(91 days)
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 analytes and their relationship with each other is intended to provide analyte concentration profiles that may aid in screening newborns for metabolic disorders.
Each NeoBase 2 Non-derivatized MSMS kit contains reagents for 960 assays. The kit is designed to be used with NeoBase 2 Non-derivatized Assay Solutions consisting of Neo MSMS Flow Solvent and NeoBase 2 Extraction Solution and NeoBase 2 Succinylacetone Assay Solution.
- NeoBase 2 Internal Standards - 1 vial
- NeoBase 2 Controls Low, High - 3 filter paper cassettes (Whatman, no. 903) containing 3 spots of each level per cassette
- Microplate, U-bottomed - 20 plates
- Adhesive microplate covers - 20 sheets
- Barcode labels for the plates - 30 pcs (10 different barcodes, 3 pcs of each)
- Lot-specific quality control certificate
This kit contains components manufactured from human blood. The source materials have been tested by FDA-approved methods for hepatitis B surface antigen, anti-hepatitis C and anti-HIV 1 and 2 antibodies and found to be negative.
Instruments used: - . QSight® 210 MD Screening System is comprised of:
- QSight® 210 MD Mass Spectrometer
- QSight® HC Autosampler MD
- QSight® Binary Pump MD ●
- Simplicity™ 3Q MD Software
- PerkinElmer MSMS Workstation software ●
Here's a breakdown of the acceptance criteria and the study that proves the device meets them, based on the provided text.
Note: This document describes a medical device for newborn screening. Performance is demonstrated through analytical studies (precision, sensitivity, linearity, interference) and screening performance data comparing it to a predicate device, rather than human-in-the-loop studies common for AI/imaging devices. Thus, several sections typically relevant to AI-based devices (e.g., number of experts, adjudication methods, MRMC studies) are not applicable here.
Acceptance Criteria and Device Performance
The acceptance criteria for the NeoBase 2 Non-derivatized MSMS kit are implied by the comprehensive analytical and screening performance evaluation studies conducted. The goal is to demonstrate that the device performs equivalently to the legally marketed predicate device (NeoBase 2 Non-derivatized MSMS kit, K173568) and provides reliable results for newborn screening.
The reported device performance below highlights key analytical and screening metrics. Specific numerical acceptance criteria are not explicitly stated as hard thresholds (e.g., "CV must be < X%"), but rather the results of the studies are presented to demonstrate acceptable performance comparable to a cleared predicate.
Table of Acceptance Criteria (Implied) and Reported Device Performance
| Acceptance Criterion Area | Study Conducted / Metric | Reported Device Performance and Observation |
|---|---|---|
| Precision | Repeatability, Within-Laboratory, Between-Lot, Between-Instrument Variation (measured as SD and CV%) for all analytes. | Detailed tables provided for each analyte showing: - Repeatability CV%: Generally low (e.g., Ala: 5.3-6.5%, Arg: 3.3-6.0%, C0: 4.3-5.6%, C26:0-LPC: 5.0-16%). Higher CV% for lower concentrations (e.g., Asa Sample 1: 29%, Met Sample 1: 18%, SA Sample 1: 17%, ADO Sample 1: 19%, C26:0-LPC Sample 1: 16%). - Within-Lab CV%: Slightly higher than repeatability, still low (e.g., Ala: 6.3-9.8%, Arg: 5.0-8.3%, C0: 5.0-6.5%). - Between-lot and Between-instrument CV%: Generally low, indicating good consistency. - Total Variation CV%: Overall low, with higher CV% at lower concentrations (e.g., Asa Sample 1: 32%, Met Sample 1: 20%, SA Sample 1: 13%, ADO Sample 1: 17%, C26:0-LPC Sample 1: 23%). |
