Article. Determination of Essential, Non-Essential, and Potentially Toxic Elements in Graviola (Annona muricata L.) Pulp by ICP OES and GF AAS

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1 Article A J. Brz. Chem. Soc., Vol. 26, No. 10, , Printed in Brzil Sociedde Brsileir de Químic $ Determintion of Essentil, Non-Essentil, nd Potentilly Toxic s in Grviol (Annon murict L.) Pulp y ICP OES nd GF AAS Gislyne A. R. Kelmer, Angerson N. Nscimento nd Pedro V. Oliveir*, Instituto de Químic, Universidde de São Pulo, São Pulo-SP, Brzil Universidde Federl de São Pulo, Cmpus Didem, Didem-SP, Brzil The determintion of essentil (B, C, Co, Cu, Cr, Fe, K, Li, Mg, Mn, P, S, Se, Si, V, nd Zn), non-essentil (Al, B, Sn, Sr, nd Ti), nd potentilly toxic (As, Cd, nd P) elements in grviol pulp y inductively coupled plsm opticl emission spectrometry (ICP OES) nd grphite furnce tomic sorption spectrometry (GF AAS) is proposed. Smples were digested in closed-vessel microwve oven. The ccurcy of the proposed method ws checked y citrus leves stndrd reference nlysis. Limits of quntifiction (LOQ) were in the rnge of mg g -1 (Ti) to 22 mg g -1 (K) for ICP OES nd mg g -1 (Cd) to 0.62 mg g -1 (Se) for GF AAS. According to the recommended dietry reference intke (DRI), grviol pulp cn e considered good source of B, Cu, K, Mg, nd Mn. For ll nlysed smples the concentrtions of As, Cd, nd P were elow the LOQ. Keywords: fruit, grviol, element, emission spectrometry, tomic sorption Introduction Fruits nd nuts from the vst Brzilin territory, especilly those coming from the north nd northest of Brzil, hve een mrketed nd pplied for different purposes. They cn e consumed, used to prepre foods nd everges, used in different cosmetics formultions, nd in phrmceuticl industry. In generl, the composition of fruits nd nuts is rich in unsturted fts, proteins, fires, sterols, phytochemicls, nd essentil nd nonessentil elements. 1-4 Some of these hve stimulted the interest of the scientific community due to the high concentrtions of essentil elements (e.g., selenium in the Brzil nut) or sustnces with nti-cncer ctivity (e.g., grviol). 5 However, for mny fruits nd nuts the elementl composition remins prcticlly unknown. Besides other nutrients, informtion out elementl composition is importnt to orgnise chemicl composition tles, to ssist the lnce of diets, nd to increse the commercil vlue of these products. 6,7 Grviol (Annon murict L.) is fruit originting from the tropicl regions of South Americ nd Centrl Americ. The fruit hs drk green rind nd romtic white pulp. The incresing demnd for the fruit hs *e-mil: pvolivei@iq.usp.r een ttriuted to its orgnoleptic chrcteristics. These cn e consumed in ntur or used to prepre juices, ice crem, jms, jellies, crems, nd yogurts. 8 Identified in the composition of grviol were lkloids, mides, steroids, flvonoids, muricins, cetogenins, nd neurotoxic sustnces Among these groups of sustnces, the cetogenins hve een highlighted due to the vriety of iologicl ctions, such s cytotoxic, ntiprsitic, nd pesticidl. 12 Grviol hs een widely used in populr medicine nd is indicted for diverse purposes such s scesses, ntiprsites, sthm, hypertension, soothing, nd tumours, nd hs een mrketed in vrious dosge forms (powders, liquids, nd cpsules). There is some exggertion out grviol s power in curing some cncers. However, studies hve een crried out to decipher wht is myth nd wht is relity in this context Recent studies hve demonstrted the ction of the cetogenins s nti-cncerous gents tht inhiit the nicotinmide denine dinucleotide (NADH) oxidse of the plsm memrnes of cncer cells. 