K Number
K141928
Date Cleared
2014-11-25

(132 days)

Product Code
Regulation Number
862.3200
Reference & Predicate Devices
Predicate For
N/A
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP AuthorizedThirdpartyExpeditedreview
Intended Use

cobas c Acetaminophen Gen.2 assay:
The cobas c Acetaminophen Gen.2 assay is an in vitro diagnostic test for the quantitative determination of acetaminophen in serum and plasma for use in the diagnosis of acetaminophen overdose in serum and plasma on Roche/Hitachi cobas c systems.

ACET2 calibrator:
The ACET2 calibrator is for use in the calibration of the Acetaminophen Gen.2 Roche assay.

Device Description

The cobas c Acetaminophen Gen.2 assay is based on a homogeneous enzyme immunoassay technique used for the quantitative analysis of acetaminophen in human serum and plasma.
Reagents are packaged in a cassette labeled with their instrument positioning R1 (Reagent 1) and R2 (Reagent 2).
R1 contains anti-acetaminophen antibody (sheep polyclonal), G6P, NAD, bovine serum albumin, preservatives and stabilizers.
R2 contains acetaminophen labeled with bacterial G6PDH, Tris buffer, preservatives, bovine serum albumin, and stabilizers.
The ACET2 calibrator contains a known quantity of acetaminophen. The cobas c 501 analyzer dilutes the ACET2 calibrator on-board the analyzer with NaCl diluent, in order to create five concentration levels, and level 1 is water. This results in a six-level calibrator set, and the calibrator set is then used to establish a standard curve. The ACET2 calibrator contains acetaminophen, phosphate buffer, and preservatives.

AI/ML Overview

The provided document describes the analytical performance of the cobas c Acetaminophen Gen.2 assay and ACET2 Calibrator. It outlines various studies conducted to demonstrate the device's characteristics, which can serve as acceptance criteria.

Here's the requested information:

1. A table of acceptance criteria and the reported device performance

Acceptance CriteriaReported Device Performance
Precision (Repeatability)
Control 1 SD (µg/mL)0.4 µg/mL
Control 2 SD (µg/mL)0.9 µg/mL
Control 3 SD (µg/mL)2 µg/mL
Human Serum 1 SD (µg/mL)0.2 µg/mL
Human Serum 2 SD (µg/mL)1.7 µg/mL
Human Serum 3 SD (µg/mL)4 µg/mL
Human Serum 4 SD (µg/mL)4 µg/mL
Human Serum 5 SD (µg/mL)4 µg/mL
Precision (Intermediate Precision)
Control 1 SD (µg/mL)0.5 µg/mL
Control 2 SD (µg/mL)1.0 µg/mL
Control 3 SD (µg/mL)3 µg/mL
Human Serum 1 SD (µg/mL)0.3 µg/mL
Human Serum 2 SD (µg/mL)1.9 µg/mL
Human Serum 3 SD (µg/mL)4 µg/mL
Human Serum 4 SD (µg/mL)5 µg/mL
Human Serum 5 SD (µg/mL)6 µg/mL
Method Comparison to Predicate (Deming Regression Weighted)
Slope (y = mx + b)1.02
Y-intercept (y = mx + b)-0.699 µg/mL
Correlation Coefficient (r)0.997
Linearity (Serum)
Linear Regression Equation (y = mx + b)y = 1.014x - 0.248
Pearson correlation coefficient (R)0.998963
In-range percentage recovery (between 5 and 200 µg/mL)94% - 98% (for spiked concentrations of 24-240 µg/mL or more broadly, 92-98% for plasma and 94-98% for serum)
Linearity (Plasma)
Linear Regression Equation (y = mx + b)y = 1.011x - 0.193
Pearson correlation coefficient (R)0.999010
In-range percentage recovery (between 5 and 200 µg/mL)90% - 95% (for spiked concentrations of 24-240 µg/mL)
Detection Limits
Limit of Blank (LoB)1.5 µg/mL
Limit of Detection (LoD)3 µg/mL
Limit of Quantitation (LoQ)5 µg/mL
Analytical Specificity - Cross Reactivity
% cross reactivity for listed compounds (e.g., Acetaminophen cysteine, glucuronide, mercapturate, sulfate, Cysteine, N-Acetylcysteine, Phenacitin)Generally low, with most noted as "not detectable" or under 1%
Analytical Specificity - Endogenous Substances (Interference)
Lipemia (L index)No significant interference up to an L index of 400 (corresponds to ~600-672 L index reported depending on acetaminophen concentration)
Hemolysis (H index)No significant interference up to an H index of 800 (actual reported values are around 926-1025 H index)
Bilirubin (I index)No significant interference up to an I index of 30 for conjugated and unconjugated bilirubin (actual reported values are around 42-63 I index)
Recovery for endogenous interferenceRecovery within ± 1 µg/mL for ~5 µg/mL acetaminophen; recovery within ± 10% for ~30 µg/mL acetaminophen
Analytical Specificity - Common Drugs (Interference)
Difference in recovery of acetaminophen≤ 10 µg/mL: ≤ ± 1 µg/mL; > 10 µg/mL: 100 ± 10 %
Matrix Comparison (Anticoagulants)
For samples ≤ 10 µg/mL, deviation from serum recovery≤ ± 1 µg/mL
For samples > 10 µg/mL, deviation from serum recovery≤ ± 10%
Serum vs. Li-heparin (Passing/Bablok)$y = 0.989x + 0.089$, r = 0.998
Serum vs. K2-EDTA (Passing/Bablok)$y = 1.000x – 0.000$, r = 0.998
Serum vs. K3-EDTA (Passing/Bablok)$y = 0.992x - 0.163$, r = 0.998

