NeoBase 2 Non-derivatized MSMS Kit

K173568 · Wallac Oy, Subsidiary of Perkinelmer, Inc. · NQL · Sep 4, 2018 · Clinical Chemistry

Device Facts

• Record ID: K173568
• Device Name: NeoBase 2 Non-derivatized MSMS Kit
• Applicant: Wallac Oy, Subsidiary of Perkinelmer, Inc.
• Product Code: NQL · Clinical Chemistry
• Decision Date: Sep 4, 2018
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1055
• Device Class: Class 2
• Attributes: Pediatric

Intended Use

The NeoBase™ 2 Non-derivatized MSMS kit is intended for the measurement and evaluation of amino acid, succinylacetone, free carnitine, acylcarnitine, nucleoside and lysophospholipid concentrations (Table 1) with a tandem mass spectrometer from newborn heel prick blood specimens dried on filter paper. Quantitative analysis of these analytes and their relationship with each other is intended to provide analyte concentration profiles that may aid in screening newborns for metabolic disorders.

Device Story

The NeoBase 2 Non-derivatized MSMS kit is an in vitro diagnostic reagent system used with a tandem mass spectrometer to analyze dried blood spot (DBS) samples from newborns. The kit facilitates the quantitative measurement of various amino acids, carnitines, succinylacetone, nucleosides, and lysophospholipids. By measuring these analyte concentrations and their interrelationships, the device generates profiles that assist healthcare providers in screening for metabolic disorders. The process involves sample preparation from heel prick blood specimens on filter paper, followed by tandem mass spectrometry analysis. The output is used by clinical laboratory professionals to identify potential metabolic abnormalities, aiding in early clinical decision-making and patient management.

Clinical Evidence

Clinical performance was evaluated in two CLIA-certified state laboratories comparing the NeoBase 2 kit to the predicate NeoBase kit. Studies included 37 amino acid disorder, 26 fatty acid oxidation disorder, 28 organic acid condition, and 8 ADA-SCID/X-ALD confirmed positive specimens. Performance was assessed using 99th, 99.5th, and low-percentile cut-offs. Results demonstrated agreement between the subject and predicate devices in detecting confirmed positive specimens across all disorder categories.

Technological Characteristics

Reagent kit for tandem mass spectrometry (MS/MS) analysis of dried blood spots. Measures amino acids, carnitines, succinylacetone, nucleosides, and lysophospholipids. Analytical methods follow CLSI guidelines (EP5-A3, EP06-A, EP7-A2, EP9-A3, EP25-A). System requires tandem quadrupole detector (TQD).

Indications for Use

Indicated for newborn screening of metabolic disorders using heel prick blood specimens dried on filter paper. Measures amino acids, succinylacetone, free carnitine, acylcarnitines, nucleosides, and lysophospholipids to aid in identifying metabolic conditions.

Regulatory Classification

Identification

A newborn screening test system for amino acids, free carnitine, and acylcarnitines using tandem mass spectrometry is a device that consists of stable isotope internal standards, control materials, extraction solutions, flow solvents, instrumentation, software packages, and other reagents and materials. The device is intended for the measurement and evaluation of amino acids, free carnitine, and acylcarnitine concentrations from newborn whole blood filter paper samples. The quantitative analysis of amino acids, free carnitine, and acylcarnitines and their relationship with each other provides analyte concentration profiles that may aid in screening newborns for one or more inborn errors of amino acid, free carnitine, and acyl-carnitine metabolism.

Special Controls

*Classification.* Class II (special controls). The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: Newborn Screening Test Systems for Amino Acids, Free Carnitine, and Acylcarnitines Using Tandem Mass Spectrometry.” See § 862.1(d) for the availability of this guidance document.

Predicate Devices

• PerkinElmer NeoBase Non-derivatized MSMS reagent kit (K093916)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA logo on the right. The FDA logo is in blue and includes the letters "FDA" followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in a stacked format.

