← Product Code [DIO](/submissions/TX/subpart-d%E2%80%94clinical-toxicology-test-systems/DIO) · K152232

# Quest Diagnostics HairCheck-DT (Cocaine) (K152232)

_Quest Diagnostics Incorporated · DIO · Nov 18, 2016 · Clinical Toxicology · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/TX/subpart-d%E2%80%94clinical-toxicology-test-systems/DIO/K152232

## Device Facts

- **Applicant:** Quest Diagnostics Incorporated
- **Product Code:** [DIO](/submissions/TX/subpart-d%E2%80%94clinical-toxicology-test-systems/DIO.md)
- **Decision Date:** Nov 18, 2016
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 862.3250
- **Device Class:** Class 2
- **Review Panel:** Clinical Toxicology

## Indications for Use

The Quest Diagnostics HairCheck-DT (Cocaine) test system utilizes an Enzyme Linked Immunosorbent Assay (ELISA) for the qualitative detection of cocaine in head hair samples through the measurement of cocaine and cocaine metabolites for concentrations at or above 300 pg/mg hair. This test system has not been evaluated for use with hair specimens from locations other than the head. It is an in vitro diagnostic device intended exclusively for in-house professional use and is not intended for sale to anyone. The HairCheck-DT (Cocaine) test system was evaluated in two distinct study populations; individuals known to be chronic drug abusers, and individuals proclaiming to be drug-free. The Quest Diagnostics HairCheck-DT (Cocaine) test system provides only a preliminary analytical test result. To confirm a presumptive screen positive result, a more specific alternate chemical method, such as gas chromatography-mass spectrometry (GC-MS) or Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS), must be used. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are obtained.

## Device Story

Device is an in vitro diagnostic ELISA for qualitative cocaine detection in head hair. Input: 10mg minced head hair sample. Process: Hair is washed with methanol, extracted in acidified methanol (0.5% TFA) at 62°C, sonicated, dried under nitrogen, and reconstituted in phosphate buffer. Extract is added to microtiter plates coated with mouse anti-cocaine monoclonal antibody; HRP-conjugated cocaine competes for binding sites. After washing, TMB substrate is added; color development is inversely proportional to analyte concentration. Output: Absorbance measured at 450/620 nm via automated plate reader. Used in professional laboratory settings. Results are preliminary; positive screens require confirmation via GC-MS or LC-MS/MS. Clinical decision-making relies on professional judgment. Benefits include screening for cocaine use in chronic abusers or drug-free populations.

## Clinical Evidence

No clinical studies performed. Bench testing included precision/reproducibility studies (n=5 donor samples), analytical specificity (143 compounds tested), and cosmetic treatment interference (60 positive/60 negative samples). Method comparison with GC/MS (n=100) showed concordance; discordant results were attributed to dye interference or near-cutoff variability.

## Technological Characteristics

Solid-phase microtiter plate ELISA. Materials: Mouse anti-cocaine monoclonal antibody, HRP-cocaine conjugate, TMB substrate, 1N H2SO4 stop solution. Energy source: Automated microplate reader (450/620 nm). Connectivity: Standalone laboratory instrument. Sterilization: Not applicable (reagent kit). Software: None (automated plate reader processing).

## Regulatory Identification

A cocaine and cocaine metabolite test system is a device intended to measure cocaine and a cocaine metabolite (benzoylecgonine) in serum, plasma, and urine. Measurements obtained by this device are used in the diagnosis and treatment of cocaine use or overdose.

## Special Controls

*Classification.* Class II (special controls). A cocaine and cocaine metabolite test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (*e.g.,* programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).

## Predicate Devices

- Quest Diagnostics HairCheck-DT (Cocaine) (k023626)

## Submission Summary (Full Text)

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE

A. 510(k) Number:
k152232

B. Purpose for Submission:
New device

C. Measurand:
Cocaine and cocaine metabolites in hair

D. Type of Test:
Qualitative Enzyme Linked Immunosorbent Assay (ELISA)

E. Applicant:
Quest Diagnostics, Inc.

F. Proprietary and Established Names:
Quest Diagnostics HairCheck-DT (Cocaine)

G. Regulatory Information:
1. Regulation section:
21 CFR §862.3250, Cocaine and Cocaine Metabolite Test System
2. Classification:
Class II
3. Product code:
DIO – Enzyme Immunoassay, Cocaine and Cocaine Metabolites

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4. Panel:
Toxicology (91)

H. Intended Use:

1. Intended use(s):
See indications for use below.

2. Indication(s) for use:
The Quest Diagnostics Hair Check-DT (Cocaine) test system utilizes an Enzyme Linked Immunosorbent Assay (ELISA) for the qualitative detection of cocaine in head hair samples through the measurement of cocaine and cocaine metabolites for concentrations at or above 300 pg/mg hair. This test system has not been evaluated for use with hair specimens from locations other than the head. It is an in vitro diagnostic device intended exclusively for in-house professional use and is not intended for sale to anyone.

The Hair Check-DT (Cocaine) test system was evaluated in two distinct study populations; individuals known to be chronic drug abusers, and individuals proclaiming to be drug-free.

