(161 days)
Not Found
No
The device description and performance studies focus on a lateral flow immunochromatographic assay, which is a traditional chemical detection method. There is no mention of AI, ML, image processing, or any computational analysis that would suggest the use of these technologies.
No
The device is described as a diagnostic immunoassay for qualitative detection of substances in oral fluid, not for treating any condition.
No
Explanation: The device is an immunoassay for the qualitative detection of COT and THC in oral fluid, providing only preliminary results. It is intended to detect the presence of substances, not to diagnose a disease or condition. The text explicitly states that an alternative laboratory test must be used to confirm the results, indicating it's a screening tool rather than a diagnostic one.
No
The device description clearly states it is an "immunochromatographic assay that uses a lateral flow system," which is a physical test strip, not software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states it's for the "qualitative detection of COT in oral fluid" and "qualitative and simultaneous detection of Marijuana (THC) and Cotinine in human oral fluid." This involves testing a sample taken from the human body (oral fluid) to gain information about a person's health or condition (presence of drugs).
- Device Description: The description confirms it's an "immunochromatographic assay" that uses a "lateral flow system for the qualitative detection of cotinine in human oral fluid." This is a common type of in vitro diagnostic test.
- Anatomical Site: The sample is taken from "Oral fluid," which is a human biological specimen.
- Performance Studies: The document details various performance studies, including comparison studies against a reference method (LC/MS/MS), which are standard for evaluating the performance of IVD devices.
- Predicate Devices: The mention of predicate devices (K141320 and K033601) which are also IVDs (Enzyme Immunoassays for oral fluid) further supports that this device falls under the IVD category.
In summary, the device meets the definition of an In Vitro Diagnostic because it is intended for use in vitro (outside the body) for the examination of specimens derived from the human body to provide information for diagnostic purposes (in this case, detecting the presence of drugs).
N/A
Intended Use / Indications for Use
The Healgen® Accurate Oral Fluid Drug Test COT is a lateral flow chromatographic immunoassay for the qualitative detection of COT in oral fluid at the cut-off concentration 30 ng/mL.
This assay provides only a preliminary result. An alternative laboratory test must be used to confirm the results provided by this drug test. Gas chromatography/mass spectrometry (GC/MS) is the preferred method confirmation test.
The Healgen® Accurate Oral Fluid Drug Test is a competitive binding lateral flow immunochromatographic assay for the qualitative and simultaneous detection of Marijuana (THC) and Cotinine in human oral fluid at the cutoff concentrations listed below and their metabolites.
| Test | Calibrator | Cut-off (ng/mL) |
|---|---|---|
| Cotinine (COT) | (-) Cotinine | 30 |
| Marijuana (THC) | Delta-9-Tetrahydrocannabinol | 40 |
This assay provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography mass spectrometry (GC/MS) and liquid chromatography mass spectrometry (LC/MS) are the preferred confirmatory methods.
Product codes (comma separated list FDA assigned to the subject device)
LDJ, MKU
Device Description
The Healgen® Accurate Oral Fluid Drug Test COT is immunochromatographic assay that uses a lateral flow system for the qualitative detection of cotinine in human oral fluid. The Healeen® Accurate Oral Fluid Drug Test immunochromatographic assay for the qualitative and simultaneous detection of Marijuana (THC) and Cotinine in human oral fluid. The tests are the first step in a two-step process. The second step is to send the sample for laboratory testing if preliminary positive results are obtained.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
oral fluid
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Over-The-Counter Use (21 CFR 801 Subpart C), Prescription Use (Part 21 CFR 801 Subpart D)
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Precision-Reproducibility-Cut-Off studies: Samples were prepared by spiking cotinine or marijuana in negative oral fluid samples. Each cotinine or marijuana concentration was confirmed by LC/MS/MS. All sample aliquots were blindly labeled by the person who prepared the samples and didn't take part in the sample testing. For each concentration, tests were performed two runs per day for 25 days per device lot in a randomized order.
For Cotinine (COT):
- Samples tested at the following LC-MS concentrations (ng/mL): 0, 7.4, 15.4, 22.6, 30.7, 37.9, 45.9, 52.9, 60.6. These correspond to -100% cut off, -75% cut off, -50% cut off, -25% cut off, cut off, +25% cut off, +50% cut off, +75% cut off, and +100% cut off, respectively.
- For each concentration and lot, 50 tests were performed (e.g., 50-/0+ means 50 negative results and 0 positive results).
