DETERMINATION OF HEAVY METALS IN MEAT, INTESTINE, LIVER,EGGS, AND CHICKEN USING NEUTRON ACTIVATION AND ATOMICABSORPTION SPECTROMETRY

Surtipanti S*., Suwirma S*., Yumiarti S*., and Yune Mellawati*

ABSTRACT

DETERMINATION OF HEAVY METALS IN MEAT, INTESTINE, LIVER, EGGS,AND CIIICKEN USING NEUTRON ACTIVATION AND ATOMIC AIJSORPTION SPEC­TROMETRY. The elements As, Cd, Co, Cr, Fe, Hg, Ni, Pb, Sb, Se and Zn in meat, intestine, andliver of cow and goat, as well as in broilcr, local breed chicken and eggs had been determined usingNeutron Activation Analysis and Atomic Absorption Spectrometry. The mercury was dctermined aileI'bcing scparated using radiochemical separation. The results showed that concentration of the essentialelements studied i.e. Cr,Cu,Fe,Zn,Co, and Ni were higher in liver and intestine than in the meat, butstill in the normal range, while the toxic elcmennts As, Cd, and Ph were undectable in all samples.

ABSTRAK

PENETUAN LOGAM BERAT DALAM DAGING, t,JSUS, HATI, AYAM DANTELOR, DENGAN ANALISIS AKTIVASI DAN SI'EKTROMETER SERAPAN ATOM. Pen en­tuan logam As, Cd, Co, Cr, Cu, Fe, Hg, Ni, Sb, Se dan Zn dalam daging hati dan usus kambing dansapi, serta daJam, ayam broiler, ayam kampung, dan telor telah dilakukan dengan menggunakan anali­sis aktivasi neutron instrumental dan spektrometer serapan atom. Hg ditentukan setelah dipisahkandengan cara pemisahan radiokimia. Hasil analisis menujukkan bahwa logam esnsial Co, Cr, Cu, Fe,Ni, dan Zn dalam contoh hati dan usus kadarnya Icbih tinggi dibandingkan dalam daging, tetapi masihdalam batas normal, sedangkan logam berat beraclln As, Cd, dan Pb tidak terdeteksi dalam semuacontoh.

INTRODUCTION

Metallic elements are found in all living organisms where they playa varietyof roles. They may be structural, components of control mechanisms, (e.g. in neversand muscles). and above all, enzyme activator, or components of redoxs systems.Some metals are essential (Co, Cr, Cu, Fe, Ni, Zn). and the others are nonessential(As, Cd, Hg, Pb). The deficiency of essential elements result in impairment ofbiological function, but when present in excess, essential elements may become atoxin as well. Non-essential elements, when enter into the body will cause toxiceffects. The major route of entry of most elements into the body is through the diet,

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The source of metals in the invironment are the combustion of foss11 fuels,

rnininl indu~tric~,w~~tudi~DmrrLrrnddomntic ~rWrrYL.frrrrniny andforntry Jimcontribute to the metal content in the environment because of ferlizers and pesticideused (l). Meat (Iamb, beef), intestine, liver, eggs, and chicken are some kind offoodstuffs, in which the metals present are mainly originated from the feed eaten bythe animals.

The objective of this investigation is to know whether the level of metallicelements in meat, liver, intestine, eggs, broiler, and local breed chickens for humanconsumption have exceeded the maximum permissible level specified in internationalrule.

The interesting elements to be analysed are As, Cd, Co, Cr, Cu, Fe, Hg,Ni, Pb, Sb, Se, and Zn. The Cd, Cu, Ni, and Pb were determined using atomicabsorption spectrometry while the others using neutron activation analysis. Themercury (Hg) was determined after being separated from selenium (Se), usingradiochemical separation (2-4).

MATERIALS AND METHODS

Samples and Standard Preparation. The samples analyzed in this studywere meat (Iamb, beef), intestine, eggs, broiler, and local breed chicken. All thesamples were obtained from the local market in Jakarta. During the study about 63samples were collected, consisting of about 1-2 kg each. The samples were broughtto the laboratory in plastic bags. There were cleaned and washed with demineralizedwater as before cooking, then cut with a stainless steel knife, and dried in an oven at

65°C for 48 hou,rs discontinuously until the weight constant. The moisture contentsof the samples are shown in Table I. The dried samples were then powdered. About200 - 500 mg of dried samples were put in polyethylene vials. The standard wasprepared by dropping 5ul aliquot of freshly prepared standard solution (1000 ppm)of each elements on filter paper Whatman 42 (5). The vials containing the samples,standard, and reference standard were put in aluminium container. The referencestandard used was orchad leaves NBS SRM-1571 and fish flesh MA-A-2 (TA)obtained from IAEA.