5 Inductively coupled plsm opticl emission spectrometry (ICP OES) is multi-element technique tht llows the determintion of mjor, minor, nd trce elements in complex food mtrices. 16,17 When n xil view configurtion is used, the detection limits decrese in comprison to rdil view ICP OES. Actully, the comintion of xil view

2 1982 Determintion of Essentil, Non-Essentil, nd Potentilly Toxic s in Grviol (Annon murict L.) Pulp J. Brz. Chem. Soc. nd chrge-injection device (CID) detector improves the sensitivity for the mjority of elements. However, this sensitivity gin is not enough to determine some elements t trce level, demnding more sensitive techniques. Grphite furnce tomic sorption spectrometry (GF AAS) is well-recommended technique for trce element determintions, hving excellent sensitivity nd selectivity in diverse mtrices. The simultneous GF AAS (SIMAAS) used in this work cn detect simultneously up to six elements t trce nd ultr-trce levels. The simultneous cpilities provide dvntges for the GF AAS, such s reduced cost of nlysis nd incresed nlyticl frequency compred to the nlyticl monoelement techniques. 18 Nevertheless, the cpility of multielement determintion in comprison with the ICP OES, is considerly reduced nd restricted to only six elements. The complex orgnic composition of grviol pulp requires well-prepred smple tretment to gurntee the conversion of the interest elements in free inorgnic forms, voiding chemicl nd spectrl interferences during nlysis y ICP OES nd GF AAS methods. Normlly, the conversion of solid smple in cler solution involves digestion with strong nd concentrted cids, lone or comined with oxidnt regents, such s H 2 O 2. HNO 3 is the cid most commonly used for the digestion of orgnic mtrices due to its simple mnipultion, esy purifiction, nd high oxidtive cpcity, especilly t high pressures nd tempertures. Procedures involving the use of diluted cids in smple preprtion y digestion in closed vessels hve een successfully pplied to otin lower lnk vlues, improving detection limits, reducing costs nd wste, nd providing higher nlyst security. Additionlly, the procedure using diluted cid mixture voids excessive dilution of the finl solution, improving the cpcity to detect low concentrtions of elements in the smple. 17,19 Considering the importnce of grviol s constituent of severl foods nd the fct tht the elementl composition is prcticlly unknown, especilly in micronutrients, the present work proposes methods for the determintion of Al, B, B, C, Co, Cu, Cr, Fe, K, Li, Mg, Mn, P, S, Si, Sn, Sr, Ti, V, nd Zn y ICP OES nd As, Cd, P, nd Se y simultneous GF AAS in grviol pulp, following digestion using diluted oxidnt mixture in closed-vessel microwve oven. Experimentl Instrumenttion An ICP OES, model icap 6300 Duo (Thermo Fisher Scientific, Cmridge, Englnd), equipped with xilly nd rdilly viewed plsm, CID detector, n echelle polychromtor for mesurements from to nm, nd rdiofrequency source of MHz ws used. Argon t % (v/v) (Air Liquide Brsil S/A, São Pulo, Brzil) ws used to purge the opticl polychromtor nd to form the plsm. The smple introduction system ws composed