IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (3) 2011 Evaluation o f Copper , Zinc , Manganese , and Magnesium Levels in Newborn Jaundice in Baghdad E. J. Hasan Departme nt of Science, College of Basic Education, Al- Mustansiriah Unive rsity Abstract Trace elements concentrations are frequently reported to be a good indicator for diagnosis and p rognosis of some disease. The levels of copp er, zinc, man ganese, and magnesium wer e measured in seru m of two group . Group one contains (52) newborn jaundice p atients while the second group contains (20) healthy newborn used as controls. In this st udy the concentration of Cu, Zn, M g, and M n were measured using atomic absorption sp ectrop hotometer (AAS). It has been found that z inc level is sign ificantly lower in newborn jaundice patients comp ared with normal subjects at (p <0.05) , also the level of Cu, M g, and M n were significantly high er in newborn jaundice compared with control group . Keywords: Jaundice; Zinc; Cop p er; M agnesium; M anganese Introduction Jaundice occurs when the b lood contains excessive amounts of bilirubin, the dep osition of this high ly insoluble substance color ed the skin and the whites of the eyes y ellow. Jaundice is p robably the most common sy mptom in neonatal p eriod[1,2]. Temp orary jaundice is observed dur ing the first week of life in app roximately 60% of term infants and 80% of p reterm infants[3,4]. Trace elements are necessary for normal function and are therefore associated with morbid deficiency st ates. Those elements are found in body tissues at levels in the micrograms p er gr am of tissue or less. So me elements are essential trace elements (like Fe and Zn) and some are not essential (like Cu and M n) but have well defin ed evidence in human metabolism[5]. Trace elements are essential micronutrients for growt h, development, and maintenance of healthy tissues. The role of trace elements in body metabolism is of p rime imp ortance. Their deficiency causes diseases, whereas their p resence in excess may result in toxicity to human life [6]. Zinc (Zn) is essential component of many metalloenzy mes involved in v irtually all asp ects of metabolism. Zinc is an integral comp onent of nearly 300 enzy me in different sp ecies of all p hy la[7,8]. Important zinc-containing metaloenzy mes in humans including carbonic anhydrase , alk aline p hosp hates, RNA and DNA p oly merases, thy midine kinase, carbo xy p eptidases, and alcohol d ehydrogenase. Zinc st abilizes the st ructures of p roteins and nucleic acids, p articip ates in transp ort p rosses[9]. Copp er (Cu) may p resent in biological IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (3) 2011 system in both the +1 and +2 valence st ates, the major functions of copp er metalloproteins involve oxidation –reduction, most known copp er – containing enzy me bind and react dir ectly with molecular o xy gen[10]. Cop p er is an integr al comp onent of many metalloenzy mes, includin g ceru loplasmin, cytochrome c o xidase, sup eroxide d ismutase, dop amine – β– hy droxy lase, ascorbate oxidase, ly sy l oxidase, and ty rosinase[11,12]. M agnesium (M g) cataly zes or activates more than 300 enzy mes in the body . M agnesium acts as an essential cofactor for enzy mes concerned with cell r esp iration, glycoly sis, and trans-membrane transp ort of other cations such as calcium and sodium[13]. M agnesim can effected enzy me activity by bindin g the active site of the enzy me (p y ruvate kinase, enolase), by ligand bindin g (AT P- requiring enzy me), by causing conformational chan ges during the catalytic p rocess (Na- K- ATPase), and by p romoting aggregation of multienzy me comp lexes[7,14]. M anganese (M n) is p resent in biological sy stems bound to p