2. Sample sized used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective)

  • Precision (Repeatability and Intermediate Precision): The study was conducted according to CLSI EP5-A2 requirements using human samples and controls. "n = 84" is mentioned for repeatability. "2 aliquots per run, 2 runs per day, 21 days" for intermediate precision. The document does not specify the country of origin or whether the data was retrospective or prospective.
  • Method Comparison to Predicate: Sample size (n) = 105 human serum samples. The document does not specify the country of origin or whether the data was retrospective or prospective.
  • Linearity: One batch of reagent, one run, samples measured in triplicate. Two separate dilution series (serum and Li-Heparin plasma) with thirteen levels each. The document does not specify the country of origin or whether the data was retrospective or prospective.
  • Detection Limits (LoB, LoD, LoQ):
    • LoB: One analyte-free sample tested in n=5 on two analyzers with three reagent batches for six runs per day across three days.
    • LoD: Five low-analyte samples spiked with acetaminophen measured in singlicate on two analyzers with three reagent batches for six runs per day across three days.
    • LoQ: A low-level sample set of six measured in two aliquots using three reagent batches on one analyzer over at least six days.
      The document does not specify the country of origin or whether the data was retrospective or prospective.
  • Analytical Specificity (Cross-Reactivity, Endogenous Substances, Common Drugs): Tested using sample pools (at two target concentrations of acetaminophen) and spiked samples. The specific number of individual samples for each compound/substance is not given, but refers to "two sample pools" or "serum sample pools". The document does not specify the country of origin or whether the data was retrospective or prospective.
  • Matrix Comparison: 60 tubes collected per anticoagulant (Lithium-heparin, K2-EDTA, K3-EDTA). This suggests 60 samples for each, so 180 total samples. The document does not specify the country of origin or whether the data was retrospective or prospective.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience)

This product is an in vitro diagnostic test for the quantitative determination of acetaminophen. The "ground truth" for such assays is typically established by reference methods or spiking known concentrations of analyte into samples. The document does not mention the involvement of medical experts (like radiologists) for ground truth establishment, as it's a quantitative chemical assay, not an image-based diagnostic. It uses USP reference standards for traceability, which represents a highly controlled and recognized standard for chemical concentration.

4. Adjudication method (e.g. 2+1, 3+1, none) for the test set

Not applicable. Adjudication methods like "2+1" are typically used in studies involving human interpretation (e.g., radiology for diagnostic accuracy) where there might be disagreement among readers, and a third reader is brought in to resolve discrepancies. This document describes the performance of a quantitative chemical assay, where the output is a numerical value.

5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

Not applicable. This is not an AI-assisted diagnostic device, nor does it involve human readers interpreting cases. It is a standalone in vitro diagnostic assay.

6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done

Yes, this is a standalone device. The entire document describes the performance of the assay itself (cobas c Acetaminophen Gen.2 assay and ACET2 calibrator) on the Roche/Hitachi cobas c 501 analyzer, which processes samples and provides quantitative results without human interpretation or algorithm assistance as part of the primary diagnostic function.

