Utilizing ELISA-plate based immunopurification and liquid chromatography-tandem mass spectrometry for the urinary detection of short- and long acting human insulin analogues

Judák, Péter; Eenoo, Peter Van; Deventer, Koen

doi:10.1016/j.jpba.2018.02.024

Cited by 21 publications

(23 citation statements)

References 25 publications

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“…This method showed a high sensitivity for quantitative analysis of insulin degludec, but is susceptible to endogenous insulin interference. Judák, Van Eenoo, and Deventer (2018) developed and validated an LC–MS/MS method for quantifying insulin degludec in human urine. This method uses immune capture to process a 15 mL urine sample to achieve an LLOQ of 10 pg/mL.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous quantitative determination of liraglutide and insulin degludec in rat plasma by liquid chromatography–tandem mass spectrometry method and its application

Zhai

Dong

et al. 2020

Biomedical Chromatography

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A simple, fast and high‐throughput LC–tandem mass spectrometry method was developed and validated to simultaneously measure liraglutide and insulin degludec in rat plasma. After protein precipitation, plasma samples were subjected to gradient elution using an InertSustain Bio C18 column with 1000/20/1 water/acetonitrile/formic acid (v/v/v) and 1000/1 acetonitrile/formic acid (v/v) as the mobile phase. The method was validated from 1.00 to 500 ng/mL of liraglutide and insulin degludec. Further, the extraction recovery from the plasma was 41.8%–49.2% for liraglutide and 56.5%–69.7% for insulin degludec. Intra‐ and inter‐day precision of liraglutide was 3.5%–9.4% and 8.4%–9.8%, respectively, whereas its accuracy was between −12.6% and −1.3%. Intra‐ and inter‐day precision of insulin degludec was 5.2%–13.6% and 11.8%–19.1%, respectively, showing an accuracy between −3.0% and 9.9%. As a result, the method was successfully applied to a pharmacokinetics study of liraglutide and insulin degludec following a single‐dose subcutaneous administration to rats.

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Section: Introductionmentioning

confidence: 99%

Simultaneous quantitative determination of liraglutide and insulin degludec in rat plasma by liquid chromatography–tandem mass spectrometry method and its application

Zhai

Dong

et al. 2020

Biomedical Chromatography

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“…Similarly to our previous study 3 all peptide stock standard solutions were prepared at 0.1 mg/mL using a mix of H 2 O (A), ACN (B), HCOOH (C) (A:B:C, 60/40/0.02 v/v/v) and were stored at −20°C. Working solutions (10 ng/mL, 20 ng/mL) were prepared using the same mixture and also contained BSA at a concentration of 5 μg/mL.…”

Section: Methodsmentioning

confidence: 99%

“…Insulin and its synthetic analogs are among the group of peptidic compounds indicated on the World Anti‐Doping Agency (WADA) List of prohibited substances 1 . Therefore, several mass spectrometric (MS) assays have been developed to detect its misuse in the past decade 2–7 …”

Section: Introductionmentioning

confidence: 99%

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Urinary detection of rapid‐acting insulin analogs in healthy humans

Judák

Coppieters

Lapauw

et al. 2020

Drug Testing and Analysis

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Human insulin and its synthetic analogs are considered as life‐saving drugs for people suffering from diabetes mellitus. Next to the therapeutic use, scientific and non‐scientific literature (e.g. bodybuilding forums; antidoping intelligence and investigation reports) indicate that these prohibited substances are used as performance enhancing agents. In the present report, the development and validation of a sensitive analytical strategy is described for the urinary detection of three rapid‐acting insulin analogs (Lispro, Aspart, Glulisine). The method is based on sample purification by the combination of ultrafiltration and immunoaffinity purification and subsequent analysis by nano‐flow liquid chromatography coupled to high resolution mass spectrometry. Next to the results on different validation parameters (LOD: 10 pg/mL; recovery: 25–48%; matrix effect: −3‐(−8) %), data on urinary elimination times, which were obtained in the frame of an administration study with the participation of healthy volunteers, are presented. The determined detection windows (~9 hours) are expected to help to evaluate current routine analytical methods and aim to aid doping authorities to set appropriate target windows for efficient testing.

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“…The detection of peptidic recombinant hormones can be challenging, especially if the structure of the exogenous hormone is similar to or identical to the endogenous hormone. Their analysis is generally performed with immuno‐affinity or ELISA, often coupled with mass‐spectrometry 65,70 . The specificity of the detection methods is the strength, but also the limitation, of these approaches.…”

Section: Current Challenges Of Screening For Anabolic Substancesmentioning

confidence: 99%

A role for metabolomics in the antidoping toolbox?

Narduzzi

Audran²,

Bizec

et al. 2020

Drug Testing and Analysis

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Evidence of the continuous rise of novel doping agents and novel doping strategies calls for the development of more accurate multi‐target screening methods. Direct multi‐target screening approaches are restricted to the targeted substances and their turnover. The development of effective “indirect” screening methods requires a priori a deep understanding of the metabolism of the substance. The biological passport has been demonstrated to be very effective, but it is limited to about 20 indirect parameters. The standard antidoping analytical methods are hence targeted and do not aim directly to identify unknown substances. Also, the detection of doping agents is limited by the excretion of the substance. This study considers metabolomics for the screening of performance enhancing hormone abuse by athletes, based on the following pieces of evidence: (1) hormones have a strong influence on human metabolism, changing several parameters in many tissues, organs, and bio‐fluids; (2) metabolomics has been demonstrated to be a very accurate tool to depict the metabolic status of several organisms, tissues, and for several human diseases, including hormone deficiencies; (3) metabolomics has been demonstrated to be able to distinguish hormone‐treated animals from controls in many species, without the need for a priori knowledge of the metabolism for the specific substance. The literature shows that metabolomics could be an appropriate tool to detect hormone abuse, keeping in mind the strength and the limitation of such an approach.

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Utilizing ELISA-plate based immunopurification and liquid chromatography-tandem mass spectrometry for the urinary detection of short- and long acting human insulin analogues

Cited by 21 publications

References 25 publications

Simultaneous quantitative determination of liraglutide and insulin degludec in rat plasma by liquid chromatography–tandem mass spectrometry method and its application

Simultaneous quantitative determination of liraglutide and insulin degludec in rat plasma by liquid chromatography–tandem mass spectrometry method and its application

Urinary detection of rapid‐acting insulin analogs in healthy humans

A role for metabolomics in the antidoping toolbox?

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