Drug Testing Procedures

Sample Log-in

A chain-of-custody form is submitted to the racetrack for the set of samples on each race date to be completed by the veterinarian. The completed form is then returned to Truesdail with the samples and our staff maintains a copy. When samples arrive, Truesdail’s personnel check all seals and sample numbers against the chain-of-custody document and log the samples into our computerized Laboratory Information Management System (LIMS). From sample receipt to the issuance of the final report, all blood samples are retained in Truesdail’s secured refrigerator and all urine samples are retained in Truesdail’s secured freezer.

Extraction

The aliquot of either blood or urine is next subjected to different extraction protocol to isolate the drugs. There are some differences to be noted in blood versus urine samples. Many drugs are removed from circulation in the blood through a metabolic process, thus we tend to find higher levels of parent drug for some in the blood and higher levels of metabolites of other drugs in the urine. Because we expect to find many drugs to have been metabolized, urine samples are subjected to enzyme hydrolysis prior to extraction. Hydrolyzed urine is isolated and separated by solid phase extraction into acidic/neutral and basic fractions covering the same type of drugs as indicated for blood samples.

Screening

The extracted samples are screened using UHPLC/HRMS systems. Truesdail screens against a target list of drug compounds of interest to the racing community to be sought in each sample. We began with setting up our target list to include the total list of compounds required by the American Graded Stakes Committee and the list of compounds in ARCI’s (Association of Racing Commissioners International) controlled therapeutic medication schedule. We have also included drugs targeted in human Olympic athletes, which could be used on horses. Our target list includes both parent drugs and known drug metabolites. Currently our target list has over 1,800 compounds and is still growing. Although immunoassay testing is decreasing in importance, there are a few drugs that do not extract well and could easily be missed in direct instrumental screens. Several immunoassays kits are used to supplement our direct instrumental screens.

Confirmatory Analysis

Samples found to be suspect from screening by UHPLC/HRMS or immunoassay are subjected to further testing to confirm the drugs identity and for drugs with thresholds to provide quantitation. Most confirmations are done by liquid chromatography coupled to triple quadrupole mass spectrometry (LC/MS/MS), a few compounds may still be confirmed by GC/MS. Multipoint calibrations are done to quantify drugs with established thresholds and duplicate determinations are made to provide an estimate of uncertainty.

Sample Custody

Our chain-of-custody procedures have withstood all legal challenges in our more than 65 years of performing drug testing. Whenever a sample is handled, a log entry is recorded to document this activity. Blood and urine samples are opened, an aliquot is removed for analysis, the containers resealed with security tape and initialed and dated by the person who removed the test portions to maintain chain-of-custody. Any subsequent removal of test portions are also completed with this tape-resealing procedure and documentation.

Truesdail’s testing laboratory is separated from our main facility via locked, key card access doors. Only authorized, trained personnel are allowed access to the lab. Our Drug Testing labs are further segregated and under a secondary key card access, allowing only Drug Testing lab personnel access to the lab, sample storage, and sample log-in areas of our Drug Testing lab areas. All Truesdail employees wear identification badges with their photo that grant access into these controlled areas. All visitors to Truesdail are required to sign in/out, wear a designated visitor badge, and have an escort when in the facility at all times. Truesdail’s facility is located within a mix-use campus with 24/7 security patrols and camera monitors for additional after hours security.

In addition to physical security, Truesdail’s instrument computers are fragmented from our main network system onto a lab only, non-externally connected network. Data from these systems can only be accessed via the physical computers within the Drug Testing Lab area. Data is backed up weekly and kept on separate, physical, fire resistant hard drives.