11lslrultlmlal Neulron Acfivalioll Analysis (lNNA). The container contain­ing samples and standars were irradiated for 36 hours in. TRIG A MARK IIREACTOR, at flux 10"n cm-2 sec-I. After irradiation, the containers were cooled

for 2 days for short half life radionuelide, and 2 weeks for long half life radionu­elide. The samples and standard were counted using a 4096 channels Nuclear Data

62 Multi Channel Analyzer, provided with a high pure Germanium (Ge-Hp) detec­tor. The system has resolution FWHM 3,75 keY for the 1332 keY peak of 6()Cogamma ray. The counting time was 900 seconds. Nuclear reaction of the elements

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analyzed using INNA, namely As, Co, Cr, Fe, Hg, Sb, Se, and Zn can be seen inTable 2 (6).

Radiochemical Separation. The irradiated samples were digested withHN03 in a distilling flask, and heated until white fume evolved. For complete diges­

tion of organic materials, 30 % H202 was added. After cooling to room tempera­ture, the solution was diluted to 20 ml, the pH of the solution was 2-4. The samples

solution was added with 1 mg of Hg as carrier, and 4 ml of 8 x 10-4M Pb(DDC)2solution, and then shaked for 10 minutes. The organic phase containing Hg(DDC},

was separated and counted using a Multi Channel Analyzer at gamma ray energy of279 keY for 203Hg.

Determination by Atomic Absorption Spectrometry. About 5-10 g of driedsamples were put in a distilling flask and digested with a mixture of concentrated

H2S04 and HN03 (1 : 3 v/v). A small amount of 30 % H202 was added to eliminatenitrogen dioxides. The digestion process was continued until the solution becameclear. The samples were transfered into volumetric flask and diluted to 50 ml withdemineralized water. Lead, cadmium and copper were excited using oxygen aceth­ylene flame. The absorption of the elements were compared with the standardabsorption.

RESULTS AND DISCUSSION

The results of determination of As, Cd, Co, Cr, Cu, Hg, Ni, Pb, Se, and Znin all samples analyzed are shown in Teble 3. The accuracy of the method of ana­lyzed used was compared to IAEA standard reference materials orchad leaves SRM­1571 and fish tlesh MA-A-2(TA) (Table 4).

The photopeaks of SICr at 320 keY, 7SSe at 264 keY, 76As at 559 keY, and60Co at 1173 keY were able to determine without any interference from other iso­topes.

The Hg was determined without any interference after being separated fromselenium by radiochemical separation. The photopeak of 203Hg was determine at 279keY.

The elements As, Cd, and Pb were undetectable, as their concentration were

lower then the detection limit (Table 5) while Ni was found only in few samples,i.e. in liver and intestine of cow and in local breed chicken. The other elements, i.e.Co, Cr, Cu, Fe, Hg, Sb, Se, and Zn were found in all samples.

The essential elements Co, Cr, Cu, Fe, and Zn were found higher in liver,intestine, and local breed chicken. These elements usually accumulate in liver,kidney, muscle, and other organ in intestine, because liver has a function to transferthe elements to the whole body (7).

Chromium is also an essential trace elem~nt like Zn and Cu. Cr is required

by mammalian. The main physiological role of this element is a cofactor for insulin

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at the insulin responsive cell membrane (7). The concentration of Cr in various food

Is generally ~JJlgJg wet we~ght (I), while the concentration of Cr in the samplesanalyzed were 0.035-0.45 Jlg/g wet weight.

Cooper, is mainly stored in liver, kidney and muscle molecular weightprotein and ceroplasmin, which is produced in the liver, and also function as anoxidase. The concentration of Cu in food is generally 20-50 Jlg/g wet weight (8),while the concentration of Cu in samples analyzed were 0.09-26.85 Jlg/g wetweight.

Iron concentrations in kidney, liver, beef, ham, and egss are generally 30­150 mg/kg, wet weight (I), while the concentration of Fe in the samples analyzerwere 5.33-37.24 Jlg/g wet weight.

Zinc is necessary for the function of various mammalian enzymes. The.concentration of Zn in food is around 10-50 Jlg/g wet weight (8), while the concen­tration of Zn in samples analyzed were found to be 0.86-27.64 Jlg/g wet weight.