| Analytical Sensitivity | Lowest measurable analyte concentrations (Limit of Quantitation, LOQ). | Specific analytical sensitivity limits in µmol/L are provided for each analyte. (e.g., Ala: 3.66 µmol/L, Arg: 0.64 µmol/L, C0: 0.18 µmol/L, SA: 0.24 µmol/L). Some entries indicate <LOQ for percentiles in screening performance tables. |
| Linearity | Linear range (lower and upper limits, µmol/L) for all analytes. | Specific linear ranges defined for each analyte (e.g., Ala: 163-1450 µmol/L, Arg: 1.84-359 µmol/L, C0: 7.80-407 µmol/L, SA: 0.24-88.2 µmol/L, C26:0-LPC: 0.22-7.08 µmol/L). |
| Interference | Evaluation of potential interfering substances (endogenous and exogenous). | A list of 12 substances were not found to interfere. Identified Interferents: Sarcosine (Ala), Creatine (Ala, Glu, Leu), L-Asparagine (Orn), L-Lysine (Arg, Gln, Glu), L-Glutamic acid (Met), L-Methionine sulfone (Tyr), Verapamil metabolite D617 (ASA), L-Ornithine (Pro), Albumin (ADO), Intralipid (C24:0-LPC), Chlorhexidine digluconate (C24:0-LPC, C26:0-LPC), Hemoglobin (SA, C24:0-LPC, ADO, Val), Hematocrit (Arg). Note on Interferents: The report concludes that most identified interferences are unlikely to impact routine testing due to physiological concentration ranges or by incorporation into user-established cut-offs. Specific caution provided for Benzocaine (Phe), C5 isomer pivalylcarnitine (C5), C8 isomer valproylcarnitine (C8), and cross-reactivity of isomers/isobars (e.g., C3DC/C4OH, C16:1OH/C17). |
| Reproducibility | Between-Site Reproducibility (measured as SD and CV%) across 2 external sites and 1 internal site. | Detailed tables provided for each analyte showing: - Within-lab CV%: Generally low (e.g., Ala: 5.4-5.9%, Arg: 4.2-6.0%, C0: 5.0-5.7%, C26:0-LPC: 7.9-24%). - Between-lab CV%: Generally low (e.g., Ala: 1.0-3.5%, Arg: 1.2-3.8%, C0: 0.66-2.3%). - Reproducibility CV%: Overall low (e.g., Ala: 5.2-6.5%, Arg: 5.2-6.3%, C0: 5.1-6.1%). Higher CV% for lower concentrations (e.g., Asa Sample 1: 28%, SA Sample 1: 22%, C26:0-LPC Sample 1: 54%). |
| Screening Performance | Comparison of newborn population distributions and positive case detection against predicate device (TQD) using defined percentiles. | Population Distribution (n=2530): Mean, Median, 1st, 10th, 99th, and 99.5th percentiles are provided for all 47 analytes, demonstrating the device's ability to measure concentrations across a typical newborn population range. Confirmed Positive Specimens: - Amino Acid Disorders (AA): QSight detected either all 19 (99th percentile) or 18/19 (99.5th percentile) confirmed cases that were also detected by TQD. - Fatty Acid Oxidation (FAO): QSight detected all 9 (99th/99.5th percentile) or all 4 (low percentile) confirmed cases that were also detected by TQD. - Organic Acid Condition (OA): QSight detected all 16 (99th/99.5th percentile) confirmed cases that were also detected by TQD. - ADA-SCID: QSight detected both 2 confirmed cases (99th/99.5th percentile) that were also detected by TQD. - X-ALD: QSight detected both 2 confirmed cases (99th/99.5th percentile) that were also detected by TQD, with notes on borderline samples. |
Study Details
-
Sample Size and Data Provenance:
- Test Set (Screening Performance): 2530 routine newborn screening specimens.
- Country of Origin: United States (one routine screening laboratory in the US).
- Retrospective/Prospective: The nature of "routine screening specimens" usually implies retrospective use of collected samples for evaluating a new method, but explicit detail (e.g., prospective collection specifically for this study) is not given. However, the study focuses on comparing the new device (QSight) to the predicate (TQD) on these samples, suggesting an evaluation of existing data or new data collected in a routine setting.