For THC:
- Samples tested at the following LC-MS concentrations (ng/mL): 0.00, 9.5, 20.7, 30.2, 40.8, 51.4, 59.2, 68.3, 81.5. These correspond to -100% cut off, -75% cut off, -50% cut off, -25% cut off, cut off, +25% cut off, +50% cut off, +75% cut off, and +100% cut off, respectively.
- For each concentration and lot, 50 tests were performed.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Analytical Performance
Precision-Reproducibility-Cut-Off
- Study Type: Precision-Reproducibility-Cut-Off studies.
- Sample Size: 50 tests per concentration per device lot, across 3 lots.
- Key Results:
- For Cotinine: The device showed 100% agreement for concentrations at -100% to -50% and +50% to +100% of the cutoff. At -25% cutoff, 48-50 out of 50 samples were negative. At the cutoff concentration (30.7 ng/mL), 23-24 negative and 26-27 positive results were observed across the 3 lots (out of 50 total). At +25% cutoff, 48-49 out of 50 samples were positive.
- For THC: The device showed 100% agreement for concentrations at -100% to -50% and +50% to +100% of the cutoff. At -25% cutoff, 47-49 out of 50 samples were negative. At the cutoff concentration (40.8 ng/mL), 24-28 negative and 22-26 positive results were observed across the 3 lots (out of 50 total). At +25% cutoff, 47-49 out of 50 samples were positive.
- Verified cut-off values: cotinine 30 ng/mL, marijuana 40 ng/mL.
Stability
- Study Type: Real time stability study.
- Key Results: Devices are stable at 4-30 ℃ for 24 months.
Interference
- Study Type: Testing with potential interfering substances and food items.
- Key Results: Over 60 compounds (e.g., Acetaminophen, Ibuprofen, Caffeine, etc.) showed no interference for cotinine and THC at 100μg/mL. Various food items (e.g., methanol cough drops, coffee, toothpaste, alcohol) showed no interference at a concentration of 5%. Hemoglobin showed no interference at 10mg/dL.
Specificity
- Study Type: Cross-reactivity testing with drug metabolites and other components.
- Key Results:
- COT:
- (-) Cotinine: 100% at 30 ng/mL
- S(-)-Nicotine: 1% at 3000 ng/mL
- Trans-3-hydroxycotinine: 300% at 10 ng/mL
- (+)-anabasine: 0.002% at 1,350,000 ng/mL
- (+/-)-nornicotine: 0.02% at 150,000 ng/mL
- Niacinamide: 100,000 ng/mL
- THC:
- △9-Tetrahydrocannabinol: 100% at 40 ng/mL
- △8-Tetrahydrocannabinol: 50% at 80 ng/mL
- 11-nor-△9-THC-9 COOH: 1000% at 4 ng/mL
- 11-hydroxy-△9-THC: 89% at 45 ng/mL
- Cannabinol: 20% at 200 ng/mL
- Cannabidiol (CBD): 1.8% at 2,200 ng/mL
- 11-Nor-△9-THC-carboxy-glucuronide: 66.7% at 60 ng/mL
- (+)-11-nor-9-carboxy-△9-THC: 80% at 50 ng/mL
- 11-nor-A8-THC-9-COOH: 200% at 20 ng/mL
- Numerous other THC metabolites and similar compounds showed varying degrees of cross-reactivity.
- COT:
Effect of Oral fluid pH
- Study Type: Testing oral fluid samples with varying pH levels.
- Key Results: Oral fluid samples with pH 4 to 9 spiked with target drugs at 50% below and 50% above Cut-Off levels showed all positive results for samples at and above +50% Cut-Off and all negative for samples at and below -50% Cut-Off.
Drug Recovery Study
- Study Type: Recovery studies for drugs stored in devices at different temperatures.
- Key Results: Over 90% recoveries observed for both cotinine and marijuana in Healgen devices. Oral fluid samples can be stored in the device at -20°C for at least 90 days and can be shipped overnight for LC-MS confirmation.
Comparison Studies
- Study Type: Method comparison studies against LC/MS/MS results.
- Sample Size: 427 samples for COT, 126 samples for THC. Performed at three testing sites with one operator at each site.
- Key Results (COT):
- Drug-Free (112 samples): 100% correct results (all negative).
- Less than Half the Cutoff Concentration (93 samples): 100% correct results (all negative).
- Near Cutoff Negative (40 samples): 88% correct results (5 positive, 35 negative).
