Skip to main content

Advertisement

Chemical and Biological Technologies in Agriculture
Chemical and Biological Technologies in Agriculture Cover Image
  • Research
  • Open Access

Evaluation of antioxidant activity and total phenol content of Gracilaria bursa-pastoris harvested in Nador lagoon for an enhanced economic valorization

  • †1,
  • †2Email author,
  • 3,
  • 4,
  • 1,
  • 1,
  • 5,
  • 1 and
  • 3
†Contributed equally
Chemical and Biological Technologies in Agriculture20174:28

https://doi.org/10.1186/s40538-017-0110-z

©  The Author(s) 2017

  • Received: 29 May 2017
  • Accepted: 3 October 2017
  • Published:

Abstract

Background

The Gracilaria bursa-pastoris is an abundant alga that is responsible for the eutrophication in Nador lagoon in Morocco. Therefore, the lagoon is a deposit of bioresources whose economic value can be enhanced. Antioxidative properties of seaweed extracts have been studied in several geographic regions, but only a few studies have been performed on tropical seaweed species, and no research has been done to determine antioxidant activity and valorization of the indigenous algae in Nador lagoon in Morocco.

Methods

The present study evaluated the antioxidant potential of methanolic and aqueous extracts from Gracilaria bursa-pastoris by measuring the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and total content of phenolic, flavonoids and flavonols content in each extract.

Results

This study showed that this macroalgae contained a high total phenolic content (142.26 and 79.43 mg GAE g−1 extract) for ethanolic and aqueous extracts, respectively. All extracts exhibited a DPPH radical scavenging activity, and the methanolic extracts demonstrated a great antioxidant potential with very low value of EC50 (0.085 mg/mL), significantly equivalent to EC50 (0,028 mg/mL) of commercial antioxidant ascorbic acid. These results suggest that the macroalgae have a great antioxidant potential which could be considered for future applications in medicine, food production or cosmetic industry in Morocco.

Conclusions

This work showed that the ethanolic extract was rich in phenolic compounds. This extract exhibited the highest antioxidant activity which may be explained by the presence of a large quantity of phenolic compounds. The Gracilaria bursa-pastoris can be utilized as a source of natural antioxidant compounds.
Graphical Abstract image

Keywords

  • Antioxidant activity
  • Macroalgae
  • Gracilaria bursa-pastoris
  • DPPH

Background

Since the year 2000, the seaweeds have attracted attention in the search for bioactive compounds to develop new drugs and healthy foods [1, 2]. In particular, these organic resources are a very important and commercially valuable resource for food, fodder, soil conditioners, pharmaceuticals, and production of green energy [3, 4]. Marine macroalgae are a valuable food resource which contains low calories, and they are rich in vitamins, minerals, proteins, polysaccharides, steroids, and dietary fibers [5–7]. In 2004, Ahn et al. has confirmed the presence of bioactive compounds such as vitamins, minerals, dietary fiber, and polysaccharides in some seaweeds.

On the other side, some authors have found other special bioactive compounds such as alkaloids, terpenes, bromophenols, phlorotannins, ascorbic acid, tocopherols, and carotenoids, in algae which have antioxidant potential [8–13]. These observations suggest that these bioactive compounds can have strong antioxidant properties.

The search for natural antioxidant compounds has gained considerable attention and the number of publications on antioxidants and oxidative stress has nearly quadrupled [14]. Because the antioxidant compounds play an important role against various diseases (Chronic inflammation, atherosclerosis, cancer, and cardiovascular disorders) and aging biochemical processes, this explains their considerable commercial potential in medicine, food production, and the cosmetic industry [15, 16]. Currently, the utilization of antioxidants from natural sources is accentuated because the consumers prefer natural products and they are afraid about the potential toxic effects of synthetic antioxidants [17]. Most countries have limited the use of synthetic antioxidant compounds in the food products because of their side effects [18]. Thus, it is essential to develop and utilize effective natural antioxidant molecules.