of cyclonic-type spry chmer nd concentric neuliser. The injector tue dimeter of the torch ws 2.0 mm. The instrumentl prmeters selected for nlysis re shown in Tle 1. Tle 1. Instrumentl prmeters for multi-elementl determintions y ICP OES Power / W 1250 Neuliser Spry chmer concentric cyclonic Outer gs flow / (L min -1 ) 12 Intermedite gs flow / (L min -1 ) 0.50 Neuliser gs flow / (L min -1 ) 0.57 Pump rottion / (ml min -1 ) 1 Anlyticl wvelength / nm (xil view) Al (I) Fe (II) Si (I) B (I) K (I) Sn (I) B (II) Li (I) Sr (II) C (II) Mg (I) Ti (II) Co (II) Mn (II) V (II) Cr (I) P (I) Zn (I) Cu (I) (I): Atomic line; (II): ionic line. S (I) A SIMAAS, model SIMAA-6000 (PerkinElmer Life nd Anlyticl Sciences, Shelton, CT, USA), equipped with longitudinl Zeemn-effect ckground corrector, echelle opticl rrngement, stndrd trnsversely-heted grphite tomizer (THGA) tue with integrted pyrolyticlly coted pltform nd solid-stte detector ws used for the simultneous determintion of As, Cd, P, nd Se. An AS-72 utosmpler (PerkinElmer) ws used for tking nd delivering the nlyticl solutions from polypropylene cups to the grphite tue. Argon t % (v/v) (Air Liquide Brsil S/A) ws used oth s protective nd purge gs. The instrumentl prmeters of SIMAA-6000 re shown in Tle 2. The digestion of the smples nd stndrd reference mteril were crried out in closed-vessel microwve oven (Multiwve 3000, Anton Pr, Austri), using diluted oxidnt mixture. This instrument is equipped with 16 fluoropolymer vessels nd cermic vessel jcket tht support mximum temperture of 240 C nd pressure

3 Vol. 26, No. 10, 2015 Kelmer et l Tle 2. Instrumentl operting conditions nd THGA heting progrm for simultneous determintion of As, Cd, P, nd Se y SIMAAS l / nm Lmp I / ma Slit / nm As EDL Cd EDL P HCL Se EDL of 4 MP. The internl temperture nd pressure were monitored continuously in only one controlling vessel, using sensor-protecting glss tue tht enters directly into contct with the digested solution. The externl temperture is controlled in ll vessels using n infrred (IR) sensor tht mesures ech digestion vessel through ports in the rotor se. A knife mill Grindomix GM 200 (Retsch, Hn, Germny) ws used to homogenise the grviol pulp smples for 5 min. Regents nd smples THGA heting progrm Step T / C Rmp / s Hold / s Ar flow / (ml min -1 ) Drying Drying Pyrolysis Atomiztion Clening EDL: Electrodeless dischrge lmp; HCL: hollow cthode lmp; THGA: trnsversely-heted grphite tomizer; injection temperture: 30 C; pipette speed: 100%. All solutions were prepred with nlyticl-regent grde nd high-purity deionised wter, with finl resistivity of 18.2 MW cm, provided y Milli-Q wter purifiction system (Millipore, Bedford, MA, USA). Anlyticl grdes of 65% (m/m) HNO 3 nd 30% (m/m) H 2 O 2 (Merck, Drmstdt, Germny) were used to prepre the oxidnt mixture for smple nd stndrd reference mteril (SRM) digestion. Titrisol stndrd reference solutions contining 1000 ± 2 mg L -1 of Al, As, B, B, C, Cd, Co, Cu, Cr, Fe, K, Li, Mg, Mn, P, P, S, Se, Si, Sn, Sr, Ti, V, nd Zn (Merck) were diluted to prepre the nlyticl reference solutions in 0.1% (v/v) HNO 3. The nlyticl curves were prepred with the following reference solutions: (i) lnk 0.1% (v/v) HNO 3 ; (ii) µg L -1 of Cd, 5-40 µg L -1 of P, µg L -1 of As nd Se; (iii) mg L -1 of Sr; (iv) mg L -1 of Al, B, B, Co, Cu, Fe, Mn, Ti, V, nd Zn; (v) 1-50 mg L -1 of Cr, Li, nd