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- Publisher
- Hindawi Publishing Corporation
- ISSN
- 2090-8865
- eISSN
- 2090-8873
- DOI
- 10.1155/2022/7890856
- Publisher site
- See Article on Publisher Site
Abstract
Hindawi Journal of Analytical Methods in Chemistry Volume 2022, Article ID 7890856, 16 pages https://doi.org/10.1155/2022/7890856 Research Article Trace Elements and Heavy Metal Contents in West Algerian Natural Honey 1 2,3 1 4 Dalila Bereksi-Reguig, Salim Bouchentouf , Hocine Allali, Agnieszka Adamczuk , 5 6 Graz˙yna Kowalska , and Radosław Kowalski Department of Chemistry, Faculty of Sciences, Abou Bekr Belka¨ıd University, P.O. Box 119, Tlemcen 13000, Algeria Doctor Tahar Moulay University of Saida Algeria, BP 138 Cit´e EN-NASR, Sa¨ıda 20000, Algeria Laboratory of Natural and Bioactive Substances (LASNABIO), Department of Chemistry, Faculty of Sciences, Abou Bekr Belkaıd University, P.O. Box 119, Tlemcen 13000, Algeria Institute of Agrophysics, Polish Academy of Sciences, Do´swiadczalna 4, Lublin 20-290, Poland Department of Tourism and Recreation, University of Life Sciences in Lublin, 15 Akademicka Street, Lublin 20-950, Poland Department of Analysis and Food Quality Assessment, University of Life Sciences in Lublin, 8 Skromna Str., Lublin 20-704, Poland Correspondence should be addressed to Radosław Kowalski; radoslaw.kowalski@up.lublin.pl Received 16 November 2022; Revised 5 December 2022; Accepted 15 December 2022; Published 30 December 2022 Academic Editor: Alessandro Buccolieri Copyright © 2022 Dalila Bereksi-Reguig et al. Tis is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Analysis of trace elements and heavy metals in honey is essential for honey quality and safety and also monitoring environmental pollution. Tis study aimed to evaluate the composition of thirty-seven honey samples of diferent botanical origins (14 multiforal and 23 uniforal) obtained from beekeepers located in the west region of Algeria. Inductively coupled plasma-mass spectrometry (ICP-MS) and atomic absorption spectroscopy (AAS) methods were used to determine the levels of 19 elements in honey (K, Na, Ca, Mg, Mn, Cu, Fe, Zn, V, Cr, Co, As, Ru, Rh, Cd, W, Pt, Au, and Pb). Ru, Rh, Pt and, Au were not detected in any of the tested honey samples. Te most abundant minerals were K, Ca, Na, and Mg ranging within 153.00–989.00 mg/kg, 33.10–502.00 mg/kg, 13.30–281.00 mg/kg, and 20.80–162.00 mg/kg, respectively. Fe, Mn, Zn, and Cu were the most abundant heavy metals while Pb, V, Cr, W, Co, and Cd were the lowest ones (<1 mg/kg) in the honey samples surveyed. Several honey types, lavender, rosemary, mild white mustard, thyme, milk thistle, carob tree, orange tree, Euphorbia, Eucalyptus, camphor, jujube tree, sage, and harmal, were studied, and the statistical analysis was carried out using principal component analysis (PCA) and hierarchical cluster analysis (HCA) techniques to evaluate the data. Te results showed that the analyses of mineral content were sufcient to determine the foral origin and their variability may be related to geochemical and geographical diferences. On other hand, all elements detected were at levels below safe thresholds. Contaminants can be anthropogenic or natural and may 1. Introduction pollute the soil with diferent fractions in many ways [6]. Bees are one of the most useful insects for humans: they Pollutants have been shown to be absorbed by plants pollinate our crops and produce honey [1, 2]. Nowadays, they growing in contaminated soil and have higher levels in their play a new role in indicating pollution and the quality of the tissue compared to other plants grown in control soils [7, 8]. environment, through produced honey [3, 4]. Te bee forages Tere is a close relationship between beehive products, for several kilometers in search of nectar; on its way, its body notably honey, and plants, which means that honey inherits accumulates particles of metals and pollutants present in the various characteristics and biological properties of plants environment which it transports to the hive [5]. with respect to their respective botanical sources and 2 Journal of Analytical Methods in Chemistry very scarce. Te present study aims to identify major and growing surface [9]. Due to this relationship, undesirable compounds or residues can be found in honey if plants or trace elements contained in diferent samples of honey collected in west Algeria. Besides physicochemical methods, soil have been exposed to these substances [10]. Among the most hazardous contaminants for human health and the we used principal component analysis (PCA) and hierar- environment which can be found in honey, we can cite heavy chical clusters analysis (HCA) techniques to diferentiate metals [11, 12]. Tey are found naturally in the Earth’s crust, Algerian honey, from diferent botanical origins, according but their presence in the environment has increased due to to their mineral content and the choice of elements with high anthropogenic activity [13]. For several decades, it has a higher discriminating power. been shown that exposure to metals implies serious damage [14]. Teir bioaccumulation and amplifcation in living 2. Materials and Methods systems lead to undeniable efects and diseases [15, 16]. 2.1. Honey Samples. A set of thirty-seven Apis mellifera Qualitative and quantitative analyses of heavy metals in intermissa honey samples (500 g each), namely, S1–S37, was honey, in addition to the classical approach, would allow collected in 2017 and 2018 from eight geographical provinces a reliable and reproducible system to establish the botanical (Tlemcen, Ain Temouchent, Sidi Bel Abbes, Mostaganem, origin [17] and geographic traceability [18]. Te environ- Mascara, Tiaret, Naama, ˆ and Bechar) in west Algeria (Fig- mental aspect of a region has also been assessed using ure 1). Table 1 shows the botanical and geographical origins minerals and heavy metals present in honey [19]. Several of the honey samples studied. Samples were taken directly studies have been conducted to establish the relationship from beekeepers with a guaranteed origin, stored in airtight between pollution in a region and the presence of con- plastic containers, and then kept in a refrigerator at 4 C until taminants in honey [20–22]. Te main elements found in ICP-MS and AAS analysis processes. diferent kinds of honey throughout the world, considered as a profling parameter, can be divided into two groups according to the literature: major elements or gold macro- 2.2. Reagents and Solution. High-purity deionized water elements (Na, K, Ca, Mg, P, S, and Cl) and trace elements acquired by passing distilled water through a water purif- including heavy metals (Al, Cu, Pb, Zn, Mn, Cd, Tl, Co, Ni, cation system (demineralizer HLP 20, Hydrolab, Poland) Rb, Ba, Be, Bi, U, V, Fe, Pt, Pd, Te, Hf, Mo, Sn, Sb, La, I, Sm, was used to prepare the solutions. Certifed single-element Tb, Dy, Sd, T, Pr, Nd, Tm, Yb, Lu, Gd, Ho, Er, Ce, Cr, As, B, standard solutions (1000 mg/L) used to prepare the cali- Br, Cd, Hg, Se, and Sr) [20, 22–28]. Te latter can afect the bration curve were of the highest purity grade (99.999%) and nervous system, kidney, liver, and respiratory functions. were supplied by Ultra Scientifc (North Kingstown, RI, Some metals, such as cadmium, arsenic, nickel, and chro- USA). Other reagents were of analytical grade unless oth- mium, are carcinogenic [29]. Exposure to heavy metals is erwise stated. Honey samples were digested with Suprapur implicated in more severe pathologies like multiple sclerosis grade nitric acid (HNO 65% m/m, Merck, Germany). A and neurodegenerative diseases (Alzheimer’s and Parkin- recovery test was performed using a single-element solution son’s disease) [29–31]; they could even play a role in trig- and two reference materials, namely, Standard Reference gering psychological and neurological disorders such as Materials (SRMs) Tomato Leaves and Pine Needles (Pinus autism [32]. It is also important to mention that the presence taeda) (SRM 1573a and SRM 1575a, resp., National Institute of certain minerals in honey is essential for the evaluation of of Standards and Technology (NIST), Gaithersburg, USA). its nutritional and curative quality [33]. Besides minerals and trace elements, honey is composed of various carbo- hydrates, polysaccharides, oligosaccharides, favonoids, vi- 2.3. Instrumentation. Elemental analysis was carried out by tamins, minerals, waxes, aroma compounds, pollen grains, using 820-MS inductively coupled plasma quadrupole mass pigments, and enzymes which also contribute to its profling spectrometer (ICP-MS; Varian, Mulgrave, Australia) and authentication [34]. In Algeria, honey is considered as equipped with an SPS3 autosampler (Varian, Australia) and a precious product because of its high nutritional value as a MicroMist nebulizer type (Varian). A MARS Express well as its therapeutic virtues widely popular traditionally microwave mineralizer system (CEM, Matthews NC, USA) [35, 36]. Te relatively mild climate in Algeria and abundant of Tefon reaction vessels was used in the digestion pro- varied fora allow the production of a variety of honey [37]. cedures. Te reaction vessels were cleaned using 10 mL of According to the Algerian Center for Quality Control and concentrated nitric acid before each digestion. Te multi- Packaging (CACQE), national honey production was esti- elemental determination was also determined by the atomic mated in 2011 at 33,000 quintals and in the same year absorption spectroscopy (AAS) method using a spectrome- 150,000 tonnes of honey, mainly from China, India, and ter SpectrAA 280 FS with autosampler SPS3 (Varian, Saudi Arabia, were imported (CACQE, https://www.cacqe. Australia), which was equipped with a deuterium lamp, org/). Te growing interest in honey, which is considered as hollow cathode lamp for each element, and an air-acetylene a noble substance, requires vigilance to its authenticity and burner. originality to preserve national production, avoiding every Appropriate results in terms of accuracy and sensitivity, sort of fraud. Based on multiple searches done on the low cost, and quickness make AAS a suitable procedure for PubMed database using diferent keywords, it is noteworthy determining the concentrations of alkaline and earth alka- that the number of studies on the physicochemical prop- line elements in the honey samples investigated. Tables 2 and erties of Algerian honey, particularly in the west region, is 3 show the instrumental parameters for ICP-MS. Journal of Analytical Methods in Chemistry 3 Mediterranean Sea Morocco Tl: Tlemcen SB: Sidi Bel Abbes AT: Ain-Temouchent M: Mostaganem Mas: Mascara O: Oran Sai: Saida Figure 1: Locations of the sampling points in west Algeria. 2.4. Analytical Determination. Trace element and heavy determination of V, Cr, Co, As, Ru, Rh, Cd, W, Pt, Au, and metal determination was performed by the inductively Pb of the honey samples, performed with an inductively coupled plasma-mass spectrometer (ICP-MS) and atomic coupled plasma-mass spectrometer equipped with a con- absorption spectroscopy (AAS) using the operating condi- centric nebulizer, a quartz torch with quartz injector tube, tions and emission wavelength lines listed in Tables 2 and 3. and cyclonic spray chamber (Table 2). Te concentration of Glass and plastic material was cleaned and kept in 10% (v/v) K, Na, Ca, Mg, Mn, Cu, Fe, and Zn ions was determined by nitric acid solution for at least two days. Te material was the AAS method using a spectrometer SpectrAA 280 FS with then rinsed three times with high-purity deionized water autosampler SPS3 (Table 3). To avoid sample ionization before being used. Approximately 0.5 g of each honey during potassium analysis, Schinkel bufer solution (mixture sample was digested with 10 mL of 65% HNO (v/v) in contents: 10 g/L cesium chloride and 100 g/L lanthanum Tefon vessels. Te sealed vessels were put into the micro- chloride) was used. Each sample was measured in triplicate wave mineralizer MARS Express (CEM, USA). A blank by AAS and ICP-MS detection. digest was carried out in the same way. Te microwave mineralization was performed stepwise at 400 W and 363 K, at 800 W and 393 K, and at 1600 W and 483 K. Te cooled 2.5. Quality Control. Te analytical quality was controlled by digestion solution was then diluted to 50 mL using high- means of certifed reference materials: NIST-1573a Tomato purity deionized water. Tis solution was fnally used for the Leaves and NIST-1575a Pine Needles. Te certifed reference 4 Journal of Analytical Methods in Chemistry Table 1: Geographical origins of honey samples from western regions of Algeria. Season/year of Region Sample Floral type Scientifc name Family Location harvest Tlemcen S1 Lavender Lavandula vera D.C. Lamiaceae Sidi Djillali Summer 2018 S2 Rosemary Rosmarinus ofcinalis L. Lamiaceae Sidi Djillali Spring 2018 S3 Multiforal Multiforal — Sidi Djillali Spring 2018 S4 Multiforal Multiforal — Sidi Djillali Summer 2017 S5 Multiforal Multiforal — El Aricha Summer 2017 S6 Mild white mustard Sinapis alba L. Brassicaceae A¨ın Fezza Summer 2017 S7 Tyme Tymus vulgaris L. Lamiaceae