The Nador lagoon, also called Marchica lagoon, is the only lagoon ecosystem in the Moroccan coast facing the Mediterranean Sea . It covers an area of 115 km2 with water depths from 3 to 8 m. [19, 20]. The daily discharges of domestic, agricultural, and industrial waste into the lagoon stimulate the proliferation of algae called “harmful algal blooms” [21]. Gracilaria bursa-pastoris is an abundant alga that is responsible for the eutrophication in this lagoon [3]. The choice of this alga (Gracilaria bursa-pastoris) was based on its potential source for the extraction of new natural medicines, its important role for industrial and biotechnological applications due to its content of phycocolloids, the main source of agar α-(1,4)-3,6-anhydro-l-galactose and β-(1,3)-d-galactose with little esterification in cell wall, the content of bioactive metabolites like the primary compound with antibiotic activity acrylic acid [22]. Therefore, the lagoon is a deposit of bioresources whose economic value can be enhanced.

Antioxidative properties of seaweed extracts have been studied in several geographic regions, but only a few studies have been performed on tropical seaweed species and no research has been done to determine antioxidant activity and valorization of the indigenous algae in Nador lagoon in Morocco [23–26].

Faced with the challenge to valorize this natural wonder, it is necessary that total phenolic content, total antioxidant capacity (in soluble of lipid and water), total protein, total carbohydrate, and vitamin (A, C, and E) contents were determined in Gracilaria bursa-pastoris, in order to identify the new resources of natural antioxidant molecules. The present study evaluated the antioxidant potential of methanolic and aqueous extracts from Gracilaria bursa-pastoris from Nador lagoon in Mediterranean Moroccan costs by measuring the 2,2-diphenyl-1- picrylhydrazyl (DPPH) radical-scavenging activity and total content of phenolic, flavonoids, and flavonols content in each extract.

Methods

Drugs and chemicals

1,1-Diphenyl-2-picrylhydrazyl (DPPH•), potassium ferricyanide (K3Fe3 + (CN)6), trichloroacetic acid (TCA), ferric chloride (FeCl3), β-carotene, linoleic acid, Tween-40, Folin–Ciocalteu’s reagent, ammonia solution (NH3), sulfuric acid (H2S04), hydrochloric acid (HCl), picric acid, acetic anhydride, chloroform, Fehling’s solutions A and B, sodium carbonate, Aluminum chloride (AlCl3) and bovine blood, ethanol, methanol, acetone, and hexane were obtained from Merck (Darmstadt, Germany). Gallic acid, Quercetin, Tannic acid, Butylated hydroxyanisole (BHA), and Butylated hydroxytoluene (BHT) are used as positive standards. Authentic standards were purchased from Sigma-Aldrich, Fluka, and Merck. All other chemicals used were of analytical grade.

Collection and preparation of algal extracts

Red algae Gracilaria bursa-pastoris was collected from Nador lagoon in May 2015 (Fig. 1). The algal species was identified by Professor N. Elkhiati. In the laboratory, the algae was carefully cleaned, washed several times with tap water, distilled water, then dried by oven at 40 °C, ground, and kept in dark (Fig. 2). Macroalgae plant was lyophilized and milled into powder before extraction. Lyophilized samples (10 g) of Gracilaria powder were macerated with 50 mL of ethanol. The mixture was stirred under ambient temperature for 4 h or refluxed at 40 °C for 2 h. After filtration, the solvent was concentrated on a rotary evaporator under reduced pressure at 40 °C. The aqueous extracts were prepared by the same method using distilled water and extracts were concentrated by drying at 40 °C and stored at − 20 °C. For each extract, different concentrations (0.02, 0.04, 0.08, 0.16, and 0.32 mg/mL) were prepared from the algae powder and then tested for antioxidant activity.
Fig. 1
Fig. 1

Map of Nador lagoon indicating the collecting sites in Mediterranean Moroccan costs [3]

Fig. 2
Fig. 2

Gracilaria bursa-pastoris

Phytochemicals compounds

Determination of total phenolics contents

The total phenolic contents were determined by the Folin–Ciocalteu method by following three steps [27]. In the first step, we mixed 6 µL of ultrapure and 500 µL of Folin–Ciocalteu reagent with 100 µL of the dissolved extract. After 1 min, in the second step, we added 1.5 mL of Na2CO3 (20%) and the volume was adjusted to 10 mL with ultrapure water. In the third step, after 30 min of incubation at room temperature, we measured the absorbance at 760 nm. Gallic acid was used for the calibration curve. All the results were expressed as Gallic acid equivalents (mg GAE/g dry weight of the extracts) according to the following equation:
$$Y = 0.0053X - 0.0030 \, \left( {R^{2} = 0.995} \right).$$
(1)

Y: The absorbance. X: The concentration of gallic acid in mg/mL.

