The Psychemedics homogeneous enzyme immunoassay (HEIA) for amphetamines in hair is an enzyme immunoassay system for the preliminary qualitative detection of methamphetamine in human head and body hair using a methamphetamine calibrator at 3 ng methamphetamine/10 mg hair or 5 ng methamphetamine/10 mg hair for the purpose of identifying methamphetamine use, and for the preliminary qualitative detection of amphetamine in human head and body hair using an amphetamine calibrator at 3 ng amphetamine/10 mg hair for the purpose of identifying amphetamine use.

This is an in vitro diagnostic device intended exclusively for Psychemedics use only and is not for sale to anyone. The Psychemedics HEIA for amphetamines in hair provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Liquid Chromatography/Mass Spectrometry/Mass Spectrometry (LC/MS/MS) is the preferred confirmatory method.

Device Description

The test consists of two parts; a pre-analytical hair treatment procedure (to extract amphetamines from the solid hair matrix to a measurable liquid matrix) and the screening assay, the Psychemedics Amphetamines HEIA. The screening portion of the test system is based on competition for antibody binding sites between drug in the measurable liquid matrix and drug-labeled recombinant glucose-6-phosphate dehydrogenase (G6PDH). As the antibody binds labeled G6PDH, enzyme activity decreases. In the presence of drug, enzyme activity increases in direct proportion to the drug concentration. Active enzyme reduces nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose 6-phosphate (G6P), resulting in an absorbance change that is measured spectrophotometrically.

The Psychemedics Amphetamines HEIA consists of reagents R1 (anti-methamphetamine monoclonal antibody with substrate) and R2 (methamphetamine labeled recombinant G6PDH) for the detection of methamphetamine, and reagents R1 (anti-amphetamine monoclonal antibody with substrate) and R2 (amphetamine labeled recombinant G6PDH) for the detection of amphetamine.

AI/ML Overview

The provided document describes the performance of the Psychemedics Homogeneous Enzyme Immunoassay (HEIA) for Amphetamines in Hair. This device is an in vitro diagnostic device intended for preliminary qualitative detection of methamphetamine and amphetamine in human hair.

Here's an analysis of the acceptance criteria and study data:

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state formal acceptance criteria values (e.g., "sensitivity must be > X%", "specificity must be > Y%"). Instead, it presents precision data and comparison studies against a confirmatory method (LC/MS/MS) to demonstrate acceptable performance. The acceptance is implied by the robust precision and high agreement with the confirmatory method, as well as satisfactory performance with cosmetic treatments and lack of cross-reactivity/interference from other substances.

However, we can infer some criteria from the presented data. The device's performance is summarized in its ability to correctly classify samples as positive or negative compared to LC/MS/MS and its precision at various concentration levels.

Inferred Acceptance Criteria vs. Reported Performance:

Feature	Acceptance Criteria (Implied)	Reported Device Performance
Precision (Intra-Assay)	Consistent classification (all negative below cutoff, all positive above cutoff) at various concentration levels (e.g., -100%, -75%, -50%, -25%, +25%, +50%, +75%, +100% relative to cutoff).	Methamphetamine (3 ng/10 mg & 5 ng/10 mg calibrator), Amphetamine (3 ng/10 mg calibrator): - At -100%, -75%, -50%, -25% of cutoff: All 8 replicates per level were correctly identified as Negative. - At +25%, +50%, +75%, +100% of cutoff: All 8 replicates per level were correctly identified as Positive. This indicates perfect intra-assay precision for the tested ranges.
Precision (Inter-Assay)	Consistent classification (all negative below cutoff, all positive above cutoff) at various concentration levels (e.g., -100%, -75%, -50%, -25%, +25%, +50%, +75%, +100% relative to cutoff) over multiple runs/days.	Methamphetamine (3 ng/10 mg & 5 ng/10 mg calibrator), Amphetamine (3 ng/10 mg calibrator): - At -100%, -75%, -50%, -25% of cutoff: All 80 replicates per level were correctly identified as Negative. - At +25%, +50%, +75%, +100% of cutoff: All 80 replicates per level were correctly identified as Positive. This indicates perfect inter-assay precision for the tested ranges.
Agreement with LC/MS/MS	High concordance between HEIA results (positive/negative) and LC/MS/MS results across the dynamic range, with minimal false positives or negatives, particularly near the cutoff. Acknowledgment and explanation of expected discordant samples due to washing protocols should be provided.	Methamphetamine (3 ng/10 mg calibrator): - 53 samples negative by HEIA were < 1.5 ng/10 mg (below 50% cutoff) by LC/MS/MS. - 2 positive HEIA samples were between 1.5-2.99 ng/10 mg by LC/MS/MS. - 19 positive HEIA samples were between 3.0-4.50 ng/10 mg by LC/MS/MS. - 35 positive HEIA samples were > 4.50 ng/10 mg by LC/MS/MS. - 5 negative HEIA samples were between 1.5-2.99 ng/10 mg by LC/MS/MS (near cutoff). -> Overall: High agreement. Discordant results (2 samples) were explained by the washing protocol for LC/MS/MS that is not applied to initial HEIA screening, leading to lower LC/MS/MS values after washing. Methamphetamine (5 ng/10 mg calibrator): - 45 samples negative by HEIA were < 2.5 ng/10 mg (below 50% cutoff) by LC/MS/MS. - 2 positive HEIA samples were between 2.5-4.99 ng/10 mg by LC/MS/MS. - 6 positive HEIA samples were between 5.0-7.50 ng/10 mg by LC/MS/MS. - 37 positive HEIA samples were > 7.50 ng/10 mg by LC/MS/MS. - 4 negative HEIA samples were between 2.5-4.99 ng/10 mg by LC/MS/MS (near cutoff). -> Overall: High agreement. Discordant results (2 samples) were explained by the washing protocol. Amphetamine (3 ng/10 mg calibrator): - 42 samples negative by HEIA were < 1.5 ng/10 mg (below 50% cutoff) by LC/MS/MS. - 2 positive HEIA samples were between 1.5-2.99 ng/10 mg by LC/MS/MS. - 15 positive HEIA samples were between 3.0-4.50 ng/10 mg by LC/MS/MS. - 31 positive HEIA samples were > 4.50 ng/10 mg by LC/MS/MS. - 6 negative HEIA samples were between 1.5-2.99 ng/10 mg by LC/MS/MS (near cutoff). -> Overall: High agreement. Discordant results (2 samples) were explained by the washing protocol.
Cross-Reactivity	Limited significant cross-reactivity with common structurally similar compounds and no cross-reactivity with a wide range of other tested compounds at specified cutoffs.	Methamphetamine Assays (3 ng/10 mg & 5 ng/10 mg calibrators): - MDMA, Para-Methoxy Methamphetamine, 1R, 2S Ephedrine, and MDEA showed some cross-reactivity (81-10% at 3 ng/10 mg; 83-10% at 5 ng/10 mg) requiring higher concentrations to be equivalent to the cutoff. - R-Methamphetamine showed low cross-reactivity (2%). - D-Amphetamine, L-Amphetamine, and MDA showed <1% cross-reactivity. - Many other drugs and common substances listed (e.g., Lisinopril, Atropine, Bupropion, Ibuprofen, Caffeine) showed no cross-reactivity. Amphetamine Assay (3 ng/10 mg calibrator): - MDA showed 100% cross-reactivity (expected, as it's a metabolite/related compound). - Phentermine showed 25% cross-reactivity. - D-Methamphetamine and MDMA showed <1% cross-reactivity. - Many other drugs and common substances listed showed no cross-reactivity.
Interference	No significant interference from a broad range of common substances.	Numerous compounds listed (e.g., Anhydroecgonine Methyl Ester, Atropine, Bupropion, Ecgonine, Meprobamate, Acetaminophen, Nicotine, Ibuprofen, Diazepam, Alprazolam) were shown to have no interference in the methamphetamine and amphetamine assays.
Cosmetic Treatment Effects	Cosmetic treatments (perm, dye, shampoo, relaxer) should not alter the test result (negative samples remain negative, positive samples remain positive).	Methamphetamine: - 5 negative samples remained negative after treatment. - 4 positive samples remained positive after treatment. Amphetamine: - 9 negative samples remained negative after treatment (dye, shampoo, relaxer, perm). - 4 positive samples remained positive after treatment. -> Overall: Cosmetic treatments did not affect the results.
Sample Shipping Stability	Samples should remain stable (positive remain positive) after typical storage and shipping conditions for a reasonable period.	- 9 methamphetamine-positive samples remained positive for approximately 8 months after storage and two coast-to-coast shipments. - 9 amphetamine-positive samples remained positive for approximately 6 months after storage and two coast-to-coast shipments. -> Overall: Demonstrated sufficient stability.
Recovery	Sufficient recovery of the target analytes during the extraction procedure.	- Recovery of methamphetamine in the phosphate buffer extraction was approximately 100% complete after 3 hours. - Recovery of amphetamine in the phosphate buffer extraction was approximately 100% complete after 3 hours. -> Overall: Satisfactory recovery.

2. Sample Sizes and Data Provenance for the Test Set

The device uses multiple test sets for different aspects of its performance evaluation:

Precision Studies:
- Methamphetamine (3 ng/10 mg calibrator): 8 replicates per level (intra-assay) and 80 replicates per level (inter-assay) for a total of 8 levels (4 negative, 4 positive).
  - Intra-assay: 8 samples/level * 8 levels = 64 samples.
  - Inter-assay: 80 samples/level * 8 levels = 640 samples.
- Methamphetamine (5 ng/10 mg calibrator): Same as above (64 intra-assay, 640 inter-assay samples).
- Amphetamine (3 ng/10 mg calibrator): Same as above (64 intra-assay, 640 inter-assay samples).
- Data Provenance: Spiked negative hair samples with LC/MS/MS validated calibrator and control solutions. This suggests controlled laboratory samples. No specific country of origin is mentioned, but "workplace setting" is mentioned for the comparison studies.
Comparison Studies (HEIA vs. LC/MS/MS):
- Methamphetamine (3 ng/10 mg calibrator): 114 individual hair samples. 58 negative and 56 positive initially identified by the test device.
- Methamphetamine (5 ng/10 mg calibrator): 94 individual hair samples. 49 negative and 45 positive initially identified by the test device.
- Amphetamine (3 ng/10 mg calibrator): 96 individual hair samples. 48 negative and 48 positive initially identified by the test device.
- Data Provenance: Hair samples collected anonymously from a "workplace setting." These appear to be retrospective real-world samples. The document does not specify a country of origin.
Cross-Reactivity & Interference Studies: Not specified, but likely involved a series of controlled laboratory experiments using specific concentrations of the compounds.
Cosmetic Treatment Studies:
- Methamphetamine: 5 methamphetamine-negative head hair samples, 4 methamphetamine-positive head hair samples.
- Amphetamine: 9 amphetamine-negative head hair samples, 4 amphetamine-positive head hair samples.
- Data Provenance: Not explicitly stated but likely controlled laboratory experiments using human hair.
Sample Shipping Stability: 9 methamphetamine-positive samples, 9 amphetamine-positive samples.
Recovery: Not specified, likely laboratory experiments.

