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Urine Adulteration and Substitution Prior to Drugs of Abuse Testing

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Abstract

Adulteration and substitution (of urine by other fluids) are practices conducted by individuals who undergo urine drug testing for the purpose of invalidating results of such tests. Drug testing laboratories that are certified by the Substance Abuse and Mental Health Services Administration will soon be required to perform tests for the presence of these adulterants and substitution. Mandatory tests include urine creatinine, specific gravity, pH, and analysis for oxidizing adulterants. This article will describe the current practices of urine adulteration and substitution, and summarize the new proposed guidelines for the laboratory tests that are used for detecting adulterants.

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... Urine substitution is achieved through the replacement of a urine specimen with that of drug-free urine [27], some nonurine liquids such as water or saline [28], or commercially available synthetic urine that possesses the same essential characteristics of human urine, eg, correct pH, specific gravity, and creatinine concentration [29]. Urine substitution can be facilitated by the use of some sophisticated devices including the Butt Wedge™, the Urinator™, and the Whizzinator™, or in extreme cases, by catheterization [1,27,29]. ...
... Hypochlorite was found to directly affect immunoassay reagents, resulting in erroneous and in some cases false negative test results for FPIA assay [82], EMIT assay [75], CEDIA assay [60], and KIMS assay [83]. Hypochlorite has also been found to cause degradation of opiate analytes in urine, which were confirmed by GC-MS analysis [28,29]. ...
... Nitrite successfully masked the detection of BZE by immunoassay screening methods, but did not interfere with GC-MS confirmation as reported in a study by Tsai et al. [76]. Nitrite-containing commercial products such as Klear ® and Whizzies ® were reported as effective masking agents for the detection of opiates [28,29,39]. Potassium nitrite, under acidic environment (pH < 7), was shown to be effective in masking the detection of 6-MAM by the CEDIA immunoassay [44]. ...
Chapter
Urine drug testing plays an important role in monitoring licit and illicit drug use for both medico-legal and clinical purposes. One of the major challenges of urine drug testing is adulteration, a practice involving manipulation of a urine specimen with chemical adulterants to produce a false negative test result. This problem is compounded by the number of easily obtained chemicals that can effectively adulterate a urine specimen. Common adulterants include some household chemicals such as hypochlorite bleach, laundry detergent, table salt, and toilet bowl cleaner and many commercial products such as UrinAid (glutaraldehyde), Stealth® (containing peroxidase and peroxide), Urine Luck (pyridinium chlorochromate, PCC), and Klear® (potassium nitrite) available through the Internet. These adulterants can invalidate a screening test result, a confirmatory test result, or both. To counteract urine adulteration, drug testing laboratories have developed a number of analytical methods to detect adulterants in a urine specimen. While these methods are useful in detecting urine adulteration when such activities are suspected, they do not reveal what types of drugs are being concealed. This is particularly the case when oxidizing urine adulterants are involved as these oxidants are capable of destroying drugs and their metabolites in urine, rendering the drug analytes undetectable by any testing technology. One promising approach to address this current limitation has been the use of unique oxidation products formed from reaction of drug analytes with oxidizing adulterants as markers for monitoring drug misuse and urine adulteration. This novel approach will ultimately improve the effectiveness of the current urine drug testing programs.
... Sodium hydroxide is a caustic strong base. It causes change to alkaline pH in urine samples, then it effects the drug binding and solubility and produces false negative urine results [13]. ...
... It was suggested that GC-MS confirmation test results aren't effected by non-oxidizing adulterans. Adding sodium hydrosulfide or sulfamic acid to the GC-MS method can help remove excess oxidizing additive and may prevent further oxidation of unchanged opium analytes in the sample, thus the accuracy of GC-MS results is less affected by oxidizing adulterans [13]. ...
... Sodium hydroxide is a caustic strong base. It causes change to alkaline pH in urine samples, then it effects the drug binding and solubility and produces false negative urine results [13]. ...
... It was suggested that GC-MS confirmation test results aren't effected by non-oxidizing adulterans. Adding sodium hydrosulfide or sulfamic acid to the GC-MS method can help remove excess oxidizing additive and may prevent further oxidation of unchanged opium analytes in the sample, thus the accuracy of GC-MS results is less affected by oxidizing adulterans [13]. ...
... These results were in agreement with Uebel and Wium, (2002) who reported that drano is very effective in masking cannabis detection by immunoassay techniques. Also in consistent with Bronner et al., (1990) and Wu, (2003) who stated that hand soap detergent adulteration has caused false-negative results across a variety of drug assays using the cloned enzyme donor immunoassay (CEDIA) including screens for THC, amphetamine, barbiturates, cocaine, opiates, and PCP. Effect of different adulterants on THC-COOH detection and quantification by GC-MS: ...
... Another reagent, acidified potassium dichromate, will give a deep blue color change that fades over time when exposed to urine containing stealth (Caitlin et al., 2007). (Wu, 2003) described a simple fluorometric method for the detection of glutaraldehyde in urine. When 0.5 mL of urine was heated with 1.0 ml of a 7.7-mmol/L concentration of potassium dihydrogen phosphate (pH3.0) ...
