ICP-MS/MS-Based Ionomics: A Validated Methodology to Investigate the Biological Variability of the Human Ionome

Journal of Proteome Research

Volumn 16, Issue 5, 2017, Pages 2080-2090

  Konz, Tobias ^a   Migliavacca, Eugenia ^a   Dayon, Loïc ^a   Bowman, Gene ^a   Oikonomidi, Aikaterini ^b   Popp, Julius ^b   Rezzi, Serge ^a  

^a NESTLÉ RESEARCH CENTER   (Switzerland)

^b LAUSANNE UNIVERSITY HOSPITAL   (Switzerland)

Author keywords

ionome; ionomics; multielemental profiling; triple quadrupole ICP MS

Indexed keywords

ALUMINUM; BARIUM ION; CADMIUM; CALCIUM; CALCIUM ION; CERULOPLASMIN; CESIUM ION; CHROMIUM; COBALT; COPPER; FERRITIN; IODINE; IRON; LEAD; MAGNESIUM; MANGANESE; MERCURY; MOLYBDENUM; NITROGEN; PHOSPHATE; PHOSPHORUS; POTASSIUM; RUBIDIUM ION; SELENIUM; STRONTIUM; SULFUR; TIN; TITANIUM; ZINC; ZINC ION; ELEMENT; ION; METAL;

AGE; AGED; ANALYTIC METHOD; ARTICLE; BONE REMODELING; FEMALE; HIGH THROUGHPUT SCREENING; HUMAN; IONOME; LIMIT OF DETECTION; LIMIT OF QUANTITATION; MALE; METABOLOMICS; PRIORITY JOURNAL; QUADRUPOLE MASS SPECTROMETRY; QUANTITATIVE ANALYSIS; SEX DIFFERENCE; TANDEM MASS SPECTROMETRY; VALIDATION STUDY; CHEMISTRY; MIDDLE AGED; PRINCIPAL COMPONENT ANALYSIS; PROCEDURES; SERUM; SEX FACTOR;

AGE FACTORS; AGED; ELEMENTS; FEMALE; HUMANS; IONS; MALE; METALS; METHODS; MIDDLE AGED; PRINCIPAL COMPONENT ANALYSIS; SERUM; SEX FACTORS; TANDEM MASS SPECTROMETRY;

EID: 85019011786     PISSN: 15353893     EISSN: 15353907     Source Type: Journal    
DOI: 10.1021/acs.jproteome.7b00055     Document Type: Article

References (73)

