Analysis of growth hormone releasing hormone and its analogs in urine using nano liquid chromatography coupled with quadrupole/orbitrap mass spectrometry.
The study presents a validated method for detecting GHRH and its analogs in urine with limits of detection as low as 0.5 ng/mL, which is important for anti-doping compliance.
Where it sits
this study against the rest of the cjc-1295 corpusSummary and findings
This study developed a method for analyzing growth hormone-releasing hormone (GHRH) and its analogs, including CJC-1295, in urine using nano liquid chromatography coupled with quadrupole/orbitrap mass spectrometry. The method aimed to meet World Anti-Doping Agency (WADA) requirements for screening and confirmation analyses. The limits of detection achieved were ≤ 0.5 ng/mL.
Abstract
Growth hormone-releasing hormone (GHRH) and its synthetic analogs are considered performance-enhancing substances and are therefore prohibited by the World Anti-Doping Agency (WADA). The analysis of GHRH and its analogs in urine presents significant analytical challenges due to their inherent in vivo instability, rapid renal clearance, and low urinary concentrations. The present study aimed to develop a robust nano-LC quadrupole/orbitrap mass spectrometry (nano-LC-Q/Orbitrap MS) method for both screening and confirmation analyses of GHRH and its synthetic analogs (sermorelin/CJC-1293, tesamorelin, and CJC-1295) and the primary metabolite of sermorelin in urine, in accordance with WADA requirements. The sample preparation workflow was systematically investigated. Existing solid-phase extraction (SPE) protocols were compared, and two additional commercially available SPE cartridges were evaluated. Within the SPE step, the influence of various washing and elution solvent strengths on peptide recovery was also systematically examined. The effectiveness of different cleanup solvents during the ultrafiltration step was further assessed. Based on these evaluations, a refined approach was developed, incorporating an initial ultrafiltration step followed by SPE. The proposed method was fully validated according to WADA guidelines, assessing key parameters such as selectivity, reliability, limits of detection (LOD), carryover, limits of identification (LOI), robustness, autosampler stability, and matrix effects. The validation results confirmed the method's suitability and robustness for anti-doping testing. Achieved LODs (≤ 0.5 ng/mL) and LOIs (0.5-0.75 ng/mL) demonstrated sufficient sensitivity for effective detection and confirmation analysis of the target peptides in urine.
Background
This paper addresses the analysis of growth hormone releasing hormone (GHRH) and its analogs, which are critical in the regulation of growth hormone secretion. Previous studies have focused on the physiological roles of GHRH, but there is limited information on the analytical methods for detecting these peptides in biological samples. This study is significant as it employs advanced techniques to improve the detection and quantification of GHRH in urine, potentially aiding in pharmacokinetic studies.
Methods
Not reported in abstract.
Results
Not reported in abstract.
Interpretation
Not reported in abstract.
Key findings
- Not reported in abstract.
Limitations
- Not reported in abstract.