Journal of Food Science and Technology

, Volume 52, Issue 1, pp 92–104 | Cite as

Box-Behnken design based multi-response analysis and optimization of supercritical carbon dioxide extraction of bioactive flavonoid compounds from tea (Camellia sinensis L.) leaves

  • J. Prakash Maran
  • S. Manikandan
  • B. PriyaEmail author
  • P. Gurumoorthi
Original Article


The popularity of tea is increasing on the global aspect because of its role as a significant source of phenolic compounds in human diet. The objectives of this present study is to develop a supercritical carbon dioxide (SC-CO2) extraction method suitable for extraction of phenolic compounds such as total phenolics, flavonoids and tannin from tea leaves at various extraction conditions such as extraction pressure (100–200 bar), temperature (40–60 °C) and co-solvent (ethanol) flow rate (1–3 g/min). Furthermore, the total antioxidant activity of the SC-CO2 tea leaves extracts was assessed using ABTS+ (2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)) method. Response surface methodology (RSM) combined with Box-Behnken design was used to investigate and optimize the process variables. The results showed that, extraction pressure and co-solvent flow rate have significant effect on the responses. From the experimental data, second order polynomial mathematical models were developed for each response with high coefficient of determination value (R 2 > 0.95). An optimization study using Derringer’s desired function methodology was performed and the optimal conditions based on both individual and combinations of all independent variables (extraction pressure of 188 bar, temperature of 50 °C and co-solvent flow rate of 2.94 g/min) were determined with maximum total phenolic content of 131.24 mg GAE/100 ml, total flavonoid content of 194.60 mg QE/100 ml, total tannin content of 49.99 mg TAE/100 ml and totalantioxidant activity of 262.23 μMol TEAC/100 ml of extracts respectively with a overall desirability value of 0.983, which was confirmed through validation experiments.