September 4, 2018

Wallac Ov, Subsidiary of PerkinElmer, Inc. Anne Marie Whalen Director, Regulatory Affairs 940 Winter Street Waltham, MA 02451

Re: K173568

Trade/Device Name: NeoBase 2 Non-derivatized MSMS Kit Regulation Number: 21 CFR 862.1055 Regulation Name: Newborn screening test system for amino acids, free carnitine, and acylcarnitines using tandem mass spectrometry Regulatory Class: Class II Product Code: NQL Dated: August 24, 2018 Received: August 27, 2018

Dear Anne Marie Whalen:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's

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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/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

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/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). 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 (http://www.fda.gov/DICE) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

# Kellie B. Kelm -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) K173568

Device Name NeoBase™ 2 Non-derivatized MSMS kit

#### Indications for Use (Describe)

The NeoBase™ 2 Non-derivatized MSMS kit is intended for the measurement and evaluation of amino acid, succinylacetone, free carnitine, acylcarnitine, nucleoside and lysophospholipid concentrations (Table 1) with a tandem mass spectrometer from newborn heel prick blood specimens dried on filter paper. Quantitative analytis of these analytes and their relationship with each other is intended to provide analyte concentration profiles that may aid in screening newborns for metabolic disorders.

Table 1. Analytes measured by the NeoBase 2 Non-derivatized MSMS kit.

#### ANALYTE NAME

ABBREVIATION

#### Amino acids

| Alanine | Ala |
|---------------------------------------------------|----------------|
| Arginine | Arg |
| Argininosuccinic acid | Asa |
| Citrulline | Cit |
| Glutamine\Lysine (*) | Gln\Lys |
| Glutamic acid | Glu |
| Glycine | Gly |
| Leucine\Isoleucine\Hydroxyproline (*) | Leu\Ile\Pro-OH |
| Methionine | Met |
| Ornithine | Orn |
| Phenylalanine | Phe |
| Proline | Pro |
| Tyrosine | Tyr |
| Valine | Val |
| Carnitines | |
| Free carnitine | C0 |
| Acetylcarnitine | C2 |
| Propionylcarnitine | C3 |
| Malonylcarnitine\3-Hydroxy-butyrylcarnitine (*) | C3DC\C4OH |
| Butyrylcarnitine | C4 |
| Methylmalonyl\3-Hydroxy-isovalerylcarnitine (*) | C4DC\C5OH |
| Isovalerylcarnitine | C5 |
| Tiglylcarnitine | C5:1 |
| Glutarylcarnitine\3-Hydroxy-hexanoylcarnitine (*) | C5DC\C6OH |
| Hexanoylcarnitine | C6 |
| Adipylcarnitine | C6DC |
| Octanoylcarnitine | C8 |
| Octenoylcarnitine | C8:1 |
| Decanoylcarnitine | C10 |

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| Decenoylcarnitine | C10:1 |
|----------------------------------------------|-------------|
| Decadienoylcarnitine | C10:2 |
| Dodecanoylcarnitine | C12 |
| Dodecenoylcarnitine | C12:1 |
| Tetradecanoylcarnitine (Myristoylcarnitine) | C14 |
| Tetradecenoylcarnitine | C14:1 |
| Tetradecadienoylcarnitine | C14:2 |
| 3-Hydroxy-tetradecanoylcarnitine | C14OH |
| Hexadecanoylcarnitine (Palmitoylcarnitine) | C16 |
| Hexadecenoylcarnitine | C16:1 |
| 3-Hydroxy-hexadecanoylcarnitine | C16OH |
| 3-Hydroxy-hexadecenoylcarnitine\ | |
| Heptadecanoylcarnitine (*) | C16:1OH\C17 |
| Octadecanoylcarnitine (Stearoylcarnitine) | C18 |
| Octadecenoylcarnitine (Oleylcarnitine) | C18:1 |
| Octadecadienoylcarnitine (Linoleylcarnitine) | C18:2 |
| 3-Hydroxy-octadecanoylcarnitine | C18OH |
| 3-Hydroxy-octadecenoylcarnitine | C18:1OH |
| 3-Hydroxy-octadecadienoylcarnitine | C18:2OH |
| | |
| Ketones | |
| | |
| Succinylacetone | SA |
| | |
| Nucleotides | |
| | |
| Adenosine | ADO |
| 2'-deoxyadenosine | D-ADO |
| | |
| Lysophospholipids | |
| | |
| C24:0 lysophosphatidylcholine | C24:0-LPC |
| C26:0 lysophosphatidylcholine | C26:0-LPC |
| | |

