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• W4291291256 abstract "To prove the involvement of insulins in intoxication is challenging in postmortem cases. Two insulins assays were developed and validated, quantitatively in blood and qualitatively in tissues. The aim is to share feedback from 4 years of these methods’ application, in clinical and forensic contexts. This retrospective study focused on cases for which insulins analysis was requested in these last 4 years. The studied parameters were: context of the request, samples transfer and analysis intervals, type and concentration results. For postmortem cases, samples were collected during autopsy according to a homemade protocol consisting in cardiac and peripheral blood, liver, kidney, muscle, injection site (IS). For clinical cases, samples were serum collected during hospitalization. Assay allows detection of human insulin and analogs: aspart, glulisine, lispro, detemir, degludec, glargine and M1. Blood samples assay consisted in 3 steps: protein precipitation, filtration and immunopurification before analysis on LC-HRMS (Bottinelli. Talanta 2021:225). Steps of cutting and ultrasonication in acidic ethanol were added for tissue samples (Bottinelli. Int J Legal Med 2021;135:1813–22). Sixty-six cases were analyzed between January 2018 and March 2022: 59 were postmortem cases for which insulin use was suspected (history of diabetes, insulin material found at corpse discovery place…) and 7 were clinical cases for which insulins analysis was requested for a diagnosis of severe hypoglycemia. Treatment by insulins was known in 23 cases and 13 cases were not diabetic. A total of 10 clinical sera, 94 postmortem bloods (n = 53 peripheral; n = 41 cardiac), 16 IS, 20 muscles, 33 livers and 33 kidneys were analyzed. At least one insulin was identified in 29 cases (43%) corresponding to 8 positive sera (80%), 10 IS (62%), 7 muscles (35%), 12 cardiac bloods (29%), 11 peripheral bloods (21%), 7 kidneys (21%), and 4 livers (12%). In 7 cases, insulin administration was characterized by analysis of tissues as blood analysis was negative. In most postmortem cases, insulins were detected when postmortem interval was less than 72 h. Moreover, 50% of samples were positive when blood is centrifuged and freeze less than 24 h after sampling. All insulins were identified at least one time, but aspart (27%) and glargine/M1 (11%) were the most represented. For aspart, postmortem blood concentrations ranged from < 1.0 ng/mL (LOQ) to 432 ng/mL (mean: 50.1 ng/mL), with the highest measured for two non-diabetic people. Insulins intoxications are underestimated as underlined by numerous authors. Since the application of the new LC-HRMS methods, numerous cases of insulins use and intoxication were revealed. Preparation of postmortem samples is the crucial point requiring several steps with specific material. The expensive and time-consuming methods lead to a non-systematic perform, but only when data from autopsy or enquiries suggest possible insulin administration. Initially not dedicated to clinical cases, methods allowed to elucidate origins of severe hypoglycemia in various situations: Münchausen, degludec spontaneous release, suicide attempt, self-medication. The major detection of aspart and glargine is consistent with the prescription trends in France. More degludec detection was expected but was limited either because it was less implicated in intoxications or due to matrix effects. Autopsy realization within 72 h after death, centrifugation/freezing of blood samples within 24 h after sampling and analyses of several matrices including blood and tissues, seem to be crucial conditions to maximize chances for highlighting presence of insulins in postmortem cases. Even if the “simple” detection of an insulin is informative in numerous cases, interpretation of concentration would be relevant. However, insulins displayed a large range concentrations as numerous parameters could have an influence such as diabetes status, type of analog, metabolism, hemolysis… Define a therapeutic or toxic range seems complex and further studies are required. This could be possible with the increase of interest for this problematic in medico-legal research teams." @default.
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• W4291291256 date "2022-09-01" @default.
• W4291291256 modified "2023-10-18" @default.
• W4291291256 title "Analysis of human insulin and its therapeutic analogs by LC-HRMS: 4 years feedback and perspectives" @default.
• W4291291256 doi "https://doi.org/10.1016/j.toxac.2022.06.049" @default.
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