Illicit medicine abuse continues to be a major public health issue. According to the 2013 national Survey on drug Abuse and also Health, an approximated 24.6 million Americans period 12 years and older were existing illicit medicine users—9.4% that the U.S. Population. Marijuana to be the most generally abused illicit drug, adhered to by cocaine, heroin, and hallucinogens.

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Federal guidelines specify an adulterated specimen together a pee specimen containing one of two people a substance the is no a normal constituent or one endogenous substance at a concentration the is not a normal physiological concentration. Pre-employment screening programs typically do no involve direct supervision of specimen collection, therefore employment candidates may attempt to cheat drug trial and error by adulterating specimens. This renders it necessary for laboratories to recognize pre-analytically any such adulterated specimens.

Ways the Cheating a medicine Test

Usually people shot to cheat drug testing by three different ways: substituting their urine with synthetic urine or drug-free urine purchased from a clandestine source; drinking a commercially easily accessible product to flush the end drugs; or adding an adulterant in vitro to the to pee specimen after collection.

Synthetic to pee is difficult to detect because it has comparable pH, creatinine, and certain gravity to typical urine. Details tests are needed to recognize compounds that are normal ingredient of person urine yet not uncovered in man-made urine, such as cortisol.Commercially available products that adulterate urine or flush out drugs can be divide under two broad categories. The very first includes fluids or tablet computers that, together with drinking big amounts of water, dilute urine. Common assets are Absolute decoding XXL drink, absolute Carbo Drinks, all set Clean drug Detox Drink, rapid Flush Capsules, and also Ready Clean gel Capsules.

The 2nd category of commodities is in vitro urinary adulterants that are included to urine ~ collection. Examples incorporate Stealth (peroxidase and peroxide), Klear (nitrite), Clean ADD-IT-ive (glutaraldehyde) and also Urine lucky (pyridinium chlorochromate ). In addition, iodine is a solid oxidizing agent and also may potentially destroy abused drugs, specifically marijuana metabolites (2). Research also indicates the papain through intrinsic ester hydrolysis ability could substantially reduce the concentration of 11-nor-9-carboxy-9-tetrahydrocannabinol (THC-COOH), a metabolite of marijuana, if added to the pee specimen in vitro (3).

Household Chemicals as Urinary Adulterants

Would-be medicine test cheaters might try adulterating your specimens with family chemicals, but most deserve to be detect by specimen verity testing. Both collection sites and also laboratories have actually at your disposal a number of mechanisms to detect possibly invalid specimens. The temperature, for instance, need to be within 90.5–98.9°F. The particular gravity should be between 1.005–1.030, and also pH should be between 4.0–10.0. The creatinine concentration must be 20–400 mg/dL. However, some drug experimentation laboratories consider a creatinine concentration of 15 mg/dL as the lower end cutoff. One common adulterant, sodium chloride, constantly produces a details gravity higher than 1.035 if added at a concentration important to develop a false-negative result.

Unfortunately, specimen integrity trial and error doesn’t detect every adulterants. Because that example, that won’t choose up adulteration the urine with Visine eye drops, isopropanol, or other urinary adulterants. However, efficient spot tests and also special pee dipsticks are easily accessible (See Table 1).

Flushing, Detoxification Agents, and also Diuretics

Flushing and also detoxification agents are generally advertised together effective way of passing drug tests. Numerous of these assets contain caffeine or various other diuretics to increase the calculation of urine, as well as sugar and also natural or artificial flavoring agents. The target is to produce diluted urine so the concentrations of abused drugs and or metabolites fall below the encourage cutoff concentrations.

Cone et al. Evaluated the result of excess liquid ingestion on false-negative marijuana and cocaine urine test outcomes by studying the ability of normally Clean natural Tea, goldenseal root, and also hydrochlorothiazide to reason false negative results. Volunteers drank one gallon the water, natural tea, or take it hydrochlorothiazide 22 hrs after cigarette smoking marijuana cigarettes or intranasal administration of cocaine. Your creatinine levels dropped listed below the cutoff 2 hrs after intake of excessive fluid. Marijuana and cocaine metabolite levels (as measured by both enzyme multiplied immunoassay method and also fluorescence polarization immunoassay ) diminished significantly and frequently switched from positive to an unfavorable in subjects after consuming 2 quarts the fluid. Even excess water was reliable in diluting a urine specimen to reason false an unfavorable results, although natural tea diluted to pee faster contrasted to water alone (5).