- Near Cutoff Positive (32 samples): 88% correct results (28 positive, 4 negative).
- High Positive (150 samples): 100% correct results (all positive).
- Discordant Results (COT):
- 5 samples were positive by device but had LC/MS results below cutoff (23.43 - 29.32 ng/mL).
- 4 samples were negative by device but had LC/MS results above cutoff (30.38 - 32.64 ng/mL).
- Key Results (THC):
- Drug-Free (35 samples): 100% correct results (all negative).
- Less than Half the Cutoff Concentration (13 samples): 100% correct results (all negative).
- Near Cutoff Negative (7 samples): 71.4% correct results (2 positive, 5 negative).
- Near Cutoff Positive (10 samples): 60% correct results (6 positive, 4 negative).
- High Positive (61 samples): 100% correct results (all positive).
- Discordant Results (THC):
- 2 samples were positive by device but had LC/MS results below cutoff (38.3 - 38.7 ng/mL).
- 4 samples were negative by device but had LC/MS results above cutoff (40.6 - 47.0 ng/mL).
Layuser Studies
- Study Type: Layuser performance studies based on product insert.
- Sample Size: 362 layusers. Performed at three testing sites.
- Key Results (COT-only):
- Drug-Free (95 samples): 100% correct results (all negative).
- Less than Half the Cutoff Concentration (39 samples): 100% correct results (all negative).
- Near Cutoff Negative (35 samples): 89% correct results (4 positive, 31 negative).
- Near Cutoff Positive (39 samples): 87% correct results (34 positive, 5 negative).
- High Positive (154 samples): 100% correct results (all positive).
- Discordant Results (COT):
- 4 samples were positive by device but had LC/MS results below cutoff (23.54 - 29.89 ng/mL).
- 5 samples were negative by device but had LC/MS results above cutoff (30.26 - 38.64 ng/mL).
- Other Results: All layusers found the device instruction easy to understand and follow. The package insert had a Flesch-Kincaid reading Grade Level of 7.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Not explicitly stated as Sensitivity, Specificity, PPV, NPV. Instead, "The percentage of correct results" is provided.
Comparison Studies (COT):
- Drug-Free: 100% correct (Negative)
- Less than Half Cutoff: 100% correct (Negative)
- Near Cutoff Negative: 88% correct
- Near Cutoff Positive: 88% correct
- High Positive: 100% correct (Positive)
Comparison Studies (THC):
- Drug-Free: 100% correct (Negative)
- Less than Half Cutoff: 100% correct (Negative)
- Near Cutoff Negative: 71.4% correct
- Near Cutoff Positive: 60% correct
- High Positive: 100% correct (Positive)
Layuser Studies (COT):
- Drug-Free: 100% correct (Negative)
- Less than Half Cutoff: 100% correct (Negative)
- Near Cutoff Negative: 89% correct
- Near Cutoff Positive: 87% correct
- High Positive: 100% correct (Positive)
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 862.3870 Cannabinoid test system.
(a)
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 includedelta -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.(b)
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).
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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
March 17, 2023
Healgen Scientific LLC % Joe Shia Director LSI International Inc. 504 East Diamond Ave. Suite I Gaithersburg, MD 20877
Re: K223162
Trade/Device Name: Healgen® Accurate Oral Fluid Drug Test, Healgen® Accurate Oral Fluid Drug Test COT Regulation Number: 21 CFR 862.3870 Regulation Name: Cannabinoid Test System Regulatory Class: Class II Product Code: LDJ, MKU Dated: February 12, 2023 Received: February 13, 2023
Dear Joe Shia:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's
1
requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Paula
Caposino -S
Digitally signed by Paula
Caposino -S
Date: 2023.03.17 13:12:26
-04'00'
Paula Caposino, Ph.D. Acting Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
2
Indications for Use
510(k) Number (if known)
K223162
Device Name Healgen® Accurate Oral Fluid Drug Test COT
Indications for Use (Describe)
The Healgen® Accurate Oral Fluid Drug Test COT is a lateral flow chromatographic immunoassay for the qualitative detection of COT in oral fluid at the cut-off concentration 30 ng/mL.
This assay provides only a preliminary result. An alternative laboratory test must be used to confirm the results provided by this drug test. Gas chromatography/mass spectrometry (GC/MS) is the preferred method confirmation test.
