(38 days)
The Psychemedics Microplate EIA for Amphetamine is an enzyme immunoassay (EIA) for the preliminary qualitative detection of amphetamine in human head and body hair using an amphetamine calibrator at 3 ng /10 mg hair cutoff for the purpose of identifying amphetamine use. This is an in vitro diagnostic device intended exclusively for Psychemedics use only. Psychemedics has not performed an evaluation of reproducibility at different laboratories.
The Psychemedics Microplate EIA amphetamine assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas or Liquid Chromatography/Mass Spectrometry (GC/MS or LC/MS/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.
The test consists of two parts; a pre-analytical hair treatment procedure (to convert the solid matrix of hair to a measurable liquid matrix) and the screening assay, the Psychemedics Microplate EIA for Amphetamine. The screening portion of the test system consists of ( 1 ) microplate wells coated with multiple antigens including methamphetamine conjugated to bovine serum albumin (BSA), monoclonal mouse anti-amphetamine, rabbit anti-mouse secondary antibody conjugated to HRP (horseradish peroxidase), substrate [3, 3', 5, 5' tetramethylbenzidine (TMB)], HCl to acidify (and stop the reaction), and wash buffer for washing the plates. Absorbance in the wells is read with a microplate reader.
Here's a breakdown of the acceptance criteria and the study details for the Psychemedics Microplate EIA for Amphetamine in Hair, based on the provided 510(k) summary:
1. Table of Acceptance Criteria and Reported Device Performance
The document doesn't explicitly state "acceptance criteria" with numerical targets in a structured table for diagnostic accuracy (e.g., sensitivity, specificity). Instead, the performance is demonstrated through various studies. The primary comparison is against LC/MS/MS as the confirmatory method. The closest we get to performance criteria are successful precision, cross-reactivity, interference, and environmental contamination studies, as well as substantial equivalence to the predicate device.
Implicit Acceptance Criteria and Reported Device Performance:
| Performance Aspect | Acceptance Criteria (Implicit from Study Design) | Reported Device Performance |
|---|---|---|
| Precision | Demonstrate consistent results across intra-assay and inter-assay conditions for negative, cutoff, and various concentration levels relative to the cutoff. | Intra-Assay: 15/15 positive or negative results for each level (including -100%, -75%, -50%, -25% for negative; +25%, +50%, +75%, +100% for positive). Inter-Assay: 75/75 positive or negative results for each level (including -100%, -75%, -50%, -25% for negative; +25%, +50%, +75%, +100% for positive). All results were concordant with expected outcomes. |
| Comparison to LC/MS/MS (Diagnostic Accuracy) | Demonstrate substantial equivalence to LC/MS/MS for identifying amphetamine use, with minimal discordant results. | Out of 180 samples: - True Negatives: 38 (EIA Negative, LC/MS/MS Negative) - False Positives: 14 (EIA Positive, LC/MS/MS Negative) - Discordant within Negative Range: - EIA Positive, LC/MS/MS < half cutoff: 2 - EIA Positive, LC/MS/MS Near Cutoff Negative: 17 - Discordant within Positive Range: - EIA Negative, LC/MS/MS < half cutoff: 17 - EIA Negative, LC/MS/MS Near Cutoff Negative: 11 - True Positives: 59 (EIA Positive, LC/MS/MS High Positive) - True Positives (Near Cutoff): 22 (EIA Positive, LC/MS/MS Near Cutoff Positive) |
| Cross-Reactivity | Identify known cross-reactants and show no interaction with a broad panel of non-target compounds. | Significant Cross-Reactivity: Chloramphetamine (79%), MDA (120%), PMA (100%), Phentermine (17.6%). Low/No Cross-Reactivity: l-amphetamine (1.1%), MDMA (0.5%), PMMA (0.5%), Phenylpropanolamine (< 0.3%), Pseudoephedrine (< 0.3%), etc. 140 other compounds showed no cross-reactivity. |
| Interference | Demonstrate no interference from common substances at relevant concentrations. | 119 compounds tested at +/-50% of the cutoff showed no interference. |
| Cosmetic Treatments | Maintain accurate results for both negative and positive samples after exposure to common cosmetic hair treatments. | Negative Samples: All 20 samples remained negative after bleach, permanent wave, dye, relaxer, and shampoo treatments. Positive Samples: None of the 12 samples became negative (by EIA or LC/MS/MS) after any cosmetic treatment. |
| Environmental Contamination (Wash Procedure Effectiveness) | Successfully differentiate between internal drug incorporation and external contamination, with contaminated samples identified as negative by the wash criterion. | For samples soaked in 500 ng amphetamine/mL water: All 11 samples were determined to be Negative by the aqueous wash criterion. For samples soaked in 500 ng amphetamine/mL saline: All 11 samples were determined to be Negative by the aqueous wash criterion. Similar results were observed with the 90% ethanol wash procedure for both water and saline soaked samples. |
| Storage & Shipping Stability | Maintain stable drug detection over prolonged storage and shipping conditions. | Storage: Average results after 1 year (ambient) were 109% of original. Shipping: Average results after 2 coast-to-coast shipments were 105% of original. |
| Calibrator & Control Stability | Calibrators and controls maintain stability for a specified period. | Stability was shown to be 9 months, with ongoing studies for 1-year stability. |
| Recovery | Demonstrate effective recovery of amphetamine from hair. | Recovery ranged from 100% to 110% in a 2-hour incubation, determined by LC/MS/MS. |
2. Sample Size Used for the Test Set and Data Provenance
-
Test Set Sample Size:
- Comparison Testing (EIA vs. LC/MS/MS): 180 samples (comprising both head and body hair).