The Quest Diagnostics Hair Check-DT (Cocaine) test system provides only a preliminary analytical test result. To confirm a presumptive screen positive result, a more specific alternate chemical method, such as gas chromatography - mass spectrometry (GC/MS) or Liquid Chromatography-Tandem Mass Spectrometry (LC/MS/MS) must be used. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are obtained.

3. Special conditions for use statement(s):
The Quest Diagnostics HairCheck-DT (Cocaine) ELISA provides only a preliminary result. Clinical consideration and professional judgment must be applied to any drug of abuse test result, particularly in evaluating a preliminary positive result. To obtain confirmed analytical results a more specific alternate method such as gas chromatography/mass spectrometry (GC/MS) or Liquid Chromatography-Tandem Mass Spectrometry (LC/MS/MS) must be used.

4. Special instrument requirements:
The device is for use with an automated microplate reader capable of measuring at 450 and 620 nm.

Confirmation testing was performed using an Agilent GC/MS system.

2

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I. Device Description:

The assay consists of two parts; a pre-analytical hair treatment procedure (to extract cocaine from the solid hair matrix to a measurable liquid matrix), and the screening assay. The screening assay is an Enzyme-Linked ImmunoSorbent Assay (ELISA).

Kit Components

Each kit Quest Diagnostics HairCheck-DT (Cocaine) contains enough reagents to make 4,800 determinations

Reagents

- 50 x 96 well micro strip plates (12 x 8), coated with mouse anti-Cocaine monoclonal antibody.
- 2 x 4 mL of enzyme concentrate conjugate (horseradish peroxidase conjugated to cocaine) in a proprietary buffer containing stabilizing agents and thimerosal.
- 2 x 400 mL of enzyme diluent as a proprietary buffer containing stabilizing agents and thimerosal.
- 1 x 500 mL of substrate containing tetramethylbenzidine (TMB) and hydrogen peroxide in a citrate buffer containing stabilizers.
- 1 x 1000 mL of concentrated wash solution with surfactants and thimerosal as a preservative; Dilute 1:10 with deionized water prior to use.
- 1 x 500 mL of acid stop solution containing 1 N H₂SO₄.

Calibrators and Controls for Screening Assay

- 100 mL of 1X HairCheck-DT (Cocaine) Negative Control containing 0 pg cocaine/mg hair (No need to dilute, provided at working concentration)
- 5 mL of 40X HairCheck-DT (Cocaine) Low Control containing 6,000 pg cocaine/mg hair (Dilute 1:40 with Negative Control to reach 1X Low Control working concentration of 150 pg cocaine/mg hair)
- 5 mL of 40X HairCheck-DT (Cocaine) Cutoff Calibrator containing 12,000 pg cocaine/mg hair (Dilute 1:40 with Negative Control to reach 1X calibrator working concentration of 300 pg cocaine/mg hair)
- 5 mL of 40X HairCheck-DT (Cocaine) High Control containing 18,000 pg cocaine/mg hair (Dilute 1:40 with Negative Control to reach 1X High Control working concentration of 450 pg cocaine/mg hair)

J. Substantial Equivalence Information:

1. Predicate device name(s):

Quest Diagnostics HairCheck-DT (Cocaine)

2. Predicate 510(k) number(s):

k023626

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3. Comparison with predicate:

|  Item | Candidate Device Quest Diagnostics Hair Check-DT (Cocaine) | Predicate Device Quest Diagnostics HairCheck-DT (Cocaine) (k023626)  |
| --- | --- | --- |
|  Similarities  |   |   |
|  Intended Use | For the qualitative detection of cocaine in head hair samples through the measurement of cocaine and cocaine metabolites | Same  |
|  Methodology | ELISA | Same  |
|  Cutoff | 300 pg cocaine/mg hair | Same  |
|  Specimen type | Head hair | Same  |
|  Competitive Enzyme-conjugate | HRP - Cocaine | Same  |
|  Differences  |   |   |
|  Antibody | Mouse anti-Cocaine monoclonal antibody | Rabbit anti-Cocaine polyclonal antibody  |
|  Extraction method | Acidified Methanol (0.5% Trifluoroacetic acid)(TFA) | Methanol  |
|  Control levels | High Control: 450 pg cocaine/mg hair Low control: 150 pg cocaine/mg hair | High Control: 600 pg cocaine/mg hair Same  |
|  Calibrators | Cutoff Calibrator containing 300 pg cocaine/mg hair | Same  |
|  Sample Size | Sample Preparation modified to 10 mg of Hair used for extraction then reconstituted with 0.3 mL phosphate buffer | Sample Preparation 20 mg of Hair used for extraction then reconstituted with 0.6 mL phosphate buffer  |
|  Measurement Wavelength | Assay absorbance measured at 450 nm with a reference wavelength of 620 nm. | Assay absorbance was measured at 450 nm with a reference wavelength of 630 nm.  |
|  Kit Configuration | Kit Configuration modified to = 50 microplate kit | Kit Configuration = 5 microplate kit  |

K. Standard/Guidance Document Referenced (if applicable):

None referenced

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5

L. Test Principle:

The test consists of two parts; a pre-analytical hair treatment procedure (to extract Cocaine from the solid hair matrix to a measurable liquid matrix), and the screening assay. The screening assay is an Enzyme-Linked ImmunoSorbent Assay (ELISA).