Cobalt in man is required in the form of Cobalt-containing vitamin B12. Cois widely distributed in the animal organ in relatively high concentration (0.10-0.25Jlg/g) in liver, kidney, bone, spleen, and other glandular tissue (7). The concentra­tion of Co in the samples analyzed were still in the normal range. The higher Coconcentration in eggs and broiler is probably originated from the feed of the poultry.

Nickel has been proved to be essential element for poultry, pig; and ratunder experimental conditions. Ni is poorly absorbed from ordinary (7). AbsorbedNi will be accumulated in liver, kidney, and lung (I). Data of Ni in foodstuffs arescarce.

Selenium is an essential element for sheep, cattle, and poultry. Deficiency ofthis element will cause a serious problem. The highest concentration of Se in animalare found in liver, kidney, brain, and muscle. The concentration of Se in foodproducts is influenced by its origin and processing. Se may be found in seafood,kidney, meat, and sometimes it is found in rice and grain. The concentration of Sein the samples analyzed were < 2 Jlg/g wet weight.

Antimon is considered as a non-essential element, and usually it is depositedin kidney and liver. The data of concentration of Sb in foodstuffs are scarce (I). Theconcentration of Sb in the samples analyzed were higher in liver, intestine than inthe meat.

CONCLUSION

The concentration of toxic elements and essential elements in beef, lamb,

liver, intestine, chicken, and eggs samples analyzed were lower than the respectiveMPC's. Therefore, it can be concluded that the samples analyzed are relatively notcontaminated by the toxic and essential metallic elements. The concentration of Co,Cr, Cu, Hg, Sb, Se, and Zn were still in natural normal range.

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ACKNOWLEDGEMENT

The study is a part of work supported by IAEA under Research ContractNo. 4494/R3/RB. The outhors express their thanks to Dr. Haryoto, Miss TiniTjintarsih ans staff of PPTN Bandung for irradiating the samples. We are alsograteful to Mr(s). Suripto and Maryoto for their technical assistance.

REFERENCE

I. FRIBERG, L., GUNNAR, F., NORDBERG, and VELIMER, B. Handbook onToxicology of Metals, Elsevier/Northe Holland Biomedical Press (1979).

2. HANI, N.M., WAI, C.M., and WILLEM, S.H., Dithiocarbamate extraction oftrace amounts of selenium from biological samples for neutron activation analy­sis, Journal of Nuclear Chemistry 104 I (1986) 19.

3. LO, J.M., WEI, J.C., YANG, M.H., and YEH, S.J., Preconcentration ofmercury with lead diethyldithiocarbamat for neutron activation analysis ofbiological and environmental samples, Journal of Radioanalytical Chemistry 721-2 (1982) 57.

4. YUMIARTI, S., MELLAWATI, Y., and SURTIPANTI, S., "Penentuan logamberat dan beracun dalam contoh ikan dengan analisis pengaktifan neutron".Reaktor Nuklir dan Penggunaannya (Ris. Seminar Bandung, 1988), PPTN ­BATAN, Bandung (1988).

5. AHMAD, S.M., CHAUDHARY, M.S., MANNAN, A., and QURESHU, I.H.,Determination of toxic elements in tea leaves by instrumental neutron activationanalysis, Journal of Radioanalytical Chemistry 78 2 (1983) 375.

6. CROUTNAMEL, C.E., Applied Gamma Ray Spectrometry, Pergamon Press(1970) 733.

/~. IAEA, Elemental Analysis of Biological Materials (Technical Report Series No./ 197), IAEA, Vienna (1980).8. REILLY, c., Metal Contamination of Food, Applied Science Publisher, London

(1980).

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0000

Table 1. Moisture contents of the samples

samples Moisture contents,%

Cow

Meat 75Liver

70Intestine

78

Goat

Meat 75Liver

70Intestine

82

Poultry

Broiler 74Local breed chicken

80

Eggs

2

Table 2. Nuclear data for the elements investigation

Elements

Nuclear reactionEnergy,keYTl/2

Se

74 75264128.4days202se (n,D)20~e

Hg50Hg (n'~)51 Hg

27946daysCr

75cr (n,'0) 76cr32027.8days

AsAs (n,'OJ ~s

55926.6hoursSb

121Sb (n 6)12 Sb5642.76 days64 '65 Zn 59zn (n'~)60Zn1115245days

CoCo (n,~) Co11735.2years

00\0

Table 3a. Concentration of heavy metal element in meat, intestine pnd liver of cow,

goat, and in broiler, local breed chicken end eggs in ~g/g wet weight

Sarrple CoCrCuFeHg-- Cow :Meat

0.12 :!:0.030.16:!:0.140.44 :!:0.337.59 :!:4.700.001:!:0.0005

Liver

0.13 :!:0.030.25 :!:0.0810.64 :!:6.4616.64 :!:6.460.002:!:0.0006

Intestine

0.10 :!:0.040.10 :!:0.060.25 :!:0.0825.82 :!:6.140.0004 :!:0.0002

Goat :