Beni Snous Spring 2018 S8 Milk thistle Silybum marianum (L.) Gaertn. Asteraceae Beni Snous Summer 2018 S9 Multiforal Multiforal — Oued Chouly Autumn 2017 S10 Carob tree Ceratonia siliqua L. Fabaceae Oued Chouly Autumn 2017 S11 Tyme Tymus vulgaris L. Lamiaceae Beni Mester Spring 2017 S12 Carob tree Ceratonia siliqua L. Fabaceae Beni Ghazli Spring 2017 S13 Multiforal Multiforal — Oued es Safsaf ˆ Summer 2018 S14 Multiforal Multiforal — Sebaa Chioukh Spring 2017 S15 Multiforal Multiforal — Hennaya Summer 2017 S16 Orange tree Citrus sinensis L. Rutaceae Remchi Spring 2017 S17 Multiforal Multiforal — Hona¨ıne Spring 2018 S18 Milk thistle Silybum marianum (L.) Gaertn. Asteraceae Hona¨ıne Summer 2018 Ain Temouchent S19 Multiforal Multiforal — Oulhaça El Gherarba Spring 2018 S20 Multiforal Multiforal — Beni Ghanem Summer 2018 S21 Multiforal Multiforal — Bouzedjar Spring 2018 Sidi Bel Abbes S22 Euphorbia Euphorbia L. Euphorbiaceae Ras El Ma Spring 2017 S23 Milk thistle Silybum marianum (L.) Gaertn. Asteraceae Telagh Spring 2017 S24 Multiforal Multiforal — Lamtar ˆ Spring 2017 S25 Eucalyptus Eucalyptus globulus Labill. Myrtaceae Sidi Brahim Spring 2017 Mostaganem S26 Camphor Cinnamomum camphora L. Lauraceae Sidi Ali Autumn 2017 S27 Eucalyptus Eucalyptus globulus Labill. Myrtaceae Mostaganem Summer 2017 S28 Orange tree Citrus sinensis L. Rutaceae Bouguirat Spring 2017 Mascara S29 Rosemary Rosmarinus ofcinalis L. Lamiaceae Djebel Stamboul Spring 2017 Tiaret S30 Multiforal Multiforal — Tiaret Spring 2018 Naama ˆ S31 Multiforal Multiforal — Aın Sefra Spring 2017 S32 Jujube tree Ziziphus lotus L. Rhamnaceae A¨ın Sefra Spring 2017 S33 Jujube tree Ziziphus lotus L. Rhamnaceae Aın Ben Khelil Spring 2017 S34 Sage Salvia ofcinalis L. Lamiaceae Naama ˆ Spring 2017 S35 Harmal Peganum harmala L. Zygophyllaceae Mecheria Spring 2017 Bechar S36 Multiforal Multiforal — Djebel Antar Winter 2017 S37 Mild white mustard Sinapis alba L. Brassicaceae Oued Zouzfana Spring 2017 Table 2: Operating ICP-MS conditions. Radio frequency power (W) 1370 Plasma gas fow rate (L/min) 18 Auxiliary gas fow rate (L/min) 1.70 Spray chamber T ( C) 2 Nebulizer gas fow (L/min) 1 Number of replicates 3 Journal of Analytical Methods in Chemistry 5 Table 3: Operating AAS conditions. Elements Na Mg K Ca Mn Fe Cu Zn Fuel fow (L/min) 2 2 2 2 2 2 2 2 Lamp current (mA) — 4 — 10 5 5 4 5 Wavelength (nm) 589.0 202.6 766.5 422.7 279.5 248.3 324.8 213.9 Slit width (nm) 0.2 1.0 0.2 0.5 0.2 0.2 0.5 1.0 Air fow (L/min) 10 10 10 10 10 10 10 10 materials were prepared according to the instructions of the investigation showed a wide range of K contents manufacturer. Honey samples and quality control samples (153.00–989.00 mg/kg) accounting for 80.08% of the total (blind samples and certifed reference materials) were de- minerals present in honey from Bouzedjar (S21). Following termined in triplicate, and the average was given as the fnal our data, similar amounts of K in honey samples were result. Table 4 presents the validation parameters obtained previously reported from the Azilal and Beni Mellal prov- inces in Moroccan (256–1023 mg/kg) [39] and Tunisian during analysis. For Au, Pt, Rh, Ru, and W, the recovery parameter was obtained from fortifcation by using certifed (172.48–976.75 mg/kg) honey [40]. Overall, the concentra- tions of K in west Algerian honey were higher than those single-element standard solutions. reported from Turkey (1.18–268 mg/kg) [20], the West Bank in Palestine (42.80–585.00 mg/kg) [41, 42], and Jableh and 2.6. Statistical Method. Principal component analysis (PCA) Tartous provinces in the western part of Syria (38.2–174 mg/ and hierarchical cluster analysis (HCA) were performed kg) [43], but the levels were lower than those reported for with the XLSTAT 2014.5.03 software for Microsoft Excel Portugal (117.55–2590.60 mg/kg) [44, 45], Italy (Addinsoft, Bordeaux, France) to classify and discriminate (237–6520 mg/kg) [23, 25], Libya (1120.1–1980.6 mg/kg) the honey samples. [46], and Spain (1615–3770 mg/kg) [47–52]. Eucalyptus honey showed the second highest concentration (987.00 mg/ kg, S27) of K among the honey samples investigated while 3. Results and Discussion carob tree honey contained the lowest amount (153.00 mg/ kg, S12). High concentrations of K were also present in the 3.1. Results of Minerals in Honey Samples. Te concentra- tions of trace element and heavy metals found in the honey mild white mustard (946.00 mg/kg, S6), lavender (808.00 mg/kg, S1), and milk thistle (802.00 mg/kg, S23) samples collected from thirty-seven locations situated in west Algeria are given in Tables 5 and 6. Validation pa- honey when compared to the other honey samples studied. Te concentration of K in the present study is also much rameters of the analytical procedure such as: limit of de- tection (LOD), limit of quantifcation (LOQ), precision, higher than the three Citrus spp. (Citrus) honey samples from Syria (38.2–174 mg/kg) [43]. Tis discrepancy may be accuracy, and uncertainty budget are listed in Table 4. A due to the geographical variation in the sources of honey detailed validation of the analytical procedure was per- [52]. A high mean value (113.92± 95.38 mg/kg) of Ca was formed including the performance parameters limit of de- observed among the investigated west Algerian honey tection (LOD) and limit of quantifcation (LOQ); precision, samples with the concentrations ranging within accuracy, and an uncertainty budget (Table 4). (33.10–502.00 mg/kg) which were higher than those of Precision and percentage recovery of the analytical procedure were determined by using randomly selected honey samples reported for Morocco (19.71–200.1 mg/kg) [39], Tunisia (113.85–221.07 mg/kg) [40], Portugal honey samples individually spiked with known spiked concentrations of the trace elements and heavy metals (6.24–134.35 mg/kg) [44, 45], Spain (11.69–218.5 mg/kg) [47–49, 52], France (8.90–130.90 mg/kg) [53, 54], Italy studied (K, Na, Ca, Mg, Mn, Cu, Fe, Zn, V, Cr, Co, As, Ru, Rh, Cd, W, Pt, Au, and Pb) and used as positive controls. Te (<43–283 mg/kg) [23, 25], Turkey (<0.001–4.5 mg/kg) [20], Palestine (44.50–150.70 mg/kg) [41, 42], Syria (43.3–118 mg/ mean percentage recoveries of the determined trace ele- kg) [43], Greece (15.22–65.93 mg/kg), Cyprus ments and heavy metals ranged between 74.51% (Co) and (23.66–143.47 mg/kg), and Egypt (44.79–112.10 mg/kg) [26] 117.32% (Fe) (Table 4), which point out good accuracy, (Tables 7 and 8). Besides, honey samples collected from precision, and validity of the method employed. Te results of mineral content in honey were