WUNDER TEST

K102428 · Hien Helen Nguyen · LDJ · Apr 18, 2012 · Clinical Toxicology

Device Facts

• Record ID: K102428
• Device Name: WUNDER TEST
• Applicant: Hien Helen Nguyen
• Product Code: LDJ · Clinical Toxicology
• Decision Date: Apr 18, 2012
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3870
• Device Class: Class 2

Indications for Use

A competitive binding immunoassay used for the qualitative determination of Cannabinoids in human urine. The device is visually read and intended for over-the-counter single use. The test has a cutoff of 50 ng/mL of Cannabinoids. The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.

Device Story

Wunder Test is a qualitative, visually-read immunochromatographic lateral flow assay for detecting 9-COOH-11-norΔ⁹-THC in human urine. Device consists of a nitrocellulose membrane strip coated with THC-BSA conjugate and a wicking pad with anti-THC monoclonal antibody-colloid gold conjugate. User submerges absorbent tip into urine sample; sample migrates across membrane. If THC is absent, anti-THC conjugate binds to THC-BSA, forming a visible colored test band (negative result). If THC is present at ≥50 ng/mL, it saturates antibody binding sites, preventing test band formation (positive result). A control band confirms proper flow. Intended for OTC home use; results are preliminary and require GC/MS confirmation. Provides rapid screening to assist in identifying potential cannabinoid presence.

Clinical Evidence

No clinical data provided; device relies on analytical performance characteristics typical of competitive binding immunoassays for drug screening.

Technological Characteristics

Immunochromatographic lateral flow assay. Components: nitrocellulose membrane, THC-BSA conjugate, anti-THC monoclonal goat anti-mouse antibody-colloid gold conjugate, mouse IgG control. Form factor: dipstick with absorbent tip. Storage: 4-30°C. Shelf life: 2 years. Qualitative visual readout. No electronic components or software.

Indications for Use

Indicated for qualitative detection of Cannabinoids in human urine for over-the-counter (OTC) use. Cutoff concentration: 50 ng/mL. Results are preliminary; requires confirmatory testing (GC/MS preferred).

Regulatory Classification

Identification

A cannabinoid test system is a device intended to measure any of the cannabinoids, hallucinogenic compounds endogenous to marihuana, in serum, plasma, saliva, and urine. Cannabinoid compounds include delta-9-tetrahydrocannabinol, cannabidiol, cannabinol, and cannabichromene. Measurements obtained by this device are used in the diagnosis and treatment of cannabinoid use or abuse and in monitoring levels of cannabinoids during clinical investigational use.

Special Controls

*Classification.* Class II (special controls). A cannabinoid 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).

Submission Summary (Full Text)

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

A. 510(k) Number:
k102428

B. Purpose for Submission:
New device

C. Measurand:
Cannabinoids

D. Type of Test:
Qualitative visually read immunochromatographic assay

E. Applicant:
Hien Helen Nguyen

F. Proprietary and Established Names:
Wunder Test

G. Regulatory Information:
1. Regulation section:
21 CFR 862.3870, Cannabinoid Test System
2. Classification:
Class II
3. Product code:
LDJ
4. Panel:

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Toxicology (91)

## H. Intended Use:

1. Intended use(s):

See indications for use below.

2. Indication(s) for use:

A competitive binding immunoassay used for the qualitative determination of Cannabinoids in human urine. The device is visually read and intended for over-the-counter single use. The test has a cutoff of 50 ng/mL of Cannabinoids.

The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.

3. Special conditions for use statement(s):

This assay is intended for over-the-counter use

4. Special instrument requirements:

Not applicable, this is a visually-read single-use device

## I. Device Description:

Qualitative immunoassay device intended to detect 9-COOH-11-norΔ⁹-THC, a major metabolite of cannabinoids in human urine at a cutoff level of 50 ng/mL. The kit includes a sterile 60 mL urine cup, sterile individually packaged testing strip, package insert, a custody and control form, and a biohazard bag.