- Precision Studies:
- Intra-Assay: 15 replicates per level (Negative, Cutoff +/- concentrations), totaling 120 samples (8 levels * 15 replicates).
- Inter-Assay: 75 replicates per level, totaling 600 samples (8 levels * 75 replicates).
- Cosmetic Treatment Studies: 20 negative samples + 12 positive samples per treatment type (bleach, permanent wave, dye, relaxer, shampoo) for "before and after" comparison. This implies at least 32 distinct samples, each tested multiple times.
- Environmental Contamination Studies:
- Aqueous Wash: 11 samples (water-soaked) + 11 samples (saline-soaked)
- Ethanol Wash: 10 samples (water-soaked) + 13 samples (saline-soaked)
- Storage & Shipping Stability: 21 amphetamine-positive samples for each study (storage and shipping).
- Cross-reactivity: MDA, d-amphetamine, PMA, Chloramphetamine, Phentermine, l-amphetamine, MDMA, PMMA, Phenylpropanolamine, Pseudoephedrine, IR, 2S Ephedrine, S,S Pseudoephedrine, Phenylethylamine, MDEA, L-methamphetamine, Ranitidine, Fenfluramine, Mephentermine, Phenmetrazine, Phendimetrazine, Metanephrin (total of 21 named), plus 140 other compounds. Each would have been tested.
- Interference: 119 compounds.
-
Data Provenance: The document does not explicitly state the country of origin or whether the data was retrospective or prospective. Given it's a 510(k) submission to the FDA, it is expected to be from studies conducted under appropriate quality systems, likely in the US, but this is not confirmed. It appears to be prospective data collection for the performance studies described.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The ground truth for the comparison testing (diagnostic accuracy) and other analytical performance studies was established using LC/MS/MS (Liquid Chromatography/Mass Spectrometry/Mass Spectrometry) or GC/MS (Gas Chromatography/Mass Spectrometry) as the "preferred confirmatory method."
These methods are highly sensitive and specific analytical techniques widely considered the gold standard for drug detection and quantification. Therefore, the "experts" in establishing this ground truth are the analytical instrumentation and the laboratory personnel qualified to operate and interpret results from LC/MS/MS/GC/MS systems. No human expert consensus was used to establish the ground truth in the clinical sense, but rather the objective biochemical analysis provided by these confirmatory methods.
4. Adjudication Method for the Test Set
No human adjudication method (like 2+1 or 3+1) was explicitly mentioned. The "ground truth" was established purely by the results of the LC/MS/MS or GC/MS confirmatory methods. Discordant results were analyzed and presented. For instance, the table of discordant results shows EIA positive results where LC/MS/MS found 0 amphetamine but identified other substances like phentermine or methamphetamine, suggesting potential cross-reactivity for the EIA.
5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study was Done
No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done. This would typically apply to scenarios where human readers interpret medical images or complex data, and the AI's role is to assist human interpretation. This device is an in vitro diagnostic (IVD) assay designed for laboratory use, yielding a preliminary qualitative result, not for direct human interpretation in the same way an imaging AI might be. Therefore, the concept of "human readers improving with AI vs. without AI assistance" does not directly apply here. The device provides a preliminary analytical result that then requires chemical confirmation (LC/MS/MS).
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) was Done
Yes, a standalone (algorithm only) performance study was done. The entire performance summary, including precision, comparison to LC/MS/MS, cross-reactivity, interference, cosmetic treatments, environmental contamination, and stability, assesses the performance of the "Psychemedics Microplate EIA for Amphetamine" assay itself, working in a laboratory setting. The results (Positive/Negative) are generated by the assay system, read by a microplate reader, without human interpretive input that would alter the primary output of the device as an "algorithm." The results are then further interpreted as "preliminary" and require confirmatory methods.
7. The Type of Ground Truth Used
The type of ground truth used was objective chemical confirmatory methods, specifically Gas or Liquid Chromatography/Double Mass Spectrometry (GC/MS or LC/MS/MS). These methods are considered the definitive analytical standard for identifying and quantifying drug substances.