3. Number of Experts and Qualifications for Ground Truth

The document does not describe the use of human experts to establish ground truth in the traditional sense of image interpretation or clinical diagnosis. The ground truth for this device is based on analytical chemistry results.

Ground Truth Establishment: The ground truth for the comparison studies is established using Liquid Chromatography/Mass Spectrometry/Mass Spectrometry (LC/MS/MS), which is the "preferred confirmatory method" for drug testing in hair. This is an objective chemical analysis method, not reliant on expert interpretation of observational data.

4. Adjudication Method for the Test Set

No adjudication method (like 2+1 or 3+1 consensus) is applicable or mentioned. The ground truth is established by a quantitative chemical analysis (LC/MS/MS). Discrepancies between the HEIA and LC/MS/MS results are analyzed and explained by the difference in sample preparation (washing for LC/MS/MS, no washing for HEIA screening), rather than resolved by expert consensus.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No MRMC study was done or is applicable for this type of in vitro diagnostic device. This device is an automated immunoassay for preliminary detection of substances, not an imaging or interpretive diagnostic tool that involves human readers' performance with and without AI assistance.

6. Standalone (Algorithm Only) Performance

The "standalone" performance is essentially what is reported. The Psychemedics HEIA itself is the "algorithm" (the immunoassay system). Its performance is directly compared to the gold standard (LC/MS/MS) without human intervention in the interpretation of the HEIA signal. The entire document focuses on the performance of the device itself.

7. Type of Ground Truth Used

The primary ground truth used is objective chemical analysis via Liquid Chromatography/Mass Spectrometry/Mass Spectrometry (LC/MS/MS). For precision studies, it involved spiking negative hair with known concentrations of analytes, confirmed by LC/MS/MS validated calibrators and control solutions.

8. Sample Size for the Training Set

The document does not explicitly describe a "training set" in the context of machine learning, as this is an immunoassay device, not an AI/ML algorithm that is 'trained.' The device is a chemical system with reagents. Its method development and optimization would involve various experiments, but these are not referred to as a "training set" in the common AI/ML terminology.

9. How Ground Truth for the Training Set Was Established

Since there isn't a "training set" in the AI/ML sense, this question is not directly applicable. The development and calibration of the immunoassay reagents (anti-methamphetamine monoclonal antibody, methamphetamine labeled recombinant G6PDH, etc.) would involve analytical methods to ensure their specificity and sensitivity. The calibrators and control materials are prepared using drug stocks purchased from commercial vendors with certificates of analysis (Page 10), which serves as the reference for established concentrations.

Summary

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Image /page/0/Picture/0 description: The image shows 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 the FDA logo, which 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.

December 15, 2021

Psychemedics Corporation Neil Stowe Principal Scientist 5832 Uplander Way Culver City, California 90230

Re: K210212

Trade/Device Name: Psychemedics Homogeneous Enzyme Immunoassay for Amphetamines in Hair Regulation Number: 21 CFR 862.3610 Regulation Name: Methamphetamine Test System Regulatory Class: Class II Product Code: LAF, DKZ Dated: September 13, 2021 Received: September 15, 2021

Dear Neil Stowe:

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

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statutes and regulations administered by other Federal agencies. You must comply with all the Act's 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 (QS) 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 medical devices and radiation-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,

Marianela Perez-Torres, Ph.D. Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality (OPEQ) Center for Devices and Radiologial Health (CDRH)

Enclosure

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Indications for Use

510(k) Number (if known) K210212

Device Name

Psychemedics Homogeneous Enzyme Immunoassay for Amphetamines in Hair

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)
Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)	X

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This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of
SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is:	K210212
Submitted By:	Psychemedics Corporation5832 Uplander WayCulver City, CA 90230TEL: 310 216 7776FAX: 310 216 6662
Submission Contact:	Neil Stowe
Date Prepared:
Device Trade Name:	Psychemedics Homogeneous Enzyme Immunoassay for Amphetamines in Hair
Predicate Device:	Psychemedics Microplate EIA for Methamphetamine in Hair, K111927;Psychemedics Microplate EIA for Amphetamine in Hair, K130811
Product Code:	LAF (Methamphetamine Test System), DKZ (Amphetamine Test System)
Device/Classification Name:	21 CFR 862.3610 Methamphetamine Test System, Classification II, 21 CFR862.3100 Amphetamine Test System, Classification II
Intended Use:	The Psychemedics homogeneous enzyme immunoassay (HEIA) foramphetamines in hair is an enzyme immunoassay system for the preliminaryqualitative detection of methamphetamine in human head and body hair using amethamphetamine calibrator at 3 ng methamphetamine/10 mg hair or 5 ngmethamphetamine/10 mg hair for the purpose of identifying methamphetamineuse, and for the preliminary qualitative detection of amphetamine in human headand body hair using an amphetamine calibrator at 3 ng amphetamine/10 mg hairfor the purpose of identifying amphetamine use. This is an in vitro diagnosticdevice intended exclusively for Psychemedics use only and is not for sale toanyone.The Psychemedics HEIA Amphetamines Assay provides only a preliminaryanalytical test result. A more specific alternative chemical method must be usedin order to obtain a confirmed analytical test result. LiquidChromatography/Mass Spectrometry/Mass Spectrometry (LC/MS/MS) is thepreferred confirmatory method.
Device Description:	The test consists of two parts; a pre-analytical hair treatment procedure (toextract amphetamines from the solid hair matrix to a measurable liquid matrix)and the screening assay, the Psychemedics Amphetamines HEIA. The screeningportion of the test system is based on competition for antibody binding sitesbetween drug in the measurable liquid matrix and drug-labeled recombinant
	glucose-6-phosphate dehydrogenase (G6PDH). As the antibody binds labeledG6PDH, enzyme activity decreases. In the presence of drug, enzyme activityincreases in direct proportion to the drug concentration. Active enzyme reducesnicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose6-phosphate (G6P), resulting in an absorbance change that is measuredspectrophotometrically
	The Psychemedics Amphetamines HEIA consists of reagents R1 (anti-methamphetamine monoclonal antibody with substrate) and R2(methamphetamine labeled recombinant G6PDH) for the detection ofmethamphetamine, and reagents R1 (anti-amphetamine monoclonal antibodywith substrate) and R2 (amphetamine labeled recombinant G6PDH) for thedetection of amphetamine.
	Sample Collection and Stability: A sample of hair should be cut as close as possible to the skin. The hair is placedin a V-shaped aluminum foil sample holder with the root end of the hairprotruding beyond the slanted edge of the foil. The aluminum foil is crimpedaround the sample, securing the hair specimen firmly into place within the foil.The hair sample, crimped within the foil, is placed in a sample acquisition cardenvelope and the envelope is sealed with a tamper-evident seal. Hair specimensare kept at ambient temperature in a secure location until they are shippedwithout refrigeration to the laboratory. Stability of amphetamines in hairsamples stored at room temperature has been shown for at least seven months.Amphetamines in samples shipped coast-to-coast twice was stable.
Materials Required:	Hair sample collection kit, HEIA for Amphetamines, automated clinicalchemistry analyzer, LC/MS/MS for confirmation.