1. Alberts, B.; Johnson, A.; Lewis, J.; et al., The chemical components of a cell. In Mol. Biol. Cell, 4 th ed.; Garland Science: New York, 2002; pp 47-68.
2. Chellan, P.; Sadler, P. J. The elements of life and medicines Philos. Trans. R. Soc., A 2015, 373 (2037) 1-56 10.1098/rsta.2014.0182
3. Tchounwou, P. B.; Yedjou, C. G.; Patlolla, A. K.; Sutton, D. J. Heavy metal toxicity and the environment EXS 2012, 101, 133-164 10.1007/978-3-7643-8340-4-6
4. Kaput, J.; Ordovas, J. M.; Ferguson, L.; van Ommen, B.; Rodriguez, R. L.; Allen, L.; Ames, B. N.; Dawson, K.; German, B.; Krauss, R.; Malyj, W.; Archer, M. C.; Barnes, S.; Bartholomew, A.; Birk, R.; van Bladeren, P.; Bradford, K. J.; Brown, K. H.; Caetano, R.; Castle, D.; Chadwick, R.; Clarke, S.; Clement, K.; Cooney, C. A.; Corella, D.; Manica da Cruz, I. B.; Daniel, H.; Duster, T.; Ebbesson, S. O.; Elliott, R.; Fairweather-Tait, S.; Felton, J.; Fenech, M.; Finley, J. W.; Fogg-Johnson, N.; Gill-Garrison, R.; Gibney, M. J.; Gillies, P. J.; Gustafsson, J. A.; Hartman, J. L. t.; He, L.; Hwang, J. K.; Jais, J. P.; Jang, Y.; Joost, H.; Junien, C.; Kanter, M.; Kibbe, W. A.; Koletzko, B.; Korf, B. R.; Kornman, K.; Krempin, D. W.; Langin, D.; Lauren, D. R.; Ho Lee, J.; Leveille, G. A.; Lin, S. J.; Mathers, J.; Mayne, M.; McNabb, W.; Milner, J. A.; Morgan, P.; Muller, M.; Nikolsky, Y.; van der Ouderaa, F.; Park, T.; Pensel, N.; Perez-Jimenez, F.; Poutanen, K.; Roberts, M.; Saris, W. H.; Schuster, G.; Shelling, A. N.; Simopoulos, A. P.; Southon, S.; Tai, E. S.; Towne, B.; Trayhurn, P.; Uauy, R.; Visek, W. J.; Warden, C.; Weiss, R.; Wiencke, J.; Winkler, J.; Wolff, G. L.; Zhao-Wilson, X.; Zucker, J. D. The case for strategic international alliances to harness nutritional genomics for public and personal health Br. J. Nutr. 2005, 94 (5) 623-632 10.1079/BJN20051585
5. Cominetti, O.; Nunez Galindo, A.; Corthesy, J.; Oller Moreno, S.; Irincheeva, I.; Valsesia, A.; Astrup, A.; Saris, W. H.; Hager, J.; Kussmann, M.; Dayon, L. Proteomic Biomarker Discovery in 1000 Human Plasma Samples with Mass Spectrometry J. Proteome Res. 2016, 15 (2) 389-399 10.1021/acs.jproteome.5b00901
6. Rueedi, R.; Ledda, M.; Nicholls, A. W.; Salek, R. M.; Marques-Vidal, P.; Morya, E.; Sameshima, K.; Montoliu, I.; Da Silva, L.; Collino, S.; Martin, F. P.; Rezzi, S.; Steinbeck, C.; Waterworth, D. M.; Waeber, G.; Vollenweider, P.; Beckmann, J. S.; Le Coutre, J.; Mooser, V.; Bergmann, S.; Genick, U. K.; Kutalik, Z. Genome-wide association study of metabolic traits reveals novel gene-metabolite-disease links PLoS Genet. 2014, 10 (2) e1004132 10.1371/journal.pgen.1004132
7. Nicholson, J. K. Global systems biology, personalized medicine and molecular epidemiology Mol. Syst. Biol. 2006, 2, 52
8. Wild, C. P. Complementing the genome with an ″exposome″: the outstanding challenge of environmental exposure measurement in molecular epidemiology Cancer Epidemiol., Biomarkers Prev. 2005, 14 (8) 1847-1850 10.1158/1055-9965.EPI-05-0456
9. Rappaport, S. M.; Smith, M. T. Epidemiology. Environment and disease risks Science 2010, 330 (6003) 460-461 10.1126/science.1192603
10. Salt, D. E.; Baxter, I.; Lahner, B. Ionomics and the study of the plant ionome Annu. Rev. Plant Biol. 2008, 59, 709-733 10.1146/annurev.arplant.59.032607.092942
11. Salt, D. E. Update on plant ionomics Plant Physiol. 2004, 136 (1) 2451-2456 10.1104/pp.104.047753
12. Baxter, I. Ionomics: The functional genomics of elements Briefings Funct. Genomics 2010, 9 (2) 149-156 10.1093/bfgp/elp055