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)

The ground truth for this chemical assay is primarily based on:

  • Known concentrations of analyte: For linearity, detection limits (LoD, LoQ), cross-reactivity, and interference studies, samples were spiked with known concentrations of acetaminophen or interfering substances.
  • Reference Methods / Standards: For traceability, the method has been standardized against USP reference standards.
  • Comparative Method: The performance was compared to a legally marketed predicate device (Siemens Emit® tox™ Acetaminophen Assay).

8. The sample size for the training set

This document describes the analytical validation of a chemical assay. The concept of a "training set" is typically associated with machine learning or AI models, where data is used to train an algorithm. This document details the performance testing of the device, not the development of a machine learning model. Therefore, a "training set" in the context of an AI algorithm is not applicable here.

9. How the ground truth for the training set was established

As explained above, there is no "training set" in the AI/machine learning sense for this type of in vitro diagnostic device. The analytical characteristics are established through various experiments using samples with known properties or references.

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Image /page/0/Picture/1 description: The image shows the seal of the Department of Health & Human Services (HHS) of the United States. The seal features a stylized caduceus-like symbol with three human profiles forming the body of the symbol. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the symbol.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

November 25, 2014

ROCHE DIAGNOSTICS OPERATIONS NOEL MENCIAS REGULATORY AFFAIRS PRINCIPAL 9115 HAGUE ROAD INDIANAPOLIS IN 46250

Re: K141928

Trade/Device Name: cobas c Acetaminophen Gen.2 Assay ACET2 Calibrator Regulation Number: 21 CFR 862.3030 Regulation Name: Acetaminophen test system Regulatory Class: II Product Code: LDP, DKB Dated: October 10, 2014 Received: October 14, 2014

Dear Mr. Noel Mencias:

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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education 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. 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

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/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 Industry and Consumer Education 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,

Courtney

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) K141928

Device Name cobas c Acetaminophen Gen.2 assay ACET2 Calibrator

Indications for Use (Describe)

cobas c Acetaminophen Gen.2 assay:

The cobas c Acetaminophen Gen.2 assay is an in vitro diagnostic test for the quantitative determination of acetaminophen in serum and plasma for use in the diagnosis of acetaminophen overdose in serum and plasma on Roche/Hitachi cobas c systems.

ACET2 calibrator:

The ACET2 calibrator is for use in the calibration of the Acetaminophen Gen.2 Roche assay.

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)

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Date prepared:November 18, 2014
Purpose ofsubmissionIn accordance with 21 CFR 807.87, Roche Diagnostics hereby submitsofficial notification as required by Section 510(k) of the Federal Food, Drugand Cosmetics Act of our intention to market the device described in thisPremarket Notification [510(k)].
The purpose of this premarket notification is to obtain FDA review andclearance for the cobas c Acetaminophen Gen.2 assay and Acetaminophencalibrator.
MeasurandAcetaminophen
Type of testQuantitative homogeneous enzyme immunoassay method
ApplicantNoel Mencias, Regulatory Affairs ConsultantRoche Diagnostics9115 South Hague RoadIndianapolis, IN 46250Telephone: (317) 521-3172Fax: (317) 521-2324Email: noel.mencias@roche.com
Candidatedevice namesProprietary name: cobas c Acetaminophen Gen.2 assayCommon name: Acetaminophen Gen.2 assayShort name: ACET2
Proprietary name: ACET2 calibratorCommon name: Acetaminophen calibratorShort name: ACETC
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RegulatorvProduct CodeClassificationRegulationPanel
informationLDPClass II21 CFR 862.3030(Acetaminophen testsystem)Toxicology
DKBClass II21 CFR 862.3200 (Clinicaltoxicology calibrator)Toxicology
Intended useand plasma on Roche/Hitachi cobas c systems.Acetaminophen Gen.2 Roche assay.The cobas c Acetaminophen assay is intended for use as an vitro diagnostictest for the quantitative determination of acetaminophen overdose in serumThe ACET2 calibrator is intended for use in the calibration of the
Indications foruseThe cobas c Acetaminophen Gen.2 assay is intended for use as an in vitro testfor the quantitative determination of acetaminophen overdose in serum andplasma on Roche/Hitachi cobas c systems. Acetaminophen is a widely usedanalgesic and antipyretic found in a number of over-the-counter andprescription products. When consumed in overdose quantities, acetaminophenmay cause severe liver and kidney damage, or death. The patient may havefew or no symptoms early after acute overdose of acetaminophen. The onlyreliable early diagnostic indicator is provided by a quantitative measurementof the serum acetaminophen level.The ACET2 calibrator is intended for use in the calibration of theAcetaminophen Gen.2 Roche assay. The calibrator is prepared to contain aknown quantity of acetaminophen in buffer. The calibrator is used toestablish a standard curve from which the quantity of acetaminophen inunknown specimens can be determined.
Specialconditions foruseFor prescription use only
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Special instrument requirementsFor use on the Roche/Hitachi cobas c clinical chemistry analyzer
Candidate device descriptionThe cobas c Acetaminophen Gen.2 assay is based on a homogeneous enzyme immunoassay technique used for the quantitative analysis of acetaminophen in human serum and plasma.Reagents are packaged in a cassette labeled with their instrument positioning R1 (Reagent 1) and R2 (Reagent 2). R1 contains anti-acetaminophen antibody (sheep polyclonal), G6P, NAD, bovine serum albumin, preservatives and stabilizers.R2 contains acetaminophen labeled with bacterial G6PDH, Tris buffer, preservatives, bovine serum albumin, and stabilizers. The ACET2 calibrator contains a known quantity of acetaminophen. The cobas c 501 analyzer dilutes the ACET2 calibrator on-board the analyzer with NaCl diluent, in order to create five concentration levels, and level 1 is water. This results in a six-level calibrator set, and the calibrator set is then used to establish a standard curve. The ACET2 calibrator contains acetaminophen, phosphate buffer, and preservatives.
Predicate deviceRoche Diagnostics claims substantial equivalence to the Siemens Emit ® tox™ Acetaminophen Assay and the Siemens Emit ® tox™ Acetaminophen Calibrators which were both cleared in K002974.