This assay requires a sample of head hair (approximately 120 strands) that is cut as close as possible to the scalp, preferably from the back of the head at the crown. The hair sample is treated with a pre-analytical treatment to extract cocaine from the hair into a liquid sample. The liquid sample is added to a well of the microplate and enzyme conjugate is added, followed by incubation. During this phase the enzyme-labeled drug conjugate competes with drug in the liquid sample for a limited number of binding sites on the mouse antibody-coated micro wells. A wash solution is applied to remove any unbound materials. Enzyme substrate solution containing a chromogen is added. The reaction is stopped with an acid and the absorbance is read using a plate reader at 450 nm and a background reading is also taken at 620 nm. Color intensity is inversely proportional to the amount of drug present in the sample.

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

Reproducibility

The reproducibility of the extraction protocol was tested using five cocaine positive donor hair specimens. Three, 10 mg aliquots of each donor sample of hair were extracted and measured in one ELISA run with %CV calculated. The results are summarized in the table below.

|  Hair sample | Cocaine pg/mg by GC/MS | Extraction replicate | Mean OD | CV% | ELISA Pos/Neg  |
| --- | --- | --- | --- | --- | --- |
|  1 | 560 | 3 | 0.811 | 0.6% | 3/0  |
|  2 | 18078 | 3 | 0.025 | 2.3% | 3/0  |
|  3 | 2926 | 3 | 0.116 | 7.8% | 3/0  |
|  4 | 7698 | 3 | 0.059 | 3.5% | 3/0  |
|  5 | 2320 | 3 | 0.231 | 5.2% | 3/0  |

Precision

Nine aliquots were taken from the negative hair matrix pool (liquid extract of negative hair samples) and spiked with cocaine at the following levels: 0 ng/mL (0 pg/mg), 2.5 ng/mL (75 pg/mg), 5 ng/mL (150 pg/mg), 7.5 ng/mL (225 pg/mg), 10 ng/mL (300 pg/mg), 12.5 ng/mL (375 pg/mg), 15 ng/mL (450 pg/mg), and 17.5 ng/mL (525 pg/mg), 20 ng/mL (600 pg/mg) representing 0, 25, 50, 75, 100, 125, 150,

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175, 200 % of the calibrator cutoff value. The aliquots were then tested in 5-replicates on each of ten different days using the Quest Diagnostics HairCheck-DT (Cocaine) ELISA. Results are summarized below.

|  % Relative to | 0% | 25% | 50% | 75% | 100% | 125% | 150% | 175% | 200%  |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  Target pg/mg | 0 | 75 | 150 | 225 | 300 | 375 | 450 | 525 | 600  |
|  Negative Count | 50 | 50 | 50 | 50 | 25 | 3 | 0 | 0 | 0  |
|  Positive Count | 0 | 0 | 0 | 0 | 25 | 47 | 50 | 50 | 50  |
|  % Negative | 100% | 100% | 100% | 100% | 50% | 6% | 0% | 0% | 0%  |
|  % Positive | 0% | 0% | 0% | 0% | 50% | 94% | 100% | 100% | 100%  |

b. Linearity/assay reportable range:

Not applicable

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

Traceability

Calibrator and controls are prepared from Certified Reference Material Cerilliant C-008 Cocaine 1.0 mg/mL in Acetonitrile.

Stability

Shelf-life: The accelerated and real-time stability support the shelf-life of 6 months for the reagent kit (including calibrators and controls), stored at 2-8°C.

Open vial stability: The study support an open vial claim of 30 days, stored at 2-8°C for all kit components, including the cutoff calibrator and controls.

Hair Specimen Stability: Established in k023626.

Shipping study:

In order to demonstrate the stability of cocaine in hair specimens during the shipping process, 56 hair samples- twenty eight (28) confirmed positive cocaine hair specimens, of which three specimens (3) have pre-shipping quantitative GC/MS results in the near cutoff positive concentration range (300 to 449 pg/mg Cocaine) and twenty eight (28) confirmed negative cocaine hair specimens, of which three (3) have pre-shipping quantitative GC/MS results in the near cutoff negative concentration range (150 to 299 pg/mg Cocaine) were shipped to eight different geographical locations and the temperatures tracked during shipping. By enclosing an electronic temperature sensor within each shipment of hair samples, we were able to retrieve a range of temperatures collected every 20 minutes during the shipping process. In order to mimic potential shipping extreme temperatures, the hair was first

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cold shocked at -15°C for 15 hours and then heat shocked at +47°C for 6 hours prior to shipping.

After the boxes were returned to facility, ELISA screening and GC/MS confirmation were performed on each sample and compared to the non-shipped results for the same samples. The cocaine negative samples that were shipped maintained their negative status both for ELISA screening and GC/MS with respect to cocaine levels. One of the borderline negative samples originally screened positive prior to shipping and all three of the borderline negative samples screened positive after shipping due to variability near the cutoff of the ELISA screening method. The cocaine positive samples that were shipped maintained their positive status for both the ELISA screening and the GC/MS confirmation with respect to cocaine levels.