... In the current research, dilution was the most common procedure of tampering of urine samples from both pre-employment/workplace and the suspected specimens settings. Diluting urine sample is often the easiest way to make a false negative drug test result [15] samples delivered to toxicology laboratory for drug abuse testing were diluted (creatinine <4 mmol/L) [16]. ...
... In fact, it is known that a large intake of fluids before the urine analysis reduces drugs of abuse levels, as well as urine creatinine levels, which allow possible counterfeit samples to be identified (5,14,39). In our population, the percentage of diluted samples was 3.8%, and positive/non-negative samples had lower creatinine than the overall average (97.6 vs.126.72 mg/dL; p-value=0.0002), ...
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Aim: Currently, procedures that identify the drugs 'destroyed' in adulterated urine specimens are very limited. This study aimed to determine the effect of pyridinium chlorochromate (PCC) on routine opiate assays and identify reaction products formed. Results/methodology: Opiate-positive urines adulterated with PCC (20 and 100 mM) were analyzed using CEDIA(®) immunoassay and GC-MS. Urine and water samples spiked with 6-monoacetylmorphine, morphine and its glucuronides (10 µg/ml) and PCC (0.02-100 mM) were monitored with LC-MS, and the products characterized. Conclusion: PCC significantly decreased the abundance of morphine, codeine and IS. Adulterated water and urine samples containing 6-monoacetylmorphine, morphine and morphine-3-glucuronide yielded morphinone-3-glucuronide, 7,14-dihydroxy-6-monoacetylmorphine, 7,8-diketo-6-monoacetylmorphine and 7,8-diketo-morphine (tentative assignment). Reaction pathways may be different in the two matrices.
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Manipulations of urine sample by urine substitution, urine dilution, and urine adulteration with highly oxidative chemicals to escape detection in doping control analysis have been reported in the past. Adulteration of urine with oxidising chemicals such as potassium permanganate, cerium ammonium nitrate, pyridinium chlorochromate etc can lead to considerable changes in the endogenous steroidal profile parameters and thus mask the abnormality in steroidal profile following steroid abuse. In this study we have identified the formation of two stable oxidation products on reaction of potassium permanganate with testosterone, an important endogenous urinary steroid. Isolation and characterisation of these oxidation products were performed using chromatography and spectroscopy and the products were elucidated as 4α,5α-dihydroxytestosterone and 4β,5β-dihydroxytestosterone. Formation of these two molecules in human urine after adulteration with potassium permanganate has been demonstrated by liquid chromatography-mass spectrometry (LC-MS) analysis. The products 4α,5α-dihydroxytestosterone and 4β,5β-dihydroxytestosterone have not been previously reported in urine and hence has the potential to be included in routine drug testing program for monitoring possible testosterone abuse and permanganate adulteration of urine.
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Drug testing programs are established to help achieve a drug-free work environment, promote fair competition in sport, facilitate harm minimization and rehabilitation programs, better manage patient care by clinicians and service law enforcement authorities. Urine remains the most popular and appropriate testing matrix for such purposes. However, urine is prone to adulteration, where chemicals, especially oxidizing chemicals, are purposely added to the collected urine specimens to produce a false-negative test result. This article will describe the effect of various popular oxidizing adulterants on urine drug test results, the countermeasures taken by laboratories in dealing with adulterated urine samples and the prospect of developing more robust and economical methods to combat urine adulteration in the future.
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Stealth is an adulterant used to avoid detection of drug abuse. The product does have an effect on the ability to detect several drugs of abuse, including the opiates morphine and codeine. It has previously been shown that low concentration (2500 ng/mL morphine) samples adulterated with Stealth tested negative by both Roche OnLine and Microgenics CEDIA immunoassays, but those spiked with higher concentrations (6000 ng/mL of codeine and morphine glucuronide) were positive. Initial results showed confirmation analysis was also sometimes negatively impacted by this adulterant. Urine samples were spiked with 6000 ng/mL of codeine and/or morphine glucuronide to assess the effect of Stealth. Each individual sample was split into separate aliquots. One aliquot of each was adulterated with Stealth following package directions. The samples were then tested by immunoassay and gas chromatography-mass spectrometry (GC-MS). The control and adulterated aliquots were positive by both immunoassays. Results of GC-MS analysis of the Stealth-adulterated aliquots following standard procedures using deuterated internal standards proved unsuccessful in several cases. In 4 of 12 cases (33%), neither the drugs nor internal standards were recovered despite repeated attempts. In one other sample, recovery was dramatically reduced, making accurate quantitation impossible, whereas the unadulterated aliquots of the same samples posed no problem with recovery. Addition of sodium disulfite to the aliquots prior to extraction allowed recovery of the drugs and internal standards from all samples. Analysis of the samples showed the concentration of morphine and codeine decreased in some by as much as 17 and 30%, respectively. In other cases, there was essentially no difference in the concentration seen before and after adulteration, with or without disulfite treatment. Unless the initial concentration of opiate is near the cutoff, samples containing opiates are likely to be immunoassay positive, it is important to consider this procedure as an option for samples that screen positive but the opiates and their respective internal standards are not recovered for GC-MS analysis.