13. Yoshida, S.; Date, Y.; Akama, M.; Kikuchi, J. Comparative metabolomic and ionomic approach for abundant fishes in estuarine environments of Japan Sci. Rep. 2014, 4, 7005 10.1038/srep07005
14. Eide, D. J.; Clark, S.; Nair, T. M.; Gehl, M.; Gribskov, M.; Guerinot, M. L.; Harper, J. F. Characterization of the yeast ionome: a genome-wide analysis of nutrient mineral and trace element homeostasis in Saccharomyces cerevisiae Genome Biol. 2005, 6 (9) R77 10.1186/gb-2005-6-9-r77
15. Muller, A.; Wenzel, M.; Strahl, H.; Grein, F.; Saaki, T. N.; Kohl, B.; Siersma, T.; Bandow, J. E.; Sahl, H. G.; Schneider, T.; Hamoen, L. W. Daptomycin inhibits cell envelope synthesis by interfering with fluid membrane microdomains Proc. Natl. Acad. Sci. U. S. A. 2016, 113 (45) E7077-E7086 10.1073/pnas.1611173113
16. Sun, L.; Yu, Y.; Huang, T.; An, P.; Yu, D.; Yu, Z.; Li, H.; Sheng, H.; Cai, L.; Xue, J.; Jing, M.; Li, Y.; Lin, X.; Wang, F. Associations between ionomic profile and metabolic abnormalities in human population PLoS One 2012, 7 (6) e38845 10.1371/journal.pone.0038845
17. Huang, Y.; Zhu, M.; Li, Z.; Sa, R.; Chu, Q.; Zhang, Q.; Zhang, H.; Tang, W.; Zhang, M.; Yin, H. Mass spectrometry-based metabolomic profiling identifies alterations in salivary redox status and fatty acid metabolism in response to inflammation and oxidative stress in periodontal disease Free Radical Biol. Med. 2014, 70, 223-232 10.1016/j.freeradbiomed.2014.02.024
18. Zhao, T.; Chen, T. L.; Qiu, Y. P.; Zou, X. Y.; Li, X.; Su, M. M.; Yan, C. H.; Zhao, A. H.; Jia, W. Trace Element Profiling Using Inductively Coupled Plasma Mass Spectrometry and its Application in an Osteoarthritis Study Anal. Chem. 2009, 81 (9) 3683-3692 10.1021/ac900311q
19. Cai, L.; Chen, T. L.; Yang, J. L.; Zhou, K. J.; Yan, X. M.; Chen, W. Z.; Sun, L. Y.; Li, L. L.; Qin, S. Y.; Wang, P.; Yang, P.; Cui, D. H.; Burmeister, M.; He, L.; Jia, W.; Wan, C. L. Serum trace element differences between Schizophrenia patients and controls in the Han Chinese population Sci. Rep. 2015, 5, 15013 10.1038/srep15013
20. Susuki, S.; Ando, Y.; Sato, T.; Nishiyama, M.; Miyata, M.; Suico, M. A.; Shuto, T.; Kai, H. Multi-elemental analysis of serum and amyloid fibrils in familial amyloid polyneuropathy patients Amyloid 2008, 15 (2) 108-116 10.1080/13506120802006013
21. Gonzalez-Dominguez, R.; Garcia-Barrera, T.; Gomez-Ariza, J. L. Characterization of metal profiles in serum during the progression of Alzheimer's disease Metallomics 2014, 6 (2) 292-300 10.1039/C3MT00301A
22. Malinouski, M.; Hasan, N. M.; Zhang, Y.; Seravalli, J.; Lin, J.; Avanesov, A.; Lutsenko, S.; Gladyshev, V. N. Genome-wide RNAi ionomics screen reveals new genes and regulation of human trace element metabolism Nat. Commun. 2014, 5, 3301 10.1038/ncomms4301
23. Begerow, J.; Turfeld, M.; Dunemann, L. New horizons in human biomonitoring of environmentally and occupationally relevant metals-sector-field ICP-MS versus electrothermal AAS J. Anal. At. Spectrom. 2000, 15 (4) 347-352 10.1039/A909056H
24. Liu, X. B.; Piao, J. H.; Huang, Z. W.; Zhang, S. Q.; Li, W. D.; Tian, Y.; Yang, X. G. Determination of 16 Selected Trace Elements in Children Plasma from China Economical Developed Rural Areas Using High Resolution Magnetic Sector Inductively Coupled Mass Spectrometry J. Anal. Methods Chem. 2014, 2014, 975820 10.1155/2014/975820
25. Heitland, P.; Koster, H. D. Biomonitoring of 37 trace elements in blood samples from inhabitants of northern Germany by ICP-MS J. Trace Elem. Med. Biol. 2006, 20 (4) 253-262 10.1016/j.jtemb.2006.08.001