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Comparison with Predicate Devices:

Item	Proposed Device	Psychemedics Methamphetamine Assay, K111927
Indications/Intended Use	The Psychemedics HEIA is an enzyme immunoassay system for the preliminary qualitative detection of methamphetamine in human head and body hair using a methamphetamine calibrator of 3 ng methamphetamine/10 mg hair or 5 ng methamphetamine/10 mg hair for the purpose of identifying methamphetamine use. This is an in-vitro diagnostic device intended exclusively for Psychemedics use only and is not for sale to anyone. The enzyme immunoassay system provides only a preliminary analytical test result. To confirm a presumptive screen positive result, a more specific alternate chemical method such as LC/MS/MS (liquid chromatography/mass spectrometry/mass spectrometry) must be used. Clinical consideration and professional judgement must be applied to the interpretation of any drug-of-abuse test result.	The Psychemedics Microplate EIA for Methamphetamine is an enzyme immunoassay (EIA) for the preliminary qualitative detection of methamphetamine in human head and body hair samples using a methamphetamine calibrator at 5 ng/10 mg hair cutoff for the purpose of identifying methamphetamine use. This product is intended exclusively for in-house professional use and not for sale to anyone. The test is not intended for over-the-counter sale to non-professionals. The Psychemedics EIA Methamphetamine Assay provides only a preliminary analytical test result. To confirm a presumptive screen positive result, a more specific alternate chemical method such as LC/MS/MS (liquid chromatography/mass spectrometry/mass spectrometry) must be used. Clinical consideration and professional judgement must be applied to the interpretation of any drug-of-abuse test result.
Product Code	LAF	LAF
Measurand	Methamphetamine	Methamphetamine
Test System	Psychemedics Homogeneous Enzyme Immunoassay for Amphetamines in Hair	Psychemedics Microplate EIA for Methamphetamine in Hair
Sample Matrix	Human Hair	Human Hair
Method of Measurement	Automated Clinical Chemistry Analyzer at 340 nm	Microplate Reader at 450 nm
Type of Test	Enzyme Immunoassay	Enzyme Immunoassay
Extraction Method	Acidic aqueous buffer	Patented Digestion Method
Confirmation Method	LC/MS/MS	LC/MS/MS
Item	Proposed Device	Psychemedics AmphetamineAssay, K130811
Indications/Intended Use	The Psychemedics HEIA is anenzyme immunoassay system forthe preliminary qualitative detectionof amphetamine in human head andbody hair using an amphetaminecalibrator of 3 ng/10 mg hair for thepurpose of identifying amphetamineuse. This is an in vitro diagnosticdevice intended exclusively forPsychemedics use only and is notfor sale to anyone.The enzyme immunoassay systemprovides only a preliminaryanalytical test result. To confirm apresumptive screen positive result,a more specific alternate chemicalmethod such as LC/MS/MS (liquidchromatography/massspectrometry/mass spectrometry)must be used. Clinicalconsideration and professionaljudgement must be applied to theinterpretation of any drug-of-abusetest result.	The Psychemedics Microplate EIAfor Amphetamine is an enzymeimmunoassay (EIA) for thepreliminary qualitative detection ofamphetamine in human head andbody hair samples using anamphetamine calibrator at 3 ng/10mg hair cutoff for the purpose ofidentifying amphetamine use. Thisis an in vitro diagnostic deviceintended exclusively forPsychemedics use only.Psychemedics has not performedan evaluation of reproducibility atdifferent laboratories.The Psychemedics Microplate EIAamphetamine assay provides only apreliminary analytical test result. Amore specific alternative chemicalmethod must be used in order toobtain a confirmed analyticalresult. Gas or LiquidChromatography/Double MassSpectrometry (GC/MS orLC/MS/MS) is the preferredconfirmatory method. Clinicalconsideration and professionaljudgement should be exercisedwith any drug of abuse test result,particularly when the result is positive.
Product Code	DKZ	DKZ
Measurand	Amphetamine	Amphetamine
Test System	Psychemedics HomogeneousEnzyme Immunoassay forAmphetamines in Hair	Psychemedics Microplate EIA forAmphetamine in Hair
Sample Matrix	Human Hair	Human Hair
Method of Measurement	Automated Clinical ChemistryAnalyzer at 340 nm	Microplate Reader at 450 nm
Type of Test	Enzyme Immunoassay	Enzyme Immunoassay
Extraction Method	Acidic aqueous buffer	Patented Digestion Method
Confirmation Method	LC/MS/MS	LC/MS/MS