8. The Sample Size for the Training Set
The document does not explicitly mention a "training set" sample size or a training process in the context of machine learning or AI. This is an immunoassay (EIA) device, which is a biochemical test, not typically an AI/machine learning algorithm that requires a training set in the conventional sense. The "training" of such assay systems primarily involves establishing reagents, calibrators, controls, and protocols based on known chemical properties and optimization, rather than ingesting a large dataset of results for algorithmic learning.
9. How the Ground Truth for the Training Set Was Established
Since this is an immunoassay and not an AI/ML algorithm, the concept of establishing ground truth for a "training set" as understood in AI development does not apply directly. The development of an immunoassay involves:
- Selection and optimization of antibodies (e.g., monoclonal mouse anti-amphetamine)
- Optimization of reagent concentrations (e.g., BSA conjugates, HRP, substrate)
- Establishing cutoff values (e.g., 3 ng/10 mg hair) through extensive analytical testing and comparison to reference methods (like LC/MS/MS) using known positive and negative samples, as well as samples with varying concentrations.
- Validation of the entire assay system to ensure it performs as intended across various parameters (precision, specificity, etc.), which is what the provided "Summary of Performance Testing" details.
Therefore, the "ground truth" during the development and validation of this EIA assay would have been established using analytically confirmed amphetamine concentrations in hair samples, verified by methods such as LC/MS/MS, to define the assay's performance characteristics and cutoff.
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510 K SUMMARY
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: K130811
Submitted By:
Psychemedics Corporation
5832 Uplander Way Culver City, CA 90230 TEL: 310 216 7776 FAX: 310 216 6662
Submission Contact:
Virginia Hill March 22, 2013
DKZ
Date Prepared:
Device Trade Name:
Predicate Device:
Product Code:
Device Classification/Name:
Intended Use:
Assay Description:
21 CFR 862.3100, Enzyme Immunoassay, Amphetamine; Classification II;
Psychemedics Microplate EIA for Amphetamine in Hair
Cozart EIA Amphetamine Oral Fluid Microplate Kit, K033743
The Psychemedics Microplate ELA for Amphetamine is an enzyme immunoassay (EIA) for the preliminary qualitative detection of amphetamine in human head and body hair using an amphetamine calibrator at 3 ng /10 mg hair cutoff for the purpose of identifying amphetamine use. This is an in vitro diagnostic device intended exclusively for Psychemedics use only. Psychemedics has not performed an evaluation of reproducibility at different laboratories.
The Psychemedics Microplate EIA amphetamine assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas or Liquid Chromatography/Double Mass Spectrometry (GC/MS or LC/MS/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.
The test consists of two parts; a pre-analytical hair treatment procedure (to convert the solid matrix of hair to a measurable liquid matrix) and the screening assay, the Psychemedics Microplate EIA for Amphetamine. The screening portion of the test system consists of ( 1 ) microplate wells coated with multiple antigens including methamphetamine conjugated to bovine serum albumin (BSA), monoclonal mouse anti-amphetamine, rabbit anti-mouse secondary antibody conjugated to HRP (horseradish peroxidase), substrate [3, 3', 5, 5' tetramethylbenzidine (TMB)], HCl to
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acidify (and stop the reaction), and wash buffer for washing the plates. Absorbance in the wells is read with a microplate reader.
Sample Collection & Stability: A sample of hair should be cut as close as possible to the skin. The hair is placed in a V-shaped aluminum foil sample holder with the root end of the hair protruding beyond the slanted edge of the foif. The aluminum foil is crimped around 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 card envelope and the envelope is sealed with a tamper-evident seal. Hair specimens are kept at ambient temperature in a secure location until they are shipped without refrigeration to the laboratory. Stability of amphetamine in hair samples stored at room temperature has been shown for at least one year. Amphetamine in samples shipped coast-to-coast twice was stable.
Materials required:
Hair sample collection kit, Microplate ElA for Amphetamine, Microplate washer and reader, LC/MS/MS for confirmation.