- Analytical Studies (Precision, Linearity, Interference):
- Precision: 80 determinations (40 plates x 2 replicates) for repeatability and within-laboratory; 75 determinations (15 plates x 5 replicates x 3 lots) for between-lot; 50 determinations (10 plates x 5 replicates x 2 QSights) for between-instrument.
- Reproducibility: 75 determinations (5 plates x 5 working days x 5 replicates) across 3 labs (1 internal, 2 external).
- Linearity, Sensitivity, Interference: Not explicitly quantified by sample size in the same way as precision. These studies typically use a controlled set of samples spiked with known concentrations or interfering substances.
- Test Set (Screening Performance): 2530 routine newborn screening specimens.
-
Number of Experts and Qualifications for Ground Truth:
- This device is a quantitative assay for measuring analyte concentrations, not an AI/imaging device requiring expert interpretation of images.
- The "ground truth" for the screening performance study are confirmed positive specimens for various metabolic disorders (Amino Acid, Fatty Acid Oxidation, Organic Acid, ADA-SCID, X-ALD). These confirmations are typically established through gold standard clinical diagnostic methods (e.g., genetic testing, enzyme assays, or further biochemical analysis), not through expert panel review of the screening test results themselves. The document states "confirmed positive specimens," implying these are definitive diagnoses.
- Therefore, the concept of "experts" establishing ground truth for the test set, in the way a radiologist reads an image, does not directly apply here.
-
Adjudication Method for the Test Set:
- Not applicable as this is a quantitative diagnostic assay. "Adjudication" typically refers to resolving disagreements among human readers/interpreters, which is not the primary function of this device or study design. The study compares the new device's quantitative results and detection rates against those of a predicate device and known confirmed positive cases.
-
Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study:
- Not applicable. This is not a study comparing human readers with and without AI assistance. It is a comparison of two analytical instruments/kits (the new NeoBase 2 on QSight vs. predicate NeoBase 2 on TQD).
-
Standalone Performance (Algorithm-only without human-in-the-loop):
- Yes, this study is a standalone performance evaluation. The device (NeoBase 2 kit run on QSight MD Screening System) provides quantitative measurements of analytes. The performance tables directly show the device's analytical capabilities (precision, sensitivity, linearity, reproducibility) and its ability to detect known positive cases in a patient population without human interpretation as part of the core measurement. Human expert judgment comes in for confirming the disease in patient specimens (the ground truth), but not in interpreting the raw output of the device itself.
-
Type of Ground Truth Used:
- Confirmed positive specimens: For the screening performance study, the ground truth was based on specimens from individuals clinically confirmed to have the targeted metabolic disorders (Amino Acid Disorders, Fatty Acid Oxidation, Organic Acid Conditions, ADA-SCID, X-ALD). This likely involved definitive diagnostic tests beyond initial screening, such as genetic testing, enzyme activity assays, or more specific quantitative metabolite analysis. This is a form of outcomes data/definitive diagnosis.
-
Sample Size for the Training Set:
- This is a medical device for quantitative measurement, not a machine learning or AI model that requires a "training set" in the conventional sense (i.e., data used to teach an algorithm to learn patterns). The device's performance validation relies on analytical studies to demonstrate its accuracy, precision, and consistency, and then clinical performance by comparing its results to a predicate device and confirmed clinical cases. Therefore, the concept of a training set as understood in AI/ML is not relevant here.
-
How Ground Truth for the Training Set Was Established:
- Not applicable, as there is no "training set" in the context of an AI/ML model for this type of device. The device's performance is intrinsically linked to its chemical reagents and mass spectrometry technology, which are developed and calibrated through standard analytical chemistry procedures.
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February 7, 2020
PerkinElmer Inc. Eva Nalian Sr. Manager Regulatory Affairs 940 Winter Street Waltham, MA 02451
Re: K193103
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: NOL Dated: November 7, 2019 Received: November 8, 2019
Dear Eva Nalian:
We have reviewed vour 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
{1}------------------------------------------------
statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely.
