Fully Automated Blood Spot Testing Detects 46 Peptide Doping Agents Including Growth Hormone Peptides

Automated dried blood spot sample preparation enabled detection of 46 peptide and non-peptide doping agents by LC-HRMS, with GHRP-2/GHRP-6 confirmed in real athlete samples.

Lange, Tobias et al.·Analytical and bioanalytical chemistry·2020·n/a (analytical method)method development
RPEP-04922Method developmentn/a (analytical method)2020RETHINKTHC RESEARCH DATABASErethinkthc.com/research

Quick Facts

Study Type
method development
Evidence
n/a (analytical method)
Sample
N=N/A (method validation)
Participants
Dried blood spot samples including authentic post-administration specimens

What This Study Found

The method combined robotic-assisted near-infrared hematocrit measurement with fully automated strong cation exchange solid-phase extraction. This enabled detection of 46 lower molecular mass (<2 kDa) peptide and non-peptide doping agents by LC-HRMS.

Target analytes included agonists of the gonadotropin-releasing hormone receptor, ghrelin receptor, growth hormone receptor, and antidiuretic hormone receptor. Several glycine derivatives of GHRPs, designed to evade current testing, were also covered.

Validation met WADA guidelines with LODs between 0.5 and 20 ng/mL. Proof of concept was demonstrated with authentic post-administration samples containing GHRP-2 and GHRP-6. The method also worked with blood collected by microneedle devices from the upper arm.

Key Numbers

46 analytes; LODs 0.5-20 ng/mL; WADA-validated; GHRP-2/GHRP-6 confirmed in real samples; microneedle DBS compatible

How They Did This

This was an analytical method development study. Researchers used robotic near-infrared spectroscopy for hematocrit measurement, automated SPE for sample preparation, and LC-HRMS for detection. Validation followed WADA guidelines. Real post-administration samples and microneedle-collected blood were tested.

Why This Research Matters

Dried blood spots are increasingly used in doping control because they are easy to collect and transport. This automated method makes peptide doping detection faster and more reliable, while also catching designer peptides that were specifically made to avoid current tests.

The microneedle compatibility means athletes could be tested with a nearly painless arm device instead of traditional blood draws.

The Bigger Picture

Dried blood spots are easy to collect and transport, making doping testing more accessible worldwide. Automating the analysis removes human error and increases throughput, strengthening anti-doping enforcement especially for peptide drugs that are increasingly abused in sports.

What This Study Doesn't Tell Us

The method focuses on lower molecular mass peptides (<2 kDa) and would not detect larger protein-based doping agents like EPO or growth hormone directly.

Long-term stability of all 46 analytes in dried blood spots was not comprehensively assessed.

Questions This Raises

  • ?Can this method be extended to larger peptide doping agents?
  • ?How stable are these analytes in dried blood spots during shipping?
  • ?Will microneedle blood collection devices improve athlete compliance?

Trust & Context

Key Stat:
46 agents detected from dried blood spots with full automation — including confirmed GHRP-2 and GHRP-6 in real doping control samples
Evidence Grade:
Strong evidence as a validated analytical method with real-world confirmation in actual doping control samples.
Study Age:
Published in 2020. DBS-based doping detection continues to expand with improved sensitivity.
Original Title:
Fully automated dried blood spot sample preparation enables the detection of lower molecular mass peptide and non-peptide doping agents by means of LC-HRMS.
Published In:
Analytical and bioanalytical chemistry, 412(15), 3765-3777 (2020)
Database ID:
RPEP-04922

Evidence Hierarchy

Meta-Analysis / Systematic Review
Randomized Controlled Trial
Cohort / Case-Control
Cross-Sectional / ObservationalSnapshot without intervening
This study
Case Report / Animal Study
What do these levels mean? →

Frequently Asked Questions

What are growth hormone releasing peptides (GHRPs)?

GHRPs like GHRP-2 and GHRP-6 stimulate growth hormone release. Athletes misuse them for muscle building and recovery. They are banned by WADA but difficult to detect because they are used in small amounts.

Why use dried blood spots for doping tests?

DBS requires only a finger prick (much easier than drawing blood), the spots are stable for shipping, and collection can happen anywhere without specialized equipment. This makes testing more accessible worldwide.

Read More on RethinkPeptides

Cite This Study

RPEP-04922·https://rethinkpeptides.com/research/RPEP-04922

APA

Lange, Tobias; Thomas, Andreas; Walpurgis, Katja; Thevis, Mario. (2020). Fully automated dried blood spot sample preparation enables the detection of lower molecular mass peptide and non-peptide doping agents by means of LC-HRMS.. Analytical and bioanalytical chemistry, 412(15), 3765-3777. https://doi.org/10.1007/s00216-020-02634-4

MLA

Lange, Tobias, et al. "Fully automated dried blood spot sample preparation enables the detection of lower molecular mass peptide and non-peptide doping agents by means of LC-HRMS.." Analytical and bioanalytical chemistry, 2020. https://doi.org/10.1007/s00216-020-02634-4

RethinkPeptides

RethinkPeptides Research Database. "Fully automated dried blood spot sample preparation enables ..." RPEP-04922. Retrieved from https://rethinkpeptides.com/research/lange-2020-fully-automated-dried-blood

Access the Original Study

Study data sourced from PubMed, a service of the U.S. National Library of Medicine, National Institutes of Health.

This study breakdown was produced by the RethinkPeptides research team. We analyze and report published research findings without making health recommendations. All interpretations are based solely on the published abstract and study data.