| Item | Proposed Device | Cozart EIA AmphetaminesOral Fluid Microplate Kit |
|---|---|---|
| Indications/Intended use | The Psychemedics Microplate EIAfor Amphetamine is an enzymeimmunoassay (EIA) for thepreliminary qualitative detection ofamphetamine in human head andbody hair using an amphetaminecalibrator at 3 ng /10 mg hair cutofffor the purpose of identifyingamphetamine use.The Psychemedics Microplate EIAamphetamine assay provides only apreliminary analytical test result. Amore specific alternative chemicalmethod must be used in order toobtain a confirmed analytical result.Gas or LiquidChromatography/MassSpectrometry/Mass Spectrometry(GC/MS or LC/MS/MS) is thepreferred confirmatory method. | Cozart EIA Amphetamines OralFluid Microplate Kit is aqualitative test for amphetaminein oral fluid. It is intended forqualitative detection ofamphetamine in human oral fluidat 100 ng/mL. The test isintended for professional use. Itis not intended for over-the-counter sales to non-professionals. It provides onlypreliminary analytical testresults. A more specific alternatechemical method must be used inorder to obtain a confirmedresult. GC/MS is the preferredconfirmatory method. |
| Product Code | DKZ | DKZ |
| Measurand | Amphetamine in Hair | Amphetamine in Oral Fluid |
| Test System | Psychemedics EIA forAmphetamine in Hair | Cozart EIA Amphetamines OralFluid Microplate Kit |
| Sample Matrix | Human Hair | Human Oral Fluid |
| Method of Measurement | Microplate reader, read at 450 nm | Microplate reader, read at 450 nm |
| Cutoff | 3 ng amphetamine/10 mg hair(300 pg amphetamine /mg hair) | 100 ng amphetamine/mL oral fluid |
Comparison with Predicate:
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| Type of Test | Enzyme Immunoassay | Enzyme Immunoassay |
|---|---|---|
| Extraction Method | Patented Digestion method | Not applicable |
| Confirmation Method | LC/MS/MS | GC/MS |
Summary of Performance Testing
·
The precision studies were performed by spiking negative hair with previously LC/MS/MS-validated calibrator and control spiking solutions to achieve concentrations of negative, the cutoff of 2 ng/10 mg hair, and +/-75%, +/-50%, and +/- 25% of the cutoff. Precision Studies
| Summary -Intra-Assay | Summary-Inter-Assay | ||||
|---|---|---|---|---|---|
| LEVEL | NEG | POS | LEVEL | NEG | POS |
| B 0 (-100%) | 15 | 0 | B 0 (-100%) | 75 | 0 |
| -75% | 15 | 0 | -75% | 75 | 0 |
| -50% | 15 | 0 | -50% | 75 | 0 |
| -25% | 15 | 0 | -25% | 75 | 0 |
| plus 25% | 0 | 15 | plus 25% | 0 | 75 |
| plus 50% | 0 | 15 | plus 50% | 0 | 75 |
| plus 75% | 0 | 15 | plus 75% | 0 | 75 |
| plus 100% | 0 | 15 | plus 100% | 0 | 75 |
ComparisonTesting
One hundred eighty samples comprising both head and body hair were confirmed by LC/MS/MS in parallel with testing by the Psychemedics Amphetamine EIA, with the results shown in the following table.
The studies comparing the EIA with LC/MS/MS documented the age, gender and ethnicity of subjects, hair color, and source of hair (body or head hair).
| AmphetamineEIA Test Result | Negativeby GC/MS | Less than halfthe cutoffconcentration byGC/MS | Near Cutoff Negative(Between 50% belowthe cutoff and thecutoff) | Near CutoffPositive(Between thecutoff and 50%above the cutoff) | High Positive(Greater than50% above thecutoff) |
|---|---|---|---|---|---|
| Positive | 14 | 2 | 17 | 22 | 59 |
| Negative | 38 | 17 | 11 | 0 | 0 |
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Discordant Results of Comparison Testing
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- September 19. 1. 1.
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| Discordant Results of Comparison Testing | ||
|---|---|---|
| Screening Cutoff( ng/10 mg hair) | Amphetamine EIAResult (POS/NEG) | LC/MS/MS Drug Result (ng/ 10 mg hair) |
| 3 | POS | 0 amphetamine, 16 phentermine |
| 3 | POS | 0 amphetamine, 16 phentermine |
| 3 | POS | 0 amphetamine, 17 phentermine |
| 3 | POS | 0 amphetamine, 18 phentermine |
| 3 | POS | 0 amphetamine, 19 phentermine |
| 3 | POS | 0 amphetamine, 23 phentermine |
| 3 | POS | 0 amphetamine, 23 phentermine |
| 3 | POS | 0 amphetamine, 23 phentermine |
| 3 | POS | 0 amphetamine, 40 phentermine |
| 3 | POS | 0 amphetamine, 44 phentermine |
| 3 | POS | 0 amphetamine, 49 phentermine |
| 3 | POS | 0 amphetamine, 55 phentermine |
| 3 | POS | 0 amphetamine, 65 phentermine |
| 3 | POS | 0 amphetamine, 86 phentermine |
| 3 | POS | 1.2 amphetamine, 9.6 methamphetamine |
