THE RETURN TO ZERO TOLERANCE(which never really disappeared)

andTHE ROLE OF DRUG CONTAMINATION IN

DRUG REGULATION IN THE HORSE RACING INDUSTRY

(it doesn’t have one)Steven A. Barker, MS, Ph.D.

Everett D. Besch Distinguished Professor of Veterinary MedicineProfessor, Comparative Biomedical Sciences and Director,

Analytical Systems Laboratory,Equine Medication Surveillance Laboratory and

The Laboratory for Drug Residue StudiesState Chemist, Louisiana State Racing Commission

The concept of “zero tolerance” arose from the fact that, in the past, we were lucky to be able to detect any trace at all of many of the

more serious drugs that could be used to affect performance.

It has evolved over the years, however, to become an over-simplified, “get-tough” way

to handle drug cases, requiring no further thought, consideration of the science of

pharmacology or analytical chemistry or even testimony regarding possible explanations,

such as contamination.

Zero Tolerance

“If we can find it and we can confirm it, you’re guilty of something.”

“The withdrawal times we publish are just guidelines.”

“The threshold is the limit of detection of the method.”

1. Effect of urine extracts on rats2. Thin Layer Chromatography (tlc)3. Gas Chromatography (GC)4. Liquid chromatography (LC)5. GC/Mass Spectrometry (GC/MS)6. Immunoassay7. LC/MS8. LC/MS/MS

a. LC/QToF-Ion Trapb. UPLC/IonTrap-OrbiTrapc. ?????

EVOLUTION OF DETECTION METHODS

A regular paper clip weighs about 1.0 g.Divided into 1,000 pieces, each piece weighs 1.0 mg.

(1/1000th; 10-3)Divide one piece into 1,000 pieces and each weighs 1.0 ug.

(1/1,000,000th; 10-6)Divide one ug piece into 1,000 pieces and each weighs 1.0 ng.

(1/1,000,000,000th; 10-9)Divide a 1 ng piece into 1,000 pieces and each weighs 1.0 pg

(1/1,000,000,000,000th; 10-12)Divide a 1 pg piece into 1,000 pieces and each weighs 1.0 fg

(1/1,000,000,000,000,000th; 10-15)THIS IS THE LEVEL NEW MASS SPECTROMETERS CAN DETECT

THE “LIMIT OF DETECTION OF THE METHOD” GETS LOWER EVERY YEAR

ANABOLIC STEROIDS30 day withdrawal?45 day withdrawal?60 day withdrawal?

“The limit of detection of the method”

0.025 ng in blood

THAT’S ZERO TOLERANCE!

Non-Steroidal Anti-Inflammatories(NSAIDs)

Three can be used. Only one can be used within 24 hrs prior to the race. One of the two others may be used 48 hrs before the race but must be absent from the blood sample at the

time of collection. All other NSAIDs are not permitted.

“The presence of more than one of the three approved NSAIDs, with the exception of Phenylbutazone in a concentration below 1 microgram per milliliter of serum or plasma or any unapproved NSAID in the post-race serum or plasma sample is not permitted.”

(i) Phenylbutazone (or its metabolite oxyphenylbutazone) – 5 micrograms per milliliter;

(ii) Flunixin – 20 nanograms per milliliter;

(iii) Ketoprofen – 10 nanograms per milliliter.

Absent means “undetectable”.

THAT’S ZERO TOLERANCE!

All other NSAIDs are not permitted.

THAT’S ZERO TOLERANCE!

CORTICOSTEROIDS7-DAY WITHDRAWAL

“We can test for corticosteroids out to seven

days.”

That’s the limit of detection of the method and that’s

ZERO TOLERANCE!

DRUG SAMPLE

Lagoon Water Test Barn Stall Ship-in Stallsn = 4 5 5

Naproxen (ng/g <1 1.6 1.8

<1 1.9 2.8

1.0 1.9 1.9

1.5 ND 97.5

1.7 2.6

CONTAMINATION

CONTAMINATION

Flunixin (ng/g) ND 78.8 5.829.4 39.8 3.211.7 0.8 5.115.5 251.1 ND 83.2

DRUG SAMPLE

Test Barn Stalls Ship-in Stalls Dust

n= 5 5 3

Barker, 2008

“For example, a study conducted in Sweden in 2000 showed that administration of the non-steroidal anti-inflammatory drug flunixin (Banamine), either orally or by the intravenous route, caused untreated horses subsequently placed in the same stalls to prove positive for flunixin, at significant levels, in their urine for up to 14 days after initial exposure (Norgren et al., 2000). “ Concentrations ranged from 16 to 78 ng/ml in urine. What were the concentrations in blood?How many positives will result from such contamination?