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System, Continued

Substantial The following table compares the similar features of the candidate device to Equivalence – the predicate device that was cleared in 510(k) K002974. Assay Similarities

Assay Comparison Similarities
FeaturePredicate Device:Emit® tox™ Acetaminophen AssayCandidate Device:Acetaminophen Gen.2 Assay
Intended UseThe Emit® tox™ Acetaminophen Assay is a homogeneous enzyme immunoassay intendedfor in vitro diagnostic use in thequantitative analysis ofacetaminophen in human serum orplasma.The cobas c Acetaminophen Gen.2 assay is intended foruse as an in vitro test for thequantitative determination oftoxic levels of acetaminophenin serum and plasma onRoche/Hitachi cobas csystems.
ReagentCompositionR1: sheep antibodies reactive toacetaminophen, G6P NAD, bovineserum albumin, preservatives, andstabilizersR2: acetaminophen labeled withbacterial G6PDH, Tris buffer,preservatives, bovine serumalbumin, and stabilizersSame
Reagent Shelf LifeStability2-8 °C until expiration dateSame
Test PrincipleHomogeneous enzymeimmunoassaySame
Measuring RangeUp to 200 µg/mL (1324 µmol/L)5-200 µg/mL (33.1-1324µmol/L)
TraceabilityThis method has been standardizedagainst USP reference standards.Same
Expected ValuesNormal therapeutic doses ofacetaminophen result in serumconcentrations of 10-30 µg/mL(66-199 µmol/L) in healthy adults.Same
Assay Comparison Differences
FeaturePredicate Device:Emit® tox™ AcetaminophenAssayCandidate Device:Acetaminophen Gen.2 Assay
Reagent On-BoardStabilityAnalyzer specificon-board in use and refrigeratedon the analyzer: 12 weeks
Sample TypesEDTA, heparin, citrate andoxalate/fluorideK2- or K3-EDTA, or lithiumheparinized plasma
ControlsCommercially available controlsTDM Control Set Level ITDM Control Set Level IITDM Control Set Level III
Instrument PlatformSeveral analyzersRoche/Hitachi cobas c 501
Lower Limits ofMeasurementThe sensitivity level of theEmit® tox™ AcetaminophenAssay is less than 0.25 µg/mLacetaminophen. This levelrepresents the lowestconcentration of acetaminophenthat can be distinguished from 0µg/mL with a confidence level of95%.LoB = 1.5 µg/mLLoD = 3 µg/mLLoQ = 5 µg/mL

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System, Continued

The following table compares the differences of the candidate device to the Substantial Equivalence – predicate device that was cleared in 510(k) K002974. Assay Differences