Kellie B. Kelm, Ph.D. Acting Director Division Director of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Ouality Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known)
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 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 1 | Gln\Lys |
| Glutamic acid | Glu |
| Glycine | Gly |
| Leucine\Isoleucine\Hydroxyproline 1 | Leu\lle\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 1 | C3DC\C4OH |
| Butyrylcarnitine | C4 |
| Methylmalonyl\3-Hydroxy-isovalerylcarnitine 1 | C4DC\C5OH |
| Isovalerylcarnitine | C5 |
| Tiglylcarnitine | C5:1 |
| Glutarylcarnitine\3-Hydroxy-hexanoylcarnitine 1 | C5DC\C6OH |
| Hexanoylcarnitine | C6 |
| Adipylcarnitine | C6DC |
1 Analytes in these rows are either isomers or isobars and cannot be distinguished in the tandem mass spectrometry experiment.
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Table 1 (continued): Analytes measured by the NeoBase 2 Non-derivatized MSMS kit.
| Octanoylcarnitine | C8 |
|---|---|
| Octenoylcarnitine | C8:1 |
| Decanoylcarnitine | C10 |
| 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<br>Heptadecanoylcarnitine 1 | 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 |
| 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 spectrometry experiment.
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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)
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Image /page/5/Picture/1 description: The image shows the logo for PerkinElmer. The logo features the company name in a bold, sans-serif font, with the "Perkin" part in black and the "Elmer" part in blue. Above the name is a stylized blue graphic element that resembles an abstract shape. Below the name is the company's slogan, "For the Better," also in blue.
510(k) Summary
This summary of safety and effectiveness information is supplied in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.
The assigned number is _K193103
| Submitted by: | PerkinElmer, Inc.940 Winter StreetWaltham MA 02451 |
|---|---|
| Contact Person: | Eva NalianTel: 647-633-8435 |
| Trade Name: | NeoBase 2 Non-derivatized MSMS kit |
| Common Name: | Newborn screening test system for amino acids, freecarnitine, and acylcarnitines using tandem mass spectrometry |
| Regulation: | 21 CFR 862.1055 |
| Classification: | II |
| Panel: | 75 Chemistry |
| Product Code: | NQL |
| Predicate device: | NeoBase 2 Non-derivatized MSMS kit (K173568) |
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Image /page/6/Picture/1 description: The image shows the logo for PerkinElmer. The logo consists of a blue stylized "P" shape with a triangle cut out of the top right corner. Below the symbol, the name "PerkinElmer" is written in a combination of black and gray text. Underneath the name, the phrase "For the Better" is written in a smaller font size.
1. Device Description:
Each NeoBase 2 Non-derivatized MSMS kit contains reagents for 960 assays. The kit is designed to be used with NeoBase 2 Non-derivatized Assay Solutions consisting of Neo MSMS Flow Solvent and NeoBase 2 Extraction Solution and NeoBase 2 Succinylacetone Assay Solution.
- NeoBase 2 Internal Standards - 1 vial
- NeoBase 2 Controls Low, High - 3 filter paper cassettes (Whatman, no. 903) containing 3 spots of each level per cassette
- Microplate, U-bottomed - 20 plates
- Adhesive microplate covers - 20 sheets
- Barcode labels for the plates - 30 pcs (10 different barcodes, 3 pcs of each)
- Lot-specific quality control certificate
This kit contains components manufactured from human blood. The source materials have been tested by FDA-approved methods for hepatitis B surface antigen, anti-hepatitis C and anti-HIV 1 and 2 antibodies and found to be negative.
Instruments used:
- . QSight® 210 MD Screening System is comprised of:
- QSight® 210 MD Mass Spectrometer
- QSight® HC Autosampler MD
- QSight® Binary Pump MD ●
- Simplicity™ 3Q MD Software
- PerkinElmer MSMS Workstation software ●
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Image /page/7/Picture/1 description: The image shows the logo for PerkinElmer. The logo features a stylized blue "P" shape with a pointed end, positioned above the company name. The text "PerkinElmer" is written in a bold, sans-serif font, with the "E" in "Elmer" slightly larger and in a lighter shade of blue. Below the company name, the tagline "For the Better" is written in a smaller, italicized font, also in blue.
