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Detection of insulins in postmortem tissues: an optimized workflow based on immunopurification and LC–MS/HRMS detection

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Abstract

Diabetes is a worldwide disease in perpetual expansion. Type 1 and sometimes type 2 diabetic patients require daily human insulin (HI) or analog administration. Easy access to insulins for insulin-treated diabetics, their relatives, and medical professionals can enable abuse for suicidal or homicidal purpose. However, demonstrating insulin overdose in postmortem blood is challenging. Tissue analyses are contributive, as insulins can accumulate before death or undergo only limited degradation. The present study describes an assay for HI and synthetic analogs (lispro, aspart, glulisine, detemir and degludec, glargine and its main metabolite (M1)) in liver, kidney, muscle, and injection site samples. It is based on a 5-step sample preparation (reduction of tissue sample size, homogenization, extraction, concentration, and immunopurification) associated with liquid chromatography coupled to high-resolution mass spectrometry (LC–MS/HRMS). Selectivity and limit of detection (LOD) for all target analogs were assessed in the above matrices. LOD was determined at 25 ng/g for HI and for analogs except detemir and degludec, where LOD was 50 ng/g in kidney and injection site samples and 80 ng/g in the liver and muscle. The method was applied to13 forensic cases in which insulin use was suspected.

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Acknowledgements

The authors gratefully acknowledge the staff from Medicolegal Institute of Lyon for their practical collaboration and their investment in this work and Iain McGill (freelance translator specializing in health and life sciences) for proofreading the article.

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Authors and Affiliations

  1. LAT LUMTOX Laboratory, 32 Rue du 35ème Régiment d’Aviation 69500, Bron, France

    Charline Bottinelli & Fabien Bévalot

  2. Department of Forensic Medicine, Edouard Herriot Hospital, Hospices Civils de Lyon, Lyon, France

    Nathalie Cartiser & Laurent Fanton

  3. Faculty of Medicine Lyon-Est, University of Lyon, UCBL1, Lyon, France

    Laurent Fanton

  4. Toxicology Laboratory, ISPB Faculty of Pharmacy, University of Lyon, UCBL1, Lyon, France

    Jérôme Guitton

  5. Pharmacology-Toxicology Laboratory, Lyon-Sud Hospital, Hospices Civils de Lyon, Lyon, France

    Jérôme Guitton

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  1. Charline Bottinelli
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  2. Fabien Bévalot
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  3. Nathalie Cartiser
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  4. Laurent Fanton
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  5. Jérôme Guitton
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Correspondence to Charline Bottinelli.

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All procedures performed in studies involving human participants were in accordance with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Samples were obtained from forensic autopsies and corresponded to a complete protocol of sample collection for forensic toxicological evidence. Toxicological results can be used anonymously for scientific purposes.

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The authors declare no competing interests.

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Bottinelli, C., Bévalot, F., Cartiser, N. et al. Detection of insulins in postmortem tissues: an optimized workflow based on immunopurification and LC–MS/HRMS detection. Int J Legal Med 135, 1813–1822 (2021). https://doi.org/10.1007/s00414-021-02598-9

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  • DOI: https://doi.org/10.1007/s00414-021-02598-9

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