Altitude Effect on the Properties of Honeys from the Region of Jijel (Algeria)
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Pol. J. Food Nutr. Sci., 2020, Vol. 70, No. 2, pp. 169–178 DOI: 10.31883/pjfns/118528 http://journal.pan.olsztyn.pl Original article Section: Food Quality and Functionality Altitude Effect on the Properties of Honeys from the Region of Jijel (Algeria) Aissam Bouhala1*, Salim Ouchemoukh2, Abdelhamid Moussi3, Said Beldjoudi1 1Faculty of Nature and Life Sciences, University of Mohammed Seddik Benyahia, Jijel, BP 98, Ouled Aissa, Jijel 18000, Algeria 2Faculty of Nature and Life Sciences, Applied Biochemistry Laboratory, A. Mira University, Bejaia 06000, Algeria 3Laboratory of Genetic, Biotechnology and Valorization of Bio-Resources, University of Biskra, 07000 Biskra, Algeria Key words: Jijel, coastal and mountain honeys, physicochemical properties, antioxidants, antioxidant activity, antibacterial activity Coastal and mountainous honey samples were collected from different regions in Jijel (Algeria) to evaluate their moisture content, electrical con- ductivity, ash content, pH, acidity, protein content, color parameters, antioxidants content, radical scavenging activity, reducing power, and antibacte- rial activity, to reveal the differences between coastal and mountain honeys and to determine the correlation between altitude and different parameters. The results indicate that Jijelian honeys were dark with acceptable physicochemical properties and a good bioactive potential. Escherichia coli was sensitive to Jijelian honeys while Staphylococcus aureus and Pseudomonas aeruginosa were more resistant. Coastal honeys had statistically signifi cantly higher pH, electrical conductivity, ash content, color intensity, hydroxymethylfurfural (HMF) content, and reducing power than the mountainous samples (p<0.05), while the total acidity was higher in the mountain honeys (p<0.05). The altitude was signifi cantly negatively correlated with HMF content, electrical conductivity, ash content, and pH. The correlation coeffi cients were -0.510, -0.405, -0.360, and -0.355, respectively. INTRODUCTION et al., 2007; Liu et al., 2013]. The concentration of hydro- gen peroxide, which is determined according to the level Honey is a natural product that honeybees produce from of glucose oxidase (from bees) and catalase (pollen source), some plant parts or excretions of some insects that feed on in honey mainly predicts its antimicrobial potential, how- plant sap [Karabagias et al., 2014]. More than two hundred ever, lysozyme, phenolic acids, and fl avonoids are the major components have been found in honey; it is an important non-peroxide contributing factors [Tenore et al., 2012]. On source of energy due to its high sugar content, mainly fruc- the other hand, the correlation of the color with bioactive tose (38%) and glucose (31%) [Alvarez-Suarez et al., 2010; compounds and antioxidant and antibacterial activities has Bueno-Costa et al., 2016]. Moreover, it has small amounts been revealed in other studies [Bueno-Costa et al., 2016]. of amino acids, proteins, phenolic compounds, carotenoids, In recent years, many authors have studied the physico- organic acids, ascorbic acid, enzymes, α-tocopherol, and oli- chemical and bioactive properties of honeys from different gosaccharides [Alvarez-Suarez et al., 2010]. The composi- regions in the world including Algeria [Bueno-Costa et al., tion and characteristics of honey are primarily determined 2016; Mouhoubi-Tafi nine et al., 2016; Ouchemoukh et al., by the food source (plants); however, environmental factors, 2007; Tahir et al., 2017; Tenore et al., 2012], using different processing, and storage affect its composition as well [Saxena analytical methods. et al., 2010]. Many scientists have studied the characteristics and Phenolic compounds content and antioxidant activity the properties of mono-fl oral honeys produced from different have been widely used as indicators to evaluate the charac- plants by honeybees [Alvarez-Suarez et al., 2010; Karabagias teristics and bioactive properties of honey [Tahir et al., 2017]. et al., 2014; Küçük et al., 2007; Tenore et al., 2012]. How- Honey contains a variety of phenolics, and is rich in antioxi- ever, the aim of this study was to evaluate the characteristics dants, which increases its usability potential for therapeutic (physicochemical properties, protein content, color param- purposes [Küçük et al., 2007]. In addition, several other au- eters, contents of total phenolics and total fl avonoids, DPPH thors have mentioned the antimicrobial potential of honey radical scavenging activity, reducing power, and antibacterial [Alvarez-Suarez et al, 2010; Bueno-Costa et al., 2016; Küçük activity) of honeys from the greenest region of Algeria (Jijel) from different altitudes, to determine the differences between * Corresponding Author: Phone: +213774876893; Fax: +21334454489; coastal and mountain honeys and to reveal the correlation be- E-mail: [email protected] (A. Bouhala) tween the altitude and different parameters. © Copyright by Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences © 2020 Author(s). This is an open access article licensed under the Creative Commons Attribution-NonCommercial-NoDerivs License (http://creativecommons.org/licenses/by-nc-nd/3.0/). 