found relatively low and diferent locations in the west of Libya show the highest concentrations with a range within (923.92–1117.5 mg/kg) varied over a range within 0.04–0.16% of the total com- position, which is in agreement with the composition of [46] of all the Mediterranean regions considered. It would be interesting, due to the presence of the high amount of this nectar honey [22, 38]. A total of 19 trace elements and heavy metals were determined. It should be noted that the con- mineral, to propose it in a strategy for the prevention of osteoporosis. Sage honey showed the highest concentration centrations of 19 elements were variable depending on the foral origin of honey [19]. Among them, the most abundant (502.00 mg/kg, S34) of Ca, followed by multiforal honey (377.00 mg/kg, S14), Eucalyptus honey (287.00 mg/kg, S25), elements were K, Ca, Na, and Mg with average levels and milk thistle honey (215.00 mg/kg, S23). Interestingly, (mean± SD) of 532.46± 212.69 mg/kg, 113.92± 95.38 mg/ there are some similarities between the Ca contents of some kg, 83.40± 64.85 mg/kg, and 100.83± 33.88 mg/kg, re- types of west Algerian honey and those coming from spectively. Te west Algerian honey samples in the present 6 Journal of Analytical Methods in Chemistry Table 4: Validation parameters of the analytical procedure for determination of minerals. Certifed reference material analysis Validation parameters Te result Te result Element declared by obtained in LOD LOQ Recovery (%) Uncertainty (%) the manufacturer own research 112.6 112.21 100.35 16 As (μg/kg) 0.5 1.0 39 42.10 92.64 21 Au (μg/kg) — — 2.3 4.6 109.12 26 50450 53700.23 93.95 17 Ca (mg/kg) 12.1 24.2 2500 2356.45 106.09 19 1517 1498.03 101.27 13 Cd (μg/kg) 0.1 0.2 233 276.20 84.36 16 577.3 774.82 74.51 18 Co (μg/kg) 0.1 0.2 61 62.76 97.20 20 1988 2123.20 93.63 11 Cr (μg/kg) 8.0 16.0 61 62.76 97.20 14 4.7 4.92 95.68 19 Cu (mg/kg) 0.7 1.4 2.8 3.1 90.32 25 367.5 391.09 93.97 10 Fe (mg/kg) 3.7 7.2 46 39.21 117.32 15 26760 30207.00 88.59 15 K (mg/kg) 23.2 46.4 4170 4376.00 95.29 14 12000 12096.00 99.21 17 Mg (mg/kg) 9.3 18.6 2610 2734.00 95.46 12 246.3 242.02 101.77 19 Mn (mg/kg) 0.5 1.0 488 460.50 105.97 11 136.1 157.15 86.61 14 Na (mg/kg) 6.0 12.0 63 67.89 92.80 20 Pb (μg/kg) 167 163.58 0.2 0.4 102.09 23 Pt (μg/kg) — — 3.0 6.0 107.12 24 Ru (μg/kg) — — 2.5 5.0 85.34 33 Rh (μg/kg) — — 2.2 4.4 105.23 28 V (μg/kg) 835 867.58 0.4 0.8 96.24 25 W (μg/kg) — — 4.1 8.2 88.12 35 30.94 36.49 84.79 21 Zn (mg/kg) 0.1 0.2 38 38.50 98.70 16 “a” NIST-1573a Tomato Leaves Standard Reference Materials; “b” NIST-1575a Pine Needles (Pinus taeda) Standard Reference Materials; “c” Certifed single- element standard solutions. neighboring countries. For instance, the Ca concentration of (Table 1). Te high content of both K and Na in mild white Eucalyptus honey is similar to that of carob honey from mustard honey makes it less dangerous when consumed by Rabat province in Morocco (286.01± 5.79 mg/kg) [55], while hypertensive patients. Magnesium was the fourth most milk thistle honey has a similar Ca concentration when abundant element in the present study, with contents compared to the Tunisian mint honey (221.07± 5.16 mg/kg) ranging from 20.80 to 162.00 mg/kg. Tese concentrations [40]. Te concentrations of Na in the west Algerian honey were similar to those of Italian honey (22–159 mg/kg) as samples were higher than those reported for honey from reported by Pisani et al. [23] and were higher than those Portugal (0.36–95.13 mg/kg) [45], Spain (11–84 mg/kg) [51], coming from most countries of the Mediterranean region Italy (4.8–176 mg/kg) [23, 25], and Turkey (0.48–13.1 mg/kg) (Table 7) except honey from Portugal (2.77–234.63 mg/kg) [20]. Euphorbia and Eucalyptus honey contained the highest [45] and Spain (30.00–402.00 mg/kg) [51]. Milk thistle concentrations of Na (281.00 mg/kg, S22; 275.00 mg/kg, S25, honey contained the highest concentration (162.00 mg/kg, resp.) among the honey types while thyme, lavender, and S8) of Mg among the honey samples investigated, while milk thistle honey had the lowest amounts (13.30 mg/kg, S7; multiforal honey samples S24 and S36 (20.80 mg/kg and 21.60 mg/kg, S1; 22.10 mg/kg, S8, resp.). Other types of west 42.50 mg/kg, resp.) contained the lowest. High concentra- tions of Mg were also exhibited by thyme (159.00 mg/kg, S7), Algerian honey such as orange tree, jujube tree, and mild white mustard honey samples are also rich in Na (179.00 mg/ mild white mustard (149.00 mg/kg, S6), and lavender kg, S28; 165.00 mg/kg, S32; 146.00 mg/kg, S6, resp.) when (142 mg/kg, S1) honey types, which are similar to the honey compared to the other honey samples investigated. On the from Spain (159 mg/kg) [23] and France (145 mg/kg) [54]. In other hand, it should be noted that the higher concentrations the case of Fe, its concentrations in the west Algerian honey of K and Na were exhibited by mild white mustard honey ranged from 8.48 to 59.60 mg/kg. Honey samples from the collected from A¨ın Fezza (S6) located in Tlemcen province Mediterranean countries reported lower Fe contents Journal of Analytical Methods in Chemistry 7 Table 5: Distribution data for Cr, Co, V, W, Ru, Rh, Pt, Au, As, Cd, and Pb (μg/kg) in west Algerian honey. b,c b,c b,c b,c b,c b,c b,c b,c b b,c b,c Sample Cr Co V W Ru Rh Pt Au As Cd Pb d d d d d d S1 70.11 9.01 8.62 — — — — — — 1.27 21.44 d d d d d S2 55.56 8.73 9.00 — — — — — 4.93 0.83 16.62 d d d d d S3 38.73 10.33 4.83 — — — — — 7.70 1.09 9.73 d d d d d S4 37.20 12.14 10.04 — — — — — 21.56 0.36 0.54 d d d d d S5 34.07 20.80 6.88 — — — — — 3.38 0.90 20.25 d d d d d S6 56.21 4.57 7.30 — — — — — 4.70 0.79 21.05 d d d d d S7 27.63 5.96 1.57 — — — — — 1.75 8.14 9.87 d d d d d d S8 45.89 4.73 4.94 — — — — — 18.20 0.24 — d d d d d S9 41.02 17.32 6.00 — — — — — 4.05 0.54 11.94 d d d d d S10 87.37 12.67 7.29 — — — — — 5.36 0.55 14.83 d d d d d S11 43.74 6.61 79.73 — — — — — 3.95 1.50 23.20 d d d d d S12 53.39 1.45 4.48 — — — — — 5.54 0.77 10.78 d d d d d S13 59.05 3.36 5.12 — — — — — 4.80 0.97 9.21 d d d d d S14 74.83 4.20 13.98 — — — — — 6.43 1.04 13.75 d d d d d S15 29.60 3.56 6.52 — — — — — 5.27 1.26 17.93 d d d d d S16 38.58 1.02 1.44 — — — — — 1.61 1.05 12.84 d d d d d d S17 39.84 0.36 2.10 — — — — — 1.68 — 132.73 d d d d d S18 41.39 3.29 3.42 — — — — — 6.28 1.14 15.28 d d d d d S19 55.46 5.77 13.84 — — — — — 6.31 0.84 14.39 d d d d d S20 34.15 2.82 3.23 — — — — — 5.93 0.83 10.18 d d d d S21 42.62 4.60 10.26 10.65 — — — — 6.68 3.38 89.44 d d d d d S22 39.17 8.42 4.58 — — — — — 6.23 1.43 14.23 d d d d S23 46.89 5.25 9.09 17.18 — — — — 7.73 