European Food Research and Technology

, Volume 244, Issue 3, pp 523–534 | Cite as

Investigating the in vitro hypoglycaemic and antioxidant properties of Citrus × clementina Hort. juice

  • Monica R. LoizzoEmail author
  • Mariarosaria Leporini
  • Vincenzo Sicari
  • Tiziana Falco
  • Teresa M. Pellicanò
  • Rosa Tundis
Original Paper


Citrus × clementina juice obtained from fruits collected in three different areas (flood plain, hill and coastal plain) was investigated for the chemical composition, radical scavenging properties (DPPH and ABTS tests), and α-amylase and α-glucosidase inhibitory activity. Neohesperidin (72.96–116.50 mg/100 mL), hesperidin (55.24–69.52 mg/100 mL) and narirutin (7.21–12.13 mg/100 mL) are the main flavonoids identified by HPLC analyses. In carbohydrate hydrolysing enzymes inhibitory activity tests, samples showed higher potency against α-glucosidase. Juice from hill was the most active with an IC50 value of 77.79 μg/mL. Data on the radical scavenging activity revealed the following trend of potency flood plain > coastal plain > hill. These results could help farmers to select fruits for different industrial purpose such as functional food and matrix to extract nutraceutical products.


HPLC phenolic profile Quality parameters Healthy properties PCA 



2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)


American Diabetes Association




C-Jun NH2-terminal kinase


Principal component analysis


Reactive oxygen species


Type 1 diabetes


Type 2 diabetes


Tumour-necrosis factor-α



The authors are grateful to Falco farms for supplying the samples.

Compliance with ethical standards


The study was not specifically funded.

Conflict of interest

The authors declare that there are no conflicts of interest.

Compliance with ethics requirements

Research does not involve any human participants and/or animal.


