Mots-C
Mitochondrial open reading frame of the 12S rRNA-c, MT-RNR1, Mitochondrial-derived peptide MOTS-c
Development of a mass spectrometry based detection method for the mitochondrion-derived peptide MOTS-c in plasma samples for doping control purposes.
Knoop. Andre A; Thomas. Andreas A; Thevis. Mario M
Key Findings
- A LC/MS assay for MOTS‑c in plasma was fully validated according to WADA standards, with a lower limit of detection of 100 pg/mL.
- The assay also detects four in‑vitro metabolites and two oxidation products, improving traceability of synthetic MOTS‑c.
- Endogenous MOTS‑c levels reported by ELISA (45.9‑218.5 ng/mL) were not confirmed by the more precise LC/MS method.
Practical Outcomes
- For biohackers, this work shows that common ELISA kits may give falsely high MOTS‑c readings, so relying on them for personal monitoring could be misleading. Accurate measurement currently requires sophisticated LC/MS equipment, which limits everyday testing. The detection limit (100 pg/mL) and metabolite coverage provide a benchmark for future, more accessible assays.
Summary
Scientists created a lab test that can reliably spot the MOTS‑c peptide in blood using liquid chromatography‑mass spectrometry. The test can detect very low amounts (down to 100 pg/mL) and also looks for related breakdown products. However, the levels measured by a popular ELISA kit were much higher than what the new method found, suggesting the ELISA may over‑estimate natural MOTS‑c in people.
Abstract
The mitochondrial open reading frame of 12S rRNA type-c (MOTS-c) peptide was recently discovered and described to control metabolic homeostasis through AMPK activation along with AICAR accumulation. Consequently, it appears advisable to monitor the potential use of synthetic MOTS-c in sports, and a detection method suitable for sports drug testing purposes is necessary. For the detection of MOTS-c in doping control plasma samples, a test method employing liquid chromatography and mass spectrometry (LC/MS) was developed. Following optimization, the assay was comprehensively validated and additional parameters such as the (long-term) stability and in vitro metabolism of the peptide were evaluated. In order to determine endogenous MOTS-c reference limits, the results generated by LC/MS-based detection were compared with those obtained with a commercially available enzyme-linked immunosorbent assay (ELISA). The LC/MS-based test method was fully validated for quantitative results interpretation according to the World Anti-Doping Agency's International Standard for Laboratories (WADA's ISL). It was found to be specific and sensitive, enabling a lower limit of detection (LLOD) for hMOTS-c in plasma at 100 pg/mL. Following optimization, animal MOTS-c analogues and four metabolites as well as two oxidation products were implemented. However, endogenous levels of a reference population of 20 healthy subjects studied by ELISA experiments (45.9-218.5 ng/mL) could not be confirmed by LC/MS. A mass spectrometric detection assay for MOTS-c in human plasma samples was developed and successfully validated according to WADA's ISL, providing an additional tool for future doping control purposes. Besides MOTS-c, the assay also includes four in vitro derived metabolites and two oxidation products, which might further improve the traceability of the drug. The analytical approach was compared with a commercially available ELISA, and considerable differences in measured MOTS-c levels were observed.
Study Information
pubmed
2019
2019-02-28T00:00:00.000Z
10.1002/rcm.8337
19
19