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Interpretation of drug testing results requires detailed scientific information, particularly in those cases where the question of legitimate use versus illicit use arises. Amphetamine remains a widely abused drug throughout the world, although it is also used therapeutically for weight loss, narcolepsy, and attention-deficit disorder with hyperactivity (ADHD). Treatment of ADHD using stimulant drugs is much more common now than it was in even the recent past. Increasingly, older individuals are diagnosed and treated for ADHD, and treatment often continues into adulthood. Amphetamine is commonly used for the treatment of ADHD and is available by prescription as either the d-enantiomer or a mixture of enantiomers. Although used for many years, there are no data available to describe the excretion profile of amphetamine and its enantiomers following repeated use of the drug. As a result, medical review officers (MROs) and forensic toxicologists have no direct evidence to base their decisions on when it comes to evaluation of use of these drugs. The current study was designed to determine the concentration and enantiomer excretion profile following repeated daily administration of mixed enantiomers of amphetamine. Twenty milligrams of Adderall was administered daily to five healthy subjects with all subsequent ad lib urine samples collected for at least five days following administration of the five-dose regimen. Adderall is a 3:1 mixture of d- and l-enantiomers of amphetamine salts and represents the mixed enantiomer proportion of amphetamine available in the United States through pharmaceutical channels. Peak amphetamine concentrations ranged from 5739 to 19,172 ng/mL. Samples containing > or = 500 ng/mL amphetamine (the administrative cutoff for a positive result by gas chromatography-mass spectrometry) were seen up to 60:15 (h:min) following administration of the last dose. Enantiomer analysis showed the d-enantiomer to be in excess of the l-enantiomer for as long as the drug was administered. After administration of the last dose of drug, the proportion of l-enantiomer increased over time. Not all samples that contained > or = 500 ng/mL total amphetamine were positive when tested by immunoassay because of the differing cross-reactivity of the enantiomers. This study provides the first description of the excretion of amphetamine following repeated administration of Adderall. The presence of the l-enantiomer separates this drug from other formulations composed of only the d-enantiomer (i.e., Dexedrine and much illicit amphetamine), thus readily differentiating them from Adderall use. Some illicit and medicinal amphetamine is, however, a mixture of amphetamine enantiomers. Because the enantiomers are metabolized at different rates, their proportion offers the opportunity to describe excretion versus time. Coupling this data with drug concentration makes it possible for forensic toxicologists and MROs to come to an informed decision regarding the involvement of this drug in a positive drug test result.

The amphetamine isomer ratios (l-amphetamine/d-amphetamine) in 373 urine specimens submitted for analysis over a two-year period have been determined using a chiral derivatizing agent in conjunction with a gas chromatograph fitted with a nitrogen-specific detector. All of the specimens were collected from known or suspected amphetamine abusers, some of which were prescribed dexedrine for maintenance and detoxification. The mean (+/- 1 standard deviation [SD]) l/d-amphetamine isomer ratio for 147 specimens from compliant subjects prescribed dexedrine was 15.0% (+/- 4.9%). The mean (+/- 1 SD) l/d-amphetamine isomer ratio from 165 subjects abusing illicit amphetamine was 98.5 (+/- 27.5%). The calculation of l/d-amphetamine isomer ratios in urine has been found to be a rapid method for determining the compliance of subjects prescribed dexedrine and is therefore a useful technique for the continued management of amphetamine abusers. In addition, 17 specimens of illicit amphetamine powder (assumed to be a racemic mixture) were submitted to the laboratory for analysis. Using a combination of gas chromatography with and without chiral derivatization, the powders were found to have a mean l/d-amphetamine isomer ratio of 89.2% (range 72.2% to 98.3%) and mean purity (w/w) of 21.5% (range 3.4% to 71.0%) relative to pure dl-amphetamine substance.

Seven healthy male volunteers received a single oral dose of 5 mg, 10 mg, or 20 mg of d-amphetamine. Urine was collected at 2, 4, 8, 12, 18, and 24 h post-dose. Total urine volume was measured, and pH and creatinine were determined. All specimens were analyzed by TDx Amphetamine/Methamphetamine II (TDx), Emit-d.a.u. Monoclonal Amphetamine/Methamphetamine (EM), and Emit II Amphetamine/Methamphetamine (EII) immunoassays at a cutoff value of 1000-ng/mL amphetamine. Quantitation of urinary amphetamine in all specimens was performed by gas chromatography-mass spectrometry. All urine testing results by the three immunoassays, EM, EII, and TDx, were in agreement; there were no discordant findings. Of the 42 total urine specimens collected following a 5-mg dose of amphetamine, only 8 (19%) screened positive by immunoassay. Twenty-four of 36 (67%) urine specimens yielded positive responses following a 10-mg dose, and 37 of 42 (88%) were positive by immunoassay following a 20-mg dose. These data demonstrate the present guideline for regulated forensic urine drug testing (FUDT) for amphetamine with a screening cutoff of 1000 ng/mL is too high to consistently detect the administration of a single 5-mg oral dose of d-amphetamine. There was considerable overlap of amphetamine concentrations in individual specimens following the various doses. Peak urinary amphetamine ranged from 620 to 3160 ng/mL following 5-mg doses. The time to peak concentration also varied widely at each dose, occurring in urines collected 2 to 18 h post-administration. The mean percent of dose excreted as unchanged amphetamine over 24 h at each dose ranged from 35 to 44%. The data demonstrated that amphetamine excretion increases with increasing urine flow and decreasing urine pH. Thus, a positive FUDT result for amphetamine means only that the individual was administered or self-administered amphetamine at some time prior to collection of the specimen.

The time-lapse changes of d- and i-forms in urine specimens collected in the 24 h after the oral dosing of two adult male subjects with 20 mg of racemic (dl)-ethylamphetamine (EAMP)-HCl were examined by high-performance liquid chromatography. The percentage of the excreted dose of nonmetabolized l-EAMP was larger than that of d-EAMP, and the percentage of the excreted dose of metabolized l-amphetamine (AMP) was smaller than that of d-AMP in both subjects, A and B. These differences were observed 4.5-24 h after administration of the drug. The changes in the l/d ratio of EAMP and AMP were not similar between subjects A and B, but the change in the total l/d ratio was nearly the same. The regression line for subject A was as follows: y = 0.021323x + 0.98399. The regression line for subject B was as follows: y = 0.020947x + 0.94893. These two regression lines suggested that the time lapsed after the administration to humans could be predicted. The total percent of the excreted doses of EAMP and AMP was 47.46% (the d-forms, 20.73%; the l-forms, 26.73%) for subject A and was 31.43% (the d-forms, 14.14%; the l-forms, 17.29%) for subject B. The l/d ratio was 1.29 for subject A and 1.22 for subject B, which was somewhat higher than that (1.01) of the dl-EAMP-HCl used.

Several positive methamphetamine/amphetamine urine samples were reexamined to determine the chirality of the detected drug or drugs. (S)-N-(Trifluoroacetyl)prolyl derivatives were prepared and analyzed using GC/MS. In one case, pure d-isomers of methamphetamine and amphetamine were detected. In the remainder of the samples involving both drugs, skewed racemates were detected, with the l-isomer of methamphetamine and the d-isomer of amphetamine predominating slightly over their enantiomers. In samples involving amphetamine only, 50:50 mixtures of d- and l-isomers were detected. In no instance was pure i-methamphetamine (from a Vicks inhaler) detected.