rotein in either the +2 or +3 valence st ate. M anganese is associated mainly with the formation of connective and bony tissue, with growt h and reproductive functions, and with carbohy drate and lip id metabolism[15]. The bioch emical b asis for man ganese essentiality is its function as a constituent of metalloenzy mes and as an enzy me activator[16,17]. Important manganese containin g enzy mes include arginase, p y ruvate carboxy lase, and man ganese sup eroxide dismutase in mitochondria[18]. Material and Method All blood samples were collected from the Ibn- Albalady hosp ital in Baghdad. The first group consisted of 55 blood samples of jaundice newborns (32were male and 23 females) , their ages r anged from 1-3 days. T wenty healthy volunteers were used as a second group and their ages were matched with p atients group . Blood samples are drown using 10ml sy ringes with st eel needles. A3ml b lood sample was drown from each p atient. The whole blood was immed iately transferred to p lain tube. These samples were allowed to st and at room temp erature for 10 minutes for clotting, the clots were sep arated from the wall of the tube using wooden app licator st ick. The tube was centrifuged for 10 minutes at 3000(r.p .m). The serum was then transferred to a second tube using micropipette and stored at -20 C 0 until the day of analysis. At omic Absorp tion Sp ectrop hotometer (AAS) model AA-6200 (shimadzu _ Jap an) fitted with air – acety lene flame wer e used for the determin ation of Cop p er, Zinc, M anganese, M agnesium. 1- Preparation of standard soluti ons for AAs measurements cpper Copp er stock solution (1000pp m) was p rep ared by dissolving (0.1 gm) of copp er metal in 5ml of 5 M nitric acid, and then diluted to 100ml with deionized water. Zinc Zinc st ock solution (1000pp m) was p rep ared by dissolving (0.1 gm) of zinc p ellet using concentrated hy drochloric acid, and the volume was comp leted into 100ml with deionized water. IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (3) 2011 Magnesium M agnesium st ock solution (1000pp m) was p rep ared by dissolving (0.1 gm) of magnesium metal in 5ml of 5 M nitric acid, and then diluted to 100ml with deionized water. Mangane se M anganese st ock solution (1000pp m) was p rep ared by dissolving (0.1 gm) of man ganese metal in 5ml of 5 M nitric acid, and then diluted to 100ml with deionized water. 1- Calibration curv e for A.A.S measurements 1- The st andard solutions for copp er ion were p repared by subsequent dilution st ock solution (1000ppm) of copp er and diluted to (10, 5, 1, 0.5, 0.1 pp m). 2- Iron st andard solutions were p rep ared by subsequent dilution stock solution (1000pp m) of iron to (10, 5, 1, 0.5,and 0.1 pp m). 3- The st andard solutions for magn esium were p repared by subsequent dilution st ock solution (1000ppm) of magnesium ion and diluted to (100, 50, 10, 5,and 1 ppm). 4- The standard solutions for manganese ion were p repared by subsequent dilution st ock solution (1000ppm) of manganese and diluted to (20, 15, 10 pp m). All above standard solutions were diluted to 100ml in a vo lmatric flask with deionized water. Result and Discussion A linear calibration curve was obtained extended from (0.1- 10) p p m for copper, zinc, man ganese, and magn esium as shown in the (1,2,3,4) figures The results were shown that there was significant difference in the levels of zinc, copp er, manganese, and magnesium in newborn jaundice and control group , at p value of (< 0.05 ), Zinc levels was significantly lower in jaundice p atients (0.0012 ± 0.002) comp ared with the healthy newborn ( 0.0021 ± 0.0036) . Cop p er levels was significantly higher (0.198 ± 0.341) in jaundice p atients comp ared with control group (0.238 ± 0.411). M agnesium