Sn; (vi) mg L -1 of C, K, Mg, nd Si; nd (vii) mg L -1 of P nd S. All these solutions were prepred in 0.1% (v/v) HNO 3. Grviol fruits from Northestern Brzil (Bhi Stte) were purchsed in supermrket in São Pulo City. The used smples were of Crioul vriety, with hert-shped fruit, weighing etween 1.5 nd 3 kg. The fruit hd drk green rind nd romtic white pulp. Three different fruits were used nd 3 su-smples for nlysis otined from ech one. Citrus leves SRM (SRM 1572) from the Ntionl Institute of Stndrds nd Technology (NIST, Githersurg, MD, USA) ws used to check the ccurcy of the nlyticl methods. Smple preprtion Grviol fruits were rinsed with tp nd distilled wter, nd the pulp seprted from the seeds with cermic knife. The rind nd seeds were discrded. The pulp ws homogenised in knife mill for five minutes, dried in n oven t 50 C until constnt mss, nd ground with n gte mortr nd pestle. A mss of ech smple of round 200 mg ws digested using diluted oxidnt mixture (2 ml HNO ml H 2 O ml H 2 O). The heting progrm ws performed in three steps (temperture in C; rmp in min; hold in min): 1 (140; 5; 1); 2 (180; 4; 5); nd 3 (220; 4; 10). There ws fourth step for cooling down the system through forced ventiltion for 20 min. After the digestion, the smples nd lnk solutions were trnsferred to plstic flsks nd diluted to 10 ml with deionised wter. The digestion procedure ws done in triplicte for ech of the su-smples. SRM 1572 ws prepred using the sme procedure descried for the grviol pulp. ICP OES nd SIMAAS opertion conditions The performnce of ICP OES ws evluted using n queous solution of 2.0 mg L -1 of Mg in 0.1% (v/v) HNO 3 to otin roust operting conditions sed on the evlution of the rtio Mg (II) ( nm) / Mg (I) ( nm), s previously proposed in the literture. 20 To otin roust conditions, the pplied rdio frequency power ws vried from 950 to 1350 W, the neuliser gs-flow ws vried from 0.28 to 0.85 L min -1, nd the uxiliry gs-flow ws vried from 0.5 to 1.5 L min -1. The choice of the est wvelengths for element determintions ws crried out y scns in wide rnge, using stndrd reference solutions nd grviol solution fter microwve digestion. The wvelength scns of grviol solution were compred with those otined using nlyticl reference solutions, contining 10 nd 50 mg L -1 of

4 1984 Determintion of Essentil, Non-Essentil, nd Potentilly Toxic s in Grviol (Annon murict L.) Pulp J. Brz. Chem. Soc. nlytes. The higher signl/noise rtio nd non-overlpping spectrl lines of other elements or concomitnt were the criteri used for the selection of pproprite wvelengths (Tle 1). Bckground emission ws corrected using instrument softwre. The verge etween the left nd right ckgrounds ws mesured round the element pek nd used to otin the solutions corrected emission intensity. For ICP OES, the limits of quntifiction (LOQ) were tken s 10 times the limits of detection (LOD). The estimtion of the LOD for ech element ws clculted using the ckground equivlent concentrtion (BEC) nd signl-to-ckground rtio (SBR): BEC = C rs / SBR; SBR = I rs I lnk / I lnk ; LOD = 3 BEC RSD / 100; where C rs is the concentrtion of element in the reference solution, I rs nd I lnk re the emission intensities of the element in the reference nd lnk solutions, respectively, fter ckground correction, nd RSD is the reltive stndrd devition for ten consecutive mesurements of lnk solution. 