Physico-Chemical And Vitamin Constituents Of Honey Samples Obtained From Different Agro- Ecological Zones Of Nigeria

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1 Physico-Chemicl And Vitmin Constituents Of Honey Smples Obtined From Different Agro- Ecologicl Zones Of Nigeri Adenekn, M.O. 1, Amus, N.A. 2,Owolde, E.O. 3,Ajetunmobi, T 4., Okpeze, V.E. 1, nd Diro, A.M 1. 1 Federl College of Agriculture, P.M.B. 5029, Ibdn, Nigeri 2 Olbisi Onbnjo University, Deprtment of Plnt Science nd Applied Zoology, Ago-Iwoye, Ogun Stte, Nigeri 3 Federl College of Animl Helth nd Production Technology. 4 Ntionl Horticulturl Reserch Institute of Nigeri, Ibdn Corresponding uthor e-mil: mopdenekn@yhoo.com Abstrct A totl of thirty six honey smples collected from different gro-ecologicl zones of Nigeri were investigted for their physicochemicl nd vitmin contents. The results reveled tht honey hd n verge electricl conductivity (EC) of 1.27mS/cm, moisture content, 25.33%, sh content, 0.63 %, ph 3.82 nd hygroscopicity,1.43. The colour rnged from light green to drk mber. The vlue of hydroxymethyfurl content rnged from ppm while totl cidity rnged from meq/kg. The vlue of 52.95%fructose sugr obtined from the ws significntly different from the vlues of 27.76%nd 28.90% obtined from Mngrove Forest nd Rin Forest respectively. The verge vlues of glucose nd sucrose obtined from Nigerin honey were 26.12% nd 1.10% respectively. The honey smples showed very low verge protein vlue of 8ppm. Potssium (k) hd the highest vlue of 6.34ppm followed by iron (Fe), 6.34ppm while clcium (C) hd the lest vlue of 0.40 ppm in Nigerin honey. Results of the vitmin content showed tht five vitmins were detected (Biotin, Riboflvin, Thimine, Ascorbic cid nd Folic cid) in honey obtined from different gro-ecologicl zones with scorbic cid hving the highest vlue of 0.31 ppm. Keywords Biotin, Hydroxymethylfurfurl, Electricl Conductivity, Nturl Honey, Acidity, Vitmins. INTRODUCTION Honey is nturl energetic food produced by honey bees (Apis mellifer) [32]. It is one of the most widely sought products becuse of its unique nutritionl nd medicinl properties [21]. Nturl honey is sweet substnce produced by honey bees from sugry solution of nectr nd flowers s their source of food in time of scrcity or during hrsh wether conditions. Nturl honey is composed of minly sugr nd wter, other constituents include; proteins, minerls, orgnic cids, vitmins, lipids, esters nd enzymes. Ech of these constituents ws known to hve distinctive nutritionl or medicinl properties nd the unique blend ccount for the vried different pplictions of nturl honeys [21].The precise composition of honey vries ccording to the plnt species on which the bees forge. On the verge, honey is composed of wter (17.2%), glucose (32.16%), fructose (38.4%), sucrose (1.5%), sh (0.18%) nd nitrogen (0.05%) [17]. Wter is quntittively the second most importnt component of honey, its content is criticl since it ffects the storge of honey. According to [14], only honeys with less thn 18 % wter cn be stored with little to no risk of fermenttion. He stted tht the finl wter content of honey depends on number of environmentl fctors during production such s wether nd humidity inside the hive, but lso on nectr condition nd tretment of honey during extrction nd storge. Severl investigtions hve shown tht honey contins vrying mount of minerl substnces rnging from g/100g, the min element found in honey is potssium with bout one-third of the totl minerl element [16][17][17]. The cid content of honey is reltively low, but it is importnt for the honey tste. [30] stted tht most cids re dded by the bees. The min cid in honey is gluconic cid, product of glucose oxidtion by glucose oxidse [22]. The ph of honey vries between due to their higher minerl contents [31]. The mount of mino cids nd protein constituents of honey re reltively smll, t the most 0.7 per cent. Thus, hving reltively smll nutritive effect, however, these components cn be importnt for judging the honey qulity [12]. Honey contins some detectble quntities of vitmins. [13] reported tht honey should not be considered s good source of vitmins s their concentrtions re best described in the prt per million rnges. The chrcteriztion of honeys ids understnding of its properties nd ppliction hence its use s food ingredients in humn diet. There is pucity of dt on Nigerin honeys, wheres there re lrge volume of dt on the chrcteriztion of honeys from North Americ, Europe, Austrli, Indi nd South Afric. JMESSP