0.41 40.55 d d d d d S24 30.18 3.25 6.40 — — — — — 6.28 0.98 14.95 d d d d S25 29.13 4.18 4.55 11.10 — — — — 8.19 0.86 24.57 d d d d d S26 58.86 7.82 106.51 — — — — — 3.95 0.69 9.74 d d d d d S27 27.36 3.07 4.75 — — — — — 4.92 0.90 26.52 d d d d d S28 37.06 7.04 7.68 — — — — — 7.38 0.56 11.15 d d d d d S29 27.81 1.89 0.81 — — — — — 2.70 1.05 11.91 d d d d d S30 37.61 16.64 7.65 — — — — — 2.81 2.94 91.36 d d d d d S31 62.04 17.78 27.11 — — — — — 10.41 1.14 24.70 d d d d d S32 53.73 16.95 15.35 — — — — — 5.64 0.89 21.32 d d d d d S33 50.33 12.11 11.62 — — — — — 2.90 0.99 20.95 d d d d d S34 44.25 12.94 10.24 — — — — — 4.35 1.00 74.99 d d d d S35 72.11 21.19 16.47 24.30 — — — — 8.30 0.84 16.41 d d d d d S36 55.96 1.01 44.33 — — — — — 4.95 1.02 8.21 d d d d d S37 47.23 6.74 12.37 — — — — — 3.96 1.19 27.30 Statistics d d d d Mean 46.65 7.83 13.52 15.81 — — — — 5.86 1.23 24.73 d d d d Max. 87.37 21.19 106.51 24.30 — — — — 21.56 8.14 132.73 d d d d Min. 27.36 0.36 0.81 10.65 — — — — 1.61 0.24 0.54 d d d d SD 14.43 5.82 21.05 4.93 — — — — 4.03 1.32 27.59 b,c d trace elements/heavy metals; concentrations were lower than LOQ; SD: standard deviation. (Table 7). It is to highlight that the highest Fe levels were higher than those reported for most honey samples coming found in lavender honey (59.60 mg/kg, S1), while multiforal from the Mediterranean region (Table 7). In the present (8.48 mg/kg, S24), Eucalyptus (11.30 mg/kg, S25), and milk study, the Zn concentrations ranged from 0.22 to 13.90 mg/ thistle (11.70 mg/kg, S8) honey had the lowest contents. Our kg (mean± SD 3.61± 2.35 mg/kg). Te highest Zn level was values were well below the provisional tolerable weekly 13.90 mg/kg in carob tree honey (S10), while the lowest one intake (PTWI) by body weight (5.6 mg/kg b.w.) recom- was 0.22 mg/kg in multiforal honey (S5). Te levels of Zn in mended by the Joint FAO/WHO Expert Committee on Food the west Algerian honey are lower than those from Palestine Additives (JECFA) [56]. Te concentration of Mn in the (0.13–25.20 mg/kg and 1.00–19.90 mg/kg) [41, 42], re- investigated west Algerian honey was between 1.36 and spectively, but are higher than those from Morocco 13.90 mg/kg. Among the honey samples investigated, mild (≤0.1–0.69 mg/kg) [39], Tunisia (0.42–2.06 mg/kg) [40], white mustard honey is the richest in Mn (13.90 mg/kg, S6). Portugal (0.03–3.29 mg/kg) [45], Spain (2.34–3.47 mg/kg) High concentrations of Mn were also shown by the milk [50], France (nd–1.4 mg/kg) [54], Italy (0.72–3.66 mg/kg) thistle and lavender honey (13.40 mg/kg, S18; 13.30 mg/kg, [23], Turkey (<1–237 μg/kg) [20], Greece (0.97–9.30 mg/kg), S1, resp.) as well as thyme (12.70 mg/kg, S7) honey. Te Cyprus (0.86–6.94 mg/kg), Egypt (0.55–1.68 mg/kg) [26], levels of Mn in honey samples surveyed in this study were and Syria (0.206–2.76 mg/kg) [43]. In the case of the 8 Journal of Analytical Methods in Chemistry Table 6: Distribution data for K, Na, Ca, Mg, Cu, Mn, Fe, and Zn (mg/kg) in west Algerian honey. Total a a a a b,c b,c b,c b,c Sample K Na Ca Mg Cu Mn Fe Zn minerals S1 808.00 21.60 56.20 142.00 3.66 13.30 59.60 2.62 1107.58 S2 460.00 49.20 58.50 126.00 5.70 10.80 24.50 2.39 737.53 S3 418.00 37.00 64.90 142.00 4.46 11.10 24.40 4.41 706.68 S4 451.00 30.70 73.60 92.70 9.62 7.67 26.10 3.38 695.19 S5 327.00 34.10 50.60 77.60 — 5.52 19.60 0.22 514.92 S6 946.00 146.00 104.00 149.00 5.92 13.90 23.20 3.39 1392.47 S7 241.00 13.30 33.10 159.00 5.08 12.70 22.60 5.29 492.53 S8 790.00 22.10 97.80 162.00 4.81 10.50 11.70 4.42 1104.26 S9 441.00 97.80 85.80 88.80 — 7.85 34.60 9.33 765.57 S10 667.00 64.40 205.00 120.00 7.86 10.80 57.30 13.90 1147.04 S11 589.00 104.00 102.00 79.00 3.44 7.61 33.70 4.98 924.28 S12 153.00 15.30 33.50 116.00 2.80 11.50 24.30 3.08 359.77 S13 572.00 102.00 104.00 127.00 1.66 13.70 16.30 4.99 942.24 S14 989.00 77.10 377.00 137.00 — 11.50 17.60 3.45 1613.00 S15 722.00 32.30 99.90 69.10 5.29 1.36 26.80 0.62 957.88 S16 254.00 62.30 81.10 89.20 1.95 9.26 26.00 4.37 528.41 S17 375.00 129.00 33.30 134.00 7.16 12.10 14.90 4.59 710.58 S18 526.00 68.20 53.80 133.00 — 13.40 21.60 5.20 821.65 S19 400.00 110.00 119.00 118.00 2.13 8.12 22.60 2.33 782.65 S20 247.00 26.50 70.00 112.00 — 10.20 25.10 2.29 493.45 S21 776.00 23.60 80.80 51.60 6.08 3.83 24.00 2.59 969.00 S22 675.00 281.00 100.00 99.50 6.61 7.38 18.10 3.30 1191.51 S23 802.00 71.70 215.00 55.30 4.67 1.47 26.20 3.63 1180.73 S24 425.00 156.00 101.00 20.80 — 1.93 8.48 1.87 715.40 S25 777.00 275.00 287.00 77.80 — 1.69 11.30 2.82 1433.12 S26 470.00 92.50 132.00 57.00 4.17 9.25 21.10 3.41 789.77 S27 987.00 23.60 165.00 124.00 7.25 6.55 21.80 4.75 1340.80 S28 338.00 179.00 91.00 66.80 — 6.50 19.10 2.46 703.31 S29 384.00 102.00 77.70 85.40 5.46 8.63 26.30 3.17 692.92 S30 471.00 44.30 70.70 78.40 4.88 3.45 24.10 1.44 699.04 S31 504.00 64.60 90.80 80.20 3.75 6.10 35.80 2.75 788.61 S32 442.00 165.00 117.00 92.80 3.59 4.70 38.30 3.62 867.55 S33 510.00 51.20 70.70 123.00 8.97 6.15 29.50 1.78 801.68 S34 441.00 114.00 502.00 111.00 — 3.90 25.60 3.68 1201.59 S35 460.00 66.10 71.90 98.60 2.52 7.28 29.60 2.69 739.34 S36 411.00 35.90 49.00 42.50 — 6.46 28.00 2.58 576.28 S37 452.00 97.40 90.50 92.60 2.78 5.87 15.40 1.93 758.88 Statistics Mean 532.46 83.40 113.92 100.83 4.90 7.95 25.28 3.61 871.55 Max. 989.00 281.00 502.00 162.00 9.62 13.90 59.60 13.90 1613.00 Min. 153.00 13.30 33.10 20.80 1.66 1.36 8.48 0.22 359.77 SD 212.69 64.85 95.38 33.88 1.65 3.66 10.39 2.35 290.66 a b b,c d Major elements; trace elements; trace elements/heavy metals; concentrations were lower than LOQ; SD: standard deviation. concentrations of Zn, up 91% of our honey samples are in west Algerian honey are slightly higher than those from the maximum tolerable weekly intake range (2.1–7 mg/kg Morocco (≤0.1 mg/kg) [39], Tunisia (0.12–0.34 mg/kg) [40], b.w.) [56]. Te concentrations of Cu in the west Algerian Portugal (0.00–5.35 mg/kg) [45], France (0.06–1.71 mg/kg) [53], and other Mediterranean countries (Table 7). In honey ranged from 1.66 to 9.62 mg/kg (mean± SD 4.90± 1.64 mg/kg). Te highest concentrations of Cu were in summary, Eucalyptus honey contained the highest K and Na multiforal, jujube tree, and carob tree honey (9.62 mg/kg, values. Sage honey is rich in Ca while milk thistle honey is S4; 8.97 mg/kg, S33; and 7.86 mg/kg, S10, resp.), but Cu was rich in Mg. Lavender honey contained high Fe levels while not detected in more than a quarter of the investigated the carob tree is the richest in Zn. Our study shows also that honey. Unfortunately, the levels in approximately 54% of the the mild white mustard honey contained highest