Abstract
In our study, we analysed the effect of a variety of storage conditions on the methaemoglobin (MetHb) content of blood samples obtained from altogether 110 deceased subjects with diverse causes of death, including three ‘poppers’-related fatalities. The obtained results were compared to data from blood samples of six living, healthy subjects. Results obtained from the spectrophotometric measurement of blood MetHb content suggest that storage at room temperature (RT) and storage at −20 °C result in either highly fluctuating values, as was the case for the RT samples, or values much higher than the initial MetHb concentrations when stored at −20 °C. Blood samples at 4 °C showed more stable MetHb levels, which, however, increased with up to 4 % of the initial value after only 3 weeks of storage. These factors pose a problem in forensic toxicology, especially in nitrite abuse cases, where the involvement of such substance abuse is often unknown at the time of blood sampling and thus often requires longer storage times. Nevertheless, even after the storage of blood samples over several months at 4 and −20 °C, ‘poppers’ cases still show a significantly higher MetHb concentration as compared to non-‘poppers’ samples that were stored for the same time period under identical conditions.
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References
Kosaka H, Imaizumi K, Imai K, Tyuma I (1979) Stoichiometry of the reaction of oxyhemoglobin with nitrite. Biochim Biophys Acta 581:184–188
Doyle M, Pickering R, Weert TD, Hoekstra J, Pater D (1981) Kinetics and mechanism of the oxidation of human deoxyhemoglobin by nitrites. J Biol Chem 256:12393–12398
O’Toole JB, Robbins GB, Dixon DS (1987) Ingestion of isobutyl nitrite, a recreational chemical of abuse, causing fatal methaemoglobinaemia. J Forensic Sci 32:1811–1822
Bradberry SM, Whittington RM, Parry DA, Vale JA (2008) Fatal methemoglobinemia due to inhalation of isobutyl nitrite. J Toxicol - Clin Toxic 32:179–184
Osterlol J, Goldfield D (1984) Butylnitrite transformation in vitro, chemical nitrosation reactions and mutagenesis. J Anal Tox 8:164–168
Vogt S, Hauschel M, Angerer V, Kemp J, Auwärter V (2015) Analysis of ‘poppers’ products and analytical detectability of a sinlge use of ‘poppers’. Toxichem Krimtech 82:74
Fukui A, Yamamoto Y, Matsubara K (1980) Reduction with the lapse of time of methemoglobin blood sample. Jpn J Legal Med 34:563–568
Warburg O, Kubowitz F, Christian W (1930) Über die Katalytische Wirkung von methylenblau in Lebenden Zellen. Biochem Zeitschr 227:245–271
Sleight SD, Sinha DP (1968) Prevention of methemoglobin reduction in blood samples. J Am Vet Med A 152:1521–1525
Sato K, Katsumata Y, Aoki M, Tamaki K, Yada S (1987) Long-term storage of blood samples at freezing temperatures in the presence of a cryoprotectant for hemoglobin assay. Biochem Med Metab B 37:190–196
Rodkey FL, O’Neal JD (1974) Effects of carboxyhaemoglobin on the determination of methemoglobin. Biochem Med 9:261–270
Sato K, Katsumata Y, Aoki M, Oya M, Yada S, Suzuki O (1981) A practical method for the accurate determination of methemoglobin in blood containing carboxyhemoglobin. Forensic Sci Int 17:177–184
Sato K, Tamaki K, Okajima H, Katsumata Y (1988) Long-term storage of blood samples as whole blood at extremely low temperatures for methemoglobin determination. Forensic Sci Int 37:99–104
Sato K, Tamaki K, Tsutsumi H, Okajima H, Katsumata Y (1990) Storage of blood for methemoglobin determination: comparison of storage with a cryoprotectant at −30 °C and without any additions at −80 °C or −196 °C. Forensic Sci Int 45:129–134
Ghanem AAE, Abdel-Rahman RH, Shabka OA (2012) Stability of carboxyhaemoglobin in blood samples at different periods and temperatures: a forensic and toxicological tool for diagnosis. J Clinic Toxicol 2:144. doi:10.4172/2161-0495.1000144
Lee CW, Tam JC, Kung LK, Yim LK (2003) Validity of CO-oximetric determination of carboxyhaemoglobin in putrefying blood and body cavity fluid. Forensic Sci Int 132:153–156
Dinis-Oliviera R, Carvalho F, Duarte J, Remiao F, Marques A, Santos A et al (2010) Collection of biological samples in forensic toxicology. Toxicol Mech Method 20:363–414
Shihana F, Dawson AH, Dissanayake DM (2011) Method of stabilizing blood for the determination of methemoglobin. J Clin Lab Anal 25:366–368
Jensen FB (2001) Comparative analysis of autooxidation of haemoglobin. J Exp Biol 204:2029–2033
Rowe AW, Eyster E, Kellner A (1968) Liquid nitrogen preservation of red blood cells for transfusion—a low glycerol-rapid freeze procedure. Cryobiology 5:119–128
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The authors wish to thank Ms. Birgit Övgüer for the temperature-dependent measurements of samples from population 2.
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Domingo, O., Stöver, A., Roider, G. et al. Detection of methaemoglobinaemia and its application in ‘poppers’ abuse: maintaining the right balance between reduction and autooxidation during storage. Int J Legal Med 131, 369–377 (2017). https://doi.org/10.1007/s00414-016-1410-8
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DOI: https://doi.org/10.1007/s00414-016-1410-8