2 Therefore, the objectives of this reserch work were to determine the physicochemicl nd vitmin constituents of honeys produced from different groecologicl zones of Nigeri. MATERIALS AND METHODS A totl of thirty-six (36) honey smples from six (6) different gro-ecologicl zones in Nigeri nmely (Mngrove Forest, Rin,, Southern, Northern nd ) were collected in sterile bottles from identified beekeepers. These were kept in the drk cupbord to prevent photodecomposition t mbient temperture condition until they were needed for lbortory nlyses. The honey smples collection were mde in the yer 2008 nd respectively nlyzed for physicl properties, chemicl nd vitmin constituents on the biochemistry lbortory of the Interntionl Institute of Agriculture (IITA) Ibdn, Nigeri. The colour of the honey smples were tested using the fund scle (n officil density reding generlly used in interntionl honey trde). Fifty milliliters (50 ml) of ech honey smples ws mesured into beker nd thoroughly stirred. Twenty milliliters (20 ml) ws mesured nd plced on profound scle in the lbortory. The redings obtined were compred nd the scle tbulted for ech of the honey smple. Electricl conductivity ws determined by mesuring 20 g of dry mtter of honey in 100 ml of ultr-pure wter. This ws thoroughly mixed to form solution. The electricl conductivity cell ws immersed t 20 0 C while the reding ws expressed in Millisiemens per centimeter (ms/cm) [6]. The moisture content ws determined by weighing l0g of honey smples in pre-weighed crucible, which ws then dried t C until constnt weight ws obtined [19]. Ash content ws determined by igniting t C in furnce to constnt mss. The ph ws mesured by ph meter in solution contining l0g of honey smple in 75ml of distilled wter ccording to Assocition of Officil Anlyticl [6]. Hygroscopicity ws determined by mesuring 50 ml of honey smples nd left open in the beker for bout three hours. The difference in volume of the honey smple ws computed from which the hygroscopicity ws determined. The composition of the hydroxymethylfurfurl- dehyde (HMF) of the honey smples ws determined by mesuring ten grmmes (10 g) of unheted honey smple nd dissolved in 20 ml of ultr-pure wter, which ws then trnsferred into 50 ml Volumetric flsk. Two milliliter (2 ml) of ech smple ws introduced into 2 test tubes nd 5.0 ml solution ws dded to ech tube. The blnk ws prepred by dding 1 ml brbituric cid to tube with 1 ml of ultr-pure wter. The bsorbnce of the test smple ws red ginst the blnk t 550 nm wve length using the spectrophotometer (Spectronic 20D model). The HMF ws clculted using the eqution proposed by Winker (1995). HMF (mg/100g) = Absorbnce x 100 Cell pth Length Honey cidity ws determined by the trimetric method. The ddition of 0.05N NOH, ws stopped t ph 8.50 (free cidity), immeditely volume of 10 ml of 0.05 m HCL from 10 ml to ph 8.30 (lctonic cid). Totl cidity ws obtined by dding free nd lctonic cidity [8]. Results were expressed s milliequivlent of cid per kg of honey (meg/kg) [6]. Sugr constituent of the honey smples ws obtined by using the phenol-sulphuric cid method of [2]. 10 ml of ech honey smple ws dissolved in 250 ml of distilled wter in clibrted flsk. This ws stirred thoroughly nd centrifuged to obtin superntnt solution for the nlysis. One milliliter (1 ml) of the diluted solution ws pipetted into test tube nd 1 ml of 52% phenol ws dded to ech test tube. Five mililitre (5 ml) of 96% H 2 SO 4 ws lso dded in drops. The test tube ws then llowed to stnd for 10 minutes before the content ws trnsferred into clen green-free cubettes. Stock glucose ws prepred s stndrd. The vlues of the reducing sugr (Fructose nd Glucose) present in ech honey smples were red on spectrophotometer t wve length of 490 nm. Honey smples were nlyzed for crude protein using the routine Kjeldhl nitrogen method [3]. Ten grmmes (10 g) of the homogeneous honey smple ws weighed into digestion flsk nd dissolved with 10 ml of ultr-pure wter. The diluent ws trnsferred into the volumetric flsk, while Kjeldhl ctlyst tblet (potssium sulphte) ws dded nd thoroughly shken. 