(*) Analytes in these rows are either isomers or isobars and cannot be distinguished in the tandem mass spectrometty experiment.

| 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)

### CONTINUE ON A SEPARATE PAGE IF NEEDED.

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# 510(k) Summary

This 510(k) Summary information is supplied in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510 (k) number is K173568

Date: August 24, 2018

#### Submitted by:

Wallac Oy, a subsidiary of PerkinElmer Inc. 940 Winter Street Waltham, MA 02451

Wallac Oy Mustionkatu 6 Turku Finland 20750

#### Submission Contact Person:

| Primary: | Anne Marie Whalen, PhD<br>Director, Regulatory Affairs<br>Tel:[phone]<br>Fax: [phone] |
|-------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Trade Name: | NeoBase 2 Non-derivatized MSMS kit |
| Common Name: | Newborn screening test system for amino acids, succinylacetone,<br>free carnitine, acylcarnitines, nucleosides, and lysophospholipids<br>using tandem mass spectrometry |
| Regulation: | 21 CFR 862.1055 |
| Classification: | II |
| Panel: | Clinical Chemistry (75) |
| Product Code: | NQL |
| Predicate device: | PerkinElmer NeoBase Non-derivatized MSMS reagent kit<br>(K093916) |

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## Intended Use:

The NeoBase™ 2 Non-derivatized MSMS kit is intended for the measurement and evaluation of amino acid, succinylacetone, free carnitine, acylcarnitine, nucleoside and lysophospholipid concentrations (Table 1) with a tandem mass spectrometer from newborn heel prick blood specimens dried on filter paper. Quantitative analysis of these and their relationship with each other is intended to provide analyte concentration profiles that may aid in screening newborns for metabolic disorders.

| ANALYTE NAME | ABBREVIATION |
|------------------------------------------------|----------------|
| Amino acids | |
| Alanine | Ala |
| Arginine | Arg |
| Argininosuccinic acid | Asa |
| Citrulline | Cit |
| Glutamine\Lysine1 | Gln\Lys |
| Glutamic acid | Glu |
| Glycine | Gly |
| Leucine\Isoleucine\Hydroxyproline1 | Leu\Ile\Pro-OH |
| Methionine | Met |
| Ornithine | Orn |
| Phenylalanine | Phe |
| Proline | Pro |
| Tyrosine | Tyr |
| Valine | Val |
| Carnitines | |
| Free carnitine | C0 |
| Acetylcarnitine | C2 |
| Propionylcarnitine | C3 |
| Malonylcarnitine\3-Hydroxy-butyrylcarnitine1 | C3DC\C4OH |
| Butyrylcarnitine | C4 |
| Methylmalonyl\3-Hydroxy-isovalerylcarnitine1 | C4DC\C5OH |
| Isovalerylcarnitine | C5 |
| Tiglylcarnitine | C5:1 |
| Glutarylcarnitine\3-Hydroxy-hexanoylcarnitine1 | C5DC\C6OH |
| Hexanoylcarnitine | C6 |
| Adipylcarnitine | C6DC |
| Octanoylcarnitine | C8 |
| Octenoylcarnitine | C8:1 |
| Decanoylcarnitine | C10 |
| Decenoylcarnitine | C10:1 |
| Decadienoylcarnitine | C10:2 |
| Dodecanoylcarnitine | C12 |

Table 1. Analytes measured by the NeoBase 2 Non-derivatized MSMS kit:

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| Dodecenoylcarnitine | C12:1 |
|----------------------------------------------|-------------|
| Tetradecanoylcarnitine (Myristoylcarnitine) | C14 |
| Tetradecenoylcarnitine | C14:1 |
| Tetradecadienoylcarnitine | C14:2 |
| 3-Hydroxy-tetradecanoylcarnitine | C14OH |
| Hexadecanoylcarnitine (Palmitoylcarnitine) | C16 |
| Hexadecenoylcarnitine | C16:1 |
| 3-Hydroxy-hexadecanoylcarnitine | C16OH |
| 3-Hydroxy-hexadecenoylcarnitine\ | C16:1OH\C17 |
| Heptadecanoylcarnitine¹ | |
| Octadecanoylcarnitine (Stearoylcarnitine) | C18 |
| Octadecenoylcarnitine (Oleylcarnitine) | C18:1 |
| Octadecadienoylcarnitine (Linoleylcarnitine) | C18:2 |
| 3-Hydroxy-octadecanoylcarnitine | C18OH |
| 3-Hydroxy-octadecenoylcarnitine | C18:1OH |
| 3-Hydroxy-octadecadienoylcarnitine | C18:2OH |
| Ketones | |
| Succinylacetone | SA |
| Nucleosides | |
| Adenosine | ADO |
| 2'-deoxyadenosine | D-ADO |
| Lysophospholipids | |
| C24:0 lysophosphatidylcholine | C24:0-LPC |
| C26:0 lysophosphatidylcholine | C26:0-LPC |

1 Analytes in these rows are either isomers or isobars and cannot be distinguished in the tandem mass spectroment.

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# Summary of Non-Clinical Studies:

A summary of the verification studies is presented below:

| Study | CLSI Guidance | Number of kit lots |
|--------------------------------------|---------------|---------------------------------------------------------------------------------------------------------------------------------------------------|
| Precision | EP5-A3 | Precision study 1:<br>1 Instrument, 1 kit lot<br>Precision study 2:<br>1 Instrument, 3 kit lots<br>Precision study 3:<br>2 Instruments, 1 kit lot |
| Analytical Sensitivity | N/A | 2 Instruments, 3 kit lots |
| Linearity | EP06-A | 1 Instrument, 3 kit lots |
| Interference | EP7-A2 | Mass transition overlap study:<br>1 Instrument |
| | | Paired-difference study:<br>2 Instruments, 3 kit lot |
| | | Dose-response study:<br>2 Instruments, 3 kit lot |
| Drift | N/A | 1 Instrument, 1 kit lot |
| Recovery | N/A | 1 Instrument, 3 kit lots |
| Method Comparison | EP9-A3 | 2 Instruments, 3 kit lots |
| Carry-over | N/A | 2 Instruments, 1 kit lot |
| Incubation times and<br>temperatures | N/A | 1 Instrument, 1 kit lot |
| Real Time/Transport<br>Stability | EP25-A | 2 Instruments, 3 kit lots |
| Accelerated Stability | EP25-A | 2 Instruments, 2 kit lots |
| Study | CLSI Guidance | Number of kit lots |
| In-use, On-board Stability | EP25-A | 1 Instrument, 1 kit lot |
| Sample Stability | EP25-A | 1 Instrument, 1 kit lot |

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All verification studies were successfully concluded and met the respective study's predetermined acceptance criteria.

#### Summary of Clinical Studies:

The 3044-0010U NeoBase 2 Non-derivatized MSMS kit was compared to 3040-001U NeoBase Non-derivatized MSMS kit by measuring analyte concentrations with a tandem quadrupole detector (TQD) in two CLIA-certified state laboratories responsible for routine newborn screening. Using data from routine newborn screening, the cut-offs for both methods were determined by calculating the 99.5th and 99th percentile for all analytes except C2. For C2, C3, C16, C18, C18:1 and Cit the low cut-off applied is based on 1st percentile. For C0, the low cut-off applied is based on 10th percentile. Please note that the cut-off values used in evaluating screening performance only apply to these studies.