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Performance Testing Summary: Precision studies were performed by spiking negative hair with previously LC/MS/MS validated calibrator and control solutions to achieve concentrations of negative, the cutoff calibrator of 3 ng methamphetamine/10 mg hair, 5 ng methamphetamine/10 mg hair, or 3 ng amphetamine/10 mg hair, and +/- 75%, +/-50% and +/-25% of the cutoff calibrator.

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Methamphetamine Intra-Assay and Inter-Assay Precision Summary, 3 ng
Methamphetamine/10 mg Hair Calibrator
Summary Intra-Assay Precision		Summary Inter-Assay Precision
Level	NEG POS	Level NEG POS
-100%	8 0	-100% 80 0
-75%	8 0	-75% 80 0
-50%	8 0	-50% 80 0
-25%	8 0	-25% 80 0
+25%	0 8	+25% 0 80
+50	0 8	+50 0 80
+75%	0 8	+75% 0 80
+100%	0 8	+100% 0 80

Methamphetamine Intra-Assay and Inter-Assay Precision Summary, 5 ng
Methamphetamine/10 mg Hair Calibrator
Summary Intra-Assay Precision		Summary Inter-Assay Precision
Level	NEG	POS	Level	NEG	POS
-100%	8	0	-100%	80	0
-75%	8	0	-75%	80	0
-50%	8	0	-50%	80	0
-25%	8	0	-25%	80	0
+25%	0	8	+25%	0	80
+50	0	8	+50	0	80
+75%	0	8	+75%	0	80
+100%	0	8	+100%	0	80

Amphetamine Intra-Assay and Inter-Assay Precision Summary
Summary Intra-Assay Precision		Summary Inter-Assay Precision
Level NEG POS -100% 8 0 -75% 8 0 -50% 8 0 -25% 8 0 +25% 0 8 +50 0 8 +75% 0 8 +100% 0 8																													Level NEG POS -100% 80 0 -75% 80 0 -50% 80 0 -25% 80 0 +25% 0 80 +50 0 80 +75% 0 80 +100% 0 80

Cross Reactivity Summary:

The cross reactivity of the following metabolites and amphetamines structural analogs was evaluated by determining the minimum concentration that would result approximately equivalent to the 3.0 ng methamphetamine/10 mg hair and 5.0 ng methamphetamine/10 mg cutoffs.

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Cross Reactivities, Methamphetamine 3 ng/10 mg Calibrator, Structurally Related
Compounds and Metabolites
Compound	% CrossReactivity	Concentration Equivalent to 3.0 ngMethamphetamine/10 mg Hair
MDMA	81	3.7
Para-Methoxy Methamphetamine	50	6.0
1R, 2S Ephedrine	20	15
MDEA	12	25
R-Methamphetamine	2	150
D-Amphetamine	<1	>300
L-Amphetamine	<1	>300
MDA	<1	>300
Para-methoxy Amphetamine	<1	>300

Cross Reactivities, Methamphetamine 5 ng/10 mg Calibrator, Structurally RelatedCompounds and Metabolites
Compound	% CrossReactivity	Concentration Equivalent to 5.0 ngMethamphetamine/10 mg Hair
MDMA	83	6
Para-Methoxy Methamphetamine	50	10
1R, 2S Ephedrine	20	25
MDEA	10	50
R-(-)-Methamphetamine	2	175
D-Amphetamine	<1	>500
L-Amphetamine	<1	>500
MDA	<1	>500
Para-methoxy Amphetamine	<1	>500

The following compounds were shown to have no cross reactivity in the methamphetamine assay using the 3 ng or 5 ng methamphetamine/10 mg hair calibrator.

Lisinopril Dihydrate, Atropine, Bupropion, Cotinine, Cannabinol, Chlorpheniramine Maleate, O-Desmethylvenlafaxine, Desipiramine, Doxylamine Succinate, 1S, 2R-Ephedrine, Amitriptyline, Dextromethorphan, Lidocaine, Methocarbamol, Nordoxepin, Pentazocine, Phenylephrine, Triamterene, Naproxen, Nicotine, Nortriptyline, Propoxyphene, R,R-Pseudoepedrine, Thioridazine, Cis-Tramadol, Venlafaxine HCl, 8-(-)11-nor-9carboxy-delta 9 THC, 11-nor-9-carboxy-delta 9 THC, Amoxicillin, Propanolol, Promethazine, Phenmetrazine, Phendimetrazine, Benzocaine, Dimenhydrinate, Metanephrin, Carbamazepine, Diazepine, Nordiazepam, Oxazepam, Acetominophen, Caffeine, Dyphylline, Methaqualone, Theophylline, Amphetamine, Cetrizine DiHCl, Imipramine, Phencyclidine, Phenylpropanolamine, Amitryptiline, Bupivacaine HCl, Lidocaine, Methocarbamol, Nordoxepin, Pentazocine, Pheynylephrine, Triamterene, Ethosuximide, Alpha-methyl-alpha-propyl succinimide, Metharbital, Barbital, Mehsuximde, Phensuximide, N-Normethsuximide, Mephyton, Ethotoin, Mephobarbital, PEMA, Phenobarbital, Methyl PEMA, 10, 11