20 ml of concentrted H 2 SO 4 ws dded nd fixed into the digester. The flsk ws cooled nd the digest trnsferred into 100 ml volumetric flsk. Five drops of bromocresol (indictor) nd 75 cm of ultrpure wter were dded. Ten milliliters (10 ml) of the digest ws pipetted into the Kjeldhl distilltion flsk nd titrted with 0.05 N of HCL, while percentge totl nitrogen ws clculted using the [3] eqution. The percentge crude protein ws obtined for ll the honey smples by multiplying the totl nitrogen by 6.25 for ech of the honey smple. The honey smples were nlyzed for minerl elementl content by using the tomic spectrophotometer (AAS) nd Flme Photometer ccording to [7]. Ten (10 g) of ech of the smples ws weighed nd dissolved in ultr-pure wter nd mixed thoroughly. The solution ws stirred for 15 minutes on mechnicl stirrer t 1550 rpm. A solution of prloric cid nd nutric cid were dded nd mixed thoroughly. This homogenous solution ws dispensed into the AAS in order to determine the concentrtion of Clcium (C), Iron (Fe) nd Potssium (K) t different wve lengths. A stndrd ws prepred for ech of these elements. The vitmins present in honey smples being nlyzed were determined by titrimetry method in ccordnce with the method of [28] nd [7]..Biotin, JMESSP

3 Riboflvin, Thimine, Absorbic cid nd Folic cid were determined from the honey smples obtined from different gro-ecologicl zones of Nigeri. Dt generted were sttisticlly nlysed, using the Sttisticl Appliction System [29]. The dt were subjected to Anlysis of Vrince (ANOVA) while the mens were seprted with the id of Duncn Multiple Rnge Test (DMRT) t p RESULTS AND DISCUSSION The physicl properties of the smples of honey obtined from different gro-ecologicl zones of Nigeri re summrized in Tble 1. The colour rnged from light green to drk green nd completely drk (Mngrove Forest ). The electricl conductivity (EC) vlues vried from ms/cm. The lowest vlue ws obtined from honey smples from Northern nd ws significntly different from the highest vlue of EC obtined from Mngrove of Nigeri. The EC is good criterion relted to botnicl origin of honey nd this is often used in routine honey control insted of the sh content. The electricl conductivity my lso be explined by tking into ccount the sh nd cid content of honey which reflects the presence of ions nd orgnic cid, the higher their content, the higher the resulting conductivity [18]. The men vlue of 1.27 ms/cmof EC obtined from Nigerin honey is closer to the minimum vlue of 1.95 ms/cm of EC reported for Liptov honey by [24]. The percentge wter content is qulity prmeter which is used to estimte shelf-life, stling rte nd crystlliztion rte of honey smples. The moisture content of honey smples investigted vried from 21.13% in honeys to 30.28% in Mngrove Forest honeys (Tble 1). There ws significnt difference in the vlues of moisture content (28.98 %) of honey obtined from Rin nd the vlues (23.02 %) of moisture content of the honey smple obtined from Northern nd % of moisture content obtined from Southern honeys. The moisture content vrition cn be explined by the composition nd florl origin of honey smples. An increse in moisture content of honey is lso indictive of dultertion. The vlues of sh content vried in the rnge of g/100g. The highest vlue of sh content (0.79 g/100g) ws obtined from honey smples from the Mngrove Forests nd this ws significntly different from the vlues of 0.64, 0.69 nd 0.57 g/100g obtined from Rin, Southern nd Northern respectively (Tble 1). Ash represents the direct mesure of inorgnic residues fter honey crboniztion. The men sh vlue of 0.63 g/100g obtined from Nigerin honey is different from the vlue of 0.50 g/100g obtined from honey from Ibdn s reported by [19]. This vribility in sh content cn be explined by the florl sources of the honey [28]. There ws significnt difference in the ph vlues of honey smples obtined from different gro-ecologicl zones of Nigeri (p 0.05).There ws no significnt difference between the vlue of ph of 3.78 nd 3.74 obtined from the Rin nd respectively but ws significntly different when compred with the vlue of 3.65 nd 4.27 obtined from nd Northern respectively (Tble 1). The men vlue of ph (3.87) obtined for Nigerin honey is closer to the men vlue of 3.96 obtined by [15] for Nigerin honey. The hygroscopicity of honey obtined in Nigeri showed tht it is strong hygroscopic compound nd tht it is n importnt chrcteristic in the processing nd storge of honey. The men vlue of 1.43 of hygroscopicity