Rapid quantification of insulin degludec by immunopurification combined with liquid chromatography high-resolution mass spectrometry

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, easy-to-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.

High‐throughput seminal fluid identification by automated immunoaffinity mass spectrometry

“…In recent years, hybrid approaches combining affinity purification and targeted MS have been developed. Termed immunoaffinity mass spectrometry, these strategies have been utilized in forensic toxicology for postmortem insulin analysis, by the pharmaceutical industry during clinical development, as well as for medical diagnostics [20][21][22]. This approach is fundamentally different from the more traditional immunochromatographic assays that are currently employed in the workflow of many caseworking laboratories.…”

Effects of Aerobic Exercise on Systemic Insulin Degludec Concentrations in People with Type 1 Diabetes

“…Quantification of IDeg was performed using a highly specific assay combining immunopurification with liquid chromatography high-resolution mass spectrometry (LC-HRMS) 8 . Methodological details are provided in the supplementary appendix.…”

“…We, therefore, applied a highly specific mass-spectrometric assay [8] with the objective to explore the effect of exercise on systemic IDeg concentrations in adults with type 1 diabetes.…”

“…5,6 However, robust quantification of insulin concentrations during exercise conditions will improve our understanding of exercise-associated metabolism, thereby informing treatment recommendations related to exercise, in particular prediction models and algorithms. 7 We, therefore, applied a highly specific mass-spectrometric assay 8 with the objective to explore the effect of exercise on systemic IDeg concentrations in adults with type 1 diabetes.…”