levels was significantly higher in p atient (1.48 ± 2.96) than the control group (1.293 ± 1.829). M anganese levels was app eared to be significantly higher (0.0277 ± 0.0391) in jaundice p atients comp ared with control (0.015 ± 0.031). As shown in table (1) . Zinc and magn esium are very imp ortant co-factors for a lot of enzy me sy stems and p lay an imp ortant role in the sy nthesis of nucleic acids [19,20]. Zinc p revents the lip id depolarization of the cell membran es and hyp ozincemia may modulate the erythrocyte membran e. It may result in deficient sy nthesis of assorted enzy mes that p lay a role in the bilirubin metabolism[21], Hyp ozincemia may also cause st ructural defects in the ery throcyte membran es, resulting in hemolysis [22]. An elevated level of serum copp er is documented in liver d isease like cirrhosis , obst ructive jaund ice and cholestasis [23]. In newborn with jaundice , the high serum copp er may be of intracellular (erythrocy te ) origin [24]. M anganese absorbed from food is avidly taken up by the liver and d istributed to tissues bound to transferring and albumin[5].The serum levels of magnesium and man ganese beco me high , depending on the existence of mild hemo lysis in newborn , the fact that these elements are high d epend on being high in erythrocy tes, esp ecially reticulocytes than in serum concentrations (25), and may depend on passing into t he serum as a result of hemolysis. IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (3) 2011 Re ferences 1. Knudsen, A.( 1996). Prediction and non-invasive assessment of n eonatal jaundice in the term healthy newborn infant. Acta Paediatr.85(4):393-397. 2. M artin, D.W.; M ay es, P.A. ;Rodwell, V.W. (1981). Harp ers review of biochemistry . 18 ed, M aruzen asia pte. Ltd. Singapore . p(319). 3. Phy llis, A.D.; William, D.R. and Davit, K.S.( 1995). Neonatal jaundice-what now? Clin Pediatr . 34(2):103-107. 4. Jeffrey M aisels, M. (2006). What’s in a Name? Phy siologic and Pathologic Jaundice: The Conundrum of Defining Normal Bilirubin Levels in the Newborn. Pediatrics 118(2): 805-807. 5. Rahil-Khazen, R.; Bolann, B.J. and My king, A.(2002). M ulti-element analysis of trace element levels in human autop sy tissues by using inductively coupled atomic emission sp ectrometry technique (ICP-AES). J T race Elem M ed Biol 16(1): 15–25. 6. Hashmi,D. R.; Ismail, S. and Shikh, G.H. (2007). Assessment of the level of trace metals in commonly edible vegetables locally available in the markets of Karachi city . Pak. J. Bot. 39(3): 747-751. 7. Berthold, K.; Olivier, G. ; Joanne,H.; Kathrin, K. and Raanan, S.(2005). Iron, M inerals and trace elements. Journal of pediatric Gastroenterology and Nutrition. 41(2):S39-S46. 8. M ansi, K. ; Aburjai, T.; Barqawi, M . and Naser H. (2009). Copp er and Zinc st atus in Jordanian p atients with β-Thalassemia major treated with defero xamine. Journal of Biolo gical Sciences 4(5): 566-572. 9. Zlotkin, S.H. and Buchanan, B.E.( 1988). Amino acid intake and urinary zinc excretion in newborn infants receivin g total p arenteral nutrition. Am J Clin Nut r. 48(2):330–334. 10. Ostermeier, C.; Iwata ,S.; M ichel, H.( 1996). Cytochrome c oxidase. Curr Op in Struct Biol. 6(4) :460–466. 11. Ashraf, S.; Javed,M , T. ; Abbas, N.; Butt, M . A. and Hameed ,S. (2001). Levels of serum Cu, Fe and Zn along with serum total p rotein and fractions in diseased malnourished children of 4-12 y ears of age. M edical Journal of Islamic Academy of Sciences 14(1): 25−33. 12. Schulp is, K. H.; Karakonstantakis, T.; Gavrili, S.; Cost alos, C. ; Roma, E. and Pap assotiriou I. (2004). Serum Copp er Is Decreased in Premature Newborns and Increased in Newborns with Hemolytic Jaundice. Clinical Chemistry 50(7): 1253-1256. 