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- By
- Thomas Tobin
- University of Kentucky
- Maxwell H. Gluck Equine Research Center
- for the
- California Association of Toxicologists
- Tempe, Arizona
- May 18th, 2002
- Last Modified: May 30th, 2002 3:00 pm
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- The less than proper medication of racing horses has a long history,
with an absolutely impeccable starting point, Greek mythology
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- Any way they can, including but not limited to the creative use of drugs
and medications
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- Classic stimulants: cocaine, caffeine, amphetamine, methylphenidate,
methamphetamine, mazindol
- Opiates: etorphine, fentanyl, morphine, buprenorphine
- Tranquilizers: “washy horses”: acepromazine, promazine, fluphenazine,
detomidine, etc.
- Bronchodilators: clenbuterol, terbutaline, atropine, ipratropium bromide
- Always illegal and usually an “inside job”
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- Morphine and synthetic analgesics
- Locomotor stimulant
- Suppress pain
- Prolongs endurance
- Potentiated by stimulants
- Reliable, classic stimulant medication: widely used pre-ELISA
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- Beta-2 agonist bronchodilators: clenbuterol, albuterol, terbutaline,
etc.
- Highly potent agents
- Presumably improve oxygen delivery to blood and musculature
- Widely used close to post prior to ELISA
- Post-ELISA: use cut back dramatically
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- Used to treat “washy” horses (“running” race in paddock)
- Allows jockey to “rate” horse in the race
- Detomidine, reserpine, acepromazine, promazine
- All legitimate therapeutic agents: rate of improper use?
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- Anabolic steroids, growth hormone, erythropoietin (EPO)
- Special food supplements
- Vitamins
- By definition an “inside job”
- With the exception of anabolic steroids, not currently a very active
area
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- “Nobbling” the English term
- Horses heavily backed “ante-post”
- Administer tranquilizer or depressant to slow or stop a horse
- Oldest form of illegal medication
- Usually considered an “outside” job
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- The oldest form of illegal medication
- In principle, always possible to medicate a horse so that it will lose
- Challenge is to treat a horse such that he goes to post but does not win
his race
- More likely where there is heavy ante-post betting
- Also possible to medicate a horse to lose so that a favored horse will
win
- Always illegal, and usually considered an “outside” job
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- Thought is that you can medicate a horse to restore “normal” performance
- No stimulant or depressant actions
- Non-steroidal anti-inflammatory agents (NSAIDs)
- Prophylaxis of EIPH: furosemide
- Corticosteroids?
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- Administer agents that will interfere with testing
- As testing improves, a less likely scenario
- Furosemide: dilution
- Polyethylene glycol: smear TLC plates
- Thiamine: interfere with UV analysis
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- Administer a drug that dilutes drugs/drug metabolites and makes them
more difficult to detect
- Diuretics generally do not affect blood levels of drugs
- Dilute out water soluble drugs/drug metabolites in equine urine
- Effect is generally up to but not greater than the diluting effect
- Rules on furosemide in the US specify dose and time of administration
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- Equine drug testing has been in place longer (since 1910), and is more
rigorous and extensive (250-2,500 agents) than any other ongoing human
drug testing endeavor
- WHAT HAPPENS
- Horsemen use drugs ® Chemists
develop tests ®
Horsemen switch to new drugs ® Chemists develop new tests; one outcomes is that:
- Neogen has a large and developing panel of Forensic Drug Tests
ad infinitum?
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- Based on post-race urine samples
- About 10% of runners sampled/500,000 samples/year worldwide
- Winners, beaten favorites, selected horses that the stewards nominate
- Usually a central university-based laboratory, testing 10-40K
samples/year
- Scope of testing very broad: 250-2,500 agents
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- Any drug given to a horse < 10 mg/horse
- Etorphine, Fentanyl, Acepromazine, Mazindol, Terbutaline,
Buprenorphine, Cocaine, Oxymorphone, Detomidine
- Not being reliably detected pre-ELISA
- Being used at will by some horsemen
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- Immunoassay program starts; Illinois Racing Board Laboratory, IDS, a
Biotech Company, and the University of Kentucky
- Sensitivity of testing increased 100-1,000 fold, virtually overnight,
for 20+ drugs
- Multiple identifications of narcotics, stimulants, tranquilizers,
bronchodilators
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- Enzyme Linked ImmunoSorbent Assay (ELISA)
- Simple, fast, high sensitivity testing
- ~1,000 times more sensitive than thin layer chromatography
- In 1985, UK started a research project that rapidly evolved into the
ELISA testing program
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- 1985: Kentucky Racing Commission requests control of high potency
narcotics
- 1986: RIA tests for fentanyl and etorphine (“Elephant juice”)
- 1987: Program develops ELISA tests
- 1988: ELISA tests dramatically effective: POLITICS
- 1989: Program reorganized – development of tests continues
- 1992: 50 ELISA tests for high potency drugs, yielding ELISA
Technologies, a division of Neogen
Corporation
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- Obtain drug or drug structure
- Derivatize drug: add COOH or NH2 group: hapten
- Link derivatized drug (hapten) to BSA
- Inject into rabbit, re-inject, wait
- Link hapten to Horse Radish Peroxidase (HRP)