Meat

0.15 :!:0.040.08 :!:0.030.31 :!:0.1823.11:!: 5.11Ud

Liver

0.16 :!:0.040.13 :!:0.0526.85 :!:9.7737.24 :!:3.290.0012 :!:0.0006

Intestine

0.13 :!:0.040.45 :!:0.020.27:!: 0.1917.72:!: 1.580.0010 :!:0.0002

Poultry:

Broi ler

0.86 :!:0.030.04 :!:0.010.13 :!:0.075 .33 :!:1.710.0005 :!:0.0002

Local breed

0.12 :!:0.020.13 :!:0.030.17 :!:0.0718.84 :!:5.300.0006 :!:0.0001

chicken Eggs

0.71 :!:0.090.17 :!:0.061.02 :!:0.5318.82 :!:7.800.0080 :!:0.0020

Ud = undetectable

The data ~epresent average of 5 replications:!: so

~

\0o

Table 3b. (Continue)

Saq>le

NiSbSeZn-- Cow :Meat

Ud0.02 :!: 0.010.09 :!: 0.0310.35 :.2.53liver

0.16:.0.130.03 = 0.010.09 :.0.0412.83 = 5.88Intestine

2.96:. 1.040.04 = 0.010.11 = 0.062.97 = 1.04

Goat :

Meat

Ud0.03 = 0.010.88 = 0.039.81 = 5.43liver

Ud0.53 = 0.300.06 = 0.0212.67 = 6.63Intestine

Ud0.01 = 0.000.05 = 0.031.97 = 0.41

Poultry:

Broiler

Ud0.10 = 0.060.05 = 0.030.86 :.0.40local breed

0.10 = 0.060.01 = 0.000.06 = 0.032.17 = 0.55chicken Eggs

Ud0.06 = 0.040.36 = 0.1727.64 =10.29

Ud = undetectableThe data represent average of 5 replications = SD

\0

Table 4. Result of analysis compared to certified value ofbiological samples NBS.SRM-1571 (Orchad leaves) andMA-A-2 (TA) (Fish) in ~g/g

NBS SRM-1 571MA-A-2(TA)Elements Certified

Result ofCertifiedResult ofvalue

analysisvalueanalysis

Se

0.090.11 ± 0.011.701.07±0.17

Hg

0.1550.21 ± 0.020.470.62±0.02Cr

2.303.09 + 0.131.301.44±0.16

12.49 :± 4.97

.As 14.30 2.602.46+1.90

Sb

--0.0050.006 ±0.0005Fe

260 ± 0.6300.00 ± 2054.0047.34± 15.72Pb

--0.58*1.1±0.10Cu

--4.00*2.7±0.25Co

--0.080.05±0.005Zn

25 ± 315.29 ± 4.2333.0025.02±2.76Cd

--0.07*0.13±0.01

* The elements were determined using AAS

\0N

Table 5. Limit of detection (LOD) for the elements analyzed

Elements LOD(/Jg/g) Method--As 0.8N.A.A.Cd

0.2A.A.S.Co

0.2N.A.A.Cu

0.5A.A.S.Cr

0.1N.A.A.Fe

1.0N.A.A.Hg

0.05N.A.A.Ni

1.0A.A.S.Pb

2.0A.A.S.Sb

0.1N.A.A.Se

0.1N.A.A.Zn

0.2N.A.A.

Elements Food**

As

1Cd

1Hg

0.03Pb

1 -55e

2Cu

20 - 50Sb

1Zn

40 - 50

\0W

Table 6. The maximum permissible concentration of elements infoodstuffs for human consumption*, in ~g/g

)JfJ~

j * Food in general excluding fish, vegetables, and beverage~ ** Other similar legislation exists in many countries also for

baby foods, animal feedstuffs, sewage sludge for agricultur­al use, and air particulates

I1•

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