IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 Evaluation of Copper , Zinc , Manganese , and Magnesium Levels in Newborn Jaundice in Baghdad E. J. Hasan Department of Science, College of Basic Education, Al- Mustansiriah University Abstract Trace elements concentrations are frequently reported to be a good indicator for diagnosis and prognosis of some disease. The levels of copper, zinc, manganese, and magnesium were measured in serum of two group. Group one contains (52) newborn jaundice patients while the second group contains (20) healthy newborn used as controls. In this study the concentration of Cu, Zn, Mg, and Mn were measured using atomic absorption spectrophotometer (AAS). It has been found that zinc level is significantly lower in newborn jaundice patients compared with normal subjects at (p<0.05) , also the level of Cu, Mg, and Mn were significantly higher in newborn jaundice compared with control group. Keywords: Jaundice; Zinc; Copper; Magnesium; Manganese Introduction Jaundice occurs when the blood contains excessive amounts of bilirubin, the deposition of this highly insoluble substance colored the skin and the whites of the eyes yellow. Jaundice is probably the most common symptom in neonatal period[1,2]. Temporary jaundice is observed during the first week of life in approximately 60% of term infants and 80% of preterm infants[3,4]. Trace elements are necessary for normal function and are therefore associated with morbid deficiency states. Those elements are found in body tissues at levels in the micrograms per gram of tissue or less. Some elements are essential trace elements (like Fe and Zn) and some are not essential (like Cu and Mn) but have well defined evidence in human metabolism[5]. Trace elements are essential micronutrients for growth, development, and maintenance of healthy tissues. The role of trace elements in body metabolism is of prime importance. Their deficiency causes diseases, whereas their presence in excess may result in toxicity to human life [6]. Zinc (Zn) is essential component of many metalloenzymes involved in virtually all aspects of metabolism. Zinc is an integral component of nearly 300 enzyme in different species of all phyla[7,8]. Important zinc-containing metaloenzymes in humans including carbonic anhydrase , alkaline phosphates, RNA and DNA polymerases, thymidine kinase, carboxypeptidases, and alcohol dehydrogenase. Zinc stabilizes the structures of proteins and nucleic acids, participates in transport prosses[9]. Copper (Cu) may present in biological IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 system in both the +1 and +2 valence states, the major functions of copper metalloproteins involve oxidation –reduction, most known copper – containing enzyme bind and react directly with molecular oxygen[10]. Copper is an integral component of many metalloenzymes, including ceruloplasmin, cytochrome c oxidase, superoxide dismutase, dopamine –β– hydroxylase, ascorbate oxidase, lysy l oxidase, and ty rosinase[11,12]. Magnesium (Mg) catalyzes or activates more than 300 enzymes in the body. Magnesium acts as an essential cofactor for enzymes concerned with cell respiration, glycolysis, and trans-membrane transport of other cations such as calcium and sodium[13]. Magnesim can effected enzyme activity by binding the active site of the enzyme (pyruvate kinase, enolase), by ligand binding (ATP- requiring enzyme), by causing conformational changes during the catalytic process (Na- K- ATPase), and by promoting aggregation of multienzyme complexes[7,14]. Manganese (Mn) is present in biological systems bound to protein in either the +2 or +3 valence state. Manganese is associated mainly with the formation of connective and bony tissue, with growth and reproductive functions, and with carbohydrate and lipid metabolism[15]. The biochemical basis for manganese essentiality is its function as a constituent of metalloenzymes and as an enzyme activator[16,17]. Important manganese containing enzymes include arginase, pyruvate carboxylase, and manganese superoxide dismutase in mitochondria[18]. Material and Method All blood samples were collected from the Ibn- Albalady hospital in Baghdad. The first group consisted of 55 blood samples of jaundice newborns (32were male and 23 females) , their ages ranged from 1-3 days. Twenty healthy volunteers were used as a second group and their ages were matched with patients group. Blood samples are drown using 10ml syringes with steel needles. A3ml blood sample was drown from each patient. The whole blood was immediately transferred to plain tube. These samples were allowed to stand at room temperature for 10 minutes for clotting, the clots were separated from the wall of the tube using wooden applicator stick. 