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Dihydrocarbamazepine, Primidone, 5,5-Diphenylhydantoin, 4-Methylprimidone, Butabarbital, Amobarbital, Secobarbital, Hexobarbital, Azithromyocin Dihydrate, Glutethimide, Methaqualone, Diazepam, Chlorpromazine, Flurazepam, AM-2201, JWH-019, JWH-081, JWH-122, CP47,497 (+/-), CP47, 497 (+/-), C8 Homologue, HU-211, JWH-200, JWH-250, Ibuprofen, Ephinephrine (+/-), Norephenephrine (+/-), Metanephrine (+/-), Normetanephrine (+/-), Vanilmandelic Acid (+/-), 5-Hydroxyindole-3-acetic acid, Homovanillic acid, Alprazolam, Cimetidine, Citalopram HBr, Clonazepam, Clopidogrel bisulfate, Fluconazole, hydrochlorothiazide, Lamotrigeine, L-Thyroxine, Methylphenidate HCl, Omeprazole, Levetiracetam, Sertaline HCl, Topiramate, Zolpidem Tartrate, Zonisamide, Amlopidine Besylate Atorvastatin Calcium Salt

Cross Reactivities of Structurally Related Compounds and Metabolites
Compound	% CrossReactivity	Concentration Equivalent to 3.0 ngAmphetamine/10 mg Hair
MDA	100	3.0
Phentermine	25	12
D-Methamphetamine	<1	>300
MDMA	<1	>300
Phenmetrazine	<1	>300
1R, 2S Ephedrine	<1	>300

The following compounds were shown to have no cross reactivity in the Amphetamine assay.

Lisinopril Dihydrate, Atropine, Bupropion, Cotinine, Cannabinol, Chlorpheniramine Maleate, O-Desmethylvenlafaxine, Desipiramine, Doxylamine Succinate, 1S, 2R-Ephedrine, Amitriptyline, Dextromethorphan, Lidocaine, Methocarbamol, Nordoxepin, Pentazocine, Phenylephrine, Triamterene, Naproxen, Nicotine, Nortriptyline, Propoxyphene, R,R-Pseudoepedrine, Thioridazine, Cis-Tramadol, Venlafaxine HCl. 8-(-)11-nor-9carboxy-delta 9 THC, 11-nor-9-carboxy-delta 9 THC, Amoxicillin, Propanolol, Promethazine, Phenmetrazine, Phendimetrazine, Benzocaine, Dimenhydrinate, Metanephrin, Carbamazepine, Diazepine, Nordiazepam, Oxazepam, Acetominophen, Caffeine, Dyphylline, Methaqualone, Theophylline, Cetrizine DiHCl, Imipramine, Methamphetamine, Phencyclidine, Phenylpropanolamine, Amitryptiline, Bupivacaine HCl, Lidocaine, Methocarbamol, Nordoxepin, Pentazocine, Pheynylephrine, Triamterene, Ethosuximide, Alpha-methyl-alphapropyl succinimide, Metharbital, Barbital, Mehsuximde, Phensuximide, N-Normethsuximide, Mephyton, Ethotoin, Mephobarbital, PEMA, Phenobarbital, Methyl PEMA, 10, 11-Dihydrocarbamazepine, Primidone, 5,5-Diphenylhydantoin, 4-Methylprimidone, Butabarbital, Amobarbital, Secobarbital, Hexobarbital, Azithromyocin Dihydrate, Glutethimide, Methaqualone, Diazepam, Chlorpromazine, Flurazepam, AM-2201, JWH-019,