obtined from honey smples collected, indicted tht honey will bsorb moisture from the tmosphere t reltive humidity of bout 60%. This is in greement with the results obtined for U.S. honeys s reported by [14]. The result of hydroxymethylfurfulrdehyde (HMF) constituents of honey smples obtined from different gro-ecologicl zones of Nigeri reveled trce mounts. The men HMF vlues vried from 0.06 ppm with n overll verge of ppm obtined from Rin nd Southern nd 0.02 ppm obtined from the Mngrove Forest nd the Northern. However, these were significntly different from the men vlues of 0.05 nd 0.06 ppm obtined from the nd respectively, though not significntly different when compred with ech other (Tble 2). The HMF mesures the qulity of honey vi the formtion of 5-hydroxymethylfurfurldehyde by cid hydrolysis of its sucrose with the formtion of red colour. The men HMF obtined from honeys from the vried widely from tht collected from the tropicl rin [25]. This vrition could be s result of the type of the florl source nd level of deteriortion (Crne, 1990). The results of lctonic free nd totl cidity showed tht honey is very cidic. The men vlues of 0.22, nd 24.28meq/kg were obtined for the lctic, free nd totl cidity respectively (Tble 2). The vlues obtined for totl cidity for Nigerin honeys in this study fell within the rnge for Moroccn honey [20]. The cidity of honey contributes to its stbility ginst micro-orgnisms. The sugr found in honey is the source of energy needed for life physiologicl ctivities. Fructose nd glucose sugr re the mjor sugr found in the honeys collected from different gro-ecologicl zones of Nigeri (Tble 3). The men vlue of fructose of 52.95% obtined from honey smples collected from the ws significntly different from 27.76, nd % obtined from honey smples collected from Mngrove Forest, Rin nd Northern respectively (Tble 3). The men glucose vlue of honey collected from Nigeri showed rnge of % indicting tht fter fructose,glucose sugr is the min constituent of honey. The men vlue of % of glucose obtined from the Mngrove Forest ws significntly different from the men vlue of % JMESSP

4 nd % obtined from Rin nd honeys respectively (Tble 3). This result is in conformity with the reserch reported by [26] tht the mjority of sugr in honey re the simple sugrs, fructose nd glucose which represents % of totl sugrs found in honey. Generlly, fructose sugr is greter thn glucose sugr for honeys collected from vrious gro-ecologicl zones of Nigeri. This is in greement with the report of Crne (1990) who reported tht fructose is more bundnt thn glucose sugr constituents in U.S. honeys. The protein content detected in Nigerin honeys is in smll quntities with the highest vlue of 0.05 ppm obtined from the Rin nd ws significntly different from the men vlues of protein of nd 0.04 ppm obtined from Southern nd respectively (Tble 3). This result is in greement with the report of [4] who stted tht honey is not intended s proteinceous food, but contins series of free mino cids necessry for helth-promoting effects in humn being. The minerl elements detected in honeys collected from different gro-ecologicl zones of Nigeri showed tht minerls re in trce quntities, except Potssium (K) nd iron (Fe) which were found to occur in significnt quntities in honeys investigted. The men vlues of 0.40, 5.23 nd 6.34 ppm of Clcium (C), iron (Fe) nd Potssium (K) respectively obtined from honey smples obtined from Nigeri showed tht these minerl elements re found in trce qulities,but significntly different from one gro ecologicl zone to nother. Potssium regultes cid-lkline blnce in the blood nd is involved in the trnsmission of nerve impulses, ctivtes the functions of severl enzymes nd the musculr function of the hert. It lso hs positive effect on the function of skin nd kidney. It is the most bundnt trce element followed by Fe nd C in the honey smples investigted. This finding is in ccord with mny other studies on honey [14] [16] [17] reported tht the min element found in honey is potssium (K) with n verge of bout one-third of the totl composition of minerl elements in honey. Severl investigtions hve shown tht trce elements content of honey depends minly on the botnicl origin of the honey; hence, the different vlues of minerl elements vried from one zone to the other in Nigeri [11] [5]. Honeys obtined from Nigeri contin smll, but