Norgren et al., 2000

amphetamine median (range) after low and high doses: 15.5 (6.5-40.5) and 53.8 (34.0-83.4) ng/patch

cocaine33-3,579 ng/patch/30 min (from heat-induced sweating)43-3,799 ng/wipe

codeine 11-1,123 ng/patch/30 min (from heat-induced sweating)

fentanyl concentrations in sweat varied from 0.17 to 1.02 ng/μl of sweat

methamphetamine

median (range) after low and high doses: 63.0 (16.8-175) and 307 (199-607)ng/patch

CONCENTRATIONS OF DRUGS IN HUMAN SWEAT

Dr. Christian G. Daughton, In pressChief, Environmental Chemistry BranchNational Exposure Research LaboratoryU.S. Environmental Protection Agency

THE SOURCES OF DRUG CONTAMINATIONARE WELL ESTABLISHED

Environmental(Equine, Human, Plant Derived, Feed, Water, Dust, etc.)

Veterinary/PharmaceuticalSample Handling and

Laboratory ContaminationBacterial/Fungal

MetabolicBut the Industry continues to allow jurisdictions to ignore them.

The Industry has paid too little attention to contamination as a source of drug positives. Much more research into contamination as an explanation for drug positives must be

undertaken.

In establishing thresholds for therapeutic drugs, the potential for contamination must

be considered.

SUMMARY

The Racing Industry cannot afford to continue to permit jurisdictions to call positives on “trace level”

residues of drugs arising from contamination or from terminal residues from “old” administrations. Such

findings are pharmacologically meaningless and their prosecution is outside the intent of many legislatures to regulate drug use in the Industry. Such “positives”

serve no purpose, destroy reputations and unnecessarily damage the integrity of the sport. It is

senseless, unscientific and antiquated. The Industry must do what’s right. The Industry must

do what’s fair. Just stop the practice of “zero tolerance” enforcement.

Elimination of such “positives” could reduce the incidence of drug calls by as much as 80% and begin the process of improving horse racing’s reputation.

Steps to Take• 1. Abolish the use of the concept of “Zero Tolerance” in all racing

jurisdictions.• 2. Establish rational, scientifically based National thresholds for

therapeutic drugs that eliminate the possibility that “trace” contamination or terminal residues will be called as a positive. The threshold must not be “The limit of detection of the method,” particularly for Category 3, 4 or 5.

• 3. Investigate every case of ARCI category 1 and 2 drugs to determine if the drug detected was from human or other sources of contamination (Caffeine, Morphine, etc.). Apply the principles of pharmacology to the results.

• 4. Establish special areas for drug treatment and drug clearance to prevent contamination/re-contamination.

• 5. Remove feces, urine, standing water, etc. in and around barns in a timely manner and dispose of these materials in an area that will not permit re-contamination.

• 6. Build stalls and facilities coming into contact with horses out of materials that are easy to clean and that are non-absorbent.

• 7. Drug test all individuals coming into contact with horses. Obtain list of prescribed medications and instruct individuals in how to avoid contamination.

• 8. Maintain “retention samples” of feed, medications, supplements, etc. Record name, date and lot numbers.

• 9. Ban drug-containing foods from stall and barn areas.• 10. Have veterinarians properly certify identification of horses

(chipping) treated and turn in all syringes used for a particular horse, to be retained until samples clear.

• 11. Veterinarians should maintain “retention samples” of feed, medications, supplements, etc. Record name, date and lot numbers for drugs administered.

• 12. Avoid the use of compounded materials, herbal supplements and drugs bought at “too good to be true” prices.

• 13. Assure that test-barn personnel are wearing FRESH gloves,particularly the Veterinarian.

• 14. Establish a “right-side/left-side” administration/collection protocol for blood samples.

• 15. Establish a National Equine Drug-Positive Review Panel to examine data before a positive result is reported to assure that inadvertent positives are not prosecuted and that the rules regarding thresholds, etc., are consistently applied.

National Equine Drug-Positive Review Panel

1. Reduce/Eliminate costly litigation, hearings, etc.

2. Prevent further damage to the reputations of trainers, owners and veterinarians and the reputation of horse racing.

3. Bring consistency to the enforcement of medication rules.

4. Bring science to the enforcement of medication rules.

Racing Commissions are authorized by their legislatures to prevent the illegal use of performance enhancing drugs so as to protect the horses and participants in a race

and to protect the betting public against fraudarising from such acts.

HOWEVER,

1.Research that shows that most of the drugs regulated even have an effect on performance has never been conducted. It is assumed that they have the “potential” to affect performance based on their known pharmacology in humans or rats. Such research is difficult to conduct and is expensive.2.The ability for a drug to have an affect is based, in part, on dose and route, which significantly affects blood level. Yet, many jurisdictions continue to often base positive calls on trace levels of a drug in urine, which is, itself, significantly affected by a number of variables. 3.Trace urine and/or blood levels of most drugs, arising from contamination or as residues from older administrations, cannot possibly affect performance, endanger the participants of a race or cause a fraud to be committed against the betting public. Do such “violations” meet the authorization of the Commissions to act?

QUESTIONS?

Avery, 3 months