Type of Use (Select one or both, as applicable)
| Prescription Use (Part 21 CFR 801 Subpart D)
X Over-The-Counter Use (21 CFR 801 Subpart C)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
3
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov
"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."
4
Indications for Use
510(k) Number (if known)
K223162
Device Name
Healgen® Accurate Oral Fluid Drug Test
Indications for Use (Describe)
The Healgen® Accurate Oral Fluid Drug Test is a competitive binding lateral flow immunochromatographic assay for the qualitative and simultaneous detection of Marijuana (THC) and Cotinine in human oral fluid at the cutoff concentrations listed below and their metabolites.
| Test | Calibrator | Cut-off (ng/mL) |
|---|---|---|
| Cotinine (COT) | (-) Cotinine | 30 |
| Marijuana (THC) | Delta-9-Tetrahydrocannabinol | 40 |
This assay provides only a preliminary analytical test result. A more specific alternate chemical must be used in order to obtain a confirmed analytical result. Gas chromatography mass spectrometry (GC/MS) and liquid chromatography mass spectrometry (LC/MS) are the preferred confirmatory methods.
Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
5
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov
"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."
6
510(k) SUMMARY K223162
-
- Date: March 16, 2023 2. Submitter: HEALGEN SCIENTIFIC LLC 5213 Maple Street Bellaire, TX 77401
- Joe Shia 3. Contact person: LSI International Inc. 504 East Diamond Ave., Suite I Gaithersburg, MD 20877 Telephone: 240-505-7880 Fax: 301-916-6213 Email:shiajl(@yahoo.com
- Healgen® Accurate Oral Fluid Drug Test 4. Device Names: Healgen® Accurate Oral Fluid Drug Test COT
Classification:
| Product Code | CFR # | Panel |
|---|---|---|
| LDJ | 21 CFR, 862.3870 Cannabinoids Test System | Toxicology |
| MKU | 21 CFR, 862.3220 Cotinine Test System | Toxicology |
-
- Intended Use
The Healgen® Accurate Oral Fluid Drug Test COT is a lateral flow chromatographic immunoassay for the qualitative detection of COT in oral fluid at the cut-off concentration 30 ng/mL.
- Intended Use
This assay provides only a preliminary result. An alternative laboratory test must be used to confirm the results provided by this drug test. Gas chromatography/mass spectrometry (GC/MS) is the preferred method confirmation test.
The Healgen® Accurate Oral Fluid Drug Test is a competitive binding lateral flow immunochromatographic assay for the qualitative and simultaneous detection of Marijuana (THC) and Cotinine in human oral fluid at the cutoff concentrations listed below and their metabolites.
7
| Test | Calibrator | Cut-off (ng/mL) |
|---|---|---|
| Cotinine (COT) | (-) Cotinine | 30 |
| Marijuana (THC) | Delta-9-Tetrahydrocannabinol | 40 |
This assay provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography mass spectrometry (GC/MS) and liquid chromatography mass spectrometry (LC/MS) are the preferred confirmatory methods.
7. Device Description
The Healgen® Accurate Oral Fluid Drug Test COT is immunochromatographic assay that uses a lateral flow system for the qualitative detection of cotinine in human oral fluid. The Healeen® Accurate Oral Fluid Drug Test immunochromatographic assay for the qualitative and simultaneous detection of Marijuana (THC) and Cotinine in human oral fluid. The tests are the first step in a two-step process. The second step is to send the sample for laboratory testing if preliminary positive results are obtained.
-
- Substantial Equivalence Information
A summary comparison of features of the Healgen® Accurate Oral Fluid Drug Test COT and Healgen® Accurate Oral Fluid Drug Test and the predicate devices are provided in following tables.