Summary of results for positive specimens

|   | Pre-Shipping |   | Post-Shipping  |   |
| --- | --- | --- | --- | --- |
|   |  GC/MS | ELISA | GC/MS | ELISA  |
|  Positive | 28 | 28 | 28 | 28  |
|  Negative | 0 | 0 | 0 | 0  |

Summary of results for negative specimens

|   | Pre-Shipping |   | Post-Shipping  |   |
| --- | --- | --- | --- | --- |
|   |  GC/MS | ELISA | GC/MS | ELISA  |
|  Positive | 0 | 1* | 0 | 3**  |
|  Negative | 28 | 27 | 28 | 25  |

* One of the three Near Cutoff Negative specimens (as determined by GC/MS pre-shipping) screened positive by ELISA prior to shipping.

** Three of three Near Cutoff Negative specimens (as determined by GC/MS pre-shipping) screened positive by ELISA post-shipping.

d. Detection limit:

Analytical performance of the device around the cutoff is described in Section M1.a (Precision/Reproducibility) above.

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e. Analytical specificity:

Interference:

One hundred and forty three commonly used compounds, many of them prescribed medicines, and abused drugs, were tested in the study. These compounds are not pharmacologically or structurally related to Cocaine. Each compound was tested at 10,000 ng/mL (equivalent to 300,000 pg/mg of hair), in hair matrix pools at two cocaine concentrations, equivalent to 50% below (150 pg/mg) and 50% above (450 pg/mg) the Cocaine cutoff value of 300 pg/mg.

Prepared samples were run in singlet on the Quest Diagnostics Hair Cocaine ELISA. If the OD of the above cutoff pool samples spiked with compound of interest was positive (had an OD lower than or equal to the cutoff), then the tested compounded was considered to not negatively interfere. If either the below cutoff pool spiked sample or above cutoff pool spiked sample had a response that showed interference, that sample was diluted 1:2 in corresponding below cutoff pool or above cutoff pool until no interference was observed.

The lists of compounds that do not interfere are listed in the table below and are provided in the product labeling.

Compounds That Do Not Interfere with the Assay:

|  (-)-11-nor-9-Carboxy-Δ9-THC | Cannabinol | Hydrochlorthiazide | Normenperidinic Acid (4-Phenylpiperidine-4-carboxylic acid hydrochloride)  |
| --- | --- | --- | --- |
|  (+)-11-Nor-Δ9-THC-9-carboxylic acid glucuronide | Catharanthine (Ergoloid) | Hydrocodone | Normorphine  |
|  (-) Cotinine | Chlordiazepoxide | Hydrocortisone (Cortisol) | Noroxymorphone  |
|  (-) Methamphetamine | Cimaterol | Hydromorphone | O-Desmethyl-cis-tramadol HCl  |
|  (+) Isoproterenol | Clenbuterol | Ibuprofen | Oxazepam  |
|  (+) Methamphetamine | Clonazepam | Imipramine | Oxymorphone  |
|  (+) Norpseudoephedrine | Codeine | LAMPA | PCP  |

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|  (±) Ketamine | Corticosterone | Levorphanol | Penicillin G Sodium salt  |
| --- | --- | --- | --- |
|  (±) MDA [(±)-3,4-Methylenedioxy amphetamine] | Cortisone | Lidocaine | Pentazocine  |
|  (±) MDEA [(±)-3,4-Methylenedioxy ethylamphetamine] | (-)-Δ8-THC | Lorazepam | Phenothiazine  |
|  (±) MDMA [(±)-3,4-Methylenedioxy methamphetamine] | (-)-Δ9-THC | LSD | Phentermine  |
|  (±) Propanolol | Desalkyl flurazepam | Meperdine | Phenylbutazone  |
|  (±)-N-Ethylamphetamine | Desipramine | Metaproterenol hemisulfate | P-Hydroxymeth amphetamine (Isodrin)  |
|  (±)-N-Propylamphetamine | Desmethyldoxepin (cis/trans) | Methedrone (Methyoxyephedrine) | Progesterone  |
|  (±)-Phenylpropanolamine (Norephedrine) | Dexamethasone | Methoxyphenamine HCl | Promethazine HCl  |
|  (±)-Methadone | Dextromethorphan | Methylergometrine maleate | Proprionyl Promazine HCl  |
|  Nandrolone (19-Nortesterone) | Diazepam | Methylphenidate | R(-) Phenylephrine  |
|  1R,2S(-)-Ephedrine | Dihydrocodeine | Monensin Sodium Salt | R(-)-Amphetamine  |
|  1S,2R(+)-Ephedrine | Dihydroergotamine Mesylate | Morphine | R(+) Methcathinone  |
|  2-Oxo-3-hydroxy-LSD (2-Oxo-3-hydroxy-lysergic acid diethylamide) | Dihydromorphine | Morphine-3-β-D-glucuronide | R,R(-)-Pseudoephedrine  |