26. Barany, E.; Bergdahl, I. A.; Bratteby, L. E.; Lundh, T.; Samuelson, G.; Schutz, A.; Skerfving, S.; Oskarsson, A. Trace element levels in whole blood and serum from Swedish adolescents Sci. Total Environ. 2002, 286 (1-3) 129-141 10.1016/S0048-9697(01)00970-6
27. Minoia, C.; Sabbioni, E.; Apostoli, P.; Pietra, R.; Pozzoli, L.; Gallorini, M.; Nicolaou, G.; Alessio, L.; Capodaglio, E. Trace element reference values in tissues from inhabitants of the European community. I. A study of 46 elements in urine, blood and serum of Italian subjects Sci. Total Environ. 1990, 95, 89-105 10.1016/0048-9697(90)90055-Y
28. Muniz, C. S.; Fernandez-Martin, J. L.; Marchante-Gayon, J. M.; Garcia Alonso, J. I.; Cannata-Andia, J. B.; Sanz-Medel, A. Reference values for trace and ultratrace elements in human serum determined by double-focusing ICP-MS Biol. Trace Elem. Res. 2001, 82 (1-3) 259-272 10.1385/BTER:82:1-3:259
29. Vaughan, M. A.; Baines, A. D.; Templeton, D. M. Multielement analysis of biological samples by inductively coupled plasma-mass spectrometry. II. Rapid survey method for profiling trace elements in body fluids Clin. Chem. 1991, 37 (2) 210-215
30. Forrer, R.; Gautschi, K.; Lutz, H. Simultaneous measurement of the trace elements Al, As, B, Be, Cd, Co, Cu, Fe, Li, Mn, Mo, Ni, Rb, Se, Sr, and Zn in human serum and their reference ranges by ICP-MS Biol. Trace Elem. Res. 2001, 80 (1) 77-93 10.1385/BTER:80:1:77
31. Harrington, J. M.; Young, D. J.; Essader, A. S.; Sumner, S. J.; Levine, K. E. Analysis of Human Serum and Whole Blood for Mineral Content by ICP-MS and ICP-OES: Development of a Mineralomics Method Biol. Trace Elem. Res. 2014, 160 (1) 132-142 10.1007/s12011-014-0033-5
32. Muniz, C. S.; Larchante-Gayon, J. M.; Alonso, J. I. G.; Sanz-Medel, A. Multi-elemental trace analysis of human serum by double-focusing ICP-MS J. Anal. At. Spectrom. 1999, 14 (2) 193-198 10.1039/A807360K
33. Verni, E. R.; Moyano, F.; Martinez, L. D.; Lapierre, A. V.; Gil, R. A. Handling spectral interferences and matrix effects in DRC-ICPMS to assess the elemental profile in human serum samples after dissolution with formic acid J. Anal. At. Spectrom. 2013, 28 (10) 1655-1659 10.1039/c3ja50191d
34. Barany, E.; Bergdahl, I. A.; Schutz, A.; Skerfving, S.; Oskarsson, A. Inductively coupled plasma mass spectrometry for direct multi-element analysis of diluted human blood and serum J. Anal. At. Spectrom. 1997, 12 (9) 1005-1009 10.1039/A700904F
35. Krachler, M.; Irgolic, K. J. The potential of inductively coupled plasma mass spectrometry (ICP-MS) for the simultaneous determination of trace elements in whole blood, plasma and serum J. Trace Elem. Med. Biol. 1999, 13 (3) 157-169 10.1016/S0946-672X(99)80006-6
36. Roos, P. M.; Vesterberg, O.; Syversen, T.; Flaten, T. P.; Nordberg, M. Metal concentrations in cerebrospinal fluid and blood plasma from patients with amyotrophic lateral sclerosis Biol. Trace Elem. Res. 2013, 151 (2) 159-170 10.1007/s12011-012-9547-x
37. Alimonti, A.; Bocca, B.; Pino, A.; Ruggieri, F.; Forte, G.; Sancesario, G. Elemental profile of cerebrospinal fluid in patients with Parkinson's disease J. Trace Elem. Med. Biol. 2007, 21 (4) 234-241 10.1016/j.jtemb.2007.05.001
38. Sarmiento-Gonzalez, A.; Marchante-Gayon, J. M.; Tejerina-Lobo, J. M.; Paz-Jimenez, J.; Sanz-Medel, A. High-resolution ICP-MS determination of Ti, V, Cr, Co, Ni, and Mo in human blood and urine of patients implanted with a hip or knee prosthesis Anal. Bioanal. Chem. 2008, 391 (7) 2583-2589 10.1007/s00216-008-2188-4