Determination of total flavonoids contents

The flavonoids contents were estimated by the AlCl3 method [28]. We mixed 1 mL of extract solution with 1 mL of 2% methanolic AlCl3*6H2O. After 30 min, we measured the absorbance at 430 nm. Quercetin (0–20 mg/l) prepared in methanol was used as a standard. The amount of flavonoids was expressed as quercetin equivalents (mg RE/g dry weight of the extract) according to the following equation [29]:
$$Y = 0.031 \, X + 0.100 \, \left( {R^{2} = \, 0.995} \right).$$
(2)

Y: the absorbance. X: the concentration of quercetin in mg/mL.

Free radical-scavenging activity

Determination of antioxidant activity

Aqueous and ethanolic extracts were subjected to screening for their possible antioxidant activities. The antioxidant activity was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay and ascorbic acid bleaching test. DPPH can be used to accurately titrate the oxidizable groups of biomolecules [30], and has commonly been used to estimate the antioxidant capacity of complex mixtures including plant oils, many of which contain high concentrations of monoterpenes [31].

Free radical-scavenging activity experimental

The free radical-scavenging activity of aqueous and ethanolic extracts was measured according to the method of Brand-Williams et al. 1995 using the stable 2, 2-diphenyl-1-picrylhydrazyl radical (DPPH). The model of scavenging DPPH radical has been widely used for estimating the free radical-scavenging activities of plant extracts/antioxidants [1]. DPPH radical is a stable-free radical, so the antioxidants are able to reduce DPPH radical to the non-radical form DPPH-H. The DPPH-scavenging activities of antioxidants are attributed to their hydrogen-donating abilities [7] (Fig. 3). The results were expressed in percentage of inhibition of DPPH.
Fig. 3
Fig. 3

DPPH-scavenging activities of antioxidants

0.6 mL of various concentrations of the each extract was used. They are mixed with 2.4 mL of a 0.004% methanol solution of DPPH (Sigma-Aldrich). At room temperature, after waiting for 30 min incubation period, the absorbance was read at 517 nm. Scavenging effect of DPPH radical was calculated using the following equation:
$$I\% \, = \, \left[ {\left( {A_{\text{blank}} {-}A_{\text{sample}} } \right)/A_{\text{blank}} } \right] \times 100.$$
(3)

A sample: the absorbance of test extract. A blank: the absorbance of the control reaction.

The IC50 value represents the concentration of the extract required to quench 50% of DPPH under experimental conditions.

Results and discussion

Phytochemicals compounds

The total phenolic and flavonoid content of the extracts, as estimated by the Folin–Ciocalteu reagent method, were 79.43 and 142.26 (mg GAE/g extract) at 40 °C and 27.47 and 36.61 (mg Q g−1 extract) at 20 °C for aqueous and ethanolic extracts, respectively. The total flavonol contents were 59.04 and 64.87 (mg Q g−1 extract) at 40 °C and 19.22 and 49.12 (mg Q g−1 extract) at 20 °C (Table 1). These results showed that the higher content of total polyphenols was found in the ethanolic extract which constituted the major antioxidant compounds. Also, we notice that total phenolic and flavonol are the major extracts at 40 °C more than at 20 °C. It demonstrated that heating promotes the extraction of antioxidant phenolic and flavonol compounds. In contrast, flavonoid content is weakly higher at 20 °C. The temperature affects the extraction processes by varying the alga matrix properties, solvent, pressure, and time [32]. In fact, temperature increases mass transfer during extraction by destroying the alga membrane structure, thus enhancing extraction and decreasing extraction time [33].
Table 1