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. Atomic Absorption Spectrophotometer (AAS) model AA-6200 (shimadzu _ Japan) fitted with air – acetylene flame were used for the determination of Copper, Zinc, Manganese, Magnesium. 1- Preparation of standard solutions for AAs measurements cpper Copper stock solution (1000ppm) was prepared by dissolving (0.1 gm) of copper metal in 5ml of 5 M nitric acid, and then diluted to 100ml with deionized water. Zinc Zinc stock solution (1000ppm) was prepared by dissolving (0.1 gm) of zinc pellet using concentrated hydrochloric acid, and the volume was completed into 100ml with deionized water. IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 Magnesium Magnesium stock solution (1000ppm) was prepared by dissolving (0.1 gm) of magnesium metal in 5ml of 5 M nitric acid, and then diluted to 100ml with deionized water. Manganese Manganese stock solution (1000ppm) was prepared by dissolving (0.1 gm) of manganese metal in 5ml of 5 M nitric acid, and then diluted to 100ml with deionized water. 1- Calibration curve for A.A.S measurements 1- The standard solutions for copper ion were prepared by subsequent dilution stock solution (1000ppm) of copper and diluted to (10, 5, 1, 0.5, 0.1 ppm). 2- Iron standard solutions were prepared by subsequent dilution stock solution (1000ppm) of iron to (10, 5, 1, 0.5,and 0.1 ppm). 3- The standard solutions for magnesium were prepared by subsequent dilution stock solution (1000ppm) of magnesium ion and diluted to (100, 50, 10, 5,and 1 ppm). 4- The standard solutions for manganese ion were prepared by subsequent dilution stock solution (1000ppm) of manganese and diluted to (20, 15, 10 ppm). All above standard solutions were diluted to 100ml in a volmatric flask with deionized water. Result and Discussion A linear calibration curve was obtained extended from (0.1- 10) ppm for copper, zinc, manganese, and magnesium as shown in the (1,2,3,4) figures The results were shown that there was significant difference in the levels of zinc, copper, manganese, and magnesium in newborn jaundice and control group, at p value of (< 0.05 ), Zinc levels was significantly lower in jaundice patients (0.0012 ± 0.002) compared with the healthy newborn ( 0.0021 ± 0.0036) . Copper levels was significantly higher (0.198 ± 0.341) in jaundice patients compared with control group (0.238 ± 0.411). Magnesium levels was significantly higher in patient (1.48 ± 2.96) than the control group (1.293 ± 1.829). Manganese levels was appeared to be significantly higher (0.0277 ± 0.0391) in jaundice patients compared with control (0.015 ± 0.031). As shown in table (1) . Zinc and magnesium are very important co-factors for a lot of enzyme systems and play an important role in the synthesis of nucleic acids [19,20]. Zinc prevents the lipid depolarization of the cell membranes and hypozincemia may modulate the erythrocyte membrane. It may result in deficient synthesis of assorted enzymes that play a role in the bilirubin metabolism[21], Hypozincemia may also cause structural defects in the erythrocyte membranes, resulting in hemolysis [22]. An elevated level of serum copper is documented in liver disease like cirrhosis , obstructive jaundice and cholestasis [23]. In newborn with jaundice , the high serum copper may be of intracellular (erythrocyte ) origin [24]. Manganese absorbed from food is avidly taken up by the liver and distributed to tissues bound to transferring and albumin[5].The serum levels of magnesium and manganese become high , depending on the existence of mild hemolysis in newborn , the fact that these elements are high depend on being high in erythrocytes, especially reticulocytes than in serum concentrations (25), and may depend on passing into the serum as a result of hemolysis. IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 References 1. Knudsen, A.