| 3 | POS | 1.3 amphetamine, 21.8 methamphetamine |
| 3 | POS | 1.4 amphetamine, 21 methamphetamine |
| 3 | POS | 2.0 amphetamine, 20.6 methamphetamine |
| 3 | POS | 2.1 amphetamine, 39.8 methamphetamine |
| 3 | POS | 2.6 amphetamine, 31.7 methamphetamine |
| 3 | POS | 2.8 amphetamine, 24.1 methamphetamine |
| 3 | POS | 1.6 amphetamine, 27.8 methamphetamine |
| 3 | POS | 1.6 amphetamine, 31 methamphetamine |
| 3 | POS | 1.7 amphetamine, 15.6 methamphetamine |
| 3 | POS | 2.0 amphetamine, 31 methamphetamine |
| 3 | POS | 2.1 amphetamine, 25 methamphetamine |
| 3 | POS | 2.2 amphetamine, 26.8 methamphetamine |
| 3 | POS | 2.6 amphetamine, 10.8 methamphetamine |
| 3 | POS | 2.5 amphetamine, 14.2 methamphetamine |
| 3 | POS | 2.5 amphetamine, 49.8 methamphetamine |
| 3 | POS | 2.7 amphetamine, 41.9 methamphetamine |
| 3 | POS | 2.8 amphetamine, 32.6 methamphetamine |
| 3 | POS | 1.7 amphetamine, 17.5 methamphetamine |
the state of the state of the states
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Cosmetic Treatments
Twenty amphetamine-negative head hair samples were treated with bleach, 20 with permanent wave, 20 with dye, 20 with relaxer, and 20 with shampoo, and the results compared to the same samples without the treatments. In each case of the 20 samples treated with a type of cosmetic treatment, 10 samples were treated with one brand of a particular product and 10 other samples with a second brand. No significant differences in EIA results were observed for the negative hair samples before and after the treatments; all samples remained negative after the treatments.
Twelve amphetamine-positive head hair samples were treated with bleach, permanent wave, dye, relaxer, and shampoo, and the results compared to the same samples without the treatments. In each case of samples treated with a type of cosmetic treatment, 6 samples were treated with one brand of a particular product and 6 other samples with a second brand. None of the samples became negative, by either EIA or LC/MS/MS, after treatment with any of the cosmetic products.
Summary of Cross-reactivity and Interference Studies
Chloramphetamine, MDA, PMA and Phentermine showed significant cross-reactivity in the amphetamine One-hundred-forty other compounds showed no cross-reactivity in the assay. One-hundred-EIA. nineteen compounds tested for interference at +/-50% of the cutoff showed no interference in the assay.
| Compound | PercentCross-reactivity* | Expected ConcentrationEquivalent to 3 ngAmphetamine/10 mg hair |
|---|---|---|
| MDA | 120 | 2.5 |
| d-amphetamine | 100 | 3.0 |
| PMA | 100 | 3.0 |
| Chloramphetamine | 79 | 3.8 |
| Phentermine | 17.6 | 17 |
| l-amphetamine | 1.1 | 270 |
| MDMA | 0.5 | 600 |
| PMMA | 0.5 | 600 |
| Phenylpropanolamine | < 0.3 | >1000 |
| Pseudoephedrine | < 0.3 | >1000 |
| IR, 2S Ephedrine | < 0.3 | >1000 |
| S,S Pseudoephedrine | < 0.3 | >1000 |
| Phenylethylamine | < 0.3 | >1000 |
| MDEA | < 0.3 | >1000 |
| L-methamphetamine | < 0.3 | >1000 |
| Ranitidine | < 0.3 | >1000 |
| Fenfluramine | < 0.3 | >1000 |
| Mephentermine | < 0.3 | >1000 |
| Phenmetrazine | < 0.3 | >1000 |
| Phendimetrazine | <0.3 | >1000 |
| Metanephrin | < 0.3 | >1000 |
Cross-reactivity of related Compounds in Amphetamine EIA
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Environmental Contamination
Aqueous Washing of Samples Soaked in Water and Saline Solutions of Amphetamine
Contamination of head hair samples by soaking in 500 ng amphetamine /mL of water resulted in a range of amphetamine on the hair of 7.9 - 225.8 ng of amphetamine /10 mg hair before washing. After washing by the procedure described below, the amount of oxycodone remaining on the hair samples from 0.6 to 76.3/10 mg hair, with 8 of the 11 samples appearing to be positive before application of the wash criterion. After application of the wash criterion, all of these samples containing amphetamine above the cutoff were determined to be contaminated (not positive).
Contamination of head hair samples by soaking in 500 ng amphetamine/mL of saline resulted in a range of amphetamine on the hair of 2.3 - 42.8 ng of amphetamine /10 mg hair before washing. by Psychemedics hair washing procedure (below), the amount of amphetamine remaining on the hair samples ranged from 0.5 to 9.5 ng/10 mg hair, with 10 of 11 samples below the cutoff even without application of the wash criterion. The one sample that was above the cutoff was determined to be negative by the wash criterion.