9

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|  Acetaminophen (4-Acetoamidophenol) | Doxepin (cis/trans) | Morphine-6-β-D-glucuronide | S(-)-Nicotine  |
| --- | --- | --- | --- |
|  4-MeO-PCP HCl (4- | Doxylamine succinate | Nadolol | S(+)-Amphetamine  |
|  Methoxyphencyclidine HCl) | Venlafaxine hydrochloride (Effexor) | Nalbuphine | Salbutamol (Albuterol)  |
|  6-Acetylmorphine (6-Monoacetylmorphine) | Erythromycin | Nalorphine | S, S(+) Pseudoephedrine  |
|  Acebutolol HCl | Ethylmorphine | Naltrexone | Stanozolol  |
|  p-Acetophenetidin (Phenacetine) | Fenfluramine | N-Desmethyodramadol | Streptomycin Sulfate  |
|  Acetylsalicylic Acid | Fentanyl | Neomycin | Sufentanil  |
|  7-Aminoflunitrazepam | Flumethasone | N-Ethylcathione | Tetracycline HCl  |
|  Amoxicillin | Flunitrazepam | (±)-4-Methyl-N-ethyl-norephedrine HCl (N-Ethyl nor ephedrine) | Theophylline  |
|  Betamethasone | Flurazepam | Norbuprenorphine | cis-Tramadol  |
|  Buprenorphine | Furosamide | Norcodeine | Triazolam  |
|  Caffeine | Heroin | Nordiazepam | Triflupromazine HCL  |
|  Cannabidiol | Normeperidine | Tylosin Tartrate |   |
|  HMMA (4-Hydroxy-3-Methoxy |  |  |   |

Of the 143 compounds tested, seven exhibited interference. For these compounds, and for an additional 10 dyes with the potential to interfere, the concentration of the test compound that gave a similar absorbance to the  $300~\mathrm{pg / mg}$  cocaine cut-off control was determined and percent cross reactivity was calculated. The results are shown below:

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|  Compound | Positive Interference observed at or above: | Equivalent Concentration in Hair (pg/mg)  |
| --- | --- | --- |
|  Thioridazine | 2,500 ng/mL | 75,000  |
|  Trifluperazine HCl | 5,000 ng/mL | 150,000  |
|  Chlorpromazine | 10,000 ng/mL | 300,000  |
|  Trimerperazine | 10,000 ng/mL | 300,000  |
|  Prochlorperazine dimaleate | 2,500 ng/mL | 75,000  |
|  Fluphenazine dihydrochloride | 5,000 ng/mL | 150,000  |
|  Iso-LSD | 1,250 ng/mL | 37,500  |
|  Basic Yellow 40 | 7,812.5 ng/mL | 234,375  |
|  Methylene Blue | 15,625 ng/mL | 468,750  |
|  Basic Violet 16 | 31,250 ng/mL | 937,500  |
|  Basic Blue 99 | 62,500 ng/mL | 1,875,000  |
|  Basic Brown 16 | 62,500 ng/mL | 1,875,000  |
|  Ethyl Violet | 62,500 ng/mL | 1,875,000  |
|  Basic Brown 17 | 62,500 ng/mL | 1,875,000  |
|  Basic Red 51 | 125,000 ng/mL | 3,750,000  |
|  Safranin O | 125,000 ng/mL | 3,750,000  |
|  Basic Yellow 87 | 500,000 ng/mL | 15,000,000  |

## Cross-reactivity

Cocaine, structurally-related compounds, pharmacologically-related compounds, and metabolites were tested for cross-reactivity in the assay. Eleven (11) compounds, structurally related to cocaine and known cocaine metabolites, were selected for the study (see table below for the list). The cross-reactant solutions were prepared by adding the compounds to negative hair matrix. The concentrations listed below produced a result approximately equal to the cutoff calibrator.

Serial dilutions of each compound were prepared and analyzed. If the OD response of the spiked sample was positive then the spiked sample was diluted further with finer dilutions until a positive result was obtained with an OD within 5% of the cutoff value (sample OD/Cutoff OD). Cross reactivity was calculated as: (Cutoff Concentration/Lowest Cross Reactant Concentration with a Positive Result) x 100.

The results are shown in the table below:

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Cross reactivity of Cocaine ELISA with Structurally Similar Compounds

|  Compound | Cross Reactivity (%) | Tested Concentration in Negative Hair Matrix (ng/mL) | Concentration of compound (pg/mg hair) needed to produce results equivalent to 300 pg/mg of Cocaine  |
| --- | --- | --- | --- |
|  Anhydroecgonine | <0.1% | 10,00 | >300,00  |
|  Anhydroecgonine methyl ester | <0.1% | 10,00 | >300,00  |
|  Atropine | <0.1% | 10,00 | >300,00  |
|  Benzoylecgonine | 143% | 7 | 210  |
|  Cocatheylene | 125% | 8 | 240  |
|  Ecgonine | <0.1% | 10,000 | >300,000  |
|  Ecgonine methyl ester | <0.1% | 10,000 | >300,000  |
|  Meta-hydroxybenzoylecgonine | 200% | 5 | 150  |
|  Norcocaine | 1% | 1,000 | 30,000  |
|  Tropacocaine | 8% | 125 | 3,750  |
|  Cocaine | 100% | 10 | 300  |
|  Cocaine N-oxide HCl | 11% | 90 | 2,700  |

# Cosmetic treatment

A panel of sixty (60) confirmed positive cocaine hair samples and sixty (60) screened negative cocaine hair samples were either treated with one of five (5) cosmetic hair treatments or left untreated. Cocaine positive hair is defined as a hair sample confirmed by GC/MS as having greater than  $300~\mathrm{pg / mg}$  cocaine. Absorbance readings after treatment were compared to absorbance readings prior to treatment, with the resulting change and direction of change noted. The resulting changes in ELISA test results are also noted, and are included in the table(s) below. (Absorbance values are normalized to the absorbance value of the cutoff calibrator absorbance value.)

