., Wathelet, B., & Paquot, M. (2007). Effect of extraction conditions on the yield, purity and surface properties of sugar beet pulp pectin extracts. Food Chemistry, 100(4), 1356–1364.

Zheng, X., Yin, F., Liu, C., Xu, X. (2011). Effect of process parameters of microwave assisted extraction (MAE) on polysaccharides yield from pumpkin. Journal of Northeast Agricultural University (English Edition), 18(2), 79–86.