Study summary · research use only
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
Plain-language summary
Paraphrased from the published abstract below — not a verdict on whether anything works.
This analytical chemistry study (species not specified; urine samples were tested) describes an antibody-free, ultrafiltration-based nanoLC-HRMS/MS method for detecting growth hormone-releasing hormone analogues (sermorelin/CJC-1293, sermorelin metabolite, CJC-1295, and tesamorelin) in urine. The authors report limits of detection between 5 and 25 pg/mL and limits of identification between 25 and 50 pg/mL. Robustness was evaluated using 200 + preconcentrated urinary extracts. Compared with immuno-affinity purification, the method achieved higher recoveries (59 - 115%) with similar sensitivity at lower operational cost. Stability experiments reported that sermorelin and its metabolite degraded rapidly at temperatures above 4 °C and at pH values below 7. The authors state the method may be expandable to other peptide drugs and their metabolites.
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.
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