  1. 1.
    Berner L, O’Donnell J (1998) Functional foods and health claims legislation: applications to dairy foods. Int Dairy J 8:355–362Google Scholar
  2. 2.
    Milind L (2008) Nutritive and medicinal value of Citrus fruit. In: Ladaniya M (ed) Citrus fruit: biology, technology and evaluation. Academic Press, Salt Lake City, pp 501–515 (2038 S 1500E Salt Lake City UT 84105 USA El ). ISBN 8601407120555Google Scholar
  3. 3.
    Patil BS, Jayaprakasha GK, Murthy KNC, Vikram A (2009) Bioactive compounds: historical perspectives, opportunities, and challenges. J Agric Food Chem 57:8142–8160PubMedGoogle Scholar
  4. 4.
    Cutuli G, Di Martino E, Lo Giudice V, Pennisi L, Raciti G, Russo F, Scuderi A, Spina P (1985) Trattato di agrumicoltura. Edagricole, BolognaGoogle Scholar
  5. 5.
    Alfadda AA, Sallam RM (2012) Reactive Oxygen Species in Health and Disease. J Biomed Biotechnol Article ID 936486Google Scholar
  6. 6.
    Houstis N, Rosen ED, Lander ES (2006) Reactive oxygen species have a causal role in multiple forms of insulin resistance. Nature 440:944–948PubMedGoogle Scholar
  7. 7.
    Sivitz WI, Yorek MA (2010) Mitochondrial dysfunction in diabetes: from molecular mechanisms to functional significance and therapeutic opportunities. Antioxid Redox Signal 12:537–577PubMedPubMedCentralGoogle Scholar
  8. 8.
    Tiwari AK, Rao JM (2002) Diabetes mellitus and multiple therapeutic approaches of phytochemicals: present status and future prospects. Curr Sci 83:30–38Google Scholar
  9. 9.
    Tundis R, Loizzo MR, Menichini F (2010) Natural products as α-amylase and α-glucosidase inhibitors and their hypoglycaemic potential in the treatment of diabetes: an update. Mini Rev Med Chem 10:315–331PubMedGoogle Scholar
  10. 10.
    Official methods of analysis, AOAC, Association of Official Analytical Chemists. Edition 18: AOAC (2005)Google Scholar
  11. 11.
    IFU (3-17-58) International Federation of Fruit Juice Producers. Methods of analysis and microbiological methods.
  12. 12.
    Loizzo MR, Bonesi M, Pugliese A, Menichini F, Tundis R (2014) Chemical composition and bioactivity of dried fruits and honey of Ficus carica cultivars Dottato, San Francesco and Citrullara. J Sci Food Agric 94:2179–2186PubMedGoogle Scholar
  13. 13.
    Rahal A, Kumar A, Singh A (2014) Oxidative stress, prooxidants, and antioxidants: the interplay. BioMed Res Int 761264:1–19Google Scholar
  14. 14.
    Loizzo MR, Bonesi M, Menichini F, Tenuta MC, Leporini M, Tundis R (2016) Antioxidant and carbohydrate-hydrolysing enzymes potential of Sechium edule (Jacq.) Swartz (Cucurbitaceae) peel, leaves and pulp fresh and processed. Plant Foods Hum Nutr 71:381–387PubMedGoogle Scholar
  15. 15.
    Sun T, Tanumihardjo SA (2007) An integrated approach to evaluate food antioxidant capacity. J Food Sci 72:159–165Google Scholar
  16. 16.
    MIRAAF (1994) Metodi Ufficiali di analisi chimica del suolo. Roma, ItalyGoogle Scholar
  17. 17.
    SISS (1985) Metodi normalizzati analisi del suolo. Edagricole, BolognaGoogle Scholar
  18. 18.
    Embleton TM, Jones WW, Labanauskas CH, Reuther W (1973) Leaf analyses as a diagnostic tool and guide to fertilization. In: Reuther W (ed) The Citrus industry. University of California, Berkeley, pp 183–210Google Scholar
  19. 19.
    White JG, Zasoski RJ (1999) Mapping soil micronutrients. Field Crops Res 60:11–26Google Scholar
  20. 20.
    Bermejo A, Pardo J, Morales J, Cano A (2016) Comparative study of bioactive components and quality from juices of different mandarins: discriminant multivariate analysis of their primary and secondary metabolites. Agricu Sci 7:341–351Google Scholar
  21. 21.
    Al-Mouei R, Choumane W (2014) Physiochemical juice characteristics of various Citrus species in Syria. Int J Plant Soil Sci 3:1083–1095Google Scholar
  22. 22.
    Boudries H, Madani K, Touati N, Souagui S, Medouni S, Chibane M (2012) Pulp antioxidant activities, mineral contents and juice nutritional properties of Algerian Clementine Cultivars and Mandarin. Afr J Biotechnol 11:4258–4267Google Scholar
  23. 23.
    Sdiri S, Bermejo A, Aleza P, Navarro P, Salvador A (2012) Phenolic composition, organic acids, sugars, vitamin C and antioxidant activity in the juice of two new triploid late-season mandarins. Food Res Int 49:462–468Google Scholar
  24. 24.
    Xu G, Liu D, Chen J, Ye X, Ma Y, Shi J (2008) Juice components and antioxidant capacity of citrus varieties cultivated in China. Food Chem 106:545–551Google Scholar
  25. 25.
    Milella L, Caruso M, Galgano F, Favati F, Padula MC, Martelli G (2011) Role of the cultivar in choosing Clementine fruits with a high level of health-promoting compounds. J Agric Food Chem 59:5293–5298PubMedGoogle Scholar
  26. 26.
