This work describes the evaluation of metals and (metallo)proteins in vitreous humor samples and their correlations with some biological aspects in different post-mortem intervals (1-7 days), taking into account both decomposing and non-decomposing bodies. After qualitative evaluation of the samples involving 26 elements, representative metal ions (Fe, Mg and Mo) are determined by inductively coupled plasma mass spectrometry after using mini-vial decomposition system for sample preparation. A significant trend for Fe is found with post-mortem time for decomposing bodies because of a significant increase of iron concentration when comparing samples from bodies presenting 3 and 7 days post-mortem interval. An important clue to elucidate the role of metals is the coupling of liquid chromatography with inductively coupled plasma mass spectrometry for identification of metals linked to proteins, as well as mass spectrometry for the identification of those proteins involved in the post-mortem interval.61018011807Palmiere, C., Mangin, P., Postmortem chemistry update part II-Review (2012) J. Leg. Med., 126, pp. 199-215Chandrakanth, H.V., Kanchan, T., Balaraj, B.M., Virupaksha, H.S., Chandrashekar, T.N., Postmortem vitreous chemistry-An evaluation of sodium, potassium and chloride levels in estimation of time since death (during the first 36 h after death) (2012) J. For. Legal Med., 19, pp. 1-6Maeda, H., Ishikawa, T., Michiue, T., Forensic biochemistry for functional investigation of death: Concept and practical application (2011) Leg. Med., 13, pp. 55-67Barnett, J.P., Scanlan, D.J., Blindauer, C.A., Protein fractionation and detection for metalloproteomics: Challenges and approaches (2012) Anal. Bioanal. Chem., 402, pp. 3311-3322Mounicou, S., Szpunar, J., Lobinski, R., Metallomics: The concept and methodology (2009) Chem. Soc. Rev., 38, pp. 1119-1138Ugarte, M., Grime, G.W., Lord, G., Geraki, K., Collingwood, J.F., Finnegan, M.E., Farnfield, H., Osborne, N.N., Concentration of various trace elements in the rat retina and their distribution in different structures (2012) Metallomics, 4, pp. 1245-1254Thierauf, A., Musshoff, F., Madea, B., Post-mortem biochemical investigation of vitreous humor (2009) Forensic Sci. Int., 192, pp. 78-82Madea, B., Musshoff, F., Postmortem biochemistry (2007) Forensic Sci. Int., 165, pp. 165-171Sanches, L.R., Seulin, S.C., Leyton, V., Paranhos, B.A.P.B., Pasqualucci, C.A., Muñoz, D.R., Osselton, M.D., Et al., Determination of opiates in whole blood and vitreous humor: A study of the matrix effect and an experimental design to optimize conditions for the enzymatic hydrolysis of glucuronides (2012) J. Anal. Toxicol., 36, pp. 162-170Muñoz-Barús, J.I., Lendoiro, E., Cordeiro, C., Rodríguez-Calvo, M.S., Veira, D.N., Suárez-Peñaranda, J.M., López-Rivadulla, M., Applications of Tandem Mass Spectrometry (LC-MSMS) in estimating the post-mortem interval using the biochemistry of the vitreous humour (2012) Forensic Sci. Int., 223, pp. 160-164Mihailovic, Z., Atanasijevic, T., Popovic, V., Milosevic, M.B., Sperhake, J.P., Estimation of the Postmortem Interval by Analyzing Potassium in the Vitreous Humor Could Repetitive Sampling Enhance Accuracy? (2012) Am. J. Forensic Med. Pathol., 33, pp. 400-403Angi, M., Kalirai, H., Coupland, S.E., Damato, B.E., Semeraro, F., Romano, M.R., Proteomic Analyses of the Vitreous Humour-Review (2012) Mediators Inflammation, 2012, pp. 1-7Tumram, N.K., Bardale, R.V., Dongre, A.P., Postmortem analysis of synovial fluid and vitreous humour for determination of death interval: A comparative study (2011) Forensic Sci. Int., 204, pp. 186-190Margalho, C., Franco, J., Corte-Real, F., Vieira, D.N., Illicit drugs in alternative biological specimens: A case report (2011) J. For. Legal Med., 18, pp. 132-135Soltyszewski, I., Janica, A.N., Pepiński, W., Spólnicka, M., Zbiec, R., Janica, J., Vitreous humour as a potential DNA source for postmortem human identification (2007) Folia Histochem. Cytobiol., 45, pp. 135-136Lang, G.K., Ophthamology: A Pocket Textbook Atlas (2006) Thieme, 2, pp. 287-289Sussulini, A., Banzato, C.E.M., Arruda, M.A.Z., Exploratory analysis of the serum ionomic profile for bipolar disorder and lithium treatment (2011) Int. J. Mass Spectrom., 307, pp. 182-184Nowak, R., Balabanova, S., Determination of calcium and magnesium in postmortem human vitreous humor as a test to ascertain the cause and time of death (1989) Z. Rechtsmed., 102, pp. 179-183Balabanova, S., Gras, V., Forensic value of phenobarbital, calcium and magnesium determination in vitreous humor (1992) Arch. Kriminol., 189, pp. 48-55Wheeler, M.S., Butts, J.D., Hudson, P., Vitreous humor magnesium in alcoholics (1983) Am. J. Forensic Med. Pathol., 4, pp. 105-110Farmer, J.G., Benomran, F., Watson, A.A., Harland, W.A., Magnesium, potassium, sodium and calcium in post-mortem vitreous humour from humans (1985) Forensic Sci. Int., 27, pp. 1-13Yamaguchi, Y., Matsumura, T., Ichida, K., Okamoto, K., Nishino, T., Human Xanthine Oxidase Changes its Substrate Specificity to Aldehyde Oxidase Type upon Mutation of Amino Acid Residues in the Active Site: Roles of Active Site Residues in Binding and Activation of Purine Substrate (2007) J. Biochem., 141, pp. 513-524Salam, H.F.A., Shaat, E.A., Aziz, M.H.A., Sheta, A.A.M., Hussein, H.A.S.M., Estimation of postmortem interval using thanatochemistry and postmortem changes (2012) Alex. J. of Med., 48, pp. 335-344McGahan, M.C., Fleisher, L.N., A micromethod for the determination of iron and total iron-binding capacity in intraocular fluids and plasma using electrothermal atomic absorption spectroscopy (1986) Anal. Biochem., 156, pp. 397-402McGahan, M.C., Fleisher, L.N., Inflammation-induced changes in the iron concentration and total iron-binding capacity of the intraocular fluids of rabbits (1986) Graefe's Arch. Clin. Exp. Ophthalmol., 226, pp. 27-30Beazley, K.E., Nurminskaya, M., Talbot, C.J., Linsenmayer, T.F., Corneal epithelial nuclear ferritin: Developmental regulation of ferritin and its nuclear transporter ferritoid (2008) Dev. Dyn., 237, pp. 2529-2541Goralska, M., Ferrell, J., Harned, J., Lall, M., Nagar, S., Fleisher, L.N., McGahan, M.C., Iron metabolism in the eye: A review (2009) Exp. Eye Res., 88, pp. 204-215Lu, J., Stewart, A.J., Sadler, P.J., Pinheiro, T.J., Blindauer, C.A., Albumin as a zinc carrier: Properties of its high-affinity zinc-binding site (2008) Biochem. Soc. Trans., 36, pp. 1317-1321Oettl, K., Reibnegger, G., Schmut, O., The redox state of human serum albumin in eye diseases with and without complications (2011) Acta Ophthalmol., 89, pp. 174-179Fukuda, M., Fullard, R.J., Willcox, M.D., Baleriola-Lucas, C., Bestawros, F., Sweeney, D., Fibronectin in the tear film (1996) Invest. Ophthalmol. Visual Sci., 37, pp. 459-467