level of Mn, investigated honey samples are above the PTWI (3.5 mg/kg whereas jujube tree and carob tree honey comprised Cu in b.w.) for Cu established by Joint FAO/WHO [56]. Generally, high concentrations. Generally, the west Algerian honey Cu is transferred and accumulated in food under the in- samples are rich in minerals. Te latter contains a signifcant fuence of the environment and also due to human con- proportion of microelements and therefore are valuable food tributions (fertilizers and pesticides) [57]. Copper levels in products. Tese latter are a signifcant fraction of Journal of Analytical Methods in Chemistry 9 Table 7: A comparison among reports regarding mineral content in honey samples from Mediterranean regions. Morocco Morocco Tunisia Portugal Portugal Spain Spain Spain Spain Spain Present Element [38] [55] [39] [43] [44] [46] [47] [48] [49] [50] study N � 29 N � 8 N � 6 N � 38 N � 16 N � 40 N � 8 N � 60 N � 140 N � 41 27.36–87.37 μg/ Cr — — 0.02–0.32 — — — — 0.049–4.480 — — kg Co 0.36–21.19 μg/kg — nd — — — — — 0.015–0.720 — — V 0.81–106.51 μg/kg — — — — — — — — — — 10.65–24.30 μg/ W — — — — — — — — — — kg Ru ˂LOQ — — — — — — — — — — Rh ˂LOQ — — — — — — — — — — Pt ˂LOQ — — — — — — — — — — Au ˂LOQ — — — — — — — — — — As 1.61–21.56 μg/kg — — — — — — — — — — Cd 0.24–8.14 μg/kg — nd — — — — — nd–0.355 4.21–4.56 μg/kg — 46.32–31.50 μg/ Pb 0.54–132.73 μg/kg ≤0.1 nd 0.01–0.05 — — — — — — kg K 153.00–989.00 256–1023 644.02–1883.15 172.48–976.75 117.55–2590.60 42.98–1352.90 434.1–1935 545.45–5570.73 514–6785 — 1615–3770 Na 13.30–281.00 18.81–118.74 367.52–855.24 251.34–521.22 90.22–727.79 0.36–95.13 11.69–218.5 9.18–151.65 38–476 — 11–84 Ca 33.10–502.00 19.71–200.1 129.35–688.43 113.85–221.07 6.24–134.35 2.77–234.63 42.59–341.0 15.14–181.69 107–420 — 68–476 Mg 20.80–162.00 19.85–45.84 18.42–131.21 — 10.62–70.41 1.79–230.81 13.26–74.38 42.11–1078.95 19–173 — 30–402 Cu 1.66–9.62 ≤0.1 0.11–1.82 0.12–0.34 — 0.00–5.35 0.531–2.117 nd 0.547–2.300 0.74–1.88 1–7 Mn 1.36–13.90 0.28–1.74 — — — 0.00–22.62 0.133–9.471 — 3.4–45 — — Fe 8.48–59.60 1.46–13.95 0.71–4.68 0.83–3.54 — 0.18–2.68 — nd–7.07 0.71–60 2.26–4.70 0–7 Zn 0.22–13.90 ≤0.1–0.69 1.41–4.26 0.42–2.06 — 0.03–3.29 1.332–7.825 0.30–7.06 0.93–5.9 2.34–3.47 0–7 From each report, the min-max range is mentioned. All values are in mg/kg, unless stated otherwise. nd: not detected; ˂LOQ: concentrations were lower than LOQ; N: the number of honey samples. 10 Journal of Analytical Methods in Chemistry Table 8: A comparison among reports regarding mineral content in honey samples from Mediterranean regions. Spain France France Italy Italy Turkey Greece Cyprus Egypt Palestine Palestine Syria Libya Element [51] [53] [54] [24] [22] [19] [25] [25] [25] [40] [41] [42] [45] N � 25 N � 86 N � 40 N � 40 N � 51 N � 71 N � 12 N � 14 N � 8 N � 21 N � 10 N � 6 N � 8 Cr — 0.08–0.36 — 10–328 — <1 μg/kg <0.12 <0.12 <0.12 0.00–0.51 — <0.018–0.054 — Co — 0.10–0.23 — 1.0–17 2.9–30.2 — <0.03 <0.03 <0.03 — — — — V — — — <3–24 — — <0.11 <0.11 <0.11 — — — — W — — — — — — — — — — — — — Ru — — — — — — — — — — — — — Rh — — — — — — — — — — — — — Pt — — — — — — — — — — — — — Au — — — — — — — — — — — — — As — — nd–8.0 <25 2.8–11.1 — <0.08 <0.08 <0.08 — — — 0.006–0.018 Cd — 0.08–0.25 1–22 1.3–4.2 1.0–15.3 <1 μg/kg <0.05 <0.05 <0.05 — — — 0.125–0.150 Pb — 0.28–1.08 3–101 9–209 28–304 <1 μg/kg <0.08 <0.08 <0.08 — 0.51–0.94 <0.082 2.42–10.98 K 261–1380 — — 237–6520 178–4140 1.18–268 — — — 42.80–585.00 203.92–495.48 38.2–174 1120.1–1980.6 Na 256–501 — — 4.8–176 60–147 0.48–13.1 — — — 41.80–306.30 35.19–196.51 — 506.8–804.6 Ca 110–248 8.90–130.90 36–219 <43–283 159–373 <0.001–4.5 15.22–65.93 23.66–143.47 44.79–112.10 44.50–150.70 64.49–138.41 43.3–118 923.92–1117.5 Mg 37–139 3.62–68.78 33–145 6.2–148 22–159 — 20.13–109.53 9.24–80.48 12.17–22.04 12.30–46.70 20.48–54.15 — — Cu — 0.06–1.71 — 0.06–5.4 0.20–2.79 <1–929 μg/kg <0.35 <0.35 <0.35 0.00–1.52 0.61–1.22 0.616–2.21 — Mn — 0.11–42.81 1.4–12.8 0.09–2.8 0.13–16.9 <1–274 μg/kg 0.26–5.54 0.07–4.11 0.17–0.55 0.11–0.99 — 0.153–1.69 — Fe — 0.56–86.76 — <1–4.4 1.0–6.3 <1–7254.62 μg/kg 0.75–4.16 0.96–3.97 2.18–8.45 2.00–10.80 2.25–8.75 1.46–16.9 — Zn — 0.17–6.42 nd–1.4 <0.5–8.9 0.72–3.66 <1–237 μg/kg 0.97–9.30 0.86–6.94 0.55–1.68 1.00–19.90 0.13–25.20 0.206–2.76 — From each report, the min-max range is mentioned. All values are in mg/kg, unless stated otherwise. nd: not detected; ˂ LOQ: concentrations were lower than LOQ; N: the number of honey samples. Journal of Analytical Methods in Chemistry 11 Table 9: Variance estimates (eigenvalues) and cumulative per- micronutrients making honey valuable food products. Some centage of total variance (%) obtained by PCA considering 37 of these elements are vital, and others are merely desirable or honey samples and trace elements and heavy metals. benefcial. Indeed, they have signifcant roles in the acti- vation of certain enzymes; these include Mg, Fe, Mn, Zn, and Honey samples Principal components Cu, which play an important role in the metabolic trans- Eigenvalues Cumulative % formations in the human body, while K, Ca, and Na are 1 1.937 30.38 essential in building strong bones and teeth, muscle con- 2 1.607 54.33 tractions, nerve signals, regulating heartbeat, and fuid 3 1.562 54.79 balance within cells [58]. Teir defciencies play critical roles 4 1.209 60.89 in many disorders such as hypertension and osteoporosis [59]. Some honey samples studied are important sources of of V were lower than those reported by Conti et al. [25] micronutrients useful to the proper functioning of the (Table 7). Tere is currently no JECFA assigned reference human body, and therefore their consumption is highly health standard for vanadium. Chromium is abundant in the recommended, especially in case of defciency. To verify the environment in a trivalent or hexavalent state. It is found in quality of the west Algerian honey, in addition to Mn, Fe, Zn, the trivalent state, at low concentrations, in a wide range of and Cu, it is very important to assess and monitor the foods [62]. Its content in foods can be greatly afected by concentrations of other heavy metals and metalloids, which anthropic and geochemical factors. Te mean and the range are potentially toxic. Tey include arsenic, lead, chromium, values of the Cr content of honey samples investigated were cadmium, vanadium, tungsten, cobalt, ruthenium, rhodium, 46.65 and 27.36–87.37 μg/kg. Levels observed were lower platinum, and gold. In the present study, the relative con- than those reported in Tunisia (0.02–0.32 mg/kg) [40], centrations of these elements in the