The total phenolic, flavonoid, and flavonol contents in the G. bursa-pastoris extracts

Extracts

Heating (°C)

Total phenolic content

(mg GAE g−1extract)

Total flavonoid content

(mg Q g−1 extract)

Total flavonol content

(mg Q g−1 extract)

Ethanolic

20

99.62 ± 1.2

36.61 ± 0.3

49.12 ± 0.7

40

142.26 ± 0.8

32.67 ± 0.09

64.87 ± 0.05

Aqueous

20

42.2 ± 0.09

27.47 ± 0.04

19.22 ± 0.12

40

79.43 ± 0.58

15.86 ± 1.53

59.04 ± 0.09

Yildiz et al. [16] reported a low phenolic content in the Turkey G. bursa-pastoris methanolic extract (0.35 mg Gallic acid/g). This difference could be attributed to the origin of the marine algae. Much of literature refers to the plant phenolics compounds' direct antioxidant effect.

DPPH radical-scavenging and antioxidant activities of Gracilaria bursa-pastoris extracts

Results of free radical-scavenging activity of aqueous and ethanolic extracts are given in Table 2. Data in the table indicated that the radical-scavenging activities of the extracts increased with increasing concentration from 0.02 to 0.32 mg/mL. It is showed a very strong antioxidant activity of G. bursa-pastoris ethanolic extract.
Table 2

Antioxidant activity of aqueous and ethanolic extracts determined by DPPH˙ radical-scavenging assay

Concentration

mg/mL

Inhibition ratio (%)

Ethanolic extract

Aqueous extract

Ascorbic acid

20 °C

40 °C

20 °C

40 °C

0.32

74

81

61

69

90

0.16

58

67

40

47

81

0.08

42

48

30

38

73

0.04

31

37

26

27

65

0.02

25

30

18

20

42

IC50

0.096

0.085

0.176

0.167

0.028

This extract exhibited the highest activity with a very low EC50 (0.085 mg mL−1) significantly equivalent to EC50 of the commercial antioxidants tested, ascorbic acid (0.028 mg mL−1). The antioxidant activity of ethanolic extract was found nearly two times higher than that of aqueous extract. Phenolic compounds are widely distributed in seaweeds and are known to exhibit higher antioxidative activities and chemopreventive agents [34]. According to this result, it is clearly indicated that G. bursa-pastoris has a high antioxidant activity. Phenolic compounds in this marine alga are correlated with antioxidant activity and also dependent on the solvent and algal species used for extraction [35].

Basically, increase in total phenolic content may increase antioxidant properties. Phenolic compounds act as electron donors and may neutralize the unwanted reactions created by free radicals in the body. The positive correlation between polyphenolic content of algae and its antioxidant activity is well documented [36]. More recently, Jimenez-Escrig et al. [37] determined that the DPPH-free radical scavenging by brown and red algal extracts were positively correlated with the total polyphenol content of these extracts. Several studies have demonstrated a highly significant correlation between the phenolic content and the antioxidant activity in seaweed extracts, [38–41]. These results suggest that the antioxidant activity of G. bursa-pastoris extracts could be correlated with its high phenolic content.

Conclusion

This study showed that the ethanolic extract was highly rich in phenolic compounds. This extract exhibited a highest antioxidant activity which may be explained by the presence of a strong quantity of phenolic compounds. The results obtained in this study shows that G. bursa-pastoris can be utilized as a source of natural antioxidant compounds. Therefore, the G. bursa-pastoris seaweed, due its easily accessible source of natural antioxidant molecules, may become valuable for the development of therapeutic products, food supplement, and pharmaceutical applications.

Notes

Declarations

Authors’ contributions

MR, BH, and NE designed the experiments; MR, OE, and NS performed the experiments; OE, FT, MM, MR, and OZ wrote this manuscript. All authors read and approved the final manuscript.

Acknowledgements

The authors warmly thank Abdelhakim Mesfioui, Director of Nador INRH, and Ahmed Chihani a diver in Nador lagoon for all the necessary materials needed for collecting G. bursa-pastoris macroalgae.

Competing interests

The authors declare that they have no competing interests.