The Aqueous Buffer Wash Procedure
- a. Wash by Psychemedics' standard wash procedure:
- Add 2 mL of dry isopropanol and shake in waterbath for 15 minutes at 37℃ with shaking @ i. 100 -120 oscillations/minute. Remove isopropanol.
- ii. Add 2 mL of Wash Buffer (0.01 M phosphate buffer, pH 6.0, containing 0.1% BSA) and shake in waterbath for 30 minutes at 37℃ with shaking @ 100 -120 oscillations/minute. Remove Buffer.
- iii. Repeat Step ii. two more times.
- iv. Add 2 mL of Wash Buffer, and shake in waterbath for 60 minutes at 37℃ with shaking @ 100 -120 oscillations/minute. Remove Buffer.
- Repeat Step iv. one more time. Remove Buffer and save for analysis. Hair samples are now V. ready for digestion and extraction for LC/MS/MS analysis
4. Contamination Results for Amphetamine
Abbreviations used in the tables below: Hair Color: BLK, black; BRN, brown; LT, light; DK, dark; BLND, Blond. Ethnicity: As, Asian; Af-Am, African; H, Hispanic; Ca, Caucasian; Curvature: S, straight; C, curly. Contamination of Hair in 500 ng Amphetamine/mL Water-Aqueous Buffer Wash
| Sample# | Color | Curvature,Ethnicity | TotalDrugonHair* | Drug in HairafterWashing** | Drug inLastWash | Apply WashCriterion*** | POS/NEG | P, porous; VP,very porous; N,normal |
|---|---|---|---|---|---|---|---|---|
| ng amphetamine/10 mg hair | ||||||||
| 1 | MED BRN | S, Ca | 35.1 | 5.98 | 39.07 | -130.77 | NEG | VP |
| 2 | DK BRN | S, H | 225.8 | 76.36 | 253.38 | -810.47 | NEG | VP |
| 3 | BLK | S, As | 7.97 | 2.68 | 8.80 | NA | NEG | N |
| 4 | DK BRN | S, H | 19.35 | 3.64 | 18.75 | -61.99 | NEG | N |
| 5 | MED BRN | S, Ca | 13.03 | 3.52 | 14.34 | -46.67 | NEG | N |
| 6 | MED BRN | S, Ca | 26.66 | 3.11 | 21.10 | -70.74 | NEG | P |
| 7 | LT BRN | S, Ca | 23.58 | 5.36 | 21.49 | -69.86 | NEG | N |
| 8 | LT BRN | S, Ca | 30.82 | 5.65 | 37.30 | -124.90 | NEG | P |
| 9 | BLK | C, Af Am | 3.40 | 0.67 | 4.06 | NA | NEG | N |
| 10 | BLND | S, Ca | 2.71 | 0.75 | 1.96 | NA | NEG | N |
| 11 | BLND | S, Ca | 72.6 | 5.93 | 49.41 | -167.01 | NEG | P |
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*Total Amphetamine found by LC/MS/MS with unwashed hair samples.
** Wash Criterion: Hair - (3.5 x Last Wash); if Wash Criterion result is < 3.0, the result is due to contamination.
| Sample # | Color | Curvature,Ethnicity | TotalDrugonHair* | Drug in Hairafter Washing | Drug inLastWash | Apply WashCriterion** | POS/NEG | P, porous;VP, veryporous; N,normal |
|---|---|---|---|---|---|---|---|---|
| ng amphetamine/10 mg hair | ||||||||
| 1 | BRN | S, Ca | 2.85 | 0.50 | 2.77 | NA | NEG | P |
| 2 | MED BRN | S, Ca | 6.63 | 1.48 | 5.19 | NA | NEG | VP |
| 3 | BLK | S, As | 2.13 | 0.95 | 1.31 | NA | NEG | VP |
| 4 | DK BRN | S, H | 7.71 | 1.45 | 6.70 | NA | NEG | N |
| 5 | MED BRN | S, Ca | 3.38 | 1.34 | 2.69 | NA | NEG | N |
| 6 | MED BRN | S, Ca | 3.91 | 1.06 | 4.32 | NA | NEG | P |
| 7 | LT BRN | S, Ca | 6.65 | 2.09 | 9.61 | NA | NEG | P |
| 8 | BLND | S, Ca | 22.81 | 9.51 | 20.56 | -62.45 | NEG | N |
| 9 | BLK | C, AfAm | 2.38 | 0.69 | 2.63 | NA | NEG | N |
| 10 | BLND | S, Ca | 3.22 | 0.76 | 3.0 | NA | NEG | N |
| 11 | BLND | S, Ca | 10.36 | 1.36 | 6.79 | NA | NEG | P |
Contamination of Hair in 500 ng Amphetamine/mI . Saline .- Aqueous Buffer Wash
*Total Amphetamine found by LC/MS/MS with unwashed hair samples.