17,21 The LOD were converted into concentrtion (µg g -1 ), sed on the mss of 200 mg of grviol nd finl volume of 10 ml. The est operting conditions for SIMAAS were estlished in order to determine simultneously As, Cd, P, nd Se. The pyrolysis nd tomiztion curves were otined in the presence of 250 µg of W µg of Rh s the permnent chemicl modifier. 22 The heting progrm for SIMAAS is shown in Tle 2. Volumes of 10 ml of the reference solutions nd smples were delivered onto the integrted pltform of the pyrolytic grphite tue. In this cse, the LOD were clculted considering the vriility of 10 consecutive mesurements of lnk solution, ccording to 3s lnk / (s lnk = stndrd devition of the lnk nd = clirtion curve slope). The LOQ were dopted s 3 LOD. Results nd Discussion Anlyticl performnce of ICP The operting conditions of ICP OES my influence the roustness of the method, which is relted to the ility of the equipment to ccept vritions in the smple mtrix without significntly ltering the nlyticl signls nd, consequently, the results. The rtio Mg (II) / Mg (I) of the intensities of the ionic emission line (Mg II: nm) nd tomic emission line (Mg I: nm) ws proposed to evlute nd find the roustness of ICP OES. A rtio vlue ove 10 indictes tht the ICP OES should e prticulrly roust to mtrix effects. On the other hnd, rtio elow 4 corresponds to high susceptiility to the occurrence of mtrix effects during nlysis. 20 For xilly viewed plsm, the Mg (II) / Mg (I) rtio should e multiplied y fctor ε, clculted from the rtio of the vlues of ckground signls in the two wvelengths (S ckground t nm / S ckground t nm) in order to correct opticl devitions. 23 In this work, the vlue of fctor ε ws set t 1.8. Thus, roust conditions were chieved with power of 1250 W, 0.57 L min -1 of neuliser gs flow, nd 0.5 L min -1 of uxiliry gs flow, with the rtio Mg (II) / Mg (I) equl to The est wvelength nd the optimum instrumentl prmeters re shown in Tle 1. Anlyticl prmeters of the methods Prmeters of the nlyticl clirtion curves, such s liner rnge, correltion coefficient (R 2 ), nd LOQ, re presented in Tle 3. Considering smple mss of 200 mg nd finl volume of 10 ml, LOQ (10 LOD) for ICP OES were in the rnge of mg g -1 (Ti) to 3.9 mg g -1 (B) for Tle 3. Prmeters of the nlyticl clirtion curves y ICP OES nd SIMAAS Liner rnge / (mg L -1 ) R 2 LOQ / (µg g -1 ) LOQ / (mg L -1 ) Al As B B C Cd Co Cr Cu Fe K Li Mg Mn P P S Se Si Sn Sr Ti V Zn Liner rnge (µg L -1 ) for simultneous determintion y SIMAAS; R 2 : correltion coefficient; LOQ: limit of quntifiction.

5 Vol. 26, No. 10, 2015 Kelmer et l microconstituents nd in the rnge of 3.6 mg g -1 (Mg) to 22 mg g -1 (K) for mcro constituents. For SIMAAS, the LOQ (3 LOD) were in the rnge of mg g -1 (Cd) to 0.62 mg g -1 (Se). These vlues re in close ccordnce with others previously otined in our group. 17,22 The comprison etween the experimentl nd certified vlues for ll nlytes in the certified reference mteril (SRM 1572) is presented in Tle 4. To check the concordnce of the results, Student s t-test ws pplied for ll elements, except for B, Co, Cr, Li, Si, Sn, nd V, whose vlues were elow the LOD or were not determined. Where it ws possile to determine elements in the certified reference mteril, the determined vlues re in greement with the recommended vlues t confidence level of 95% (Student s t-test), except for Al, Zn, nd Cr, where the greement