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System, Continued

Substantial The following table compares the similar features of the candidate device to equivalence – the predicate device that was cleared in 510(k) K002974. calibrator similarities

Calibrator Comparison Similarities
FeaturePredicate Device:Emit® tox™ AcetaminophenCalibratorsCandidate Device:ACET2 Calibrator
Intended UseThe Emit® tox™ AcetaminophenCalibrators are intended for use withthe Emit® tox™ AcetaminophenAssay.ACET2 calibrator is for use inthe calibration of theAcetaminophen Gen.2 Rocheassay.
FormatLiquid ready-to-useSame
AnalyteAcetaminophenSame
Storage Conditions2-8°CSame
Storage and StabilityWhen stored refrigerated at 2-8°C,the Emit® tox™ AcetaminophenCalibrators are stable untilthe expiration date printed on thedropper vial label.Store at 2-8 °C. Do notfreeze.The calibrator may be useduntil the expiration date whenstored at 2-8°C.

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System, Continued

The following table compares the difference of the candidate device to the Substantial equivalence – predicate device that was cleared in 510(k) K002974. calibrator difference

Calibrator Comparison Difference
FeaturePredicate Device:Emit® tox™ AcetaminophenCalibratorsCandidate Device:ACET2 Calibrator
QuantityEmit® tox™ AcetaminophenCalibrators are a six-level set thatcontain the followingacetaminophen concentrations: 0,10, 25, 50, 100, 200 µg/mL.The ACET2 calibrator has anominal value of 200 µg/mL ofacetaminophen. The analyzerdilutes the ACET2 calibratoron-board with NaCl diluent, inorder to create fiveconcentration levels, and level1 is water. This results in a six-level calibrator set: 0.0, 10.0,30.2, 75.0, 100, 200 µg/mL.

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Test principleThe assay is based on a homogeneous enzyme immunoassay techniqueused for the quantitative analysis of acetaminophen in human serum orplasma. The assay is based on competition between drug in the sampleand drug labeled with the enzyme glucose-6-phosphate dehydrogenase(G6PDH) for antibody binding sites. Enzyme activity decreases uponbinding to the antibody, so the drug concentration in the sample can bemeasured in terms of enzyme activity. Active enzyme converts oxidizednicotinamide adenine dinucleotide (NAD) to NADH, resulting in anabsorbance change that is measured spectrophotometrically. Endogenousserum G6PDH does not interfere because the coenzyme functions only withthe bacterial (Leuconostoc mesenteroides) enzyme employed in the assay.
PrecisionPrecision was determined using human samples and controls inaccordance with the CLSI EP5-A2 requirements with repeatability (n = 84)and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). Thefollowing results were obtained on the Roche/Hitachi cobas c 501.
Repeatability Summary
SpecimenMean (µg/mL)SD (µg/mL)CV (%)
Control 115.30.42.5
Control 234.90.92.5
Control 310622.2
Human Serum 17.70.22.9
Human Serum 273.21.72.3
Human Serum 313042.7
Human Serum 416842.5
Human Serum 518442.3
Intermediate Precision Summary
SpecimenMean (µg/mL)SD (µg/mL)CV (%)
Control 115.30.53.2
Control 234.91.02.8
Control 310633.0
Human Serum 17.40.33.5
Human Serum 273.21.92.7
Human Serum 313043.2
Human Serum 416853.2

185

б

Human Serum 5

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3.0

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MethodComparison toPredicateDeviceAcetaminophen values for human serum samples obtained onRoche/Hitachi cobas c 501 analyzer (y) were compared to those determinedwith the Emit® tox™ Acetaminophen assay reagent on Olympus AU5400analyzer (x). The sample concentrations were between 5.2 and 198 µg/mL,and they were tested in singlicate.Sample size (n) = 105Deming Regression Weighted$y = 1.02x - 0.699 \frac{\text{µg}}{\text{mL}}$$r = 0.997$
LinearityLinearity was assessed according to CLSI EP6-A with one batch of reagent,in one run, and with samples measured in triplicate. Two separate dilutionseries differing by sample type (serum and Li-Heparin plasma) were preparedwith thirteen levels.The diluted samples span the measuring range including a non-zero samplebelow the measuring range and a sample above the measuring range.The linearity results support the measuring range of 5.0-200 µg/mL.Linear Regression Equation for Serum:$y = 1.014x - 0.248$ Pearson correlation coefficient (R) = 0.998963Linear Regression Equation for Plasma:$y = 1.011x - 0.193$ Pearson correlation coefficient (R) = 0.999010