39. Konz, T.; Montes-Bayon, M.; Bettmer, J.; Sanz-Medel, A. Analysis of hepcidin, a key peptide for Fe homeostasis, via sulfur detection by capillary liquid chromatography-inductively coupled plasma mass spectrometry J. Anal. At. Spectrom. 2011, 26 (2) 334-340 10.1039/C0JA00053A
40. ICH Harmonised Tripartite Guideline. Q2(R1): Validation of Analytical Procedures: Text and Methodology, http://www.ich.org/products/guidelines/quality/quality-single/article/validation-of-analytical-procedures-text-and-methodology.html.
41. Barwick, V. J.; Ellison, S. L. R. The evaluation of measurement uncertainty from method validation studies-Part 1: Description of a laboratory protocol Accredit. Qual. Assur. 2000, 5 (2) 47-53 10.1007/s007690050010
42. Barwick, V. J.; Ellison, S. L. R.; Rafferty, M. J. Q.; Gill, R. S. The evaluation of measurement uncertainty from method validation studies-Part 2: The practical application of a laboratory protocol Accredit. Qual. Assur. 2000, 5 (3) 104-113 10.1007/s007690050022
43. Van Loco, J.; Elskens, M.; Croux, C.; Beernaert, H. Linearity of calibration curves: use and misuse of the correlation coefficient Accredit. Qual. Assur. 2002, 7 (7) 281-285 10.1007/s00769-002-0487-6
44. Guidance for Industry Bioanalytical Method Validation; U.S. Dept. of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research, Center for Veterinary Medicine: Rockville, MD, 2001; p 1.
45. Vogl, J. Measurement uncertainty in single, double and triple isotope dilution mass spectrometry Rapid Commun. Mass Spectrom. 2012, 26 (3) 275-281 10.1002/rcm.5306
46. Caroli, S.; Alimonti, A.; Coni, E.; Petrucci, F.; Senofonte, O.; Violante, N. The Assessment of Reference Values for Elements in Human Biological Tissues and Fluids-a Systematic Review Crit. Rev. Anal. Chem. 1994, 24 (5-6) 363-398 10.1080/10408349408048824
47. Rodushkin, I.; Ödman, F.; Olofsson, R.; Burman, E.; Axelsson, M. Multi-element analysis of body fluids by double-focusing ICP-MS Recent Res. Devel. Pure Appl. Anal. Chem. 2001, 5, 51-66
48. Bansal, V. K. Serum Inorganic Phosphorus. In Clinical Methods: The History, Physical, and Laboratory Examinations, 3 rd ed.; Walker, H. K.; Hall, W. D.; Hurst, J. W., Eds.; Butterworths: Boston, 1990; pp 895-899.
49. Ross, A. C.; Taylor, C. L.; Yaktine, A. L.; Del Valle, H. B., Eds.; Dietary Reference Intakes for Adequacy: Calcium and Vitamin D. In Dietary Reference Intakes for Calcium and Vitamin D; National Academies Press: Washington, D.C., 2011; pp 345-362.
50. Chung, M.; Balk, E. M.; Brendel, M.; Ip, S.; Lau, J.; Lee, J.; Lichtenstein, A.; Patel, K.; Raman, G.; Tatsioni, A.; Terasawa, T.; Trikalinos, T. A. Vitamin D and calcium: a systematic review of health outcomes Evid. Rep. Technol. Assess. 2009, 183) 1-420
51. Gunter, T. E.; Buntinas, L.; Sparagna, G.; Eliseev, R.; Gunter, K. Mitochondrial calcium transport: mechanisms and functions Cell Calcium 2000, 28 (5-6) 285-296 10.1054/ceca.2000.0168
52. Peacock, M. Calcium metabolism in health and disease Clin. J. Am. Soc. Nephrol. 2010, 5 (Suppl 1) S23-30 10.2215/CJN.05910809
53. Feng, X.; McDonald, J. M. Disorders of bone remodeling Annu. Rev. Pathol.: Mech. Dis. 2011, 6, 121-145 10.1146/annurev-pathol-011110-130203
54. Warburton, D. E.; Nicol, C. W.; Gatto, S. N.; Bredin, S. S. Cardiovascular disease and osteoporosis: balancing risk management Vasc. Health Risk Manag. 2007, 3 (5) 673-689
55. Quinn, J. F.; Harris, C.; Kaye, J. A.; Lind, B.; Carter, R.; Anekonda, T.; Ralle, M. Gender effects on plasma and brain copper Int. J. Alzheimer's Dis. 2011, 2011, 150916 10.4061/2011/150916
