An antibody-free, ultrafiltration-based assay for the detection of growth hormone-releasing hormones in urine at low pg/mL concentrations using nanoLC-HRMS/MS

This work presents an ultrafiltration-based, validated method for the screening and confirmation of prohibited growth hormone-releasing hormone (GHRH) analogues (sermorelin/CJC-1293, sermorelin metabolite, CJC-1295 and tesamorelin) in urine by nanoLC-HRMS/MS. Sample preparation avoids the use of lab...

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Published in:Journal of pharmaceutical and biomedical analysis 2022-05, Vol.214, p.114726-114726, Article 114726
Main Authors: Coppieters, Gilles, Deventer, Koen, Polet, Michaël, Van Eenoo, Peter, Judák, Péter
Format: Article
Language:English
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Doping
GHRH
nanoLC-HRMS
Peptides
Ultrafiltration
Urine
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container_start_page 114726
container_title Journal of pharmaceutical and biomedical analysis
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creator Coppieters, Gilles
Deventer, Koen
Polet, Michaël
Van Eenoo, Peter
Judák, Péter
description This work presents an ultrafiltration-based, validated method for the screening and confirmation of prohibited growth hormone-releasing hormone (GHRH) analogues (sermorelin/CJC-1293, sermorelin metabolite, CJC-1295 and tesamorelin) in urine by nanoLC-HRMS/MS. Sample preparation avoids the use of laborious antibody-based extraction approaches and consists solely of preconcentration by ultrafiltration. Even in the absence of immuno-affinity purification steps, high sensitivity was still ensured as limits of detection between 5 and 25 pg/mL and limits of identification between 25 and 50 pg/mL were established. The robustness of the miniaturized chromatographic setup was evaluated through the injection of 200 + preconcentrated urinary extracts. In a comparison with immuno-affinity purification, enhanced recoveries (59 – 115%) and similar sensitivity were achieved, yet at lower operational costs. Stability experiments showed the importance of the proper handling of urine samples to avoid degradation of these peptide hormones, especially for sermorelin and its metabolite which were found to rapidly degrade at temperatures > 4 °C and pH values
doi_str_mv 10.1016/j.jpba.2022.114726
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subjects Doping
GHRH
nanoLC-HRMS
Peptides
Ultrafiltration
Urine
title An antibody-free, ultrafiltration-based assay for the detection of growth hormone-releasing hormones in urine at low pg/mL concentrations using nanoLC-HRMS/MS
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