On the road of dried blood spot sampling for antidoping tests: Detection of GHRP‐2 abuse

Abstract

Dried blood spots (DBSs) sampling is gaining support by the antidoping community because of simplicity and cost‐effective characteristics, especially in collection, transport, and storage. Nevertheless, DBS applicability demands specific studies for each of the analytes proposed for testing. Here, GHRP‐2 has been selected as a representing member of the growth hormone‐releasing peptides (GHRPs) family to provide further evidence of DBS suitability for GHRPs abuse detection in sport testing. An analytical procedure to extract GHRP‐2 and its main metabolite (AA‐3) from DBS and to detect them by liquid chromatography–tandem mass spectrometry (LC–MS/MS) has been developed. The method has been validated for the detection of GHRP‐2. Specificity and identification capabilities have been assessed in agreement with antidoping guidelines. The low AA‐3 levels found in DBS samples prevented its effective application for the determination of this metabolite. The limit of detection (LoD) for GHRP‐2 has been established at 50 pg/ml. Long‐term stability (>2 years) has been confirmed. The procedure has been successfully applied to actual DBS samples from an administration study with a single intravenous dose of GHRP‐2 (100 μg) being detected up to 4 h after drug injection. GHRP‐2 concentrations have been higher in venous blood DBS than in capillary blood DBS. Despite the observed differences, a similar detection window has been achieved independently of the type of blood used. In summary, this study provides specific evidence supporting DBS usefulness to detect GHRP‐2, and potentially other GHRPs family members, for antidoping tests.

Annual banned‐substance review: Analytical approaches in human sports drug testing 2020/2021

“…The Q/orbitrap‐based mass analyzer was operated with ESI and in targeted SIM (tSIM) mode at a resolution of 60,000 (@ m/z 200), and a single analytical run was completed within 15 min, thus representing settings compatible with established routine doping control approaches. Whether or not DBS offer added value also for testing GH releasing factors, for example, GHRPs, was assessed by Reverter‐Brachat et al 85 Despite of a LOD as low as 50 pg/ml and a proven long‐term stability of the analyte in DBS for more than 2 years, GHRP‐2 was detected in post‐injection DBS samples (100 μg GHRP‐2 bolus) for 4 h only. These results are particularly important for situations where test menus need to be decided on grounds of limited availability of resources, regardless whether the resources concern manpower or costs of the testing material and sample shipment.…”

Coordination Ion Spray for Analysis of the Growth Hormones Releasing Peptides in Urine—An Application Study

“…In the case of well-equipped high-end instruments (especially HRMS instruments), there are no issues in finding concentrations below 1 ng/mL. The main issue with such compounds is their stability and the excretion time in urine [14][15][16]. The main sample preparation technique for such compounds, like GHRP-2 and GHRP-6, is solid-phase extraction on SCX or WCX sorbents of different volumes.…”

An analytical approach for on‐site analysis of breath samples for Δ9‐tetrahydrocannabinol (THC)

“…Two promising alternatives include micro‐sampling techniques of dried blood spots (DBS) 12–14 and dried plasma spots (DPS), 15–17 which do not require venous puncture, a phlebotomist, or refrigerated sample shipments. Recent developments in this new technology have led to its acceptance in antidoping programs globally 18 . Another promising alternative is the collection of oral fluids (OF), 19,20 which is also noninvasive and easy to perform and has shown promising results over the past 10 years.…”

“…Recent developments in this new technology have led to its acceptance in antidoping programs globally. 18 Another promising alternative is the collection of oral fluids (OF), 19,20 which is also noninvasive and easy to perform and has shown promising results over the past 10 years. An additional biological sample matrix that has shown some recent promise is exhaled breath (EB).…”