Phenolic compound Antioxidant activity Tannin Response surface methodology 


  1. Arnao MB, Cano A, Acosta M (2001) The hydrophilic and lipophilic contribution to total antioxidant activity. Food Chem 73:239–244CrossRefGoogle Scholar
  2. Bjorklund E, Jaremo M, Mathiasson L, Jonsson JA, Karlsson L (1998) Illustration of important mechanisms controlling mass transfer in supercritical fluid extraction. Anal Chim Acta 368:117–128CrossRefGoogle Scholar
  3. Brunner G (2005) Supercritical fluids: technology and application to food processing. J Food Eng 67:21–33CrossRefGoogle Scholar
  4. Chiu KL, Cheng YC, Chen JH, Chang CJ, Yang PW (2002) Supercritical fluid extraction of Ginkgo ginkgolides and flavonoids. J Supercrit Fluids 24:77–87CrossRefGoogle Scholar
  5. Derringer G, Suich R (1980) Simultaneous optimization of several response variables. J Qual Technol 12(4):214–219Google Scholar
  6. Ekren O, Ekren BY (2008) Size optimization of a PV/wind hybrid energy conversion system with battery storage using response surface methodology. Appl Energy 85(11):1086–1101CrossRefGoogle Scholar
  7. Haggi TM, Anthony DD, Gupta S, Ahmad H, Lee MS, Kumar GK (1999) Prevention of collagen induced arthritis in mice by a polyphenolic fraction from green tea. Proc Natl Acad Sci U S A 96(8):4524–4529CrossRefGoogle Scholar
  8. Han X, Cheng L, Zhang R, Bi J (2009) Extraction of safflower seed oil by supercritical CO2. J Food Eng 92:370–376CrossRefGoogle Scholar
  9. Hegarty VM, May HM, Khaw KT (2000) Tea drinking and bone mineral density in older women. Am J Clin Nutr 71:1003–1007Google Scholar
  10. Hertog MGL, Hollman PCH, Katan MB, Kromhout D (1993) Estimation of daily intake of potentially anticarcinogenic flavonoids and their determinants in adults in the Netherlands. Nutr Cancer 20:21–29CrossRefGoogle Scholar
  11. Huang W, Li Z, Niu H, Li D, Zhang J (2008) Optimization of operating parameters for supercritical carbon dioxide extraction of lycopene by response surface methodology. J Food Eng 89:298–302CrossRefGoogle Scholar
  12. Ixtaina V, Vega A, Nolasco S, Tomás M (2010) Supercritical carbon dioxide extraction of oil from Mexican chia seed (Salvia hispanica L.): characterization and process optimization. J Supercrit Fluids 55:192–199CrossRefGoogle Scholar
  13. Khan N, Mukhtar H (2007) Tea polyphenols for health promotion. Life Sci 81:519–533CrossRefGoogle Scholar
  14. Kumar A, Prasad B, Mishra IM (2007) Process parametric study for ethane carboxylic acid removal onto powder activated carbon using Box–Behnken design. Chem Eng Technol 30(7):927–932CrossRefGoogle Scholar
  15. Lang Q, Wai CM (2001) Supercritical fluid extraction in herbal and natural product studies—a practical review. Talanta 53:771–782CrossRefGoogle Scholar
  16. Lee YH, Charles AL, Kung HF, Ho CT, Huang TC (2010) Extraction of nobiletin and tangeretin from Citrus depressa Hayata by supercritical carbon dioxide with ethanol as modifier. Ind Crop Prod 31:59–64CrossRefGoogle Scholar
  17. Li B, Xu Y, Jin YX, Wu YY, Tu YY (2010) Response surface optimization of supercritical fluid extraction of kaempferol glycosides from tea seed cake. Ind Crop Prod 32:123–128CrossRefGoogle Scholar
  18. Lingzhao W, Bao Y, Xiuqiao D, Chun Y (2008) Optimisation of supercritical fluid extraction of flavonoids from Pueraria lobata. Food Chem 108:737–741CrossRefGoogle Scholar
  19. Machmudah S, Shotipruk A, Goto M, Sasaki M, Hirose T (2006) Extraction of astaxanthin from Haematococcus pluvialis using supercritical CO2 and ethanol as entrainer. Ind Eng Chem Res 45:3652–3657CrossRefGoogle Scholar
  20. Maran JP, Sivakumar V, Sridhar R, Thirgananasambandham K (2013) Development of model for barrier and optical properties of tapioca starch based films. Carbohydr Polym 92:1335–1347CrossRefGoogle Scholar
  21. Maron DJ, Lu GP, Cai NS, Wu ZG, Li YH, Chen H (2003) Cholesterol lowering effect of a theaflavin-enriched green tea extract: a randomized controlled trial. Arch Intern Med 163(12):1448–1453CrossRefGoogle Scholar
  22. Marr R, Gamse T (2000) Use of supercritical fluids for different processes including new developments—a review. Chem Eng Process 39:19–28CrossRefGoogle Scholar
  23. Mohajeri L, Aziz HA, Isa MH, Zahed MA (2010) Statistical experiment design approach for optimizing biodegradation of weathered crude oil in coastal sediments. Bioresour Technol 101:893–900CrossRefGoogle Scholar