2. Intended Use:
The NeoBase 2 Non-derivatized MSMS kit is intended for the measurement and evaluation of amino acid, succinylacetone, free carnitine, 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.
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 1 | Gln\Lys |
| Glutamic acid | Glu |
| Glycine | Gly |
| Leucine\Isoleucine\Hydroxyproline 1 | Leu\lle\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 1 | C3DC\C4OH |
| Butyrylcarnitine | C4 |
| Methylmalonyl\3-Hydroxy-isovalerylcarnitine 1 | C4DC\C5OH |
| Isovalerylcarnitine | C5 |
| Tiglylcarnitine | C5:1 |
| Glutarylcarnitine\3-Hydroxy-hexanoylcarnitine 1 | C5DC\C6OH |
| Hexanoylcarnitine | C6 |
| Adipylcarnitine | C6DC |
1 Analytes in these rows are either isomers or isobars and cannot be distinguished in the tandem mass spectrometry experiment.
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Table 1 (continued): Analytes measured by the NeoBase 2 Non-derivatized MSMS kit.
| Octanoylcarnitine | C8 |
|---|---|
| Octenoylcarnitine | C8:1 |
| Decanoylcarnitine | C10 |
| 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\ | C16:1OH\C17 |
| Heptadecanoylcarnitine 1 | 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 |
| 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 spectrometry experiment.
3. Substantial Equivalency:
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Image /page/9/Picture/1 description: The image shows the logo for PerkinElmer. The logo consists of a blue abstract shape resembling a stylized "P" and a right-pointing arrow. Below the shape, the text "PerkinElmer" is written in a bold, sans-serif font. Underneath "PerkinElmer", the tagline "For the Better" is written in a smaller, italicized, blue font.
| Characteristics | Proposed Device | Predicate (K173568) |
|---|---|---|
| Intended Use/Indications for Use | The NeoBase™ 2 Non-derivatized MSMS kit isintended for the measurement and evaluation ofamino acid, succinylacetone, free carnitine,acylcarnitine, nucleoside and lysophospholipidconcentrations with a tandem mass spectrometerfrom newborn heel prick blood specimens dried onfilter paper. Quantitative analysis of these analytesand their relationship with each other is intended toprovide analyte concentration profiles that may aidin screening newborns for metabolic disorders. | Same |
| Test Principle | Analytes in sample are measured by tandem massspectrometry through analyte-specific masstransitions appropriate for each type of analyte. Theextracted analytes are measured for set timeperiods and compared to the signal intensitiesproduced by the corresponding isotope- labeledinternal standards. The concentrations aredetermined by comparing the signal intensities ofthe known standards to the measured analytes. | Same |
| Disorders Screened | Amino-, organic-, and fatty acid metabolic disorders | Same |
| Analytes Measured | Amino acids, free carnitine, acylcarnitines,succinylacetone, nucleosides, and lysophospholipids | Same |
| Instrument / SoftwarePlatform | PerkinElmer QSight 210MD Screening System: QSight® Waters TQD instrument with MassLynx v4.1HC Autosampler MD,QSight® Binary Pump MD,Simplicity™ 3Q MD Software, PerkinElmer MSMSWorkstation Software | firmware, with Waters 1525 sample pump,with Waters 2777c autosampler, with WatersNeoLynx v4.1 software and with thePerkinElmer MSMS Workstation Software |
| Sample Type | Punch from dried blood spot specimen | Same |
| Calibrators | Internal calibration using several isotopically labeledstandards, included as dried material in vials.Internal standards must be reconstituted withextraction solution prior to their use. | Same |
| Throughput | Ninety-six tests per microtiterplate. Multiple plates can be analyzed | Same |
{10}------------------------------------------------
Image /page/10/Picture/1 description: The image shows the logo for PerkinElmer. The logo consists of the company name in a bold, sans-serif font, with the words "For the Better" in a smaller, italicized font underneath. Above the company name is a blue graphic element that resembles a stylized "P" or an arrow pointing to the right. The overall design is clean and modern, conveying a sense of innovation and progress.