**Wash Criterion: Hair - (3.5 x Last Wash); if Wash Criterion result is < 3.0, the result is due to contamination.
An Alternative Wash Procedure
Studies were performed on an alternative washing method for porous samples that may lose some drug from ingestion when subjected to aqueous wash procedures. The alternative wash procedure for porous samples consists of a series of 90% ethanol washes: three 30-minute washes and two 60-minute washes. The last wash is analyzed and the value obtained is used to determine a wash criterion. The wash criterion is the multiplication of the last wash drug content by 3.5 and subtraction of this value from the drug content of the hair. If the result of the subtraction falls below the cutoff, the sample is interpreted as negative or contaminated, not positive.
For 10 head hair samples soaked in a water solution of 500 ng/mL amphetamine, after washing by the alternative ethanol procedure, the amount of amphetamine remaining on the hair samples ranged from 2.2 to 140.1 ng/10 mg hair, with 9 of the 10 samples appearing to be positive before application of the wash criterion. After application of the wash criterion, all of these samples containing amphetamine above the cutoff were determined to be contaminated (not positive).
For 13 head hair samples soaked in a saline solution of 500 ng/mL amphetamine, after washing by the alternative ethanol procedure, the amount of amphetamine remaining on the hair samples ranged from 0.9 to 14.7 ng/10 mg hair, with 8 of 13 samples below the cutoff even without application of the wash criterion. The 5 samples above the cutoff were determined to be negative by the wash criterion.
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| Sample# | Color | Curvature,Ethnicity | TotalDrugonHair* | Drug in Hairafter Washing | Drug inLastWash | Apply WashCriterion** | POS/NEG | P,porous;VP, veryporous;N,normal |
|---|---|---|---|---|---|---|---|---|
| ng amphetamine/10 mg hair | ||||||||
| 1 | BRN | S, Ca | 2.85 | 1.43 | 0.33 | NA | NEG | P |
| 2 | MED BRN | S, Ca | 6.63 | 3.44 | 1.47 | -1.71 | NEG | VP |
| 3 | DK BRN | S, H | 13.67 | 8.24 | 1.86 | 1.73 | NEG | VP |
| 4 | BLK | S, As | 2.13 | 0.93 | 0.22 | NA | NEG | N |
| 5 | DK BRN | S, H | 7.71 | 3.58 | 0.80 | 0.78 | NEG | N |
| 6 | MED BRN | S, Ca | 3.38 | 2.29 | 0.25 | NA | NEG | N |
| 7 | MED BRN | S, Ca | 3.91 | 2.42 | 0.65 | NA | NEG | P |
| 8 | LT BRN | S, Ca | 1.91 | 1.16 | 0.29 | NA | NEG | N |
| 9 | LT BRN | S, Ca | 6.65 | 2.45 | 1.81 | NA | NEG | P |
| 10 | BLND | S, Ca | 22.81 | 14.77 | 5.31 | -3.82 | NEG | N |
| 11 | Af-Am | C, AfAm | 2.38 | 1.45 | 0.31 | NA | NEG | N |
| 12 | BLND | S, Ca | 3.22 | 0.94 | 0.57 | NA | NEG | N |
| 13 | BLND | S, Ca | 10.36 | 3.45 | 1.72 | -2.57 | NEG | P |
Contamination of Hair in 500 ng Amphetamine/mL Water—90% Ethanol Wash
:
*Total Amphetamine found by LC/MS/MS with unwashed hair samples.
**Wash Criterion: Hair – (3.5 x Last Wash); if Wash Criterion result is < 3.0, the result is interpreted as possibly due to external contamination with amphetamine.
| Sample# | Color | Curvature,Ethnicity | TotalDrugon Hair* | Drug in Hairafter Washing | Drug inLastWash | Apply WashCriterion** | POS/NEG | P,porous;VP, veryporous;N,normal | |
|---|---|---|---|---|---|---|---|---|---|
| ng amphetamine/10 mg hair | |||||||||
| 1 | BRN | S, Ca | 2.85 | 1.43 | 0.33 | NA | NEG | P | |
| 2 | MED BRN | S, Ca | 6.63 | 3.44 | 1.47 | -1.71 | NEG | VP | |
| 3 | DK BRN | S, H | 13.67 | 8.24 | 1.86 | 1.73 | NEG | VP | |
| 4 | BLK | S, As | 2.13 | 0.93 | 0.22 | NA | NEG | N | |
| 5 | DK BRN | S, H | 7.71 | 3.58 | 0.80 | 0.78 | NEG | N | |
| 6 | MED BRN | S, Ca | 3.38 | 2.29 | 0.25 | NA | NEG | N | |
| 7 | MED BRN | S, Ca | 3.91 | 2.42 | 0.65 | NA | NEG | P | |
| 8 | LT BRN | S, Ca | 1.91 | 1.16 | 0.29 | NA | NEG | N | |
| 9 | LT BRN | S, Ca | 6.65 | 2.45 | 1.81 | NA | NEG | P | |
| 10 | BLND | S, Ca | 22.81 | 14.77 | 5.31 | -3.82 | NEG | N | |
| 11 | Af-Am | C, AfAm | 2.38 | 1.45 | 0.31 | NA | NEG | N | |
| 12 | BLND | S, Ca | 3.22 | 0.94 | 0.57 | NA | NEG | N | |
| 13 | BLND | S, Ca | 10.36 | 3.45 | 1.72 | -2.57 | NEG | P |
Contamination of Hair in 500 ng Amphetamine/mL Saline-90% Ethanol Wash
*Total Amphetamine found by LC/MS/MS with unwashed hair samples.
