Rapid quantification of insulin degludec by immunopurification combined with liquid chromatography high-resolution mass
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RESEARCH PAPER
Rapid quantification of insulin degludec by immunopurification combined with liquid chromatography high-resolution mass spectrometry Gemma Reverter-Branchat 1 & Michael Groessl 2 Eric E. Niederkofler 5 & Lia Bally 1
&
Christos T Nakas 3,4 & Jean-Christophe Prost 4 & Kwasi Antwi 5 &
Received: 22 June 2020 / Revised: 31 August 2020 / Accepted: 24 September 2020 # The Author(s) 2020
Abstract Insulin degludec is an ultra-long-acting insulin analogue that is increasingly being used in diabetes due to its favourable efficacy and safety profile. Thus, there is an increasing demand for a reliable and specific analytical method to quantify insulin degludec for research, pharmaceutical industry and clinical applications. We developed and validated an automated, high-throughput method for quantification of insulin degludec in human blood samples across the expected clinical range combining immunopurification with high-resolution mass spectrometry. Validation was performed according to the requirements of the US Food and Drug Administration. The method satisfyingly met the following parameters: lower limit of quantification (120 pM), linearity, accuracy (error < 5%), precision (CV < 7.7%), selectivity, carry-over, recovery (89.7–97.2%), stability and performance in the presence of other insulin analogues. The method was successfully applied to clinical samples of patients treated with insulin degludec showing a good correlation with the administered dose (r2 = 0.78). High usability of the method is supported by the small specimen volume, automated sample processing and short analysis time. In conclusion, this reliable, easyto-use and specific mass spectrometric insulin degludec assay offers great promise to address the current unmet need for standardized insulin analytics in academic and industrial research. Keywords Insulin degludec . Immunopurification . Mass spectrometry . Quantification
Gemma Reverter-Branchat and Michael Groessl contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00216-020-02971-4) contains supplementary material, which is available to authorized users. * Michael Groessl [email protected] 1
Department of Diabetes, Endocrinology, Nutritional Medicine and Metabolism, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, Switzerland
2
Department of Nephrology and Hypertension, Inselspital, Bern University Hospital, University of Bern, Freiburgstrasse, 3012 Bern, Switzerland
3
Laboratory of Biometry, School of Agriculture, University of Thessaly, 38446 Nea Ionia, Magnesia, Greece
4
University Institute of Clinical Chemistry, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, Switzerland
5
Thermo Fisher Scientific, Tempe, AZ 85280, USA
Introduction Over 200 million people with diabetes are treated with insulin and this number continues to increase globally [1]. Nowadays, synthetic insulin or insulin analogues, which have modified primary sequences compared w
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