- Substantial Equivalence Information
| Item | Device | Predicate - K033601 |
|---|---|---|
| Indication(s) | ||
| for Use | For the qualitative determination of cotinine in | |
| human oral fluid. | For the qualitative and semi- | |
| quantitative analysis of | ||
| cotinine in human oral fluid | ||
| Calibrators | Cotinine | Same |
| Methodology | Competitive binding, lateral flow | |
| immunochromatographic assays | A solid phase competitive | |
| enzyme-linked immunoassay | ||
| Type of Test | Qualitative | qualitative and semi- |
| quantitative | ||
| Specimen Type | Human Oral fluid | Same |
| Intended Use | For Over-The-Counter uses | For prescription uses |
Table 1: Features Comparison of Healgen® Accurate Oral Fluid Drug Test COT and the Predicate Device
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Table 2: Features Comparison of Healgen® Accurate Oral Fluid Drug Test and the Predicate Device
| Item | Device | Predicate - K141320 |
|---|---|---|
| Indication(s) | ||
| for Use | Same | Preliminary Drug screening test for the |
| qualitative detection of drug analytes in | ||
| oral fluid (human saliva) | ||
| For In Vitro Diagnostic Use | ||
| Methodology | Same | Competitive binding, |
| lateral flow | ||
| immunochromatographic | ||
| assays | ||
| Type of Test | Same | Qualitative |
| Specimen Type | Same | Human Oral Fluid |
9. Test Principle
The Healgen® oral fluid drug test device is an immunoassay based on the principle of competitive binding. Drugs that may be present in the oral fluid specimen compete against their respective drug conjugates for binding sites on their specific antibody.
During testing, a portion of the oral fluid specimen migrates upward by capillary action. A drug, if present in the oral fluid specimen below its cut-off concentration, will not saturate the binding sites of its specific antibody. The antibody will then react with the drug-protein conjugate and a visible colored line will show up in the test line region of the specific drug strip. The presence of drug above the cut-off concentration in the oral fluid specimen will saturate all the binding sites of the antibody. Therefore, the colored line will not form in the test line region.
A drug-positive oral fluid specimen will not generate a colored line in the specific test line region of the strip because of drug competition, while a drug-negative oral fluid specimen will generate a line in the test line region because of the absence of drug competition.
To serve as a procedural control, a colored line will always appear at the control line region, indicating that proper volume of specimen has been added and membrane wicking has occurred.
10. Performance Characteristics
-
- Analytical Performance
- Precision-Reproducibility-Cut-Off a.
Precision-Reproducibility-Cut-Off studies were carried out for samples with concentrations of -100% cut off, -75% cut off, -50% cut off, -25% cut off, cut off, +25% cut off, +50% cut off, +75% cut off and +100% cut off. These samples were prepared by spiking cotinine or marijuana in negative oral fluid samples. Each cotinine or marijuana concentration was confirmed by LC/MS/MS. All sample aliquots were blindly labeled by the person who prepared the samples and didn't take part in the sample testing. For each concentration, tests were performed two runs per day for 25 days per device lot in a randomized order. The following are summaries.
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| Results | -100%
cut off | -75%
cut off | -50%
cut off | -25%
cutoff | cut off | +25%
cut off | +50%
cut off | +75%
cut off | +100%
cut off |
|---------------|------------------|-----------------|-----------------|----------------|---------|-----------------|-----------------|-----------------|------------------|
| Device Lot | | | | | | | | | |
| LC-MS (ng/mL) | 0 | 7.4 | 15.4 | 22.6 | 30.7 | 37.9 | 45.9 | 52.9 | 60.6 |
| Lot 1 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 24-/26+ | 48+/2- | 50+/0- | 50+/0- | 50+/0- |
| Lot 2 | 50-/0+ | 50-/0+ | 50-/0+ | 48-/2+ | 23-/27+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 3 | 50-/0+ | 50-/0+ | 50-/0+ | 49-/1+ | 24-/26+ | 49+/1- | 50+/0- | 50+/0- | 50+/0- |
| THC | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Result | -100% | |||||||||
| cut off | -75% | |||||||||
| cut off | -50% | |||||||||
| cut off | -25% | |||||||||
| cutoff | cut off | +25% | ||||||||
| cut off | +50% | |||||||||
| cut off | +75% | |||||||||
| cut off | +100% | |||||||||
| cut off | ||||||||||
| Device Lot | LC-MS (ng/mL) | 0.00 | 9.5 | 20.7 | 30.2 | 40.8 | 51.4 | 59.2 | 68.3 | 81.5 |
| Lot 1: | 50-/0+ | 50-/0+ | 50-/0+ | 47-/3+ | 24-/26+ | 49+/1- | 50+/0- | 50+/0- | 50+/0- | |
| Lot 2: | 50-/0+ | 50-/0+ | 50-/0+ | 48-/2+ | 28-/22+ | 48+/2- | 50+/0- | 50+/0- | 50+/0- | |
| Lot 3: | 50-/0+ | 50-/0+ | 50-/0+ | 49-/1+ | 24-/26+ | 47+/3- | 50+/0- | 50+/0- | 50+/0- |
The following cut-off values for the candidate device has been verified.
| Calibrator | Cut-off (ng/mL) |
|---|---|
| cotinine | 30 |
| marijuana | 40 |
b. Linearity
Not applicable.
c. Stability
The devices are stable at 4-30 ℃ for 24 months based on the real time stability study at 4°C and 30°C.
d. Interference
Potential interfering substances were added to drug-free oral fluid and target drugs oral fluid with concentrations at 50% below and 50% above Cut-Off levels. These oral fluid samples were tested using three batches of the Healgen device. Compounds that showed no interference for cotinine and THC at a concentration of 100μg/mL are summarized in the following tables.
