An undergraduate forensic biochemistry laboratory experiment to detect doping in animal hair using LCMS

Main Article Content

ILTAF SHAH Syed Salman Ashraf


Doping using performance enhancing banned substances is a serious problem in almost every sport competition. Not surprisingly, the detection of these contra banned drugs is an area of active and continuous improvement and innovation by bioanalytical chemists. Additionally, most students working out in the gym and taking part in various sports need to be made aware of the doping and the health problems associated with it. Science or STEM students, in particular chemistry students, must not only be made aware of these issues, but also be taught that chemistry (and science) can provide solutions to such real-life issues. To this end, a newly developed forensic laboratory experiment is described that guides students to learn liquid chromatography mass spectrometry instrumentation (LC-MS) to detect four common doping drugs cortisol, dexamethasone, methyl prednisolone and flumethasone in camel hair samples.
In addition, the project is also designed to reinforce the importance of hair analysis as an additional sample matrix, complementary to saliva, blood and urine tests, in doping applications. In addition to learning various aspects of sample preparation, extraction, and LC-MS principles, students will also learn how to validate this method according to Food and Drugs Administration guidelines for intra and inter day precision and accuracy, recovery, stability and linearity.
This “applied forensic science” experiment was successfully implemented in a biochemistry undergraduate research course to enhance students' learning of doping issues as well as important bio-analytical and forensic biochemistry concepts. Student survey confirmed that this laboratory experiment was successful in achieving the objectives of raising awareness of doping control in students and illustrating the usefulness of chemistry in solving real-life problems. This experiment can be easily adopted in an advanced biochemistry laboratory course and taught as an inquiry-guided exercise. Such hands-on and engaging experiments should be part of undergraduate curriculum to foster deeper interest and innovation in STEM subjects to better prepare the next-generation workforce in science and technology.


Download data is not yet available.

Article Details

How to Cite
SHAH, I., & Ashraf, S. (2018). An undergraduate forensic biochemistry laboratory experiment to detect doping in animal hair using LCMS. International Journal for Innovation Education and Research, 6(1), 133-148. Retrieved from
Author Biography