:
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** Wash Criterion: Hair - (3.5 x Last Wash); if Wash Criterion result is < 3.0, the result is interpreted as possibly due to external contamination with amphetamine.
Storage and Shipping Stability
Twenty-one amphetamine-positive head hair samples were tested before and after 1 year of storage under ambient conditions. The average of the results after one year was 109% of the average of the original results. Twenty-one amphetamine-positive head hair samples were also tested before and after two coast-to-coast shippings. The samples after shipping had average results of 105% of the results of the samples before shipping.
Stability of Calibrator and Control Solutions
Psychemedics manufactures calibrators and control materials using drug stocks purchased from a commercial vendor. Each lot of drug is received with its specific certificate of analysis. The commercially obtained stock is made into the calibrators and controls to the desired concentrations. The concentrations are confirmed by LC/MS/MS. Stability of the amphetamine calibrator and control solutions was shown to be 9 months, with ongoing studies to demonstrate 1-year stability.
Recovery
Recovery of amphetamine from hair in a 2-hour incubation ranged from 100 to 110%. This was determined by LC/MS/MS measurements of extended sequential hair extractions.
Conclusion:
Comparison of results of the Psychemedics Microplate EIA for Amphetamine in Hair with LC/MS/MS confirmation showed the results to be substantially equivalent. The Psychemedics Microplate EIA for Amphetamine in Hair is substantially equivalent to the predicate, based on acceptable performance studies, including precision, specificity, interference (including cosmetic effects), and removal of external contamination.
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Image /page/9/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle with its wings spread, and the words "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" are arranged in a circular pattern around the eagle. The logo is black and white.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
May 2, 2013
Psychemedics Corp. C/O Virginia Hill 5832 Uplander Way CULVER CITY CA 90230
Re: K130811
Trade/Device Name: Psychemedics Microplate EIA for Amphetamine in Hair Regulation Number: 21 CFR 862.3100 Regulation Name: Amphetamine test system Regulatory Class: II Product Code: DKZ Dated: March 22, 2013 Received: March 25, 2013
Dear Ms. Hill:
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. 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 requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please go to http://www.fda.gov/AboutFDA/CentersOffices/CDRH/CDRHOffices/ucm115809.htm for
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the Center for Devices and Radiological Health's (CDRH's) Office of Compliance. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportalProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours,
Carol Benson -S for
Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology-Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known): K130811
Device Name: Psychemedics Microplate EIA for Amphetamine in Hair
Indications for Use:
The Psychemedics Microplate EIA for Amphetamine is an enzyme immunoassay (EIA) for the preliminary qualitative detection of amphetamine in human head and body hair using an amphetamine calibrator at 3 ng /10 mg hair cutoff for the purpose of identifying amphetamine use. This is an in vitro diagnostic device intended exclusively for Psychemedics use only. Psychemedics has not performed an evaluation of reproducibility at different laboratories.
The Psychemedics Microplate EIA amphetamine assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas or Liquid Chromatography/Mass Spectrometry (GC/MS or LC/MS/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.
Psychemedics plans to perform this test at one site. Psychemedics has not performed an evaluation of reproducibility at different sites.
Prescription Use (21 CFR Part 801 Subpart D) And/Or
Over the Counter Use X (21 CFR Part 801 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED)
Concurrence of CDRH, Office of In Vitro Diagnostics and Radiological Health (OIR)
Denise Johnson=byless=5
2013.05.01 15:53:34:00:00
Division Sign-Off Office of In Vitro Diagnostics and Radiological Health
.510(k) . k13
510(k)
§ 862.3100 Amphetamine test system.
(a)
Identification. An amphetamine test system is a device intended to measure amphetamine, a central nervous system stimulating drug, in plasma and urine. Measurements obtained by this device are used in the diagnosis and treatment of amphetamine use or overdose and in monitoring levels of amphetamine to ensure appropriate therapy.(b)
Classification. Class II (special controls). An amphetamine 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).