honey samples decreased France (0.08–0.36 mg/kg) [53], Palestine (0.00–0.51 mg/kg) in the following order: Pb> V> Cr> W> As> Co> Cd> Ru, [41], and Spain (0.049–4.480 mg/kg) [60]. On the other Rh, Pt, and Au. All of them were detected at levels <1 mg/kg. hand, very low Cr levels were found in Eucalyptus, thyme, Te resulting data, summarized in Table 5, were consistent and rosemary (27.36 μg/kg, S27; 27.63 μg/kg, S7; and with the ranges indicated for honey from other studies 27.81 μg/kg, S29, resp.) honey samples, whereas carob tree [20, 26, 50, 53, 60]. Lead is considered a strict contaminant contained the highest concentration (87.37 μg/kg, S10). and is toxic to living organisms, even at very low concen- Chromium has low toxicity in foods in part due to its low trations. Its content in honey is examined in several studies. bioavailability. Currently, there is no formal recommended Te range values of Pb content in honey samples from the dietary allowance for Cr [63]. Tungsten is found naturally on west Algerian region were 0.54–132.73 μg/kg. Pb was found Earth almost exclusively in the combined state with other at high concentrations in the multiforal honey samples elements in ores as wolframite and scheelite. Because it is (132.73 μg/kg, S17; 91.36 μg/kg, S30; and 89.44 μg/kg, S21) a rare metal and its compounds are generally inert, the and was absent in milk thistle (S8). Furthermore, the ob- efects of W on the environment are limited [64]. Few honey served concentrations of Pb were higher than those mea- samples from the west Algerian region contained quanti- sured in Morocco (≤0.1 mg/kg) [38], Tunisia (0.01–0.05 mg/ fable concentrations of W. Te highest W concentration was kg) [40], Greece (<0.08 mg/kg), Cyprus (<0.08 mg/kg), found in harmal honey (24.30 μg/kg, S35). Besides, W Egypt (<0.08 mg/kg) [26], Spain (46.32–31.50 μg/kg) [50], concentrations of approximately 90% of honey samples were and Syria (<0.082 mg/kg) [43]. However, the concentrations below LOQ. W was not investigated in others studies around of Pb in the examined honeys were lower than in the fol- the Mediterranean region. Terefore, it is not possible to lowing honeys: Palestinian (0.51-0.94 mg/kg) [42], French comment on any diferences in the level of W in honey. (3-101 mg/kg) [54], and Italian (28-304 mg/kg) [23], and (9- Arsenic is a ubiquitous element with metalloid properties. It 209 mg/kg) [25]. However, concentrations found were lower occurs naturally in organic and inorganic forms. Tis is the than those found in Palestinian (0.51–0.94 mg/kg) [42], most toxic element and is considered carcinogenic to French (3–101 mg/kg) [54], and Italian (28–304 mg/kg, [23]; humans [65]. Te major use of As compounds is in agri- 9–209 mg/kg, [25] honey. In this study, the mean level of Pb culture and forestry as pesticides and herbicides [66]. Te in honey samples is 24.73 μg/kg, which is still within the levels of As ranged from 1.61 to 21.56 μg/kg with a mean PTWI of Pb for adults (25 μ/kg b.w.) [56]. Te extremely concentration of 5.86 μg/kg (Table 5). Te highest levels of high Pb levels found in Italian honey depicted contami- As were found in the multiforal and milk thistle honey nation caused by external sources or by incorrect procedures (21.56 μg/kg, S4 and 18.20 μg/kg, S8, resp.) while its con- during honey processing as reported by Pisani et al. [23]. centration was lower than LOQ in lavender honey (S1). Also, Vanadium is a natural component of the Earth’s crust, which low concentrations were found in orange tree and thyme is widespread in nature. One of the main sources of envi- honey (1.61 μg/kg, S16 and 1.75 μg/kg, S7, resp.). Our values ronmental pollution by V comes from the combustion of for As were similar to the Libyan values (0.006–0.018 mg/kg) fossil fuels [61]. Te mean value of V content in honey [46] and were below those from Morocco (0.00–0.045 mg/ samples from the thirty-seven locations in west Algeria was kg) [67], Greece, Cyprus, and Egypt (<0.08 mg/kg) [26]. 13.52 μg/kg. Among the honey samples surveyed, camphor Very high As contents have been reported in some honey and thyme honey samples are the richest in V (106.51 μg/kg, samples from France (nd–8.0 mg/kg) [54] and Italy (<25 mg/ S26; 79.73 μg/kg, S11, resp.), whereas rosemary honey kg; 2.8–11.1 mg/kg, resp.) [23, 25], which may be from (0.81 μg/kg, S29) was the poorest one. In this study, the levels 12 Journal of Analytical Methods in Chemistry Figure 2: Dendrogram of analyzed honey samples. overwhelmingly as mineral species. Te uses of these metals environmental contamination. Up 94% of our values were below the PTWI of inorganic As (15 μg/kg b.w.) established are distributed among the chemical, electrical, jewellery, and glass industries. In order to fnd out if these metals have by JECFA [56]. Low concentrations of Co (0.36–21.19 μg/kg) were observed in the investigated west Algerian honey found their way into the west Algerian honey, their levels samples, which were lower than those previously reported were monitored in all the samples considered. In this study, for Moroccan (0.00–1.435 mg/kg) [67], French the results show that Ru, Rh, Pt, and Au were lower than the (0.10–0.23 mg/kg) [53], Spanish (0.015–0.720 mg/kg) [60], limit of quantifcation in any of the tested honey samples. In and Italian (1.0–17 mg/kg and 2.9–30.2 mg/kg, resp.) [23, 25] summary, the levels of heavy metals and metalloids were, honey. Besides, the levels of Co were similar to those generally, low and comparable with the levels in honey from published for honey coming from Greece, Cyprus, and Egypt uncontaminated regions [20, 39, 40], indicating that the west (<0.03 mg/kg) reported by Karabagias et al. [26]. Highest Algerian honey is of good quality. concentrations of Co were found in harmal honey (21.19 μg/ kg, S35), whereas the lowest concentrations were in mul- tiforal and orange tree honey (0.36 μg/kg, S17; 1.02 μg/kg, 3.2. Multivariate Statistical Methods. Te principal com- S16, resp.). Overall, our data indicate that there are low levels ponent analysis (PCA) and hierarchical cluster analysis of Co contamination in the west Algerian honey samples. (HCA) of the 37 honey samples were performed by XLSTAT Tere is currently no upper level of intake established by 2014.5.03 software for the trace elements and heavy metals WHO and JECFA for cobalt. Cadmium is a metallic element shown in Tables 5 and 6 except Ru, Rh, Pt, and Au, which that occurs naturally at low concentrations in the envi- were not detected excluded. Te variance estimate results ronment. It has been added to the