Syed Salman Ashraf, UAEU

Professor, Chemistry, College of Science


Aidman, E, A Petroczi, I Hussain, N Deshmukh, T Nepusz, M Uvacsek, M Toth, J Barker, and D Naughton. 2010. "Beyond self-report in doping: Validating declared substance use in sport with hair sample analysis." Review of. Journal of Science and Medicine in Sport 13:e19-e20.
Al Katheeri, NA, IA Wasfi, M Lambert, A Giuliano Albo, and C Nebbia. 2006. "In vivo and in vitro metabolism of dexamethasone in the camel." Review of. The Veterinary Journal 172 (3):532-43.
Al Katheeri, NA, IA Wasfi, M Lambert, and A Saeed. 2004. "Pharmacokinetics and pharmacodynamics of dexamethasone after intravenous administration in camels: effect of dose." Review of. Veterinary research communications 28 (6):525-42.
Ashraf, S Salman, Sayed AM Marzouk, Ihsan A Shehadi, and Brian M Murphy. 2010. "An integrated professional and transferable skills course for undergraduate chemistry students." Review of. Journal of Chemical Education 88 (1):44-8.
Bechshøft, Thea Østergaard, Christian Sonne, Rune Dietz, Erik W Born, Melinda A Novak, Elizabeth Henchey, and Jerrold S Meyer. 2011. "Cortisol levels in hair of East Greenland polar bears." Review of. Science of the Total Environment 409 (4):831-4.
Bell, Randy L, Lara Smetana, and Ian Binns. 2005. "Simplifying inquiry instruction." Review of. The Science Teacher 72 (7):30.
Berlyne, GM, R Shainkin-Kestenbaum, S Shany, J Finberg, and R Yagil. 1978. "Renal physiology of the dromedary camel." Review of. 日本腎臓学会誌 20 (9):1015-21.
Bryan, Heather M, Amanda G Adams, Rosemary M Invik, Katherine E Wynne-Edwards, and Judit EG Smits. 2013. "Hair as a meaningful measure of baseline cortisol levels over time in dogs." Review of. Journal of the American Association for Laboratory Animal Science 52 (2):189-96.
Calfee, Ryan, and Paul Fadale. 2006. "Popular ergogenic drugs and supplements in young athletes." Review of. Pediatrics 117 (3):e577-e89.
Cameron, David, Tracy Epton, Paul Norman, Paschal Sheeran, Peter R Harris, Thomas L Webb, Steven A Julious, Alan Brennan, Chloe Thomas, and Andrea Petroczi. 2015. "A theory-based online health behaviour intervention for new university students (U@ Uni: LifeGuide): results from a repeat randomized controlled trial." Review of. Trials 16 (1):555.
Davenport, Matthew D, Stefan Tiefenbacher, Corrine K Lutz, Melinda A Novak, and Jerrold S Meyer. 2006. "Analysis of endogenous cortisol concentrations in the hair of rhesus macaques." Review of. General and comparative endocrinology 147 (3):255-61.
De Clercq, Nathalie, Julie Vanden Bussche, Siska Croubels, Philippe Delahaut, and Lynn Vanhaecke. 2012. Plackett-Burman design as a tool to develop a generic extraction for U-HPLC-Orbitrap-MS analysis of glucocorticoids in livestock urine. Paper presented at the 14th International symposium on Advances in Extraction Technologies (ExTech 2012).
Deshmukh, Nawed, Iltaf Hussain, James Barker, Andrea Petroczi, and Declan P Naughton. 2010. "Analysis of anabolic steroids in human hair using LC–MS/MS." Review of. Steroids 75 (10):710-4.
Duclos, Martine. 2010. "Evidence on ergogenic action of glucocorticoids as a doping agent risk." Review of. The Physician and sportsmedicine 38 (3):121-7.
Elias, E, and Simy Weil. 1989. "Serum cortisol levels in camels (Camelus dromedarius) during the reproductive cycle." Review of. Comparative Biochemistry and Physiology Part A: Physiology 94 (4):787-90.
Epton, Tracy, Paul Norman, Aba-Sah Dadzie, Peter R Harris, Thomas L Webb, Paschal Sheeran, Steven A Julious, Fabio Ciravegna, Alan Brennan, and Petra S Meier. 2014. "A theory-based online health behaviour intervention for new university students (U@ Uni): results from a randomised controlled trial." Review of. BMC public health 14 (1):563.
Evans, Mildred H. 1984. "Understanding drugs: A new interdisciplinary course." Review of. J. Chem. Educ 61 (8):671.
Fourie, Nicolaas H, and Robin M Bernstein. 2011. "Hair cortisol levels track phylogenetic and age related differences in hypothalamic–pituitary–adrenal (HPA) axis activity in non-human primates." Review of. General and comparative endocrinology 174 (2):150-5.
Gow, R, S Thomson, M Rieder, S Van Uum, and G Koren. 2010. "An assessment of cortisol analysis in hair and its clinical applications." Review of. Forensic science international 196 (1):32-7.
Holt, Richard IG, Ioulietta Erotokritou-Mulligan, and Peter H Sönksen. 2009. "The history of doping and growth hormone abuse in sport." Review of. Growth Hormone & IGF Research 19 (4):320-6.
Kataria, N, and AK Kataria. 2004. "Use of blood analytes in assessment of stress due to drought in camel." Review of. Journal of camel practice and research 11 (2):129-33.
Kerr, WJ, REG Murray, BD Moore, and DC Nonhebel. 2000. "An integrated communication skills package for undergraduate chemists." Review of. J. Chem. Educ 77 (2):191.