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ELISA Results for Cocaine Positive Hair

|   | Pre-Treatment |   | Post-Treatment |   | Range of % differences in raw OD from Pre-Treatment to Post - Treatment  |
| --- | --- | --- | --- | --- | --- |
|   |  Number of Samples Positive by GC/MS | Number of Samples Positive by ELISA | Number of Samples Positive byGC/MS | Number of Samples Positive by ELISA  |   |
|  Shampoo | 12 | 12 | 12 | 12 | -48% to + 56%  |
|  Brown Dye | 12 | 12 | 12 | 12 | -59% to + 174%  |
|  Bleach | 12 | 12 | 12 | 12 | -31% to + 278%  |
|  Perm | 12 | 12 | 12 | 12 | -60% to + 27%  |
|  Relaxer | 12 | 12 | 12 | 12 | +12% to + 197%  |

ELISA Results for Cocaine Negative Hair

|   | Pre-Treatment |   | Post-Treatment |   | Range of % differences in raw OD from Pre-Treatment to Post - Treatment  |
| --- | --- | --- | --- | --- | --- |
|   |  Number of Samples Negative by GC/MS | Number of Samples Negative by ELISA | Number of Samples Negative by GC/MS | Number of Samples Negative by ELISA  |   |
|  Shampoo | 12 | 12 | 12 | 12 | -8% to + 10%  |
|  Brown Dye | 12 | 12 | 12 | 12 | -13% to + 4%  |
|  Bleach | 12 | 12 | 12 | 12 | -9% to + 5%  |
|  Perm | 12 | 12 | 12 | 12 | -7% to + 6%  |
|  Relaxer | 12 | 12 | 12 | 12 | -16% to + 4%  |

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Positive Specimens Individual Results

|  Treatment | ID | GC/MS Pre-Cosmetic Treatment pg/mg | ELISA Pre-Treat Raw OD | ELISA Pre-Treat P/N | ELISA Post-Treat Raw OD | ELISA Post-Treat P/N  |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |  Cocaine  |   |   |   |   |
|  Shampoo | P1 | 809 | 0.772 | POS | 0.976 | POS  |
|   |  P2 | 1391 | 0.505 | POS | 0.504 | POS  |
|   |  P3 | 4209 | 0.164 | POS | 0.165 | POS  |
|   |  P4 | 5927 | 0.089 | POS | 0.115 | POS  |
|   |  P5 | 804 | 0.664 | POS | 0.849 | POS  |
|   |  P6 | 929 | 0.475 | POS | 0.739 | POS  |
|  Shampoo | P7 | 793 | 0.817 | POS | 0.698 | POS  |
|   |  P8 | 984 | 0.275 | POS | 0.316 | POS  |
|   |  P9 | 1336 | 0.417 | POS | 0.222 | POS  |
|   |  P10 | 1471 | 0.617 | POS | 0.322 | POS  |
|   |  P11 | 2064 | 0.229 | POS | 0.143 | POS  |
|   |  P12 | 812 | 0.437 | POS | 0.3 | POS  |
|  Brown Dye | P13 | 951 | 0.722 | POS | 0.623 | POS  |
|   |  P14 | 968 | 0.739 | POS | 0.544 | POS  |
|   |  P15 | 1553 | 0.429 | POS | 0.368 | POS  |
|   |  P16 | 1596 | 0.366 | POS | 0.414 | POS  |
|   |  P17 | 3536 | 0.199 | POS | 0.082 | POS  |
|   |  P18 | 1306 | 0.166 | POS | 0.455 | POS  |
|   |  P19 | 1480 | 0.57 | POS | 0.389 | POS  |
|   |  P20 | 1852 | 0.322 | POS | 0.259 | POS  |
|   |  P21 | 919 | 0.499 | POS | 0.467 | POS  |
|   |  P22 | 968 | 0.125 | POS | 0.262 | POS  |
|   |  P23 | 4375 | 0.159 | POS | 0.166 | POS  |
|   |  P24 | 6948 | 0.047 | POS | 0.043 | POS  |
|  Bleach Blonde | P25 | 810 | 0.516 | POS | 0.899 | POS  |
|   |  P26 | 1315 | 0.264 | POS | 0.379 | POS  |
|   |  P27 | 3605 | 0.118 | POS | 0.16 | POS  |
|   |  P28 | 5158 | 0.043 | POS | 0.06 | POS  |
|   |  P29 | 5796 | 0.063 | POS | 0.096 | POS  |