ws t confidence level of 98%. l composition in grviol pulp Tle 5 shows the concentrtions of 24 elements determined in the grviol pulp y ICP OES nd SIMAAS. It is importnt to emphsise tht these concentrtions re relted to the nlysis of three different smples nd ech su-smple ws nlysed in triplicte. The humidity content determined fter drying in n oven t 50 C ws round 80%. Among the elements determined in the grviol pulp, K showed the highest concentrtion (3049 µg g -1 ), followed y P (993 µg g -1 ), C (874 µg g -1 ), S (683 µg g -1 ), nd Mg (481 µg g -1 ). The presence of P nd S t high concentrtions cn e relted to the presence of protein, whose concentrtions re round 0.8% (m/m). 24 A reltively high concentrtion of Al (6.7 µg g -1 ), B (8.7 µg g -1 ), B (13.6 µg g -1 ), Si (49 µg g -1 ), nd Sr (6.7 µg g -1 ), were found. These re considered non-essentil elements for humn eings. High concentrtions of B nd Sr were lso found in Brzil nut. The presence of these elements (B nd Sr) in Brzil nut nd grviol my e relted to the mount found in the soil of Northern nd Northwestern Brzil nd the physiologicl chrcteristics of the plnts. 25,26 The vlues found for ll elements nlysed y SIMAAS re elow the LOQ (0.47 µg g -1 for As, µg g -1 for Cd, 0.48 µg g -1 for P, nd 0.62 µg g -1 for Se). These results indicte tht these grviol smples re not contminted with As, Cd, nd P, nd re not good source of Se. The recoveries were otined for dditions of the elements efore digestion in the microwve oven (Tle 5). These good recoveries demonstrte the good performnce of the smple preprtion procedure nd confirm tht the determintions y ICP OES or SIMAAS were performed in the sence of mtrix-effects nd spectrl interferences. The recoveries for ICP OES rnged from 80 to 112% nd for SIMAAS from 102 to 116%, except for Al, tht presented n cceptle vlue of 60%. Even for the SRM 1572 nlysis, the recovery for Al ws not so good (71%). These results re in greement with literture tht report recoveries up to 45% for Al in spinch leves, using xil view ICP OES. 27 Tle 6 shows the content of mcro nd micronutrients in 100 g of edile grviol pulp in comprison with literture dt for this fruit. The K concentrtions found in this work re comprle with the vlues otined using the Brzilin Tle of Food Composition (TACO, Tel Brsileir de Composição de Alimentos). 24 On the Tle 4. Results of the certified reference mteril (SRM 1572, citrus leves) nlysis y ICP OES nd SIMAAS Certified vlue / Found vlue / (mg g -1 ) (mg g -1 ) Certified vlue / Found vlue / (mg g -1 ) (mg g -1 ) Al 92 ± ± 5 Mg 0.58 ± ± 0.12 As 3.1 ± ± 0.2 Mn 23 ± 2 22 ± 1 B ND c 53 ± 3 P 0.13 ± ± 0.03 B 21 ± 3 17 ± 2 P 13.3 ± ± 0.1 C 3.15 ± ± 0.88 S 0.407± ±0.007 Cd 0.03 ± ± Se d < LOQ Co 0.02 d < LOQ Si ND c 214 ± 13 Cr 0.8 ± 0.2 < LOQ Sn 0.24 d < LOQ Cu 16.5 ± ± 2 Sr 100 ± 2 90 ± 9 Fe 90 ± ± 2 Ti ND c 2.9 ± 0.7 K 1.82 ± ± 0.36 V ND c < LOQ Li ND c < LOQ Zn 29 ± 2 23 ± 3 Men ± stndrd devition (n = 3); % (m/m); c ND: not determined; d noncertified concentrtion.

