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.
Coppieters. Gilles G; Deventer. Koen K; Polet. Michaël M; Van Eenoo. Peter P; Judák. Péter P
Key Findings
- An ultrafiltration‑based prep combined with nanoLC‑HRMS/MS can detect GHRH analogues at 5‑25 pg/mL in urine.
- The new method gives recovery rates of 59‑115% and comparable sensitivity to immuno‑affinity methods, but at lower cost.
- Sermorelin and its metabolite degrade quickly in urine above 4 °C or at pH < 7, highlighting the need for proper sample handling.
Practical Outcomes
- For biohackers, the study mainly offers a way to test whether sermorelin is present in urine, which could be useful for anti‑doping checks or quality control. It does not provide new dosing guidance, safety data, or performance benefits, so its direct impact on personal health protocols is limited.
Summary
Scientists created a cheap, antibody‑free lab test that can spot tiny amounts of growth‑hormone‑releasing peptides like sermorelin in urine. The method is sensitive, works after many samples, and costs less than traditional antibody‑based tests, but it mainly helps labs detect illegal or banned substances rather than guide how to use the peptides.
Abstract
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 < 7 in accordance with earlier studies. Without the need for specific antibodies, this method may be expanded to cover emerging peptide drugs (≥ ~3 kDa), as well as their metabolites in the future to facilitate coverage for this class of prohibited substances.
Study Information
pubmed
2022
2022-03-12T00:00:00.000Z
10.1016/j.jpba.2022.114726
2
31