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LinearitySampleSpikedconc.(µg/mL)c 501 mean(µg/mL)Absolutedeviation(µg/mL)% recovery
1000-
232.4-0.6-
365.4-0.6-
42423.1-96
54845.7-95
67268.4-95
79692.1-96
8120113-94
9144139-97
10168160-95
11192188-98
12216206-95
13240232-97

Linearity (continued)

Calculation to spiked concentration levels: I inearity Statistic ith S

Linearity Statistics with Plasma

LinearitySampleSpikedconc.(µg/mL)c 501 mean(µg/mL)Absolutedeviation(µg/mL)% recovery
1000-
232.4-0.6-
365.1-0.9-
42421.7-90
54844.3-92
67265.4-91
79689.0-93
8120110-92
9144132-92
10168155-92
11192180-94
12216194-90
13240228-95

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Detection limitLoB, LoD, and LoQ studies were performed based upon CLSI EP17-A2.
LoB Protocol: One analyte-free sample was tested in n=5 with two analyzerswith three reagent batches for six runs per day across three days.
LoD Protocol: Five low-analyte samples spiked with acetaminophen weremeasured in singlicate on two analyzers with three reagent batches for sixruns per day across three days.
LoQ Protocol: A low-level sample set of six was measured in two aliquotsusing three reagent batches on one analyzer over at least six days. The LoQ isdetermined based on a total error of 20%.
The LoB, LoD, and LoQ claims represent the specifications for each.LoB claim = 1.5 µg/mLLoD claim = 3 ug/mLLoQ claim = 5 ug/mL
Analyticalspecificity-cross reactivityTwo sample pools at two target concentrations of acetaminophen, one at a lowconcentration of ~5.0 µg/mL and the second one at a high concentration of ~ 30.0µg/mL are used.The absolute bias is calculated as follows:Total Bias = Mean concentration of test sample - Mean concentration ofcontrol sampleThe cross reactivity is calculated as follows:י% cross reactivity =( (Total Bias) / (concentration of test substance) ) x 100
CompoundCompoundConcentration(ug/mL)Acetaminophen(µg/mL)% crossreactivity
Acetaminophen cysteine1006.10.5
Acetaminophen glucuronide10005.2n.d.*
Acetaminophen mercapturate3005.40.2
Acetaminophen sulfate2006.1n.d.*
Cysteine13005.8n.d.*
N-Acetylcysteine16636.3n.d.*
Phenacitin5006.70.5

*n.d. = not detectable

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Analytical
specificity-cross reactivity,
continued
CompoundCompoundConcentration(µg/mL)Acetaminophen(µg/mL)% crossreactivity
Acetaminophen cysteine10029.2-0.3
Acetaminophen glucuronide100025.4-0.1
Acetaminophen mercapturate30025.90.2
Acetaminophen sulfate20027.80.1
Cysteine130029.0n.d.*
N-Acetylcysteine166328.5n.d.*
Phenacitin50029.31.3

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Analytical specificity interference from endogenous substances

The reagent was evaluated with three endogenous substances, hemoglobin, lipids, and bilirubin for potential interference with the measurement of acetaminophen.

Data Summary

ApproximateAcetaminophenConcentration(µg/mL)no interferenceup toTriglyceridetheoreticalvalue (mg/dL)Claim as it appears in thelabeling.
Lipemia Level 15672 L index1100No significant interferenceup to an L index of 400.
Lipemia Level 230608 L index997
Lipemia Level 350492 L index810
Lipemia Level 4100418 L index698
Lipemia Level 5150413 L index682
Lipemia Level 6180411 L index678
Hemolysis Level 15926 H indexN/ANo significant interferenceup to an H index of 800.
Hemolysis Level 230936 H indexN/A
Hemolysis Level 3501009 H indexN/A
Hemolysis Level 4901008 H indexN/A
Hemolysis Level 51401016 H indexN/A
Hemolysis Level 61801025 H indexN/A
UnconjugatedBilirubin Level 1547 I indexN/A
UnconjugatedBilirubin Level 23045 I indexN/A
UnconjugatedBilirubin Level 35063 I indexN/A
UnconjugatedBilirubin Level 410063 I indexN/ANo significant interferenceup to an I index of 30 forconjugated andunconjugated bilirubin.
UnconjugatedBilirubin Level 515063 I indexN/A
UnconjugatedBilirubin Level 618063 I indexN/A
Conjugated BilirubinLevel 1545 I indexN/A
Conjugated BilirubinLevel 23046 I indexN/A
Conjugated BilirubinLevel 35044 I indexN/A
Conjugated BilirubinLevel 410047 I indexN/A
Conjugated BilirubinLevel 515050 I indexN/A
Conjugated BilirubinLevel 618042 I indexN/A