- Check rabbit serum for antibodies
- Construct and validate ELISA test
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- ELISA testing of frozen post-race samples in New Mexico
- First panel of ELISA tests puts down 40 plus trainers in New Mexico
- A problem that had been in place, in one form or another for 100, years
had been controlled
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- Probably the biggest single “across the board” increase in the
sensitivity of equine drug screening
- Stopped patterns of medication that had been in place in one form or
another for the better part of 100 years
- Also greatly increased the sensitivity of testing for therapeutic
medications
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- Offers virtually complete control of the abuse of high potency
medications
- Also offers super-sensitive testing for therapeutic medications
- Has accentuated a long-standing problem in equine medication control:
Detection of trace residues of therapeutic medications and environmental
and endogenous substances
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- 1987 -- Five ELISA tests
(but 100’s of
drugs)
- 2002 -- Neogen’s nearly 100
ELISAs, can screen for more
than 250 agents
- Many current ELISA tests are of interest in HUMAN FORENSICS and the FOOD
SAFETY INDUSTRIES
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- Alfentanil, amphetamine, amphetamine ultra, anileridine, apomorphine,
azaperone, barbiturate group, benzodiazepine group, boldenone,
bronchodilator group, bumetanide, buprenorphine, butorphanol,
caffeine/pentoxifylline, carfentanil, clenbuterol, cocaine/benzoylecgonine,
corticosteroid group, cromoglycate, dantrolene, detomidine,
dexamethasone, diprenorphine, doxapram, droperidol, ethacrynic acid,
etorphine, fentanil group, fentanyl, flunitrazepam, flunixin,
fluoxetine, fluphenazine, furosemide, glycopyrrolate, guanabenz,
haloperidol…
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- Haloperidol metabolite, hydromorphone, isoxsuprine, ketamine, ketorolac,
levallorphan, lidocaine, lofentanil, mazindol, mazindol metabolite,
meperidine, mephentermine, mepivacaine, methadone, methamphetamine/MDMA,
methocarbamol, methotrexate, methylphenidate, methylprednisolone,
nalbuphine, nandrolone, opiate group, oxymorphone/oxycodone,
pentazocine, phenylbutazone, procaine, promazine group, propanolol,
pyrilamine, reserpine, stanozolol, sufentanil, sulfamethazine, THC
metabolite, theophylline, trenbolone, triamcinolone acetonide,
tricyclics group, zomepirac
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- Alfentanil, Amphetamine, Barbiturate group, Benzodiazepine group,
Boldenone, Buprenorphine, Butorphanol,Carfentanil, Cocaine/BZE,
Etorphine, Fentanil group, Flunitrazepam, Hydromorphone, Lofentanil,
Ketamine, LSD, Meperidine, Methadone, Methylphenidate, Methamphetamine/MDMA,
Nandrolone, Opiate group, Oxymorphone/Oxycodone, Sufentanil, THC
Metabolite
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- Neogen has available a large and expanding range of high quality
forensic tests
- Largest selection of high quality forensic ELISA tests available
worldwide
- Available to Equine and Human Forensic Labs.
-AD INFINITUM?-
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- Rate of detection of performance altering agents in racing is relatively
low
- “Positives” in about 1-1,000 samples tested.
- All identifications confirmed by mass spectrometry in the primary and
the referee laboratory.
- “Top-end” of racing is actually very clean
- Thanks in part to the technology I have just described
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- ELISA testing technology is highly effective: the Worldwide Standard in
Racing
- Simple,fast,inexpensive and highly sensitive
- Technology is highly adaptable
- Single, portable “Horse Side” tests
- Inexpensive automated systems
- High end, high volume, high throughput completely automated systems
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- Limited Throughput: 10K
samples/year
5-10
tests/sample
- Medium Throughput: 20K samples/year
10-15
tests/sample
- High Throughput: 100K
samples/year
25-30
tests/sample
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- Contract to buy a certain annual volume of ELISA tests (100K tests/year,
or so)
- Make available fully automated $50K Thermolabsystems ‘DSX’ Analyzer to
perform the ELISA analyses
- Information: scottcongleton@Neogen.com
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- Contract to buy a certain annual volume of ELISA tests (300K tests/year,
or so)
- Make available fully automated $100K Tecan Genesis System to perform the
ELISA analyses
- Information: scottcongleton@Neogen.com
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- Contract to buy a certain annual volume of ELISA tests (3,000K
tests/year, or so)
Make available fully automated $200K Beckman Coulter Orca System
to perform the ELISA analyses
Information: scottcongleton@Neogen.com
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- Thanks to my longtime colleagues at Neogen Corp, for their ongoing
support in expanding and commercializing this technology
- To all the members of my research group,and especially Drs. W. Karpiesiuk, S.
Kwiatkowski and Mr. W. E. Woods.
- The Kentucky Racing Commission and the Kentucky Equine Drug Council for
support
- To the Director of the Gluck Equine Research Center and to my colleagues
there and at UK for their support over the years
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- 1/ Remifentanil in the Horse: Identification and Detection of its Major Urinary
Metabolite; Lehner et al., J. Anal. Tox. Vol 24, July/August, 2000
- 2/ “Kentucky Laboratory Takes Lead Against Horse Dopers,” Tobin, Clinical
Chemistry News, Vol. 18, Issue 1, January 1992
- 3/ Non-isotopic Immunoassay Drug Tests in Racing Horses: A Review of
their Application to Pre- and Post-Race Testing, Drug Quantitation and
Human Testing; Tobin et al., Res. Comm. Chem. Path. Pharmacol. Vol 62,
#3, P371-396, 1988
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