## J. Substantial Equivalence Information:

1. Predicate device name(s):

LifeSign Home Drug Test THC

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

k014066

3. Comparison with predicate:

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| Similarities | | |
| --- | --- | --- |
| Item | Predicate | Candidate Device |
| Intended Use | A competitive binding immunoassay used for the qualitative determination of Cannabinoids in human urine. | Same |
| Product Type | Dipstick | Same |
| Mechanism of Action | Immunochromatographic lateral flow assay with visual, qualitative screening result | Same |
| Cutoff Concentration | 50 ng/mL | Same |
| Target User Population | OTC | Same |
| Differences | | |
| --- | --- | --- |
| Item | Predicate | Candidate Device |
| Size | 4.25” x 1.2” x 0.2” | 3.6” x 0.7” x 0.1” |
| Physical Design | Plastic casing includes absorbent tip in a “sample well” | Absorbent tip exposed |
| Storage Temperature | 2 - 30°C | 4 - 30°C |

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

None were referenced

# L. Test Principle:

The dipstick end of the device, an absorbent nitrocellulose membrane strip, is submerged into a container with the urine specimen and the urine sample migrates towards a region of the dipstick that is coated with a THC-BSA conjugate. A colored anti-THC monoclonal goat anti-mouse antibody-colloid gold conjugate wicking pad is positioned at the end of the dipstick strip, and after the urine sample has migrated through the THC-BSA conjugate area, brining the THC-BSA conjugate with it, if the urine is negative for THC the THC-BSA will react with the anti-THC conjugate to form a color line in the test band region. If the urine sample contains a sufficient concentration of THC then the endogenous THC will saturate the binding sites on the anti-THC antibody conjugates, preventing the THC-BSA from binding and therefore preventing the formation of a color band in the results region of the strip.

A control band coated with monoclonal mouse IgG which binds with the antibody-colloid gold conjugate to ensure that the test is performing properly.

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M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

| Precision of the test was characterized at the sponsor's facility by two operators. Samples were commercial urine (Biorad Liquicheck Urine Toxicology Control; with THC concentrations ranging from 0 ng/mL to 140 ng/mL) that were mixed to obtain the target concentrations. Testing was performed using three different device lot numbers. Three lots (aliquots) of total 27 samples were prepared which included drug-free urine, drug-free urine spiked to 75% below cutoff, 50% below cutoff, 25% below cutoff, cutoff, 25% above cutoff, 50% above cutoff, 75% | Drug-free urine | -75% cutoff | -50% cutoff | -25% cutoff | Cutoff | +25% cutoff | +50% cutoff | +75% cutoff | +100% cutoff |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |

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| above cutoff and 100% above cutoff. Each three lots of 27 samples were assayed once a day for 20 days, yielding a total of 60 runs per sample concentration. Results are presented below. | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Concentration (ng/mL) | 0 | 12.5 | 25 | 37.5 | 50 | 62.5 | 75 | 87.5 | 100 |
| THC Negative | 60 | 60 | 60 | 60 | 4 | 0 | 0 | 0 | 0 |
| THC Positive | 0 | 0 | 0 | 0 | 56 | 60 | 60 | 60 | 60 |

A separate cutoff characterization study was performed to assess the 50 ng/mL cutoff concentration of the device. 20 devices from 3 lots were tested at each of 8 sample concentrations made from spiking commercial THC into drug-free urine. The results are below.

| | Drug-free urine | -75% cutoff | -50% cutoff | -25% cutoff | Cutoff | +25% cutoff | +50% cutoff | +75% cutoff | +100% cutoff |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Concentration (ng/mL) | 0 | 12.5 | 25 | 37.5 | 50 | 62.5 | 75 | 87.5 | 100 |
| Lot 1 THC (+/-) | 0/20 | 0/20 | 0/20 | 0/20 | 18/2 | 20/0 | 20/0 | 20/0 | 20/0 |
| Lot 2 THC (+/-) | 0/20 | 0/20 | 0/20 | 0/20 | 19/1 | 20/0 | 20/0 | 20/0 | 20/0 |
| Lot 3 THC (+/-) | 0/20 | 0/20 | 0/20 | 0/20 | 19/1 | 20/0 | 20/0 | 20/0 | 20/0 |

b. Linearity/assay reportable range:

Not applicable. The assay is intended for qualitative use.

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

All materials are traceable to the commercial suppliers from which they are purchased.