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Analyticalspecificity –interferencefromendogenoussubstances(continued)The I Index value corresponds approximately to mg/dL bilirubin.The H Index value corresponds approximately to mg/dL hemoglobin.The Triglyceride theoretical value is the calculated correlation of L index totriglyceride concentration.
All data passed the following acceptance criteria:Recovery within ± 1 µg/mL of initial value at an acetaminophen level ofapproximately 5 µg/mL and recovery within ± 10% of initial value at anacetaminophen level of approximately 30 µg/mL.

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Analytical specificityinterference from common drugs

Twenty four commonly used drugs were examined for potential interference on measurement with cobas c Acetaminophen Gen.2 assay. Drug interference testing was performed with serum sample pools at two target concentrations of acetaminophen (~ 5 and ~ 30 µg/mL). Acetaminophen concentration in all aliquots is measured in triplicate. The mean value among the triplicates for each aliquot is determined, and the percent recovery to the initial value (no drug in sample) is calculated.

DrugHighest Concentration ShownNot to Interfere with ACET2
Acetylcystein150 mg/L
Acetylsalycilic acid1000 mg/L
Ampicillin-sodium1000 mg/L
Ascorbic acid300 mg/L
Cefoxitin2500 mg/L
Cyclosporine5 mg/L
Doxycycline50 mg/L
Phenylbutazone400 mg/L
Rifampicin64 mg/L
Theophylline100 mg/L
Amitriptylline277 µg/mL
Caffeine1000 µg/mL
Codeine1.6 µg/mL
Diazepam5.1 µg/mL
Heparin5000 U
Ibuprofen500 mg/L
Levodopa20 mg/L
Methyldopa + 1.5 H2O20 mg/L
Metronidazole200 mg/L
Methionine1000 µg/mL
Phenylephrine20 µg/mL
Propoxyphene20 µg/mL
Salicylate1000 µg/mL
Secobarbital22 µg/mL

The table below summarizes the common drug interferences data:

All data passed the following acceptance criteria:

Difference in recovery of acetaminophen:

≤ 10 µg/mL: ≤ ± 1μg/mL and > 10 µg/mL: 100 ± 10 %

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MatrixcomparisonLithium-heparin, K2-EDTA, and K3-EDTA are permissible anticoagulantsfor use with this reagent because they do not interfere with recovery ofacetaminophen. In an internal study, 60 tubes were collected peranticoagulant. Plasma results were compared to serum results and percentrecovery was determined. In terms of % recovery to serum, all data passedthe following criteria:For sample concentrations ≤ 10 µg/mL, the deviation must be ≤ ± 1 µg/mL.For sample concentrations > 10 µg/mL, the deviation must be ≤ ± 10%.
anticoagulantsSample concentrationrange tested (µg/mL)Claimed MeasuringRange (µg/mL)
Li-Heparin2.6 - 187
K2-EDTA2.6 - 1875 – 200
K3-EDTA2.6 - 187
In addition, method comparisons with plasma vs serum were calculated withthe following results (Passing/Bablok):Serum vs. Li-heparin $y = 0.989x + 0.089,$ $r = 0.998$Serum vs. K2-EDTA $y = 1.000x – 0.000,$ $r = 0.998$Serum vs. K3-EDTA $y = 0.992x - 0.163,$ $r = 0.998$
ConclusionThe submitted information in this premarket notification supports a

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

§ 862.3200 Clinical toxicology calibrator.

(a)
Identification. A clinical toxicology calibrator is a device intended for medical purposes for use in a test system to establish points of reference that are used in the determination of values in the measurement of substances in human specimens. A clinical toxicology calibrator can be a mixture of drugs or a specific material for a particular drug (e.g., ethanol, lidocaine, etc.). (See also § 862.2 in this part.)(b)
Classification. Class II (special controls). 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.