Using point out Tests

When specimen integrity testing cannot detect an adulterated specimen, laboratories have the right to employ a selection of reliable spot tests.

• urine LuckWu et al. Reported that the active ingredient the “Urine Luck” was PCC, a strong oxidizing agent, which at a concentration of 100 gm/L, caused substantially decreased solution rate for all EMIT II drug screens, indicating the opportunity of false-negative results. In contrast, because that the Abbott Abuscreen test, only morphine and marijuana assays were affected, yet a false-positive result was observed v the amphetamine assays. This adulteration that urine did not alter GC/MS check of methamphetamine, benzoylecgonine, and phencyclidine, but obvious concentrations that opiates and also THC-COOH were substantially reduced.Wu et al. Likewise described a simple spot test making use of 1,5-diphenylcarbazide in methanol (10 gm/L) to detect the existence of PCC in urine, in i m sorry a reddish violet color emerged in the presence of PCC (6). Moreover, including a couple of drops of 3% family members hydrogen peroxide solution to approximately 0.5 mL of pee specimen brought about immediate breakthrough of a dark brown color and also dark brown precipitate if PCC was present in the urine. As a solid oxidizing agent, PCC could additionally liberate iodine from potassium iodide solution in acidic medium (7). Notably, several other adulterants easily accessible online save PCC.

• Nitrite comprise AgentsProducts such together “Klear,” which includes potassium nitrite, can cause interference in GC/MS confirmation of THC-COOH. However, a bisulfite action at the start of sample preparation can eliminate this problem (8).Nitrite in urine might arise in vivoin patient receiving medicines such together nitroglycerin, isosorbide dinitrate, and also nitroprusside, or as result of urinary tract infection. However, concentrations of nitrite normally are below 36 µg/mL in together specimens, when nitrite concentrations room 1,910–12,200 µg/mL in urine specimens adulterated with nitrite (9).Nitrite have the right to be conveniently detected by straightforward spot tests. Enhancement of a few drops of a nitrite-adulterated urine specimen come 0.5 mL the 1% potassium permanganate solution, complied with by enhancement of a few drops that 2N hydrochloric acid, rotate the pink permanganate equipment colorless through effervescence. One more spot check to detect nitrite supplied 1% potassium iodide solution. Adding a couple of drops that nitrite adulterated urine come 0.5 mL the potassium iodide solution, adhered to by enhancement of a couple of drops that 2N hydrochloric acid, result in instant release that iodine from the colorless potassium iodide solution. If any organic solvent, such as hexane, was included the iodine was easily transferred in the necessary layer providing the layer a distinct shade of iodine (7). Nitrite could also be recognize by diazotizing sulfanilamide and also coupling the product v N-(1-napthyl) ethylenediamine.

• StealthStealth is an adulterant which is composed of 2 vials, one include a powder (peroxidase) and another vial include a fluid (hydrogen peroxide), both added to the urine. Stealth is capable of invalidating immunoassay screening of THC-COOH, LSD, and also opiates making use of both Roche online assays and also Microgenics CEDIA assays if these drug or metabolites are current in modest concentration (125–150% of cutoff values). In addition, GC/MS confirmation might be impacted (10).Valtier and Cody described a quick spot test to detect the existence of Stealth in urine. Enhancement of 10 µL that urine to 50 µL that tetramethylbenzidine working solution, complied with by addition of 500 µL that 0.1 M phosphate buffer solution brought about the specimen to rotate dark brown. Peroxidase activity could likewise be monitored by utilizing a spectrophotometer (11). Our examination showed that if a few drops of a pee specimen adulterated with Stealth were included to potassium dichromate adhered to by a few drops the 2N hydrochloric acid, a deep blue color developed immediately, which generally faded with time.