The disorders and amount of confirmed positive specimens included in the studies 1 and 2 are listed in Tables A and B (study 1), and Tables C and D (study 2). In total, there were 37 specimens included in the group of amino acid disorders, 26 in the group of fatty acid oxidation disorders and 28 in the group of organic acid conditions. In addition, the group of other disorder contains adenosine deaminase severe combined immunodeficiency (ADA-SCID) and X-linked adrenoleukodystrophy (X-ALD) of which 8 specimens, 4 of each disorder, were included.

The results for the screening performance data including confirmed positive specimens tested with TQD in the study 1 are presented in Table A.

The screening performance for adenosine deaminase severe combined immunodeficiency (ADA-SCID) and X-linked adrenoleukodystrophy (X-ALD) was done by comparing the result of NeoBase 2 Non-derivatized MSMS kit to the clinical condition. Results of the study 1 are presented in Table B.

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| | | Table A. Agreement between NeoBase 2 Non-derivatized MSMS kit and NeoBase Non- | | | |
|--------------------------------------|--|--------------------------------------------------------------------------------|--|--|--|
| derivatized MSMS kit in the study 1. | | | | | |

| Amino acid disorders<br>(AA) | | NeoBase | | | | | |
|------------------------------|--------------------|--------------------|--------------------|-------|--------------------|--------------------|-------|
| | | | 99th percentile | | | 99.5th percentile | |
| | | Screening positive | Screening negative | Total | Screening positive | Screening negative | Total |
| NeoBase 2 | Screening positive | 621 | 70 | 132 | 452 | 41 | 86 |
| | Screening negative | 28 | 1591 | 1619 | 20 | 1645 | 1665 |
| | Total | 90 | 1661 | 1751 | 65 | 1686 | 1751 |

¹ Includes all 15 specimens confirmed positive for AA detected with both methods using 99ᵗʰ percentile cut-offs.

1 Includes all 15 specimens confirmed positive for AA detected with both methods using 99th percentile cut-offs.
2 Includes all 15 specimens confirmed positive for AA detecte

| Amino acid disorders (AA) | NeoBase | | | |
|---------------------------|--------------------|--------------------|-------|------|
| | 1st percentile | | | |
| | Screening positive | Screening negative | Total | |
| NeoBase 2 | Screening positive | 161 | 18 | 34 |
| | Screening negative | 16 | 1687 | 1703 |
| | Total | 32 | 1705 | 1737 |

1 Includes 1 specimen confirmed positive for OTCD with both methods using 1st percentile cut-offs.

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| Fatty acid oxidation (FAO) | NeoBase | | | | | | |
|----------------------------|--------------------|--------------------|-------|--------------------|--------------------|-------|------|
| | 99th percentile | | | 99.5th percentile | | | |
| | Screening positive | Screening negative | Total | Screening positive | Screening negative | Total | |
| NeoBase 2 | Screening positive | 801 | 40 | 120 | 451 | 17 | 62 |
| | Screening negative | 45 | 1581 | 1626 | 23 | 1661 | 1684 |
| | Total | 125 | 1621 | 1746 | 68 | 1678 | 1746 |

1 Includes all 10 specimens confirmed positive for FAO detected with both methods using 99th and 99.5 percentile cut-offs.

| Fatty acid oxidation (FAO) | NeoBase | | | |
|----------------------------|--------------------|--------------------|-------|------|
| | Low percentile | | | |
| | Screening positive | Screening negative | Total | |
| NeoBase 2 | Screening positive | 1732 | 29 | 202 |
| | Screening negative | 150 | 1386 | 1536 |
| | Total | 322 | 1415 | 1738 |

I "Low Percentile" reflects the positive C0 specimens detected with a 10% cut-off with other carnitine-positive samples at the 1% cut-off 2 Includes 2 specimens confirmed positive for CUD detected with both methods using low percentile cut-offs.