An accelerated study was performed to determine the shelf life of the candidate device. The study was designed based on the bracket table technique where the Wunder Test™ was stressed at 50°C for 60 days to predict stability for 2 years. The protocol and acceptance criteria was reviewed and found to be acceptable, yielding a stable shelf life at 4 - 30°C of 2 years.

Real-time stability study protocols and acceptance criteria were reviewed and found

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to be acceptable. These studies are ongoing.

## d. Detection limit:

Sensitivity of a qualitative assay may be characterized by validating performance around the claimed cutoff concentration of the assay, and demonstrating the lowest concentration of drug that is capable of consistently producing a positive result. This information appears in the precision section (please see above).

## e. Analytical specificity:

Twenty three compounds and nine biologic substances were tested for interference by testing various amounts of each compound when added to urine with at 37.5 ng/mL THC (-25% cutoff) and at 65 ng/mL (+25% cutoff). Five samples were tested for each compound by a single operator at the sponsor's facility. All test results were negative.

Several structurally related compounds were tested for cross reactivity. Various amounts of each compound (serial dilutions from an original cross-reactant concentration of 100 µg/mL) were tested in drug-free control urine and cross-reactivity was determined by when the test result changed from positive to negative. Results are expressed as cross-reactant concentration equivalent to analyte cutoff, and are shown below.

| Compound | Response equivalent to cutoff in ng/mL |
| --- | --- |
| Δ⁹-Tetrahydrocannabinol | 15,000 |
| Cannabinol | 20,000 |
| Δ⁸-Tetrahydrocannabinol | 25,000 |
| 11-nor- Δ⁹-Tetrahydrocannabinol carboxylic acid | 50 |
| Cannabidiol | 100,000 |

Several structurally unrelated compounds were tested for interference:

| Compound Concentration (100 µg/mL) | -25% of cutoff (37.5 ng/mL) negative/positive | +25% of cutoff (62.5 ng/mL) negative/positive |
| --- | --- | --- |
| Acetaminophen | NEG | POS |
| Acetylsalicylic acid | NEG | POS |
| Ampicillin | NEG | POS |
| Ethanol | NEG | POS |
| Lidocaine | NEG | POS |
| Aspirin | NEG | POS |
| Atropine | NEG | POS |

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| Benzoic acid | NEG | POS |
| --- | --- | --- |
| Oxalic acid | NEG | POS |
| Caffeine | NEG | POS |
| Methanol | NEG | POS |
| Pencicillin-G | NEG | POS |
| Ranitidine | NEG | POS |
| Salicylic acid | NEG | POS |
| | | |
| Drug compounds from Biorad Liquitox™ Urine Toxicology Control | -25% of cutoff (37.5 ng/mL) negative/positive | +25% of cutoff (62.5 ng/mL) negative/positive |
| d-Amphetamine | NEG | POS |
| Secobarbital | NEG | POS |
| Nordiazepam | NEG | POS |
| Benzoylecgonine | NEG | POS |
| Lysergic acid | NEG | POS |
| Methadone | NEG | POS |
| Methaqualone | NEG | POS |
| Morphine | NEG | POS |
| Phencyclidine | NEG | POS |
| Propoxyphene | NEG | POS |
| Nortriptyline | NEG | POS |
| | | |
| Biologic substances | -25% of cutoff (37.5 ng/mL) negative/positive | +25% of cutoff (62.5 ng/mL) negative/positive |
| Albumin | NEG | POS |
| Bilirubin | NEG | POS |
| Creatine | NEG | POS |
| Hemoglobin | NEG | POS |
| Glucose | NEG | POS |
| Vitamin (L-Ascorbic Acid) | NEG | POS |
| Uric acid | NEG | POS |
| Urine pH | NEG | POS |
| 3.0 | NEG | POS |
| 4.0 | NEG | POS |
| 5.0 | NEG | POS |
| 6.0 | NEG | POS |
| 7.0 | NEG | POS |
| 8.0 | NEG | POS |
| 9.0 | NEG | POS |
| Urine specific gravity (g/mL) | | |
| 1.010 | NEG | POS |
| 1.020 | NEG | POS |
| 1.030 | NEG | POS |

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f. Assay cut-off:

The cut-off characterization study results can be found in the precision section of this summary.