• GlutaraldehydeGlutaraldehyde containing commodities were one of the first that appeared in the industry to invalidate drugs of abuse testing. Glutaraldehyde remedies are also accessible in hospitals and clinics as a clean agent. Glutaraldehyde at a concentration that 0.75% volume can lead come false-negative screening outcomes for a cannabinoid test using the EMIT II drugs of abuse screen. Amphetamine, methadone, benzodiazepine, opiate, and cocaine metabolite tests room also affected at glutaraldehyde concentration between 1 and also 2% using EMIT II immunoassays.Wu et al. Described a an easy fluorometric method for the detection the glutaraldehyde in urine. As soon as 0.5 mL that urine to be heated with 1.0 mL that 7.7 mmol/L potassium dihydrogen phosphate (pH 3.0) saturated with diethyl-thiobarbituric acid for 1 hour at 96–98°C in a heating block, a yellow eco-friendly fluorophore arisen if glutaraldehyde to be present. Shiver the specimen v n-butanol caused the move of this adduct to the organic layer which might be regarded under lengthy wavelength UV light. Glutaraldehyde in to pee can also be estimated using a fluorometer (14).

• Zinc Sulfate: A brand-new Urinary AdulterantAlthough not widely used, zinc sulfate is an reliable urinary adulterant that can invalidate all drug tests using EMIT assay. Right now there is no suitable technique for detecting zinc sulfate in adulterated urine. Therefore, two fast spot tests come detect the visibility of zinc sulfate in urine were developed. Addition of 3–4 fall of 1N sodium hydroxide equipment to roughly 1 mL of urine containing zinc sulfate result in development of a white precipitate, which was soluble in excess salt hydroxide. In the second spot test, addition of 3–4 autumn of 1% sodium chromate solution to 1 mL of pee containing zinc sulfate complied with by enhancement of 4–5 autumn of 1N salt hydroxide led to formation of a yellow precipitate (zinc chromate) (15).

Testing pee Specimens for Adulterants

Specially designed to pee dipsticks such as AdultaCheck 4, AdultaCheck 6, or Intect 7 can be supplied to detect numerous adulterants in urine. AdultaCheck 6 detects creatinine, oxidants, nitrite, glutaraldehyde, pH, and also chromate. The Intect 7 test strip for check adulteration in urine is written of seven various pads to test because that creatinine, nitrite, glutaraldehyde, pH, details gravity, bleach, and also PCC.

Guidelines native the substance Abuse and also Mental wellness Services administration require added tests for urine specimens through abnormal physical qualities or ones that show qualities of an adulterated specimen during initial screening or confirmatory tests. A pH much less than 3 or more than 11, and nitrite concentrations higher than 500 mg/mL show the visibility of adulterants. A nitrite colorimetric check or a general oxidant colorimetric test have to be carry out to identify nitrite.

The visibility of chromium (VI) in a urine specimen likewise is indicative that adulteration in ~ a cutoff concentration of 50 mg/mL. The visibility of chromium in a pee specimen can be evidenced by a chromium colorimetric test or a general test because that the visibility of oxidant. A confirmatory test have to be performed utilizing multi-wavelength spectrophotometry, ion chromatography, atomic absorption spectrophotometry, capillary electrophoresis, or inductively combination plasma fixed spectrometry.

Elemental halogens, such as pure bromine or iodine, can also be offered as adulterants. The existence of these halogens need to be confirmed by a halogen colorimetric test or a basic test because that the visibility of oxidants. Confirmatory tests may employ multi-wavelength spectrophotometry, ion chromatography, atomic absorption spectrophotometer, capillary electrophoresis, or inductively coupled plasma fixed spectrometry.

To detect glutaraldehyde, laboratories must use a general aldehyde check or the characteristics immunoassay solution in one or more drug immunoassay tests for initial screening. Similarly, the visibility of PCC need to be evidenced using a basic test because that the visibility of oxidant and also a GC/MS confirmatory test. Finally, surfactant must be confirmed by utilizing a surfactant colorimetric test through a greater than or equal to 100 mg/mL dodecyl benzene sulfonate equivalent cutoff.


It is necessary for laboratories come detect adulterated urine in the pre-analytical step, as plenty of adulterants invalid immunoassay screening tests. Although routine specimen verity tests can detect most of the family adulterants except Visine eye drops and alcohol/isopropanol, adulterants containing strong oxidizing agents such together potassium nitrite, pyridinium chlorochromate, or Stealth need a different approach. Spot tests, specially designed pee dipsticks, also as more analytically advanced methods such together chromatographic methods, are easily accessible in the toxicology activities to recognize these adulterants. If a pee specimen is adulterated it have to be documented and also reported, however no further experimentation is necessary.

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