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| Organic acid condition<br>(OA) | | NeoBase | | | NeoBase | | |
|--------------------------------|--------------------|--------------------|--------------------|-------|--------------------|--------------------|-------|
| | | 99th percentile | | | 99.5th percentile | | |
| | | Screening positive | Screening negative | Total | Screening positive | Screening negative | Total |
| NeoBase 2 | Screening positive | 571 | 23 | 80 | 361 | 13 | 49 |
| | Screening negative | 11 | 1660 | 1671 | 5 | 1697 | 1702 |
| | Total | 68 | 1683 | 1751 | 41 | 1710 | 1751 |

1 Includes all 15 specimens confirmed positive for OA detected with both methods using 99th and 99.50 percentile cut-offs.

| | Table B. Screening performance for ADA-SCID and X-ALD using 99th and 99.500 percentile in | | | |
|----------|-------------------------------------------------------------------------------------------|--|--|--|
| study 1. | | | | |

| ADA-SCID | | 99th percentile | | | 99.5th percentile | | |
|-----------|--------------------|-----------------|--------|-------|-------------------|--------|-------|
| | | Positive | Normal | Total | Positive | Normal | Total |
| NeoBase 2 | Screening positive | 21 | 75 | 77 | 21 | 36 | 38 |
| | Screening negative | 0 | 1661 | 1661 | 0 | 1700 | 1700 |
| | Total | 2 | 1736 | 1738 | 2 | 1736 | 1738 |

1 Includes both specimens confirmed positive for ADA-SCID.

| X-ALD | 99th percentile | | | 99.5th percentile | | | |
|-----------|--------------------|--------|-------|-------------------|--------|-------|------|
| | Positive | Normal | Total | Positive | Normal | Total | |
| NeoBase 2 | Screening positive | 21 | 12 | 14 | 21 | 5 | 7 |
| | Screening negative | 0 | 1724 | 1724 | 0 | 1731 | 1731 |
| | Total | 2 | 1736 | 1738 | 2 | 1736 | 1738 |

1 Includes both specimens confirmed positive for X-ALD.

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The results for the screening performance data including retrospective confirmed positive specimens tested with TQD in the study 2 are presented in Table C.

The screening performance for Adenosine deaminase severe combined immunodeficiency (ADA-SCID) and X-linked adrenoleukodystrophy (X-ALD) was done by comparing the result of NeoBase_2 Non-derivatized MSMS kit to the clinical condition. Results of the study 2 are presented in Table D.

Table C. Agreement between NeoBase_2 Non-derivatized MSMS kit and NeoBase Nonderivatized MSMS kit in the study 2.

| Amino acid disorders (AA) | NeoBase | | | | | | |
|---------------------------|--------------------|--------------------|-------|--------------------|--------------------|-------|------|
| | 99th percentile | | | 99.5th percentile | | | |
| | Screening positive | Screening negative | Total | Screening positive | Screening negative | Total | |
| NeoBase 2 | Screening positive | 1161 | 138 | 254 | 782 | 67 | 145 |
| | Screening negative | 41 | 2353 | 2394 | 29 | 24743 | 2503 |
| | Total | 157 | 2491 | 2648 | 107 | 2541 | 2648 |

Includes all 19 specimens confirmed positive for AA detected with both methods using 99th percentile cut-offs.

4 Includes 18 specimens confirmed positive for AA detected with both method using 99.5" percentile cut-offs.
3 Includes 1 specimen confirmed positive for AA not detected with

³Includes 1 specimen confirmed positive for AA not detected with either method using 99.5ᵗʰ percentile cut-offs.

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| Amino acid disorders (AA) | NeoBase | | | |
|---------------------------|--------------------|--------------------|-------|------|
| | 1st percentile | Screening negative | Total | |
| NeoBase 2 | Screening positive | 141 | 40 | 54 |
| | Screening negative | 6 | 2571 | 2577 |
| | Total | 20 | 2611 | 2631 |

1 Includes 2 specimens confirmed positive for OTCD detected with both methods using 1* percentile cut-offs.

| Fatty acid oxidation<br>(FAO) | | NeoBase | | | | | |
|-------------------------------|--------------------|--------------------|--------------------|-------|--------------------|--------------------|-------|
| | | 99th percentile | 99.5th percentile | | | | |
| | | Screening positive | Screening negative | Total | Screening positive | Screening negative | Total |
| NeoBase 2 | Screening positive | 1601 | 83 | 243 | 1081 | 37 | 145 |
| | Screening negative | 72 | 2326 | 2398 | 54 | 2442 | 2496 |
| | Total | 232 | 2409 | 2641 | 162 | 2479 | 2641 |

1 Includes all 12 specimens confirmed positive for FAO detected with both methods using 99th and 99.50 percentile cut-offs.