( 1996). Prediction and non-invasive assessment of neonatal jaundice in the term healthy newborn infant. Acta Paediatr.85(4):393-397. 2. Martin, D.W.; Mayes, P.A. ;Rodwell, V.W. (1981). Harpers review of biochemistry . 18 ed, Maruzen asia pte. Ltd. Singapore . p(319). 3. Phyllis, A.D.; William, D.R. and Davit, K.S.( 1995). Neonatal jaundice-what now? Clin Pediatr . 34(2):103-107. 4. Jeffrey Maisels, M. (2006). What’s in a Name? Physiologic 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 Myking, A.(2002). Multi-element analysis of trace element levels in human autopsy tissues by using inductively coupled atomic emission spectrometry technique (ICP-AES). J Trace Elem Med 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, Minerals and trace elements. Journal of pediatric Gastroenterology and Nutrition. 41(2):S39-S46. 8. Mansi, K. ; Aburjai, T.; Barqawi, M. and Naser H. (2009). Copper and Zinc status in Jordanian patients with β-Thalassemia major treated with deferoxamine. Journal of Biological Sciences 4(5): 566-572. 9. Zlotkin, S.H. and Buchanan, B.E.( 1988). Amino acid intake and urinary zinc excretion in newborn infants receiving total parenteral nutrition. Am J Clin Nutr. 48(2):330–334. 10. Ostermeier, C.; Iwata ,S.; Michel, H.( 1996). Cytochrome c oxidase. Curr Opin 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 protein and fractions in diseased malnourished children of 4-12 years of age. Medical Journal of Islamic Academy of Sciences 14(1): 25−33. 12. Schulpis, K. H.; Karakonstantakis, T.; Gavrili, S.; Costalos, C. ; Roma, E. and Papassotiriou I. (2004). Serum Copper 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). Magnesium in disease: a review with special emphasis on the serum ionized magnesium. Clin Chem Lab Med 37(11):1011-1033. 14. Maggioni, A.; Orzalesi, M.; Mimouni, F.B. (1998). Intravenous correction of neonatal hypomagnesemia: effect on ionized magnesium. J Pediatr 132(4): 652–655. 15. Fok, T.F.;Chui, K.K.; Cheung, R. et al.( 2001). Manganese intake and cholestatic jaundice in neonates receiving parenteral nutrition: a randomized controlled study. Acta Pediatr. 90(9):1009–15. 16. Watts ,D. L. (1990). The nutritional relationships of manganese. Journal of Orthomolecular Medicine 5(4) : 219-222. 17. Wada, O. (2004). What are trace elements ? their deficiency and excess state. JMAJ. 47(8): 351-358. 18. Selvaraju ,R. (2009). Change in serum trace elements concentration before and after removal of gallbladder with gallstone. Journal of gastroenterology 8(1) : 1-6. 19. Selvaraju, R.; Ganapathiraman, R.; Narayanaswamy, R.; Valliappan, R. and Baskaran, R. (2009). Trace element analysis hepatitis B affected human blood serum by inductively coupled plasma atomic emission spectroscopy(ICP-AES). Romanian j. biophys.19(1): 35-42. IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 20. Pintov, S.; Kohelet, D.; Arbel, E. and Goldberg, M. (1992). Predictive inability of cord zinc, magnesium and copper levels on the development of benign hyperbilirubinemia in the newborn. Acta Paediatrica. 81(11): 868–869. 21. Levenson , C. W. (2005). Trace metal regulation of neuronal apoptosis: From genes to behavior. Physiology & Behavior 86 (3) 399 – 406. 22. Chvapil, M. (1973). New aspects in the biological role of zinc. A stabilizer of macromolecules and biological membranes. Life Sci. 13(8):1041-1049 . 23. Selvaraju, 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.; McKim, J.M. and Devereau, M.W. (1998). α-Tocopherol ameliorates oxidant injury of isolated copper 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 compared 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 Mg 1.48 ± 2.96 1.293 ± 1.829 Mn 0.0277 ± 0.0391 0.015 ± 0.031 IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 Fig. (1):Calibration curve for Fig.(2):Calibration curve for copper manganese Fig. (3):Calibration curve for Fig.(4): Calibration curve for magnesium zinc للعلوم الصرفة والتطبیقیة 2011) 3 (24 المجلد مجلة ابن الھیثم 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 من تركیزه في ) (p<0.05 وكان متوسط تركیز الخارصین في مصل دم المرضى اقل معنویا، طیف االمتصاص الذري نحاس ، حدیثي الوالدة األصحاء ةمصل دم مجموع راكیز ال المغنیسیوم ، والمنغنیز اعلى من ووكذلك وجد ان متوسط ت . األصحاء ة مصل دم مجموعتركیزهم في المنغنیز ، المغنیسیوم، النحاس، الخارصین،الیرقان : الكلمات المفتاحیة