Availability of data and materials

All data generated and analyzed during this study are included in this manuscript.

Consent for publication

All authors have approved to submit this work to Chemical and Biological Technologies in Agriculture. They declare that there is no conflict of interest in relation to the submission of the article.

Ethics approval and consent to participate

All authors approved to the ethics approval and all authors actively participated in the creation of this excellent work.

Funding

Not applicable.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Authors’ Affiliations

(1)
Ecology, Water and Environment Laboratory, Mohamed First University, Oujda, Morocco
(2)
Biochemistry and Biotechnology Laboratory, Mohamed First University, Oujda, Morocco
(3)
Applied Analytical Chemistry, Materials and Environment Laboratory, Mohamed First University, Oujda, Morocco
(4)
Department of Biology, Ain Chock Faculty, Casablanca, Morocco
(5)
Applied Biology and Pathology Laboratory, Abdelmalek Essaâdi University, Tetouan, Morocco

References

  1. Qi H, Zhao T, Zhang Q, Li Z, Zhao Z, Xing R. Antioxidant activity of different molecular weight sulfated polysaccharides from Ulva pertusa Kjellm (Chlorophyta). J Appl Phycol. 2005;17:527–34.View ArticleGoogle Scholar
  2. Van Oosten MJ, Pepe O, De Pascale S, Silletti S, Maggio A. The role of biostimulants and bioeffectors as alleviators of abiotic stress in crop plants. Chem Biol Technol Agric. 2017;4:5. doi:10.1186/s40538-017-0089-5.View ArticleGoogle Scholar
  3. El Asri O, Ramdani M, Latrach L, Haloui B, Mohamed R, elamin Afilal M. Energetic valorization of Nador lagoon algae and proposal to use it as a means of elimination of the eutrophication in this lagoon. Ecol Eng. 2017;103:236–43. doi:10.1016/j.ecoleng.2017.04.016.View ArticleGoogle Scholar
  4. Yang Y-F, Fei X-G, Song J-M, Hu H-Y, Wang G-C, Chung IK. Growth of Gracilaria lemaneiformis under different cultivation conditions and its effects on nutrient removal in Chinese coastal waters. Aquaculture. 2006;254:248–55.View ArticleGoogle Scholar
  5. Darcy-Vrillon B. Nutritional aspects of the developing use of marine macroalgae for the human food industry. Int J Food Sci Nutr (UK). 1993.Google Scholar
  6. Ito K, Hori K. Seaweed: chemical composition and potential food uses. Food Rev. Int. 1989;5:101–44.View ArticleGoogle Scholar
  7. Lahaye M. Marine algae as sources of fibres: determination of soluble and insoluble dietary fibre contents in some “sea vegetables”. J Sci Food Agric. 1991;54:587–94.View ArticleGoogle Scholar
  8. Liu M, Hansen PE, Lin X. Bromophenols in marine algae and their bioactivities. Mar Drugs. 2011;9:1273–92.View ArticlePubMedPubMed CentralGoogle Scholar
  9. Peng J, Yuan J-P, Wu C-F, Wang J-H. Fucoxanthin, a marine carotenoid present in brown seaweeds and diatoms: metabolism and bioactivities relevant to human health. Mar Drugs. 2011;9:1806–28.View ArticlePubMedPubMed CentralGoogle Scholar
  10. Sharma HSS, Selby C, Carmichael E, McRoberts C, Rao JR, Ambrosino P, Chiurazzi M, Pucci M, Martin T. Physicochemical analyses of plant biostimulant formulations and characterisation of commercial products by instrumental techniques. Chem Biol Technol Agric. 2016;3:13. doi:10.1186/s40538-016-0064-6.View ArticleGoogle Scholar