2. Comparison studies:

a. Method comparison with predicate device:

In order to characterize performance of the Wunder Test™ assay compared with GC/MS testing, 80 clinical samples were purchased from a laboratory and analyzed with each assay. Samples were unaltered and were selected with the intent to obtain adequate distribution of the targeted drug around the claimed cutoff concentration. The samples were categorized based upon the GC/MS result concentration of THC. Results were obtained by the sponsor's in-house laboratory and by an outside GC/MS laboratory, and are presented below.

| THC Metabolite | Test Positive | Test Negative |
| --- | --- | --- |
| True Negative (drug-free urine) | 0 | 20 |
| Low Negative (<50% cutoff) | 0 | 20 |
| Near Cutoff Negative (between -50% + cutoff) | 0 | 20 |
| Near Cutoff Positive (between cutoff + 50%) | 17 | 3 |
| High Positive (>50% cutoff) | 20 | 0 |

Discordant Results #1

| Cutoff Value (ng/mL) | Wunder Test™ (POS/NEG) | Drug/Metabolite GC/MS value (ng/mL) |
| --- | --- | --- |
| 50 | Negative | 55.45 |
| 50 | Negative | 50.00 |
| 50 | Negative | 50.76 |

In addition, 44 samples (40 from the positive category and 4 from the negative category above) were sent to a separate certified laboratory for confirmatory GC/MS testing, and results are presented below.

| THC Metabolite | Near Cutoff Negative (between -50% & cutoff) | Near Cutoff Positive (between cutoff & + 50%) | High Positive (>50% cutoff) |
| --- | --- | --- | --- |
| Positive | 1 | 17 | 20 |
| Negative | 3 | 3 | 0 |

Discordant Results #2

| Cutoff | Wunder Test™ | Drug/Metabolite |
| --- | --- | --- |

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| Value (ng/mL) | (POS/NEG) | GC/MS value (ng/mL) |
| --- | --- | --- |
| 50 | Positive | 48 |
| 50 | Negative | 44 * |
| 50 | Negative | 48 * |
| 50 | Negative | 47 * |

*These samples tested positive at the first GC/MS Laboratory confirmation but negative and this secondary GC/MS Laboratory confirmation testing; therefore although they are not discordant results as shown in the above Discordant Results #2 table, they were categorized as Positive samples from the first GC/MS Laboratory results (the 3 discordant results from Discordant Results #1) and are therefore listed here with the discordant results as well as above.

# b. Matrix comparison:

Not applicable. The assay is intended for only one sample matrix.

# 3. Clinical studies:

# a. Clinical Sensitivity:

Not applicable. Clinical studies are not typically submitted for this device type and matrix.

# b. Clinical specificity:

Not applicable. Clinical studies are not typically submitted for this device type and matrix.

# c. Other clinical supportive data (when a. and b. are not applicable):

A consumer study was performed with 192 lay-users. Urine samples were purchased at specific drug concentrations, or spiked to desired concentrations and distributed to volunteers at one of three locations. Volunteers were of various age, race, sex and educational backgrounds, and none had ever before used a drug test. Volunteers tested the urine samples using the Wunder Test™. Volunteers were provided with the instructional materials available in the device packaging. Testing was conducted at three separate locations. Combined results are divided into concentration categories based upon the GC/MS result of each sample, and are presented below.

| THC (ng/mL) category by GC/MS | Drug-free Urine (0) | 75% Below Cutoff (<12.5) | 50% Below Cutoff (25) | 25% Below Cutoff (37.5) | 25% Above Cutoff (67.5) | 50% Above Cutoff (75) | 100% Above Cutoff (>100) |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| 2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| 3 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |

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| Wunder Test™ Positive/Negative | 0/20 | 0 / 30 | 0 / 35 | 4 / 30 | 34 / 0 | 32 / 0 | 27 / 0 |
| --- | --- | --- | --- | --- | --- | --- | --- |

A post-test survey of volunteers showed that 100% thought that the test was easy to run, easy to read, and that result line shading does not matter (a question specifically addressed in the device instructions). A Flesh-Kincaid reading analysis resulted in a reading grade level of 7.8.

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.

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