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	JWH-081, JWH-122, CP47,497 (+/-), CP47, 497 (+/-), C8 Homologue, HU-211,JWH-200, JWH-250, Ibuprofen, Ephinephrine (+/-), Norephenephrine (+/-),Metanephrine (+/-), Normetanephrine (+/-), Vanilmandelic Acid (+/-), 5-Hydroxyindole-3-acetic acid, Homovanillic acid, Alprazolam, Cimetidine,Citalopram HBr, Clonazepam, Clopidogrel bisulfate, Fluconazole,hydrochlorothiazide, Lamotrigeine, L-Thyroxine, Methylphenidate HCl,Omeprazole, Levetiracetam, Sertaline HCl, Topiramate, Zolpidem Tartrate,Zonisamide, Amlopidine Besylate Atorvastatin Calcium Salt
Interference:	The following compounds were shown to have no interference in themethamphetamine and amphetamine assays.
	Anhydroecgonine Methyl Ester, Atropine, Bupropion, Cotinine, Cannabinol,Chlorpheniramine maleate, O-Desmethylvenlafaxine, Desipiramine,Doxylamine Succinate, 1S, 2R-Ephedrine, Naproxen, Nicotine, Nortriptyline,Propoxyphene, R,R-Pseudoephedrine, Thioridazine, Cis-Tramadol, VenlafaxineHCl, 8-(-)-11-nor-9-caroxy-delta-9 THC, 11-nor-9-carboxy-delta-9 THC,Amoxicillin, Propanolol, Promethazine, Phenmetrazine, Phendimetrazine,Benzocaine, Ecgonine, Metanephrin, Glutethimide, Meprobamate, Methyprylon,Carbamazepine, Diazepam, Nordiazepam, Oxazepam, Acetaminophen,Caffeine, Dyphylline, Methaqualone, Theophylline, Amitryptiline,Dextromethorphan, Lidocaine, Methocarbamol, Nordoxepin, Pentazocine,Phenylephrine, Triamterene, Ethosuximide, a-Methyl-a-propylsuccinimide,Metharbital, Barbital, Methsuximide, Phensuximide, N-Normethylsuximide,Mephenytoin, Ethytion, Mephobarbital, PEMA, Phenobarbital, Methyl PEMA,10, 11-Dihydrocarbamazepine, Primidone, Carbamazepine, 5,5-Diphenylhydantoin, 4-Methylprimidone, Glutethimide, Methaqualone,Chlorpromazine, Flurazepam, AM-2201, JWH-019, JWH-081, JWH-122, CP47,497 (±), CP 47, 497 (±) C8 Homologue, HU-211, JWH-200, JWH-250,Acetaminophen, Caffeine, Chlorpheniramine, Ibuprofen, Naproxen, R,R-(-)-Pseudoephedrine, Epinephrine (±), Norepinephrine (±), normetanephrine (±),Alprazolam, Cimetidine, Citalopram HBr, Clonazepam, Clopidogrel Bisulfate,Fluconazole, Hydrochlorothiazide, Lamotrigine, L-Thyroxine, MethylphenidateHCl, Omeprazole, Amlopidine Besylate, Atorvastatin Calcium Salt,Azithromycin Dihydrate, Bupivacaine HCl, Cetirizine Di-HCl, Dimenhydrinate,Lisinopril Dihydrate, Butabarbital, Amobarbital, Secobarbital, Hexobarbital,Medazepam, Lorazepam, Temazepam, Bromazepam, Carbamazepine,Levetiracetam, Metformin HCl, Phenobarbital, Phenytoin, R-(-)-PhenylephrineHCl, Sertraline HCl, Topiramate, Zolpidem Tartrate, Zonisamide, 11-nor-9-caroxy-delta-8 THC, Streptomycin Solution, Procaine, Erythromycin, PenicillinG, Mepivacaine, Vanilmandelic Acid (±), 5-Hydroxyindole-3-Acetic Acid,Homovanilic Acid
Calibrator:	Psychemedics prepares calibrators and control materials using drug stockspurchased from a commercial vendor. Each lot of drug is received with itsspecific certificate of analysis. The commercially obtained stock is made intocalibrators and controls to the desired concentrations. The concentrations are
Sample Shipping StabilityDuring Storage:	9 methamphetamine positive samples remained positive after approximately 8months in storage and after shipping twice coast-to-coast. 9 amphetaminepositive samples remained positive after approximately 6 months in storage andafter shipping twice coast-to-coast.
Recovery:	The hair sample preparation for the screening HEIA is a phosphate bufferextraction procedure. Recovery of methamphetamine in the method was shownon average to be approximately 100% complete after extraction for 3 hours.Recovery of amphetamine in the method was shown on average to beapproximately 100% complete after extraction for 3 hours.
Cosmetic Treatments:	5 methamphetamine-negative head hair samples were treated with perm, dye,shampoo and relaxer and the results compared to the same samples withouttreatments. In each case of the 5 samples treated with a type of cosmetictreatment, all samples remained negative after the treatments.4 methamphetamine-positive head hair samples were treated with perm, dye,shampoo and relaxer and the results compared to the same samples without thetreatments. In each case, the samples remained positive after the treatments.5 amphetamine-negative head hair samples were treated with dye, shampoo andrelaxer and the results compared to the same samples without treatments. 4amphetamine-negative head hair samples were treated with perm and the resultscompared to the same samples without treatments. In each case of the samplestreated with a type of cosmetic treatment, all samples remained negative afterthe treatments.4 amphetamine-positive head hair samples were treated with perm, dye,shampoo and relaxer and the results compared to the same samples without thetreatments. In each case, the samples remained positive after the treatments.
Comparison Studies:	Methamphetamine, 3 ng methamphetamine/10 mg hair calibratorSamples positive or negative for methamphetamine were identified using thePsychemedics methamphetamine microplate assay (K111927), and then testedwith the test device, the Psychemedics HEIA for methamphetamine in hair. Thetest device (assay) has been validated at the 3 ng methamphetamine/10 mg haircutoff using 114 individual hair samples collected anonymously from aworkplace setting. 58 negative samples and 56 positive samples were identifiedby the test device.The stored hair samples were then tested using Psychemedics' LC/MS/MSconfirmatory assay, to compare the Psychemedics HEIA results with theLC/MS/MS results. The studies comparing the HEIA with LC/MS/MSdocumented the source of hair (head or body) and other demographics asavailable. The comparison of the Psychemedics Methamphetamine HEIA at the3 ng/10 mg hair calibrator with LC/MS/MS is shown in the following table.

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Methamphetamine 3 ng/10 mg hair Calibrator Comparison Study
HEIA Result	Washed LC/MS/MS Result, ng Methamphetamine/10 mg hair (% of cutoff calibrator)
	< 1.5 (< 50% below cutoff)	1.50 – 2.99 (≥ 50% below cutoff to cutoff)	3.0 – 4.50 (cutoff to ≥ 50% above cutoff)	> 4.50 (> 50% above cutoff)
Positive	0	2	19	35
Negative	53	5	0	0

Methamphetamine Discordant Results at the 3 ng methamphetamine/10 mg hair calibrator: Positive HEIA/Negative LC/MS/MS

Washing of hair before confirmation: exclusion of potential environmental contamination and/or sweat-derived drug from hair analysis results can result in APPARENT discordant results. Ingested drugs are present in perspiration, which settles on the hair and requires removal if the hair analysis result is to reflect amount of drug ingested rather than exposure to the sweat-derived drug. Drug could also be present as a result of environmental contamination (e.g., powder, smoke). Hair is not washed prior to screening, as it would not be reasonable to wash hundreds of negative hair samples; i.e., samples that are negative without washing. Thus, it is expected that the washing performed before LC/MS/MS confirmation of presumptive positives removes external drug and the confirmation results may then be lower than the screening may have predicted without the consideration of sweat-derived drug and/or drug from environmental contamination.