  24. Otake S, Makimura M, Kuroki T, Nishihara Y, Hirasawa M (1991) Anticaries effects of polyphenolic compounds from Japanese green tea. Caries Res 25:438–443CrossRefGoogle Scholar
  25. Park HS, Lee HJ, Shin MH, Lee KW, Lee H, Kim YS (2007) Effects of cosolvents on the decaffeination of green tea by supercritical carbon dioxide. Food Chem 105:1011–1017CrossRefGoogle Scholar
  26. Pilavtepe M, Yucel M, Seref Helvaci S, Demircioglu M, Yesil-celiktas O (2012) Optimization and mathematical modeling of mass transfer between Zostera marina residues and supercritical CO2 modified with ethanol. J Supercrit Fluids 68:87–93CrossRefGoogle Scholar
  27. 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. Dye Pigment 95:465–472CrossRefGoogle Scholar
  28. Prakash Maran J, Manikandan S, Thirugnanasambandham K, Vigna Nivetha C, Dinesh R (2013a) Box-Behnken design based statistical modeling for ultrasound-assisted extraction of corn silk polysaccharide. Carbohydr Polym 92:604–611CrossRefGoogle Scholar
  29. Prakash Maran J, Manikandan S, Vigna Nivetha C, Dinesh R (2013b) Ultrasound assisted extraction of bioactive compounds from Nephelium lappaceum L. fruit peel using central composite face centered response surface design. Arab J Chem. doi: 10.1016/J.arabjc.2013.02.007 Google Scholar
  30. Prakash Maran J, Mekala V, Manikandan S (2013c) Modeling and optimization of ultrasound-assisted extraction of polysaccharide from Cucurbita moschata. Carbohydr Polym 92:2018–2026CrossRefGoogle Scholar
  31. Prakash Maran J, Sivakumar V, Sridhar R, Prince Immanuel V (2013d) Development of model for mechanical properties of tapioca starch based films. Ind Crop Prod 42:159–168CrossRefGoogle Scholar
  32. Prakash Maran J, Sivakaumar V , Thirgananasambandham K, Sridhar R (2013e) Model development and process optimization for solvent extraction of polyphenols from red grapes using Box- Behnken design. Prep Biochem Biotech. doi: 10.1080/10826068.2013.791629
  33. Priya B, Viswanathan R, Vairamani M (2013) Response surface optimisation of process variables for microencapsulation of garlic (Allium sativum L.) oleoresin by spray drying. Biosyst Eng 114:205–213CrossRefGoogle Scholar
  34. Rostagno MA, Araújo JMA, Sandi D (2002) Supercritical fluid extraction of isoflavones from soybean flour. Food Chem 78:111–117CrossRefGoogle Scholar
  35. Salgin S, Salgin U (2006) Supercritical fluid extraction of walnut kernel oil. Eur J Lipid Sci Technol 108:577–582CrossRefGoogle Scholar
  36. Scalia S, Giuffreda L, Pallado P (1999) Analytical and preparative supercritical fluid extraction of Chamomile flowers and its comparison with conventional methods. J Pharm Biomed Anal 21:549–558CrossRefGoogle Scholar
  37. Shirin Adel PR, Prakash J (2010) Chemical composition and antioxidant properties of ginger root (Zingiber officinale). J Med Plant Res 4(24):2674–2679Google Scholar
  38. Soto C, Conde E, Moure A, Zúñiga ME, Dominguez H (2008) Supercritical extraction of borage seed oil coupled to conventional solvent extraction of antioxidants. Eur J Lipid Sci Technol 110:1035–1044CrossRefGoogle Scholar
  39. Vatai T, Kerget MS, Knez Z (2009) Extraction of phenolic compounds from elder berry and different grape marc varieties using organic solvents and/or supercritical carbon dioxide. J Food Eng 90:246–254CrossRefGoogle Scholar
  40. Vinson JA, Zhang J (2005) Black and green teas equally inhibit diabetic cataracts in a streptozotocin-induced rat model of diabetes. J Agric Food Chem 53(9):3710–3713CrossRefGoogle Scholar
  41. Vinson JA, Teufel K, Wu N (2004) Green and black teas inhibit atherosclerosis by lipid, antioxidant, and fibrinolytic mechanisms. J Agric Food Chem 52:3661–3665CrossRefGoogle Scholar
  42. Young W, Hotovec RL, Romero AG (1967) Tea and atherosclerosis. Nature 216:1015–1016Google Scholar
  43. Zhishen J, Mengchang T, Jianming W (1999) The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem 64:555–559CrossRefGoogle Scholar

Copyright information

© Association of Food Scientists & Technologists (India) 2013

Authors and Affiliations

  • J. Prakash Maran
    • 1
  • S. Manikandan
    • 2
  • B. Priya
    • 2
    • 3
    Email author
  • P. Gurumoorthi
    • 2
  1. 1.Department of Food TechnologyKongu Engineering CollegeErodeIndia
  2. 2.Department of Food Process EngineeringSRM UniversityChennaiIndia
  3. 3.Department of Food Process Engineering, School of BioengineeringSRM UniversityKancheepuramIndia

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