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| Acetaminophen | Diacetylmorphine | Naltrexone |
|---|---|---|
| (IR,2S)-(-)Ephedrine Hydrochloride | Diazepam | Naproxen |
| Acetylcodeine | Digoxin | Niacinamide |
| Acetylsalicylic Acid | Dihydrocodeine | Nicotinamide |
| Allobarbital | Diltiazem HCl | Niordiazepan |
| Alprazolam | Diphenhydramine HCl | Noscapine |
| Amobarbital | DL-Methadone | Omeprazole |
| Ampicillin | DL-Propranolol | Papaverine |
| Apomorphine | Doxylamine | Penicillin |
| ascorbic acid | Ecgonine Hydrochloride | Pentazocine |
| Atenolol | Ecgonine methylester | Pentobarbital |
| Atropine | Estradiol | Phencyclidine (PCP) |
| Baclofen | Estrone | Phenobarbital |
| Benzocaine | Fenoprofen | Phentermine |
| Benzoylecgonine | Fluconazole | Phenylephrine |
| beta-Phenethylamine | Furosemide | Phenylpropanolamine |
| bilirubin | Gemfibrozil | Phenytoin |
| Butabarbital | Gentisic Acid | Pioglitazone HCl |
| Butalbital | Heroin hydrochloride | Prednisolone |
| Caffeine | Hexobarbital | Prednisone |
| Carbamazepine | Hydrochlorothiazide | Procainamide HCl |
| Chlordiazepoxide | Hydrocodone | Procaine HCL |
| Chlorpromazine | Hydromorphone | Promethazine |
| Cimetidine | Ibuprofen | Pseudoephedrine |
| Citalopram HBr | Imipramine | Quinidine |
| Clobazam | L-Ephedrine | Quinine HCl |
| Clomipramine | L-Methamphetamine | R,R(-)-Pseudoephedrine |
| Clonazepam | L-Thyroxine | Salicylic Acid |
| Clonidine | Lamotrigine | Sertraline HCL |
| Clopidogrel bisulfate | Levetiracetam | Simvastin |
| Clorazepate | Lidocaine | Temazepam |
| Cocaethylene | Lormetazepam | Theophylline |
| Cocaine | Meperidine | Theophylline |
| Codeine | Metformin HCl | Thiamine |
| Cortisol | Methylphenidate HCl | Topiramate |
| D-Amphetamine | Metoprolol | Valproic Acid |
| D-Methamphetamine | Metronidazole | Verapamil |
| d,l-Salbutamol | Montelukast sodium salt | Zomepirac |
| Deoxycorticosterone | Morphine Sulfate | Zonisamide |
| Delta-9-THC (except for THC test) | Nalorphine | |
| Dextromethorphan | Naloxone |
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Food items such as methanol cough drops, cough syrup, cola, mouthwash, coffee, tea, milk, sugar, chewing gum, alcohol, baking soda, salt, cranberry juice, orange juice, food coloring (red, blue, green), toothpaste, tomatoes and MSG were added in either drug-free oral fluid or oral fluid containing the target drugs with concentrations of 50% below and 50% above cutoff levels to a concentration of 5%. None of the substances showed interference. Hemoglobin showed no interference at 10mg/dL.
e. Specificity
To test specificity, drug metabolites and other components that are likely to interfere in oral fluid samples were tested using three batches of the Healgen device. The following are summaries.
| COT | ||
|---|---|---|
| (-) Cotinine | ||
| (Cut-off=30 ng/mL) | Result | |
| Positive at (ng/mL) | % Cross-Reactivity | |
| (-) Cotinine | 30 | 100% |
| S(-)-Nicotine | 3000 | 1% |
| Trans-3-hydroxycotinine | 10 | 300% |
| (+)-anabasine | 1,350,000 | 0.002% |
| (+/-)-nornicotine | 150,000 | 0.02% |
| Niacinamide | >100,000 |