Methamphetamine, 3 ng/10 mg Hair Calibrator Discordant Results
Sample	HEIA Result	UnwashedLC/MS/MSResult (ngMeth/10mg hair)	WashedLC/MS/MSResult (ngMeth/10mg hair)	Comment
1	POS	4.70	2.43	Samples are screened by HEIA without hair washing.LC/MS/MS is performed only after washing once with
2	POS	4.80	2.46	isopropanol, 3 times 30 minutes with 90% Ethanol,then 2 times 60 minutes with 90% Ethanol. Thus, it isexpected that some samples will be negative byLC/MS/MS after washing.

Methamphetamine, 5 ng methamphetamine/10 mg hair calibrator

Samples positive or negative for methamphetamine were identified using the Psychemedics methamphetamine microplate assay (K111927), and then tested with the test device, the Psychemedics HEIA for methamphetamine in hair. The test device (assay) has been validated at the 5 ng methamphetamine/10 mg hair cutoff using 94 individual hair samples collected anonymously from a workplace setting. 49 negative samples and 45 positive samples were identified by the test device.

The stored hair samples were then tested using Psychemedics' LC/MS/MS confirmatory assay, to compare the Psychemedics HEIA results with the

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LC/MS/MS results. The studies comparing the HEIA with LC/MS/MS documented the source of hair (head or body) and other demographics as available. The comparison of the Psychemedics Methamphetamine HEIA at the 5 ng/10 mg hair calibrator with LC/MS/MS is shown in the following table.

Methamphetamine 5 ng/10 mg hair Calibrator Comparison Study
HEIA Result	Washed LC/MS/MS Result, ng Methamphetamine /10 mg hair (% of cutoff calibrator)
	< 2.5 (< 50% below cutoff)	2.5 – 4.99 (≥ 50% below cutoff to cutoff)	5.0 – 7.50 (cutoff to ≥ 50% above cutoff)	> 7.50 (> 50% above cutoff)
Positive	0	2	6	37
Negative	45	4	0	0

Methamphetamine Discordant Results at the 5 ng methamphetamine/10 mg hair calibrator: Positive HEIA/Negative LC/MS/MS

Methamphetamine, 5 ng/10 mg Hair Calibrator Discordant Results
Sample	HEIA Result	UnwashedLC/MS/MSResult (ngMeth/10mg hair)	WashedLC/MS/MSResult (ngMeth/10mg hair)	Comment
1	POS	6.18	4.28	Samples are screened by HEIA without hair washing.LC/MS/MS is performed only after washing once with
2	POS	5.57	4.56	isopropanol, 3 times 30 minutes with 90% Ethanol,then 2 times 60 minutes with 90% Ethanol. Thus, it isexpected that some samples will be negative byLC/MS/MS after washing.

Amphetamine, 3 ng Amphetamine/10 mg hair calibrator

Samples positive or negative for amphetamine were identified using the Psychemedics methamphetamine microplate assay (K130811), and then tested with the test device, the Psychemedics HEIA for amphetamine in hair. The test device (assay) has been validated using 96 individual hair samples collected

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anonymously from a workplace setting. 48 negative samples and 48 positive samples were identified by the test device.

The stored hair samples were then tested using Psychemedics' LC/MS/MS confirmatory assay, to compare the Psychemedics HEIA results with the LC/MS/MS results. The studies comparing the HEIA with LC/MS/MS documented the source of hair (head or body) and other demographics as available. The comparison of the Psychemedics Amphetamine HEIA with LC/MS/MS is shown in the following table.

Amphetamine 3 ng/10 mg hair Calibrator Comparison Study
HEIA Result	Washed LC/MS/MS Result, ng Amphetamine/10 mg hair (% of cutoff calibrator)
	< 1.5 (< 50% below cutoff)	1.50 – 2.99 (≥ 50% below cutoff to cutoff)	3.0 – 4.50 (cutoff to ≥ 50% above cutoff)	> 4.50 (> 50% above cutoff)
Positive	0	2	15	31
Negative	42	6	0	0

Amphetamine Discordant Results: Positive HEIA/Negative LC/MS/MS

Amphetamine, 3 ng/10 mg Hair Calibrator Discordant Results
Sample	HEIA Result	UnwashedLC/MS/MSResult (ngAmphetamine/10mg hair)	WashedLC/MS/MSResult (ngAmphetamine/10mg hair)	Comment
1	POS	3.18	2.51	Samples are screened by HEIA without hair washing. LC/MS/MS is performed only after washing once with isopropanol, 3 times 30 minutes with 90% Ethanol, then 2 times 60 minutes with 90% Ethanol. Thus, it is expected that some samples will be negative by LC/MS/MS after washing.
2	POS	5.82	2.73

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Conclusion:

The Psychemedics HEIA for amphetamines in hair is substantially equivalent based on acceptable performance studies, including precision, specificity and interference (including cosmetic effects).

Regulation Number and Section

§ 862.3610 Methamphetamine test system.

(a)
Identification. A methamphetamine test system is a device intended to measure methamphetamine, a central nervous system stimulating drug, in serum, plasma, and urine. Measurements obtained by this device are used in the diagnosis and treatment of methamphetamine use or overdose.(b)
Classification. Class II (special controls). A methamphetamine 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).