    Dhuique-Mayer C, Caris-Veyrat C, Ollitrault P, Curk F, Amiot MJ (2005) Varietal and interspecific influence on micronutrient contents in citrus from the Mediterranean area. J Agric Food Chem 53:2140–2145PubMedGoogle Scholar
  27. 27.
    Rapisarda P, Bellomo SE, Fabroni S, Russo G (2008) Juice quality of two new mandarin-like hybrids (Citrus clementina Hort. Ex tan × Citrus sinensis L. Osbeck) containing anthocyanins. J Agric Food Chem 56:2074–2078PubMedGoogle Scholar
  28. 28.
    Rapisarda P, Pannuzzo P, Romano G, Russo G (2003) Juice components of a new pigmented citrus hybrid Citrus sinensis (L.) Osbeck × Citrus clementina Hort. ex Tan. J Agric Food Chem 51:1611–1616PubMedGoogle Scholar
  29. 29.
    Atmani D, Chaher N, Atmani D, Berboucha M, Debbache N, Boudaoud H (2009) Flavonoids in human health: from structure to biological activity. Curr Nutr Food Sci 5:225–237Google Scholar
  30. 30.
    Gattuso G, Barreca D, Gargiulli C, Leuzzi U, Caristi C (2007) Flavonoid composition of Citrus juices. Molecules 12:1641–1673PubMedGoogle Scholar
  31. 31.
    Kanaze FI, Gabrielli C, Kokkalou E, Georgarakis M, Niopas I (2003) Simultaneous reversed-phase high-performance liquid chromatographic method for the determination of diosmin, naringin and hesperidin in different Citrus fruit juices and pharmaceutical formulation. J Pharm Biomed Anal 33:243–249PubMedGoogle Scholar
  32. 32.
    Nogata Y, Sakamoto K, Shiratsuchi H, Ishii T, Yano M, Ohta H (2006) Flavonoid composition of fruit tissues of citrus species. Biosci Biotechnol Biochem 70:178–192PubMedGoogle Scholar
  33. 33.
    Aruoma OI, Landes B, Ramful-Baboolall D, Bourdon E, Neergheen-Bhujun V, Wagner KH, Bahorun T (2012) Functional benefits of citrus fruits in the management of diabetes. Prev Med 54:S12–S16PubMedGoogle Scholar
  34. 34.
    Abirami A, Nagarani G, Siddhuraju P (2014) In vitro antioxidant, anti-diabetic, cholinesterase and tyrosinase inhibitory potential of fresh juice from Citrus hystrix and C. maxima fruits. Food Sci Hum Well 3:16–25Google Scholar
  35. 35.
    Owira PM, Ojewole JA (2009) Grapefruit juice improves glycaemic control but exacerbates metformin-induced lactic acidosis in non-diabetic rats. Methods Find Exp Clin Pharmacol 31:563–570PubMedGoogle Scholar
  36. 36.
    Mollace V, Sacco I, Janda E, Malara C, Ventrice D, Colica C, Visalli V, Muscoli S, Ragusa S, Muscoli C, Rotiroti D, Romeo F (2011) Hypolipemic and hypoglycaemic activity of bergamot polyphenols: from animal models to human studies. Fitoterapia 82:309–316PubMedGoogle Scholar
  37. 37.
    Tadera K, Minami Y, Takamatsu K, Matsuoka T (2010) Inhibition of alpha-glucosidase and alpha-amylase by flavonoids. J Nutr Sci Vitaminol 52:149–153Google Scholar
  38. 38.
    Tundis R, Bonesi M, Sicari V, Pellicanò TM, Tenuta MC, Leporini M, Menichini F, Loizzo MR (2016) Poncirus trifoliata (L.) Raf.: chemical composition, antioxidant properties and hypoglycaemic activity via the inhibition of α-amylase and α-glucosidase enzymes. J Funct Foods 25:477–485Google Scholar
  39. 39.
    Kim Y, Keogh JB, Clifton PM (2016) Polyphenols and glycemic control. Nutrients 8:17. doi: 10.3390/nu8010017 PubMedCentralGoogle Scholar
  40. 40.
    Shen W, Xu Y, Lu YH (2012) Inhibitory effects of citrus flavonoids on starch digestion and antihyperglycemic effects in HepG2 cells. J Agric Food Chem 60:9609–9619PubMedGoogle Scholar
  41. 41.
    Jia S, Hu Y, Zhang W, Zhao X, Chen Y, Sun C, Li X, Chen K (2015) Hypoglycemic and hypolipidemic effects of neohesperidin derived from Citrus aurantium L. in diabetic KKA(y) mice. Food Funct 6:878–886PubMedGoogle Scholar
  42. 42.
    Seshagirirao P, Giri KV (1942) The mechanism of β-amylase inhibition by vitamin C. Proc Ind Acad Sci Sec B 16:190–204Google Scholar
  43. 43.
    Ullah A, Khan A, Khan I (2016) Diabetes mellitus and oxidative stress—a concise review. Saudi Pharm J 24:547–553Google Scholar
  44. 44.
    Russo D, Bonomo MG, Salzano G, Martelli GBG, Milella L (2012) Nutraceutical properties of Citrus clementina juices. Pharmacologyonline 1:84–93Google Scholar
  45. 45.
    Codoñer-Franch P, López-Jaén AB, Muñiz P, Sentandreu E, Bellés VV (2008) Mandarin juice improves the antioxidant status of hypercholesterolemic children. J Pediatr Gastroenterol Nutr 47:349–355PubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Monica R. Loizzo
    • 1
    Email author
  • Mariarosaria Leporini
    • 1
  • Vincenzo Sicari
    • 1
  • Tiziana Falco
    • 1
  • Teresa M. Pellicanò
    • 2
  • Rosa Tundis
    • 1
  1. 1.Department of Pharmacy, Health Science and NutritionUniversity of CalabriaRendeItaly
  2. 2.Department of AgrariaUniversity “Mediterranea” of Reggio CalabriaReggio CalabriaItaly

Personalised recommendations