WHAT is hair analysis and how does impact the equine industries? The use of prohibited substances in sports is a hot topic, with high profile cases involving Lance Armstrong and Russian athletes making headlines around the world.
Equine sports are not immune from the negative effects of prohibited substances use, having a number of its own high profile incidents. In Ireland, the principle bodies responsible for equine anti-doping is Irish Horseracing Regulatory Board (IHRB) and Horse Sport Ireland (HSI), who have significant scientific expertise and technology at their disposal.
Recently, equine anti-doping testing in Ireland has moved towards the use of equine hair analysis in a bid to improve compliance and detection rates.
Recent research
Of course, Ireland is not the only jurisdiction looking at equine hair analysis in this regard. A recent publication in the Journal of Pharmaceutical and Biomedical Analysis describes the research of Wong and co-workers from the Racing Laboratory at The Hong Kong Jockey Club. Here, the team of scientists are developing cutting edge analytical techniques in an effort to detect low levels of prohibited substances in equine hair samples.
To do this they utilised a machine called an ultra-high performance liquid chromatography (UHPLC)-tandem mass spectrometry (MS). In effect, this is two machines in one.
The first, the UHPLC, allows the scientist to separate out the mixture of different chemical components, including prohibited substances, found in the hair sample.
The second, the MS, allows the scientist to generate a unique “chemical finger print” for each individual chemical they separate, which they compare with a catalogue containing the “chemical finger prints” of prohibited substances. A match indicates the presence of the corresponding prohibited substance.
What sets the work of Wong and his team apart is the number of prohibited substances they can detect from a hair sample, as well as the low levels of substance they can detect. A total of 121 drugs and/or their metabolites could be detected in equine hair by the Hong Kong team, covering 10 classes of prohibited substances.
Very low estimated detection limits of between 0.1 and 10 pg/mg were reported and this equates to below one part per billion (or about one blade of grass in a two acre field).
One set of results from Wong’s team describes a horse that was treated with broncopulmin, which contains 16 µg/g of the prohibited substance clenbuterol (clenbuterol is a bronchodilator that opens the airways).
Twice daily oral treatment, at a dosage of five grams per 100 kg bodyweight, took place for five days around six months prior to hair sample collection. Clenbuterol was detected in the hair sample by the Racing laboratory team six months after administration had ceased and at levels below one part per billion.
Mixed impact
This level of analysis is good news in the fight against the misuse of prohibited substances, but can the innocent horse owner or trainer accidentally fall foul of testing? The answer is yes, with a number of accidental cases reported in the recent past.
In 2015, samples taken from horses ridden by international Swiss showjumper Steve Guerdat returned positives for the banned substances codeine, oripavine, and morphine. The FEI subsequently cleared Guerdat after independent laboratory tests proved that the feed used by Guerdat was contaminated with poppy seeds (the three prohibited substances in the horses’ samples was consistent with poppy seed contamination).
Another recent case in 2018 involved French racehorses, where the use of old hay resulted in a positive test result. In the hay, the scientists found the precursor of boldenone (an anabolic steroid), which is called boldione.
Bacteria in the digestive tract of the horse caused the metabolism (breakdown) of boldione and the positive test resulted.
Cutting edge testing systems are “hair” to stay and the bottom line for owners and trainers is to consult your nutritional advisor in relation to feed programmes, as some feed producers have in house analytical systems to ensure accidental contamination, like in the cases above, cannot happen.