6 1986 Determintion of Essentil, Non-Essentil, nd Potentilly Toxic s in Grviol (Annon murict L.) Pulp J. Brz. Chem. Soc. Tle 5. Determintion of essentil, non-essentil, nd potentilly toxic elements in grviol pulp (n = 3) Grviol pulp / Addition / (µg g -1 ) (mg L -1 ) Recovery / % Al 6.7 ± As < B 8.7 ± B 13.6 ± C 874 ± Cd < Co < Cr < Cu 3.9 ± Fe 8.2 ± K 3049 ± Li < Mg 481 ± Mn 6.9 ± P 993 ± P < S 683 ± Se < Si 49 ± Sn < Sr 6.7 ± Ti < V < Zn 3.3 ± Men ± stndrd devition (n = 3); mg L -1 ; <: elow the limit of quntifiction. other hnd, hlf the concentrtion ws found in the fruits cultivted in Colomi. 28 The P concentrtion is out three times higher thn those previously reported, 28 nd when compred with dt from TACO, this difference is greter. The concentrtions of C nd Mg re pproximtely doule of those found in this work, Mn nd Cu re out 10 times higher, nd Fe nd Zn re closer to those previously reported. 24,28 Severl fctors my contriute to the differences found in this study in comprison with dt previously reported, such s soil composition, climtic conditions, nd differences in genetic nd griculturl prctices. 28 Dietry reference intke (DRI) The reference vlues of nutrient intkes considered dequte in plnning nd ssessing diets for helthy people during dy, is termed the dietry reference intke (DRI). The regultions of the Food nd Drug Administrtion (FDA) estlished nutrition lelling criteri for most food. However, there is no report for grviol pulp. The DRIs considering tolerle upper intke levels per dy for some essentil elements of humn nutrition hve een estlished in the rnge: B (3 to 20 mg), C (1,000 to 3,000 mg), Cu (1 to 10 mg), Fe (40 to 45 mg), K (400 to 5,100 mg), Mg (65 to 350 mg), Mn (2 to 11 mg), P (3,000 to 4,000 mg), nd Zn (4 to 40 mg). 29 These vlues were estlished depending on the life stge of groups of people: infnts, children, mles, femles, pregnncy nd restfeeding. Tking into ccount the concentrtions of the elements shown in Tle 5 nd the mount of consumed grviol pulp s 100 g, the DRIs were clculted for ll elements considered to e nutrients. In this context, the mount of these elements ingested per dy could e 870 mg for B, 87.4 mg for C, 390 mg for Cu, 820 mg for Fe, mg for K, 48.1 mg for Mg, 690 mg for Mn, 99.3 mg for P, 68.3 mg for S, 4.9 mg for Si, nd 330 mg for Zn. For ll these elements, if 100 g of grviol re consumed, the mount of the mcro nd microconstituents re elow the DRIs. The results provide informtion tht cn e useful to evlute the contriution of grviol with mcro nd microconstituents to the recommended dily intke (RDI) vlues for dults. 30 According to the results, grviol pulp cn e considered source of Cu nd Mn, supplying 43 nd 30% of DRI per 100 g of pulp, respectively, s well s good source of Mg nd P, with contriutions from 19 to 14%, respectively. The presence of B s microconstituent should e considered crefully ecuse this element hs essentil roles in humn physiology, 31 however, the dietry reference Tle 6. Content of mcro nd micronutrients in grviol compred with literture dt Composition / (mg per 100 g edile portion) Study K P C Mg Mn Fe Cu Zn : This study; 2: reference 24; 3: reference 28.

7 Vol. 26, No. 10, 2015 Kelmer et l intke vries considerly for children ged 1 to 8 yers (3 to 6 mg per dy) nd for dults (up to 20 mg per dy). For Si there is no DRI due to lck of dt of dverse effects in ge groups from infnts to dults. Conclusions By using ICP OES nd SIMAAS, it ws possile to ccess informtion out the concentrtion of 24 elements in grviol pulp with good precision nd ccurcy. Diluted oxidnt mixture ws very efficient to digest the orgnic mtrix, voiding excessive finl dilution of the smple nd, consequently, loss of sensitivity. The