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| NeoBase | | | | |
|----------------------------|--------------------|--------------------|-------|------|
| Fatty acid oxidation (FAO) | Low percentile | | | |
| | Screening positive | Screening negative | Total | |
| NeoBase 2 | Screening positive | 1581 | 45 | 203 |
| | Screening negative | 66 | 2363 | 2429 |
| | Total | 224 | 2408 | 2632 |

1 "Low Percentile" reflects positive C0 specimens detected with a 10% cut-off with other camples at the 1% cut-off 2 Includes 3 specimens confirmed positive for FAO detected with both methods using low percentile cut-offs. CPT II was confirmed positive with both methods with each of the 99th, 99.5th and 1st percentile cut-offs.

| Organic acid condition<br>(OA) | NeoBase | | | | | | |
|--------------------------------|-----------------------|-----------------------|-------|-----------------------|-----------------------|-------|------|
| | 99th percentile | | | 99.5th percentile | | | |
| | Screening<br>positive | Screening<br>negative | Total | Screening<br>positive | Screening<br>negative | Total | |
| NeoBase 2 | Screening<br>positive | 861 | 52 | 138 | 422 | 27 | 69 |
| | Screening<br>negative | 25 | 2479 | 2504 | 12 | 25613 | 2573 |
| | Total | 111 | 2531 | 2642 | 54 | 2588 | 2642 |

1 Includes all 13 specimens confirmed positive for OA detected with both methods using 99th and 99.50 percentile cut-offs.

2 Includes 12 specimens confirmed positive for OA detected with both methods using 99.5th percentile cut-offs.

3 Includes 1 specimen confirmed positive for OA not detected with either method using 99.54 percentile cut-offs.

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| Table D. Screening performance for ADA-SCID and X-ALD using 99th and 99.5th percentile in | |
|-------------------------------------------------------------------------------------------|--|
| the study 2. | |

| ADA-SCID | | 99th percentile | | | 99.5th percentile | | |
|-----------|--------------------|-----------------|--------|-------|-------------------|--------|-------|
| | | Positive | Normal | Total | Positive | Normal | Total |
| NeoBase 2 | Screening positive | 21 | 66 | 68 | 21 | 51 | 53 |
| | Screening negative | 0 | 2563 | 2563 | 0 | 2578 | 2578 |
| | Total | 2 | 2629 | 2631 | 2 | 2629 | 2631 |

1 Includes both specimens confirmed positive for ADA-SCID.

| X-ALD | | 99th percentile | | | 99.5th percentile | | |
|-----------|-----------------------|-----------------|--------|-------|-------------------|--------|-------|
| | | Positive | Normal | Total | Positive | Normal | Total |
| NeoBase 2 | Screening<br>positive | 21 | 3 | 5 | 21 | 1 | 3 |
| | Screening<br>negative | 0 | 2626 | 2626 | 0 | 2628 | 2628 |
| | Total | 2 | 2629 | 2631 | 2 | 2629 | 2631 |

1 Includes both specimens confirmed positive for X-ALD.

#### Substantial Equivalency:

The proposed device and predicate device utilize similar design shown to produce equivalent screening performance in a clinical setting. Both devices are intended for the measurement and evaluation of multiple metabolite concentrations from newborn heel prick blood samples dried on filter paper. Quantitative analysis of these analytes and their relationship with each other is intended to provide analyte concentration profiles that may aid in screening newborns for metabolic disorders.

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### Conclusion:

The NeoBase 2 Non-derivatized MSMS kit demonstrates analytical and screening performance that supports its substantial equivalency with the predicate device, NeoBase 2 Non-derivatized MSMS kit (K093916).