13. Sanders, G.T .; Huijgen, H.J. and Sanders, R.( 1999). M agnesium in disease: a review with sp ecial emphasis on the serum ionized magnesium. Clin Chem Lab M ed 37(11):1011-1033. 14. M aggioni, A.; Orz alesi, M .; M imouni, F.B. (1998). Intravenous correction of neonatal hyp omagnesemia: effect on ionized magnesium. J Pediatr 132(4): 652–655. 15. Fok, T.F.;Chui, K.K.; Cheung, R. et al.( 2001). M anganese intake and cholestatic jaundice in neonates receivin g p arenteral nutrition: a r andomized controlled st udy . Acta Pediatr. 90(9):1009–15. 16. Watts ,D. L. (1990). The nutritional relationships of manganese. Journa l of Ort homolecular M edicine 5(4) : 219-222. 17. Wada, O. (2004). What are trace elements ? their deficiency and excess st ate. JMAJ. 47(8): 351-358. 18. Selv araju ,R. (2009). Change in serum trace elements concentration before and after removal of gallbladder with gallstone. Journal of gastroenterology 8(1) : 1-6. 19. Selv araju, R. ; Ganapathiraman, R. ; Narayanaswamy , R.; Valliapp an, R. and Baskaran, R. (2009). Trace element analysis hep atitis B affected human blood serum by inductively coupled p lasma atomic emission sp ectroscopy (ICP-AES). Romanian j. biophy s.19(1): 35-42. IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (3) 2011 20. Pintov, S.; Kohelet, D.; Arbel, E. and Goldber g, M . (1992). Predictive inability of cord zinc, magnesium and copp er levels on the development of benign hy p erbilirubinemia in the newborn. Acta Paediatrica. 81(11): 868–869. 21. Levenson , C. W. (2005). Trace metal regulation of neuronal apoptosis: From genes to behavior. Phy siology & Behavior 86 (3) 399 – 406. 22. Chvapil, M . (1973). New asp ects in the biological role of zinc. A st abilizer of macromo lecules and biological membran es. Life Sci. 13(8):1041-1049 . 23. Selv araju, R. (2009). Change in serum trace elements concentration before and after removal of gallbladder with gallstone. The internet journal of gastroenterology . 8(1) : 1-6. 24. Sokol, RJ.; M cKim, J.M. and Devereau, M .W. (1998). α-Tocop herol ameliorates oxidant injury of isolated copp er overloaded rat hepatocytes. Am J Clin Nutr. 67(5):1012–1016. 25. Ronald JE. (1971). Assessment of magnesium status. Clin Chem. 33(11) :1965-1971. Table (1): The mean values for the Cu, Zn, Mg, Mn compa red to the control group with p value < 0.05 Trace elements Mean ±SD ppm of patients Mean ±SD ppm of controls Cu 0.238 ± 0.411 0.198 ± 0.341 Zn 0.0012 ± 0.002 0.0021 ± 0.0036 M g 1.48 ± 2.96 1.293 ± 1.829 M n 0.0277 ± 0.0391 0.015 ± 0.031 IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (3) 2011 Fig. (1):Calibration curve for Fig.(2):Calibration curve for copper mangane se Fig. (3):Calibration curve for Fig.(4): Calibration curve for magne sium zinc 20) 3 (24 المجلد مجلة ابن الھیثم للعلوم الصرفة والتطبیقیة 11 المنغنیز ، والمغنیسیوم في مرضى والخارصین ، وتقییم مستویات النحاس ، الیرقان الحدیثي الوالدة في بغداد ایناس جبار حسن الجامعة المستنصریة قسم العلوم ، كلیة التربیة األساسیة ، 2011 شباط 3: البحث في استلم 2011 آیار 10: قبل البحث في الخالصة لتشخیص وتخمین بعض االمراض ، تتضمن الدراسة قیاس مستوى ة جیدا كواشفة تراكیز العناصر النزردتع المنغنیز ، والمغنیسیوم في أمصال مجموعتین من األطفال حدیثي الوالدة ، تتكون المجموعة و الخارصین ، والنحاس ، ا حدیث الوالدة طبیعیطفال) 20( حدیث الوالدة مصاب بالیرقان ، بینما تتكون المجموعة الثانیة من طفال) 52(األولى من طریقة المع المنغنیز ، والمغنیسیوم باستوصین ،الخارو تراكیز مستوى النحاس ، تحسب. سیطرة ةتم اعتمادها مجموع p وكان متوسط تركیز الخارصین في مصل دم المرضى اقل معنویا، طیف االمتصاص الذري من تركیزه في ) (0.05> المغنیسیوم ، والمنغنیز اعلى من ووكذلك وجد ان متوسط تراكیز النحاس ، حدیثي الوالدة األصحاء ةمصل دم مجموع . األصحاء ة مصل دم مجموعتركیزهم في المنغنیز ، المغنیسیوم، النحاس، الخارصین،الیرقان : الكلمات المفتاحیة