multi-elementl determintion reveled the presence of some elements tht re non-essentil (Al, B, nd Sr) for humn eings. Bsed on the vlues of DRI for infnts, children, mles, femles, pregnncy nd restfeeding, grviol pulp cn e considered good source of B, Cu, K, Mg, nd Mn. On the other hnd, grviol pulp is not source of Se, nd potentilly toxic elements, such s As, Cd, nd P. Acknowledgments The uthors re grteful to the Conselho Ncionl de Desenvolvimento Científico e Tecnológico (CNPq) nd Fundção de Ampro à Pesquis do Estdo de São Pulo (FAPESP) for finncil support. P. V. O. nd G. A. R. K. re thnkful to CNPq for the reserch nd fellowship support. References 1. Vllilo, M.; Tvres, M.; Aued-Pimentel, S.; Cmpos, N.; Moit, N. J.; Food Chem. 1999, 66, Kris Etherton, P. M.; Zho, G.; Binkoski, A. E.; Covl, S. M.; Etherton, T. D.; Nutr. Rev. 2001, 59, Knnmkumrth, S. S.; Wuilloud, R. G; Cruso J. A.; J. Agric. Food Chem. 2004, 52, Rodushkin, I.; Engström, E.; Sörlin, D.; Bxter, D.; Sci. Totl Environ. 2008, 392, Pul, J.; Gnnm, R.; Jydeep, R. M; Arul, L.; Curr. Top. Med. Chem. 2013, 13, Almeid, M. M. B.; de Sous, P. H. M.; Arrig, A. M. C.; do Prdo, G. M.; Mglhães, C. E. D. C.; Mi, G. A.; de Lemos, T. L. G.; Food Res. Int. 2011, 44, Crdoso, L. D. M.; Mrtino, H. S. D.; Moreir, A. V. B.; Rieiro, S. M. R.; Pinheiro- Snt An, H. M.; Food Res. Int. 2011, 44, Scrmento, C. D.; Fri, J. C.; Cruz, F. L. D.; Brreto, W.; Gspr, J. W.; Leite, J. B. V.; Rev. Brs. Frutic. 2003, 25, Augusto, F.; Vlente, A. L. P.; Td, E. D. S.; Rivellino, S. R.; J. Chromtogr. A. 2000, 873, Mohnty, S.; Hollinshed, J.; Jones, L.; Jones, P. W.; Thoms, D.; Wtson, A. A.; Wtson, D. G.; Gry, A. I.; Molyneux, R. J.; Nsh, R. J.; Nt. Prod. Commun. 2008, 3, Sun, S.; Liu, J.; Kdouh, H.; Sun, X.; Zhou, K.; Bioorg. Med. Chem. Lett. 2014, 24, Alli, F. Q.; Liu, X.-X.; McLughlin, J. L.; J. Nt. Prod. 1999, 62, Cssileth, B.; Oncology 2008, 22, Di, Y.; Hogn, S.; Schmelz, E. M.; Ju, Y. H.; Cnning, C.; Zhou, K.; Nutr. Cncer 2011, 63, Brlho, S. M.; Souz, M. D. S. S.; Bueno, P. C. D. S.; Guiguer, E. L.; Frinzzi-Mchdo, F. M. V.; Arújo, A. C.; Meneguim, C. O.; Silveir, E. P.; Oliveir, N. S.; d Silv, B. C.; J. Med. Food 2012, 15, Doln, S. P.; Cpr, S. G.; J. Food Compos. Anl. 2002, 15, Nozuk, J.; Vieir, E. C.; Nscimento, A. N.; Oliveir, P. V.; Food Chem. 2011, 124, Correi, P.; Nomur, C.; Oliveir, P. V.; Anlytic 2003, 5, Arújo, G. C.; Gonzlez, M. H.; Ferreir, A. G.; Nogueir, A. R. A.; Nóreg, J. A.; Spectrochim. Act, Prt B 2002, 57, Mermet, J.-M.; Poussel, E.; Appl. Spectrosc. 1995, 49, 12A. 21. Poussel, E.; Mermet, J.-M.; Spectrochim. Act, Prt B 1996, 51, Nomur, C. S.; Correi, P. R.; Oliveir, P. V.; Oliveir, E.; J. Brz. Chem. Soc. 2004, 15, Stepn, M.; Musil, P.; Poussel, E.; Mermet, J.; Spectrochim. Act, Prt B 2001, 56, ccessed in July Prekh, P.; Khn, A.; Torres, M.; Kitto, M.; J. Food Compos. Anl. 2008, 21, Gonclves, A. M.; Fernndes, K. G.; Rmos, L. A.; Cvlheiro, E. T. G.; Noreg, J. A.; J. Brz. Chem. Soc. 2009, 20, Trevisn, L. C.; Silv, F. V.; Nogueir, A. R. A.; Nóreg, J. A.; Microchem. J. 2007, 86, Leterme, P.; Buldgen, A.; Estrd, F.; Londoño, A. M; Food Chem. 2006, 95, ccessed in July dd53fc4c6735/RDC_269_2005.pdf?MOD=AJPERES ccessed in July Hunt, C. D.; J. Trce Elem. Med. Biol. 2012, 26, 157. Sumitted: Mrch 23, 2015 Pulished online: July 17, 2015 FAPESP hs sponsored the puliction of this rticle.