environment through (eigenvalues) obtained are presented in Table 9. PCA is anthropogenic activities such as Cd metal production in a very powerful pattern recognition technique that describes industrial processes or by the use of phosphate fertilizers in the variance of a large dataset of intercorrelated variables agricultural soils [68]. Te level of Cd in the west Algerian with a lesser set of independent variables [27]. Analyzing the honey samples ranged from 0.24 to 8.14 μg/kg, which was data in Table 9, it can be observed that PCA account contains lower than that reported for the honey from most of the together for 60.89% of the total variance of the dataset. Te Mediterranean countries. Te levels of Cd of half of them frst component eigenvalues 1.937 and cumulative per- are, however, higher than those from Turkey (<1 μg/kg) as centage is 30.38%, second component eigenvalues 1.607, reported by Altun et al. [20] (Table 8). Moreover, Cd was cumulative 54.33%, third component eigenvalues 1.562, present at a high concentration in thyme honey (8.14 μg/kg, cumulative 54.79% and fourth component eigenvalues S7) and below the limit of quantifcation in multiforal honey 1.209, cumulative 60.89% of the total variance. From Table 9 (S17). In 2005, the Joint FAO/WHO established a PTWI for it can be observed that PCA account contains together for Cd of 7 µg/kg b.w. [66]. It is worth noting that the average Cd 60.89% of the total variance of the dataset. Te frst com- content (1.23 μg/kg) for the examined honeys was lower than ponent eigenvalues 1.937 and cumulative % is 30.38, second the PTWI value. [66]. Ruthenium, rhodium, and platinum component eigenvalues 1.607, cumulative 54.33%, third are part of the platinum-group metals belonging to the component eigenvalues 1.562, cumulative 54.79% and group VIII transition metals. Tey are generally grouped fourth component eigenvalues 1.209, cumulative 60.89% of with gold and silver as precious metal commodities. Each of the total variance. HCA was accomplished to classify the the metals occurs naturally in its native form, and in eco- data according to the botanical origin and mineral content nomically exploitable deposits, the elements occur and to point out the similarity among diferent groups. Te Dissimilarity S22 S25 S14 S6 S27 S15 S21 S1 S8 S10 S23 S34 S12 S7 S16 S20 S11 S13 S3 S2 S35 S4 S30 S31 S18 S33 S5 S36 S24 S32 S19 S29 S26 S9 S37 S17 S28 Journal of Analytical Methods in Chemistry 13 level of similarity in which explanations are joined together a major discriminating strength based on the botanical may be used to create a dendrogram. In the present work, the origin of honey samples. Overall, our results indicated that the west Algerian honey is rich in essential minerals ben- optimal linkage distance level was 410, represented by the dashed line in Figure 2. It represents a relative measure of efcial for human health. Te levels of the trace elements and similarity among analyzed honey samples. From Figure 2, it heavy metals were below the PTWI established by JECFA. can be seen that three groups were formed, which can be Finally, it can be concluded that the west Algerian honey is identifed as follows: eleven samples were clustered in group uncontaminated and therefore represents a good indicator I (S1, S6, S8, S10, S14, S15, S21, S22, S23, S25, and S27); one for monitoring environmental pollution with metals. sample in group II (S34); and twenty-fve samples in group Moreover, the levels of mineral contents are at safe levels for III (S2, S3, S4, S5, S7, S9, S11, S12, S13, S16, S17, S18, S19, human consumption. S20, S24, S26, S28, S29, S30, S31, S32, S33, S35, S36, and S37). It can be observed that group I contains lavender, mild white Data Availability mustard, milk thistle, carob tree, Euphorbia, and Eucalyptus honey types with high mineral contents, which showed Te data used to support the fndings of this study are in- similarities to each other. Group II contains sage honey type. cluded within the article. Furthermore, group III comprises rosemary, thyme, orange tree, camphor, harmal, jujube tree, and many multiforal Conflicts of Interest honey types with low levels of minerals, which correlated to each other. Tese results show that west Algerian honey Te authors declare that there are no conficts of interest samples vary according to their geographic origins and also regarding the publication of this paper. to their mono or multiforal characteristics (Table 1). In addition, it is to underline that clear separation among the Authors’ Contributions samples was obtained from several diferent geographic locations in western Algeria. From the PCA, it can be AH was responsible for conceptualization, project admin- verifed that the characteristics that infuenced honey sample istration, and original draft preparation. AH and RK were clustering the most were major elements K, Na, Ca, Mg, and responsible for methodology and formal analysis. GK was Fe, whereas minor ones such as Cr, Co, V, W, As, Cd, and Pb responsible for software. AA and RK were responsible for were considered less important. In summary, the visuali- validation and data curation. DBR, SB, AA, GK, and RK zation of the data by HCA and PCA ofers enough in- were responsible for investigation. DBR, GK, and SB were formation to develop a classifcation method to determine responsible for resources. SB and RK were responsible for the botanical origin of honey samples considering the review and editing. AH and SB were responsible for visu- mineral element composition. alization. RK was responsible for supervision. 4. Conclusion Acknowledgments Te present chemical study provided a detailed picture of the Tis study was carried out under project no. trace element and heavy metal contents of natural honey B00L01UN130120220004 sponsored by the Ministry of from west Algeria, through the analysis of 19 elements in 37 Higher Education and Scientifc Research of the People’s types of uniforal honey (i.e., lavender, rosemary, mild white Democratic Republic of Algeria and Abou Bekr Belka¨ıd mustard, thyme, milk thistle, carob tree, orange tree, Eu- University-Tlemcen, Algeria. Te authors express their phorbia, Eucalyptus, camphor, jujube tree, sage, and harmal) thanks to the beekeepers who donated honey samples for and multiforal honey. Trace elements and heavy metals were analysis in this study. Also special thanks to Pr. M.E.A. Dib determined and measured by ICP-MS and AAS methods. In for statistical analyses. summary, K was the most abundant element. 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