MSACL 2019 EU Abstract

LC-MS-based Pharmacokinetics of Insulin Aspart Measured in Individuals with Type 2-Diabetes During Operation Closed-Loop Glucose Control

Jean-Christophe Prost (1), David Herzig (2), Joachim Dehais (2), Cédric Bovet (1), Christoph Stettler (2), Lia Bally (2)
(1) University Institute of Clinical Chemistry, Inselspital, Bern University Hospital, University of Bern, Switzerland (2) Department of Diabetes, Endocrinology, Clinical Nutrition & Metabolism, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland

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Jean-Christophe Prost (Presenter) Bern University Hospital

Relevant Financial Disclosures (within past 24 months) No relevant financial relationship(s) to disclose. Conflict mitigation NOT REQUIRED.

Introduction:
Simultaneous quantification of endogenous and synthetic insulins could benefit metabolic research and clinical practice, but the available ligand-binding immunoassays lack this capacity. Ultra high-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS) offers a mean to circumvent this analytical shortcoming. Our aim was to establish a UHPLC-HRMS assay to determine pharmacokinetics of two different insulin aspart formulations during closed-loop glucose control in patients with type 2 diabetes.

Methods:
Plasma samples were obtained from a randomized double-blind crossover trial contrasting short-term closed-loop insulin delivery with Fiasp vs. Novorapid (Fiasp and Novorapid, Novo Nordisk Pharma AG) in 15 patients with insulin-treated type 2 diabetes. All patients had residual endogenous insulin productions and had long-acting insulin on board during the experiment. Plasma samples were collected over 10 hours every 15 min for 3h post-meal intake and every 30 min at other times (n = 1000 samples). Insulin quantification was performed using immune-purification (InsuQuant Mass Spectrometric Kit, Thermo Scientific) combined with UHPLC-HRMS (Q Exactive Plus, Thermo). Insulins were enriched from 500 µL of plasma spiked with bovine insulin (internal standard) by using an automated liquid handling system. A pharmacokinetic two-compartment model was used to estimate time-to-peak plasma concentrations (tmax) and metabolic clearance rate (MCR) of insulin aspart.

Results:
Prior measurement of study samples, the approach was validated for human, aspart and glargine insulins. Based on spiking experiments with stripped serum, acceptable inter and intra-assay CVs ≤ 12 % and bias < 18.5 % were obtained between 7.5 - 960 pM for all insulins.
Estimated tmax was 68±22 min and 90±32 min for Fiasp and Novorapid, respectively (mean of the differences = -12±21 min, p=0.097). MCR was 23.3±20.9 ml/kg/min for Fiasp and 20.0±16.1 ml/kg/min for Novorapid (mean of the differences = 3.7±24.2 ml/kg/min, p=0.64). The non-significant differences in pharmacokinetics between the two insulin formulations was in line with the absence of significant closed-loop performance differences (similar glucose control with Fiasp and Novorapid).

Conclusions & Discussion:
We report here for the first time a UHPLC-HRMS- based approach for a pharmacokinetic study in patients with type 2 diabetes who were exposed to two types of insulins (long-acting insulin and insulin aspart). Simultaneous quantification of human and different synthetic insulins by UHPLC-HRMS at high throughput offers new avenues to evaluate and compare different diabetes treatments.