  11. Takamatsu S, Hodges TW, Rajbhandari I, Gerwick WH, Hamann MT, Nagle DG. Marine natural products as novel antioxidant prototypes. J Nat Prod. 2003;66:605–8.View ArticlePubMedPubMed CentralGoogle Scholar
  12. Thomas NV, Kim S-K. Potential pharmacological applications of polyphenolic derivatives from marine brown algae. Environ Toxicol Pharmacol. 2011;32:325–35.View ArticlePubMedGoogle Scholar
  13. Zubia M, Fabre MS, Kerjean V, Le Lann K, Stiger-Pouvreau V, Fauchon M, Deslandes E. Antioxidant and antitumoural activities of some Phaeophyta from Brittany coasts. Food Chem. 2009;116:693–701.View ArticleGoogle Scholar
  14. Huang D, Ou B, Prior RL. The chemistry behind antioxidant capacity assays. J Agric Food Chem. 2005;53:1841–56.View ArticlePubMedGoogle Scholar
  15. Cohen K, Nyska A. Oxidation of biological system: oxidative stress phenomena, antioxidants, redox reactions and method for their quantification. Toxicol Path. 2002;30:620–50.View ArticleGoogle Scholar
  16. Yildiz G, Vatan Ö, Celikler S, Dere Ş. Determination of the phenolic compounds and antioxidative capacity in red algae Gracilaria bursa-pastoris. Int J Food Prop. 2011;14:496–502.View ArticleGoogle Scholar
  17. Safer AM, Al-Nughamish AJ. Hepatotoxicity induced by the anti-oxidant food additive, butylated hydroxytoluene (BHT), in rats: an electron microscopical study. Histol Histopathol. 1999;14(2):391–406.PubMedGoogle Scholar
  18. Kabouche A, Kabouche Z, Öztürk M, Kolak U, Topçu G. Antioxidant abietane diterpenoids from Salvia barrelieri. Food Chem. 2007;102:1281–7.View ArticleGoogle Scholar
  19. El-Alami M, Mahjoubi R, Damnati B, Kamel S, Icole M, Taieb M. Sédimentologie et géochimie organique des sédiments superficiels de la lagune de Nador (Maroc nord-oriental). J Afr Earth Sci. 1998;26:249–59.View ArticleGoogle Scholar
  20. Erimesco P. La Mar Chica de Melilla. Bull Int Pêche Marit Maroc. 1961;7:3–11.Google Scholar
  21. Lapointe BE, Herren LW, Debortoli DD, Vogel MA. Evidence of sewage-driven eutrophication and harmful algal blooms in Florida’s Indian River Lagoon. Harmful Algae. 2015;43:82–102.View ArticleGoogle Scholar
  22. De Almeida CL, Falcão DS, Lima DM, Gedson R, Montenegro DA, Lira NS, De Athayde-Filho PF, Rodrigues LC, De Souza MD, Barbosa-Filho JM, Batista LM. Bioactivities from marine algae of the genus Gracilaria. Int J Mol Sci. 2011;12(7):4550–73.View ArticlePubMedPubMed CentralGoogle Scholar
  23. Anggadiredja J, Andyani R. Antioxidant activity of Sargassum polycystum (Phaeophyta) and Laurencia obtusa (Rhodophyta) from Seribu islands. J Appl Phycol. 1997;9:477–9.View ArticleGoogle Scholar
  24. Fallarero A, Loikkanen JJ, Männistö PT, Castañeda O, Vidal A. Effects of aqueous extracts of Halimeda incrassata (Ellis) Lamouroux and Bryothamnion triquetrum (SG Gmelim) Howe on hydrogen peroxide and methyl mercury-induced oxidative stress in GT1-7 mouse hypothalamic immortalized cells. Phytomedicine. 2003;10:39–47.View ArticlePubMedGoogle Scholar
  25. Lim SN, Cheung PCK, Ooi VEC, Ang PO. Evaluation of antioxidative activity of extracts from a brown seaweed, Sargassum siliquastrum. J Agric Food Chem. 2002;50:3862–6.View ArticlePubMedGoogle Scholar
  26. Santoso J, YOSHIE-STARK Y, Suzuki T. Anti-oxidant activity of methanol extracts from Indonesian seaweeds in an oil emulsion model. Fish Sci. 2004;70(1):183–8.View ArticleGoogle Scholar