Khalaf, Sulayman. 1999. "Camel racing in the Gulf. Notes on the evolution of a traditional cultural sport." Review of. Anthropos:85-106.
———. 2000. "Poetics and politics of newly invented traditions in the Gulf: Camel racing in the United Arab Emirates." Review of. Ethnology:243-61.
Kindlundh, Anna, Dag GL Isacson, Lars Berglund, and Fred Nyberg. 1999. "Factors associated with adolescent use of doping agents: anabolic‐androgenic steroids." Review of. Addiction 94 (4):543-53.
Kindlundh, Anna MS, Dag GL Isacson, Lars Berglund, and Fred Nyberg. 1998. "Doping among high school students in Uppsala, Sweden: A presentation of the attitudes, distribution, side effects, and extent of use." Review of. Scandinavian journal of social medicine 26 (1):71-4.
Kintz, Pascal, Marion Villain, and Vincent Cirimele. 2006. "Hair analysis for drug detection." Review of. Therapeutic drug monitoring 28 (3):442-6.
Lokken, DA. 1975. "The determination of alcohol content of beer: A general and analytical experiment with high student interest value." Review of. J. Chem. Educ 52 (5):329.
Lunsford, Suzanne K, and William Slattery. 2006. "An interactive environmental science course for education science majors." Review of. J. Chem. Educ 83 (2):233.
Majchrzak, Yasmine N, Gabriela F Mastromonaco, Wendy Korver, and Gary Burness. 2015. "Use of salivary cortisol to evaluate the influence of rides in dromedary camels." Review of. General and comparative endocrinology 211:123-30.
Mills, Kenneth V. 2015. "Biochemistry in an undergraduate writing‐intensive first‐year program: Seminar courses in drugs and bioethics." Review of. Biochemistry and Molecular Biology Education 43 (4):263-72.
Mohamed, HE. 2006. "Factors affecting cortisol status in camels (Camelus dromedarius)." Review of. Journal of Animal and Veterinary Advances 5 (4):307-9.
Petróczi, Andrea, Jorge A Vela Ocampo, Iltaf Shah, Carl Jenkinson, Rachael New, Ricky A James, Glenn Taylor, and Declan P Naughton. 2015. "Russian roulette with unlicensed fat-burner drug 2, 4-dinitrophenol (DNP): evidence from a multidisciplinary study of the internet, bodybuilding supplements and DNP users." Review of. Substance abuse treatment, prevention, and policy 10 (1):39.
Raul, Jean-Sébastien, Vincent Cirimele, Bertrand Ludes, and Pascal Kintz. 2004. "Detection of physiological concentrations of cortisol and cortisone in human hair." Review of. Clinical Biochemistry 37 (12):1105-11.
Russell, Evan, Gideon Koren, Michael Rieder, and Stan Van Uum. 2012. "Hair cortisol as a biological marker of chronic stress: current status, future directions and unanswered questions." Review of. Psychoneuroendocrinology 37 (5):589-601.
Sauvé, Brittany, Gideon Koren, Grace Walsh, Sonya Tokmakejian, and Stan HM Van Uum. 2007. "Measurement of cortisol in human hair as a biomarker of systemic exposure." Review of. Clinical & Investigative Medicine 30 (5):183-91.
Shah, Iltaf, Andrea Petroczi, Martina Uvacsek, Márta Ránky, and Declan P Naughton. 2014a. "Hair-based rapid analyses for multiple drugs in forensics and doping: application of dynamic multiple reaction monitoring with LC-MS/MS." Review of. Chemistry Central Journal 1 (8):1-10.
———. 2014b. "Hair-based rapid analyses for multiple drugs in forensics and doping: application of dynamic multiple reaction monitoring with LC-MS/MS." Review of. Chemistry Central Journal 8 (1):73.
Sid-Ahmed, Omer-Elfaroug, Ahmed Sanhouri, Badr-Eldin Elwaseela, Imad Fadllalah, Galal-Eldin Elazhari Mohammed, and Erich Möstl. 2013. "Assessment of adrenocortical activity by non-invasive measurement of faecal cortisol metabolites in dromedary camels (Camelus dromedarius)." Review of. Tropical animal health and production 45 (6):1453-8.
Stalder, Tobias, and Clemens Kirschbaum. 2012. "Analysis of cortisol in hair–state of the art and future directions." Review of. Brain, behavior, and immunity 26 (7):1019-29.
Tucci, Valerie K, Abby R O’Connor, and Lynn M Bradley. 2014. "A three-year chemistry seminar program focusing on career development skills." Review of. Journal of Chemical Education 91 (12):2071-7.
van Amsterdam, Jan, Antoon Opperhuizen, and Fred Hartgens. 2010. "Adverse health effects of anabolic–androgenic steroids." Review of. Regulatory Toxicology and Pharmacology 57 (1):117-23.
Wasfi, IA. "On the Pharmacokinetics/Pharmacodynamics, Detection Time and Metabolism of Drugs in Racing Camels: 17 Years of Experience." Review of. Keynote Presentations:32.
Wood, Ruth I. 2008. "Anabolic–androgenic steroid dependence? Insights from animals and humans." Review of. Frontiers in neuroendocrinology 29 (4):490-506.
Yesalis, Charles E, and Michael S Bahrke. 2000. "Doping among adolescent athletes." Review of. Best Practice & Research Clinical Endocrinology & Metabolism 14 (1):25-35.