170 Effect of Altitude on Honey Properties MATERIALS AND METHODS DPPH radical scavenging activity Assay of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical Samples scavenging activity (RSA) was performed according to Meda T wenty-two honey samples were collected from different et al. [2005] procedure. The 0.75 mL of each honey solu- regions in Jijel (Algeria). Half of these samples were collected tion in methanol (2.5–160 mg/mL) was mixed with 1.5 mL from regions close to the Mediterranean Sea and the other of DPPH in methanol (0.02 mg/mL). The mixture was left half from mountain regions. All these samples were produced in the dark for 15 min and then its absorbance was measured from hives placed in areas with diverse vegetation in order at 517 nm. The DPPH radical solution without the sample to get poly-fl oral honeys that are more representable of this served as the blank sample. The results were calculated based region. In addition, honeys from hives placed in vast mono- on the following formula: %Inhibition = [(blank absorbance culture fi elds were avoided. All samples were stored at 4–5oC – sample absorbance)/blank absorbance] ×100. The half in airtight glass containers until analyses. maximal inhibitory concentration (IC50) value of each honey sample was estimated from the plot of % inhibition vs. honey Physicochemical analyses concentration. The physicochemical analyses were determined according to the International Honey Commission [2009]. Moisture and ash Reducing power contents were expressed in g/100g. Acidity, electrical conductiv- The following method of reducing power (RP) deter- ity, and hydroxymethylfurfural (HMF) content were expressed mination was used [Küçük et al., 2017]: 1 mL of a honey in milliequivalents of sodium hydroxide required to neutralize solution (5.0%) was added to 2.5 mL of a phosphate buf- 1 kg of honey (meq/kg), mS/cm, and mg/kg, respectively. fer (0.2 M, pH 6.6) and 2.5 mL of 1% potassium ferric o cyanide (K3Fe (CN)6). The mixture was incubated at 50 C Protein content for 20 min. Afterwards, 2.5 mL of 10% trichloroacetic acid The protein content was analyzed according to the Brad- was added, and the mixture was centrifuged at 3000×g for ford method reported by Azeredo et al. [2003]. The absor- 10 min. The supernatant (2.5 mL) was mixed with 2.5 mL bance was measured at 595 nm (UV-1800 UV-Vis Spectropho- of distilled water and 0.5 mL of 0.1% FeCl3. The absorbance tometer from Shimadzu, Kyoto, Japan), against a standard was measured at 700 nm. Ascorbic acid (1.0 mg/mL) was solution of bovine serum albumin (0.1–1.4 mg/mL). used as a reference standard. Color analysis Antibacterial activity Color analysis was reviewed according to Ferreira et al. Agar disc diffusion assay of 100% honey concentration [2009]. Honeys in distilled water solutions of 50% (w/v) were was used against three strains of bacteria, which were Staphy- centrifuged at 3000×g for 10 min (centrifuge Model 3–16P, lococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Sigma Laborzentrifugen GmbH, Osterode, Germany). and Pseudomonas aeruginosa ATCC 27853 (Pasteur Institute The color was measured spectrophotometrically at 635 nm. of Algeria, Algeria), according to Alderman & Smith [2001]. The Pfund scale was used to classify the honeys as follows: The results were expressed in zone of growth inhibition (mm). mm Pfund = -38.70 + 371.39 ×Abs. Statistical analysis Total phenolics content All tests were performed in triplicate and the results were The following method described by Bueno-Costa et al. expressed as mean ± standard deviation. The parameters [2016] was used to determine the total phenolics content of the descriptive statistics were calculated using the Micro- (TPC): a honey solution of (0.1 g/mL) was centrifuged at soft Excel 2007 program. A one-way analysis of variance 3000×g for 10 min. Then, 0.5 mL of supernatant and 2.5 mL (ANOVA) was carried out with the STATISTICA 7.1 software of 0.2 N Folin–Ciocalteu reagent were mixed for 5 min. Af- to highlight the presence or absence of a signifi cant difference terwards, 2 mL of a sodium carbonate solution (75 g/L) between the samples of honey, which was considered statis- was added and the mixture was incubated for 2 h in dark. tically signifi cant at the level of 0.05. LSD test was used as The absorbance was measured using a spectrophotometer at post-hoc ANOVA test (p<0.05). The relationships between 765 nm. The TPC was expressed as mg gallic acid equivalent the parameters were determined by the correlation matrix per 100 g of sample (mg GAE/100 g). (p<0.01), while the comparison between means obtained for the coastal and mountain honeys was determined by Stu- Total fl avonoids content dent’s t-test using XLSTAT 2014. The total fl avonoids content (TFC) was determined accord- ing to the method described by Chaikham et al.