56. Trumbo, P.; Yates, A. A.; Schlicker, S.; Poos, M. Dietary reference intakes: vitamin A, vitamin K, arsenic, boron, chromium, copper, iodine, iron, manganese, molybdenum, nickel, silicon, vanadium, and zinc J. Am. Diet. Assoc. 2001, 101 (3) 294-301 10.1016/S0002-8223(01)00078-5
57. Vendelboe, T. V.; Harris, P.; Zhao, Y.; Walter, T. S.; Harlos, K.; El Omari, K.; Christensen, H. E. The crystal structure of human dopamine beta-hydroxylase at 2.9 A resolution Sci. Adv. 2016, 2 (4) e1500980 10.1126/sciadv.1500980
58. Johnson, M. A.; Fischer, J. G.; Kays, S. E. Is copper an antioxidant nutrient? Crit. Rev. Food Sci. Nutr. 1992, 32 (1) 1-31 10.1080/10408399209527578
59. Linder, M. C.; Hazegh-Azam, M. Copper biochemistry and molecular biology Am. J. Clin. Nutr. 1996, 63 (5) 797S-811S
60. Olivares, M.; Mendez, M. A.; Astudillo, P. A.; Pizarro, F. Present situation of biomarkers for copper status Am. J. Clin. Nutr. 2008, 88 (3) 859S-862S
61. Mineral Tolerance of Animals, 2 nd rev. ed.; National Academies Press: Washington, D.C., 2005; p xiv, p 496.
62. Haas, E. M.; Levin, B. Staying Healthy with Nutrition: The Complete Guide to Diet and Nutritional Medicine; Celestial Arts: 2006; Vol. 1, p 944.
63. Stolk, J. M.; Nowack, W. J.; Barchas, J. D.; Platman, S. R. Brain norepinephrine: enhanced turnover after rubidium treatment Science 1970, 168 (3930) 501-503 10.1126/science.168.3930.501
64. Hodkinson, C. F.; Kelly, M.; Coudray, C.; Gilmore, W. S.; Hannigan, B. M.; O'Connor, J. M.; Strain, J. J.; Wallace, J. M. Zinc status and age-related changes in peripheral blood leukocyte subpopulations in healthy men and women aged 55-70 y: the ZENITH study Eur. J. Clin. Nutr. 2005, 59 (Suppl 2) S63-S67 10.1038/sj.ejcn.1602301
65. Ezzati, M.; Lopez, A. D.; Rodgers, A. A.; Murray, C. J. L. Zinc deficiency. In Comparative Quantification of Health Risks; World Health Organization: Geneva, 2004; pp 257-279.
66. McCall, K. A.; Huang, C.; Fierke, C. A. Function and mechanism of zinc metalloenzymes J. Nutr. 2000, 130 (5S Suppl) 1437S-1446S
67. Rink, L.; Gabriel, P. Zinc and the immune system Proc. Nutr. Soc. 2000, 59 (4) 541-552 10.1017/S0029665100000781
68. Wang, X.; Zhou, B. Dietary zinc absorption: A play of Zips and ZnTs in the gut IUBMB Life 2010, 62 (3) 176-182 10.1002/iub.291
69. Fairweather-Tait, S. J.; Harvey, L. J.; Ford, D. Does ageing affect zinc homeostasis and dietary requirements? Exp. Gerontol. 2008, 43 (5) 382-388 10.1016/j.exger.2007.10.015
70. Mocchegiani, E.; Muzzioli, M.; Giacconi, R. Zinc and immunoresistance to infection in aging: new biological tools Trends Pharmacol. Sci. 2000, 21 (6) 205-208 10.1016/S0165-6147(00)01476-0
71. Malavolta, M.; Giacconi, R.; Piacenza, F.; Santarelli, L.; Cipriano, C.; Costarelli, L.; Tesei, S.; Pierpaoli, S.; Basso, A.; Galeazzi, R.; Lattanzio, F.; Mocchegiani, E. Plasma copper/zinc ratio: an inflammatory/nutritional biomarker as predictor of all-cause mortality in elderly population Biogerontology 2010, 11 (3) 309-319 10.1007/s10522-009-9251-1
72. Malavolta, M.; Piacenza, F.; Basso, A.; Giacconi, R.; Costarelli, L.; Mocchegiani, E. Serum copper to zinc ratio: Relationship with aging and health status Mech. Ageing Dev. 2015, 151, 93-100 10.1016/j.mad.2015.01.004
73. Reunanen, A.; Knekt, P.; Marniemi, J.; Maki, J.; Maatela, J.; Aromaa, A. Serum calcium, magnesium, copper and zinc and risk of cardiovascular death Eur. J. Clin. Nutr. 1996, 50 (7) 431-437