{14}

|   | P30 | 6812 | 0.073 | POS | 0.067 | POS  |
| --- | --- | --- | --- | --- | --- | --- |
|   |  P31 | 22770 | 0.034 | POS | 0.028 | POS  |
|   |  P32 | 39769 | 0.016 | POS | 0.016 | POS  |
|   |  P33 | 750 | 0.389 | POS | 0.627 | POS  |
|   |  P34 | 6920 | 0.049 | POS | 0.185 | POS  |
|   |  P35 | 1440 | 0.315 | POS | 0.511 | POS  |
|   |  P36 | 1607 | 0.345 | POS | 0.239 | POS  |
|  Perm | P37 | 1662 | 0.181 | POS | 0.23 | POS  |
|   |  P38 | 1779 | 0.249 | POS | 0.365 | POS  |
|   |  P39 | 2298 | 0.112 | POS | 0.123 | POS  |
|   |  P40 | 3497 | 0.1 | POS | 0.088 | POS  |
|   |  P41 | 4142 | 0.054 | POS | 0.087 | POS  |
|   |  P42 | 4522 | 0.11 | POS | 0.107 | POS  |
|   |  P43 | 5461 | 0.098 | POS | 0.104 | POS  |
|   |  P44 | 5698 | 0.093 | POS | 0.112 | POS  |
|   |  P45 | 5993 | 0.034 | POS | 0.039 | POS  |
|   |  P46 | 6376 | 0.072 | POS | 0.081 | POS  |
|   |  P47 | 6786 | 0.082 | POS | 0.102 | POS  |
|   |  P48 | 8315 | 0.219 | POS | 0.087 | POS  |
|  Relaxer | P49 | 9203 | 0.049 | POS | 0.067 | POS  |
|   |  P50 | 20000 | 0.021 | POS | 0.026 | POS  |
|   |  P51 | 856 | 0.619 | POS | 0.833 | POS  |
|   |  P52 | 1455 | 0.189 | POS | 0.509 | POS  |
|   |  P53 | 1555 | 0.354 | POS | 0.409 | POS  |
|   |  P54 | 1597 | 0.065 | POS | 0.107 | POS  |
|   |  P55 | 2039 | 0.245 | POS | 0.294 | POS  |
|   |  P56 | 3462 | 0.109 | POS | 0.122 | POS  |
|   |  P57 | 4491 | 0.129 | POS | 0.187 | POS  |
|   |  P58 | 8147 | 0.024 | POS | 0.043 | POS  |
|   |  P59 | 11024 | 0.03 | POS | 0.089 | POS  |
|   |  P60 | 20000 | 0.015 | POS | 0.031 | POS  |

{15}

Negative Specimens Individual Results

|  Treatment | ID | GC/MS Pre-Cosmetic Treatment pg/mg | ELISA Pre-Treat Raw OD | ELISA Pre-Treat P/N | ELISA Post-Treat Raw OD | ELISA Post-Treat P/N  |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |  Cocaine  |   |   |   |   |
|  Shampoo | N1 | 0 | 2.65 | NEG | 2.698 | NEG  |
|   |  N2 | 0 | 2.785 | NEG | 2.559 | NEG  |
|   |  N3 | 0 | 2.828 | NEG | 2.928 | NEG  |
|   |  N4 | 0 | 2.767 | NEG | 2.85 | NEG  |
|   |  N5 | 0 | 2.795 | NEG | 2.767 | NEG  |
|   |  N6 | 0 | 2.553 | NEG | 2.802 | NEG  |
|  Shampoo | N7 | 0 | 2.5 | NEG | 2.64 | NEG  |
|   |  N8 | 0 | 2.837 | NEG | 2.763 | NEG  |
|   |  N9 | 0 | 2.562 | NEG | 2.406 | NEG  |
|   |  N10 | 0 | 2.572 | NEG | 2.663 | NEG  |
|   |  N11 | 0 | 2.648 | NEG | 2.705 | NEG  |
|   |  N12 | 0 | 2.679 | NEG | 2.647 | NEG  |
|  Brown Dye | N13 | 0 | 2.703 | NEG | 2.471 | NEG  |
|   |  N14 | 0 | 2.756 | NEG | 2.583 | NEG  |
|   |  N15 | 0 | 2.41 | NEG | 2.5 | NEG  |
|   |  N16 | 0 | 2.618 | NEG | 2.326 | NEG  |
|   |  N17 | 0 | 2.678 | NEG | 2.57 | NEG  |
|   |  N18 | 0 | 2.559 | NEG | 2.375 | NEG  |
|   |  N19 | 0 | 2.81 | NEG | 2.455 | NEG  |
|   |  N20 | 0 | 2.94 | NEG | 2.468 | NEG  |
|   |  N21 | 0 | 2.731 | NEG | 2.605 | NEG  |
|   |  N22 | 0 | 2.765 | NEG | 2.427 | NEG  |
|   |  N23 | 0 | 2.772 | NEG | 2.515 | NEG  |
|   |  N24 | 0 | 2.528 | NEG | 2.526 | NEG  |
|  Bleach Blonde | N25 | 0 | 2.72 | NEG | 2.484 | NEG  |
|   |  N26 | 0 | 2.487 | NEG | 2.611 | NEG  |
|   |  N27 | 0 | 2.568 | NEG | 2.512 | NEG  |
|   |  N28 | 0 | 2.754 | NEG | 2.711 | NEG  |

