Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Apr;41(2):80-6.
doi: 10.4103/0253-7613.51347.

A simple and rapid ESI-LC-MS/MS method for simultaneous screening of doping agents in urine samples

I Madhusudhana Reddy  1 , Alka Beotra, S Jain, S Ahi
Affiliations

A simple and rapid ESI-LC-MS/MS method for simultaneous screening of doping agents in urine samples

I Madhusudhana Reddy et al. Indian J Pharmacol. 2009 Apr.

Abstract

Objective: The use of performance enhancing substances is banned in sports by the World Anti-Doping Agency (WADA). Though most prohibited substances can be detected by GC/MS, inclusion of corticosteroids and designer drugs has made it essential to detect these critical doping agents on LC/MS/MS due to their better separation and detection.

Materials and methods: A common extraction procedure for the isolation of acidic, basic and neutral drugs from urine samples was developed. A total of 28 doping drugs were analyzed on API 3200 Triple quadrupole mass spectrometer using C18 column in atmospheric pressure electrospray ionization. The mobile phase composition was a mixture of 1% formic acid and acetonitrile with gradient time period.

Results: The method developed was very sensitive for detection of 28 doping agents. The linearity was performed for each drug and the total recovery percentage ranged from 57 to 114. Limit of detection is found to be 0.5 ng/ml for carboxy finasteride and 1-5 ng/ml for other drugs. The method was successfully used to detect positive urine samples of 3-OH-stanozolol, methyl phenidate, mesocarb, clomiphene metabolite and carboxy finasteride.

Conclusion: The method developed based on controlled pH extraction method and HPLC-mass spectrometry analysis allowed better identification and confirmation of glucocorticosteroids and a few other drugs in different categories. The validated method has been used successfully for testing of 1000 In-competition samples. The method helped in detection of chemically and pharmacologically different banned drugs in urine in a single short run at a minimum required performance limit set by WADA.

Keywords: Detection limit; LC/MS/MS; glucocorticosteroids; validation.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Sample preparation flow steps
Figure 3
Figure 3
Single MRM transition of few doping drugs
Figure 2
Figure 2
LCMSMS total ion chromatogram resulting from (A) analysis of urine sample spiked with mixture of doping agents; (B) blank urine sample with internal standard
Figure 4
Figure 4
Average recoveries for extraction of doping agents
Figure 5
Figure 5
TIC and mass spectrum of 3-OH-Stanozolol from human urine excretion study sample
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Larry Bowers D. Analytical advances in detection of performance enhancing compounds. Clin Chem. 1997;43(7):1299–304. - PubMed
    1. Duntas LH, Parisis C. Doping: A challenge to the endocrinologist: A reappraisal in view of the Olympic games of 2004. Hormones. 2003;2:35–42. - PubMed
    1. Reddy MI, Beotra A, Jain S, Kaur T, Lal R. Purification of urine samples to improve detection limit of anabolic agents. Indian J Pharmacol. 2007;39:39–41.
    1. Thevis M, Schanzer W. Mass spectrometry in doping control analysis. Curr Organ Chem. 2005;9:825–48.
    1. Fouracre C, Kazlauskas R. Diuretics screening and confirmation using LC/MS/MS. Recent advances in doping analysis (9) Sport und Buch Strauβ, Koln. 2001. pp. 321–6.