THE RETURN TO ZERO TOLERANCE (which never really disappeared) and THE 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 Medicine Professor, Comparative Biomedical Sciences and Director, Analytical Systems Laboratory, Equine Medication Surveillance Laboratory and The Laboratory for Drug Residue Studies State 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 rats 2. Thin Layer Chromatography (tlc) 3. Gas Chromatography (GC) 4. Liquid chromatography (LC) 5. GC/Mass Spectrometry (GC/MS) 6. Immunoassay 7. LC/MS 8. LC/MS/MS a. LC/QToF-Ion Trap b. UPLC/IonTrap-OrbiTrap c. ????? EVOLUTION OF DETECTION METHODS

A regular paper clip weighs about 1.0 g. Divided into 1,000 pieces, each piece weighs 1.0 mg. (1/1000 th; ) Divide one piece into 1,000 pieces and each weighs 1.0 ug. (1/1,000,000 th; ) Divide one ug piece into 1,000 pieces and each weighs 1.0 ng. (1/1,000,000,000 th; ) Divide a 1 ng piece into 1,000 pieces and each weighs 1.0 pg (1/1,000,000,000,000 th; ) Divide a 1 pg piece into 1,000 pieces and each weighs 1.0 fg (1/1,000,000,000,000,000 th; ) THIS IS THE LEVEL NEW MASS SPECTROMETERS CAN DETECT THE “LIMIT OF DETECTION OF THE METHOD” GETS LOWER EVERY YEAR

ANABOLIC STEROIDS 30 day withdrawal? 45 day withdrawal? 60 day withdrawal? “The limit of detection of the method” 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!

CORTICOSTEROIDS 7-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 Stalls n = Naproxen (ng/g < < ND CONTAMINATION

Flunixin (ng/g) ND ND83.2 DRUG SAMPLE Test Barn Stalls Ship-in Stalls Dust n= 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 ( ) and 53.8 ( ) ng/patch cocaine 33-3,579 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 ( ) and 307 ( ) ng/patch CONCENTRATIONS OF DRUGS IN HUMAN SWEAT Dr. Christian G. Daughton, In press Chief, Environmental Chemistry Branch National Exposure Research Laboratory U.S. Environmental Protection Agency

THE SOURCES OF DRUG CONTAMINATION ARE WELL ESTABLISHED Environmental ( Equine, Human, Plant Derived, Feed, Water, Dust, etc. ) Veterinary/Pharmaceutical Sample Handling and Laboratory Contamination Bacterial/Fungal Metabolic But 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 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 fraud arising 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