{16}

|   | N29 | 0 | 2.853 | NEG | 2.875 | NEG  |
| --- | --- | --- | --- | --- | --- | --- |
|   |  N30 | 0 | 2.744 | NEG | 2.862 | NEG  |
|   |  N31 | 0 | 2.712 | NEG | 2.644 | NEG  |
|   |  N32 | 0 | 2.911 | NEG | 2.785 | NEG  |
|   |  N33 | 0 | 2.472 | NEG | 2.579 | NEG  |
|   |  N34 | 0 | 2.737 | NEG | 2.718 | NEG  |
|   |  N35 | 0 | 2.674 | NEG | 2.654 | NEG  |
|   |  N36 | 0 | 2.709 | NEG | 2.775 | NEG  |
|  Perm | N37 | 0 | 2.469 | NEG | 2.327 | NEG  |
|   |  N38 | 0 | 2.486 | NEG | 2.4 | NEG  |
|   |  N39 | 0 | 2.445 | NEG | 2.504 | NEG  |
|   |  N40 | 0 | 2.439 | NEG | 2.56 | NEG  |
|   |  N41 | 0 | 2.436 | NEG | 2.271 | NEG  |
|   |  N42 | 0 | 2.574 | NEG | 2.519 | NEG  |
|   |  N43 | 0 | 2.578 | NEG | 2.573 | NEG  |
|   |  N44 | 0 | 2.672 | NEG | 2.52 | NEG  |
|   |  N45 | 0 | 2.574 | NEG | 2.496 | NEG  |
|   |  N46 | 0 | 2.452 | NEG | 2.303 | NEG  |
|   |  N47 | 0 | 2.578 | NEG | 2.531 | NEG  |
|   |  N48 | 0 | 2.446 | NEG | 2.589 | NEG  |
|  Relaxer | N49 | 0 | 2.481 | NEG | 2.393 | NEG  |
|   |  N50 | 0 | 2.338 | NEG | 2.251 | NEG  |
|   |  N51 | 0 | 2.543 | NEG | 2.3 | NEG  |
|   |  N52 | 0 | 2.381 | NEG | 2.355 | NEG  |
|   |  N53 | 0 | 2.576 | NEG | 2.161 | NEG  |
|   |  N54 | 0 | 2.583 | NEG | 2.281 | NEG  |
|   |  N55 | 0 | 2.528 | NEG | 2.323 | NEG  |
|   |  N56 | 0 | 2.294 | NEG | 2.139 | NEG  |
|   |  N57 | 0 | 2.223 | NEG | 2.317 | NEG  |
|   |  N58 | 0 | 2.365 | NEG | 2.248 | NEG  |
|   |  N59 | 0 | 2.475 | NEG | 2.228 | NEG  |
|   |  N60 | 0 | 2.359 | NEG | 2.396 | NEG  |

This study agrees with the extensive published literature that use of various cosmetic hair treatments can potentially reduce the amount of drugs and drug metabolites detected in hair specimens. It is possible that a cosmetic hair treatment could cause a negative hair sample to read positive or a positive hair sample to read negative. These effects are dependent upon the nature of the hair specimen and the treatment used and independent

{17}

of the method of analysis.

f. Assay cut-off:

The Assay cut-off is 300 pg cocaine/mg hair. Analytical performance of the device around the claimed cutoff is described in the precision section M.1a above.

2. Comparison studies:

a. Method comparison with predicate device:

A total of 100 donor head hair specimens were analyzed in singlet using the candidate assay and GC/MS. The results are summarized below:

Concordance Table

|  Candidate Device Result | Negative (Less thanhalf the cutoff concentration) by confirmatory analysis | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration) by confirmatory analysis | Near Cutoff Positive (Between the cutoff and50% above the cutoff concentration) by confirmatory analysis | High Positive (greater than 50% above the cutoff concentration) by confirmatory analysis  |
| --- | --- | --- | --- | --- |
|  [Conc] by GCMS | <150 pg/mg | 150-299 pg/mg | 300-450 pg/mg | >450 pg/mg  |
|  Negative | 44 | 2 | 0 | 0  |
|  Positive | 1 | 3 | 5 | 45  |

Summary and Justification of Discordant Results between ELISA and GC/MS

|  Reference number | Specimen | Device Result | Confirmation results by GC/MS  |   |   |   |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |  | BE | COC | CE | NOR  |
|  1 | 313707BH | Pos | 0 | 0 | 0 | 0  |
|  2 | 840072CH | Pos | 0 | 155 | 0 | 0  |
|  3 | 769009BH | Pos | 0 | 190 | 0 | 0  |
|  4 | 757751BH | Pos | 80 | 272 | 0 | 0  |

{18}

(1) This sample was dyed a bright pink color. An investigation of this issue concluded that dyes within the bright pink color interfere with the immunoassay, producing false positive results. This dyes tested are included in the Interference section above.

(2) This specimen confirmed negative by GC-MS with a quantitative value of 155 pg cocaine/mg hair

(3) This specimen confirmed negative by GC-MS with a quantitative value of 190 pg cocaine/mg hair

(4) This specimen confirmed negative by GC-MS with a quantitative value of 272 pg cocaine/mg hair, but also contained 80 pg/mg of benzoylecgonine

b. Matrix comparison:
Not applicable

3. Clinical studies:
a. Clinical Sensitivity:
Not applicable
b. Clinical specificity:
Not applicable
c. Other clinical supportive data (when a. and b. are not applicable):
Not applicable

4. Clinical cut-off:
Not applicable

5. Expected values/Reference range:
Not applicable

N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

19

{19}

O. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

20

---

**Source:** [https://fda.innolitics.com/submissions/TX/subpart-d%E2%80%94clinical-toxicology-test-systems/DIO/K152232](https://fda.innolitics.com/submissions/TX/subpart-d%E2%80%94clinical-toxicology-test-systems/DIO/K152232)

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