  27. Singleton VL, Orthofer R, Lamuela-Raventós RM. [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods Enzymol. 1999;299:152–78.View ArticleGoogle Scholar
  28. Lamaison JL, Carnart A. Teneurs en principaux falvonoïdes des fleurs et des feuilles de Crataegus monogyna Jacq. et de Crataegus laevigata (Poiret) DC. en fonction de la période de végétation. Plantes Médicinales Phythér. 1991;25:12–6.Google Scholar
  29. Alimi H, Hfaiedh N, Bouoni Z, Sakly M, Ben Rhouma K. Evaluation of antioxidant and antiulcerogenic activities of Opuntia ficus indica F. inermis flowers extract in rats. Environ Toxicol Pharmacol. 2011;32:406–16. doi:10.1016/j.etap.2011.08.007.View ArticlePubMedGoogle Scholar
  30. Blois MS. Antioxidant determinations by the use of a stable free radical. Nature. 1958;181:1199–200. doi:10.1038/1811199a0.View ArticleGoogle Scholar
  31. Behrendorff JB, Vickers CE, Chrysanthopoulos P, Nielsen LK. 2,2-Diphenyl-1-picrylhydrazyl as a screening tool for recombinant monoterpene biosynthesis. Microb Cell Factories. 2013;12:76. doi:10.1186/1475-2859-12-76.View ArticleGoogle Scholar
  32. Hernandez Y, Lobo M, Gonzalez M. Factors affecting sample extraction in the liquid chromatographic determination of organic acids in papaya and pineapple. Food Chem. 2009;114:734–41. doi:10.1016/j.foodchem.2008.10.021.View ArticleGoogle Scholar
  33. Azmir J, Zaidul ISM, Rahman MM, Sharif KM, Mohamed A, Sahena F, Jahurul MHA, Ghafoor K, Norulaini NAN, Omar AKM. Techniques for extraction of bioactive compounds from plant materials: a review. J Food Eng. 2013;117:426–36. doi:10.1016/j.jfoodeng.2013.01.014.View ArticleGoogle Scholar
  34. Bravo L. Polyphenols: chemistry, dietary sources, metabolism, and nutritional significance. Nutr Rev. 1998;56:317–33.View ArticlePubMedGoogle Scholar
  35. Ramdani M, Sahnouni F, Haloui B, Roger NE, Mesfioui FA, Ramdani M. Macroepiphytes and macroalgae (Phaeophyceae, rhodophyceae and chlorophyceae) in the nador lagoon (Morocco). J Biodivers Env Sci. 2015;7:252–61.Google Scholar
  36. Horincar V-B, Parfene P, Bahrim G. Evaluation of bioactive compounds in extracts obtained from three Romanian marine algae species. Romanian Biotechnol Lett. 2011;16:71–8.Google Scholar
  37. Jiménez-Escrig A, Jiménez-Jiménez I, Pulido R, Saura-Calixto F. Antioxidant activity of fresh and processed edible seaweeds. J Sci Food Agric. 2001;81:530–4.View ArticleGoogle Scholar
  38. Karawita R, Siriwardhana N, Lee K-W, Heo M-S, Yeo I-K, Lee Y-D, Jeon Y-J. Reactive oxygen species scavenging, metal chelation, reducing power and lipid peroxidation inhibition properties of different solvent fractions from Hizikia fusiformis. Eur Food Res Technol. 2005;220:363–71.View ArticleGoogle Scholar
  39. Manian R, Anusuya N, Siddhuraju P, Manian S. The antioxidant activity and free radical scavenging potential of two different solvent extracts of Camellia sinensis (L.) O. Kuntz, Ficus bengalensis L. and Ficus racemosa L. Food Chem. 2008;107:1000–7.View ArticleGoogle Scholar
  40. Siriwardhana N, Lee K-W, Jeon Y-J, Kim S-H, Haw J-W. Antioxidant activity of Hizikia fusiformis on reactive oxygen species scavenging and lipid peroxidation inhibition. Food Sci Technol Int. 2003;9:339–46.View ArticleGoogle Scholar
  41. Velioglu YS, Mazza G, Gao L, Oomah BD. Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. J Agric Food Chem. 1998;46:4113–7.View ArticleGoogle Scholar

Copyright

© The Author(s) 2017

Advertisement