detectble quntities of vitmin. The vitmins detected include Biotin, Riboflvin, Thimine, Ascorbic cid nd Folic cid with Ascorbic cid (vitmin C) being the dominnt, followed by Folic cid in honeys investigted. The highest men vlue of biotin of 0.34 ppm obtined from ws significntly different from the lowest vlue of 0.16 ppm nd 0.20ppm obtined from the nd Mngrove honeys respectively. Biotin hs positive effect on these with seborrheic dermtitis. It is very importnt for growth nd developmentl disorders, frgile or splitting finger nils or premture geing of the skin. Riboflvin contributes to metbolism nd helps the uptke of ft, normlizes the functions of the nervous, digestive nd crdiovsculr system. It lso supports the function of the lower gstrointestinl trct (GIT) while Thimine contributes to crbohydrte metbolism, supports the functions of the nervous system, hert nd liver. Ascorbic cid (Vitmin C) contributes to ll metbolic ctivities in nimls. Folic cid lso contributes to the production of red-blood cells nd nucleic cid. It is lso essentil s it supports helthy immune system. However, honey should not be considered s good source of vitmins s their concentrtion is best described in prts per million rnge. This result grees with the report of Bker nd [10] who submitted tht honey contins trce elements of severl vitmins. The men vlue for riboflvin detected in Nigerin honey is closer to tht of the U.S. s reported by [13]. CONCLUSION The results obtined in this study showed tht physiochemicl properties of Nigerin honey compred fvourbly with honey from other countries for the period of investigtion. The colour nd other physicl chrcteristics tested vried slightly from one florl source loction to the other nd from one phytogeogrphicl zone to nother. Drk honey obtined from most of the honeys nlyzed showed tht the honey is rich in minerl constituents s reported by Aubert nd [29]. The most importnt spect of honey colour is in its mrket vlue nd the determintion of its end use. Drk honeys re most often used for industril purposes, while light honey is mrket for direct consumption. Honey is deceptively cidic s its cidity contributes to its stbility ginst micro-orgnisms nd flvor. The results showed tht Nigerin honey is very cidic with the vlues for totl cidity flling within the rnge reported by [20] for Moroccn honey. In this study, honeys obtined from different gro-ecologicl zones in Nigeri were found to hve vrious nutritionl compositions. The fructose nd glucose sugrs were found to be the predominnt sugrs in ll the honeys investigted. These results were in conformity with the rnge vlue requirement of the USDA stndrd for honey. The differences nd other constituents obtined from different groecologicl zones my be due to the composition nd florl origin of the honey. JMESSP

5 Tble 1: vlues ( of physicl properties of honey smples obtined from different gro-ecologicl zones of Nigeri Agroecologicl Colour (ms EC zone cm -1 ) 1.57 Mngrove rin Southern Northern Light green Drk mber Light green Light green Green Drk c b 1.08 e d 1.27 MC (%) b e Ash (%) c 6 b d 3.11 d f c c d 0.63 ph Hygroscopicity d c 4.17 b d with the sme letters in the sme column re EC Electricl conductivity MC - Moisture content Tble 2: vlues ( of chemicl properties of honey smples obtined from different groecologicl zones of Nigeri Agroecologicl zone Mngrove rin Southern Northern Hydroxymethyl furfurl Lctose cidity (m eq kg -1 ) c f d b Free cidity (m eq kg - 1 ) e c d d b Totl cidity (m eq kg - 1 ) e d c e b with the sme letters in the sme column re Tble 3: vlues ( of sugr, protein nd minerl constituents of honey smples obtined from different gro-ecologicl zones of Nigeri Agroecologi cl zone Mngro ve rinfore st Souther n Norther n Fructo Gluco Sucro se (%) se (%) se (%) b b b b b b d c c d Ns Prote in (ppm ) Ns Clciu m c b b 0.33 e 0.35 d 0.40 Iron (pp m) c e d d b 5.23 Potssi um f c b e d 6.34 with the sme letters in the sme column re Tble 4: vlues of vitmins ( obtined in honey smples obtined from different gro-ecologicl zones of Nigeri Agroecologicl zone Mngrove rin Southern Northern Biotin d 0.12 d Riboflvin c Thimine d Ascorbic cid d Folic cid with the sme letters in the sme column re REFERENCES [1] A. E. Jeffrey nd C. M. Echzrret. Medicl uses of honey. Rev. Blomed, 7: JMESSP

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