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.
4. Summary of the Studies:
Precision:
The precision was determined in accordance with CLSI document EP05-A3. The repeatability and withinlaboratory variation for NeoBase 2 Non-derivatized MSMS kit is based on 80 determinations: 40 plates measured over 20 working days, each plate having 2 replicates per sample. One QSight was used. Betweenlot variation is based on 75 determinations: 15 plates measured over five working days using three kit lots, each plate having 5 replicates per sample. One TQD was used. Between-instrument variation is based on altogether 50 determinations: 10 plates were measured with two QSights over 5 working days, each plate having 5 replicates per sample. The results are presented in the table below.
Repeatability, within-laboratory, between-lot, between-instrument and total variation determined for the NeoBase 2 Non-derivatized MSMS kit using QSight:
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 161 | 11 | 6.5 | 17 | 9.8 | 4.1 | 2.8 | 19 | 12 | 26 | 16 |
| 2 | 361 | 21 | 5.5 | 27 | 7.1 | 6.2 | 1.8 | 17 | 4.6 | 32 | 9.0 |
| 3 | 414 | 24 | 5.4 | 30 | 6.8 | 4.2 | 1.0 | 3.4 | 0.84 | 30 | 7.3 |
| 4 | 518 | 28 | 5.3 | 34 | 6.3 | 15 | 3.1 | 0.04 | 0.01 | 37 | 7.2 |
Ala
Arg
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | |||
| 1 | 7.5 | 0.45 | 6.0 | 0.62 | 8.3 | 0.47 | 6.8 | <0.01 | <0.01 | 0.78 | 10 | |
| 2 | 23 | 1.3 | 5.7 | 1.7 | 7.3 | 0.11 | 0.49 | <0.01 | <0.01 | 1.7 | 7.6 | |
| 3 | 69 | 3.4 | 4.9 | 3.7 | 5.4 | 1.7 | 2.6 | <0.01 | <0.01 | 4.1 | 5.9 | |
| 4 | 157 | 5.2 | 3.3 | 8.0 | 5.0 | 5.2 | 3.5 | 1.1 | 0.67 | 9.6 | 6.1 |
{11}------------------------------------------------
Image /page/11/Picture/1 description: The image shows the logo for PerkinElmer. The logo features a blue stylized "P" with a triangle cut out of the top right corner, positioned above the company name "PerkinElmer" in a bold, sans-serif font. Below the company name is the tagline "For the Better" in a smaller, italicized font, also in blue.
| Asa1 | |||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||||
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||||
| 1 | 0.31 | 0.06 | 29 | 0.09 | 43 | <0.01 | 0.74 | 0.03 | 14 | 0.10 | 32 | ||
| 2 | 2.2 | 0.14 | 6.7 | 0.24 | 11 | 0.07 | 3.1 | 0.10 | 4.3 | 0.27 | 12 | ||
| 3 | 8.1 | 0.39 | 5.0 | 0.76 | 9.9 | 0.44 | 5.3 | 0.66 | 7.8 | 1.1 | 14 | ||
| 4 | 21 | 0.74 | 3.6 | 1.7 | 8.1 | 0.95 | 4.5 | 1.7 | 7.8 | 2.6 | 12 | ||
| 5 | 57 | 2.3 | 3.8 | 5.6 | 9.2 | 2.1 | 4.6 | 5.7 | 8.7 | 8.2 | 14 |
¹ Asa is measured as a total concentration of Asa and its anhydrides.
Cit
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 10 | 1.1 | 12 | 1.2 | 12 | 0.33 | 3.0 | <0.01 | 0.01 | 1.2 | 12 |
| 2 | 68 | 3.3 | 5.0 | 4.3 | 6.5 | 2.1 | 3.1 | 1.2 | 1.7 | 5.0 | 7.3 |
| 3 | 202 | 10 | 5.1 | 11 | 5.3 | 5.4 | 2.7 | <0.01 | <0.01 | 12 | 5.9 |
| 4 | 470 | 27 | 5.8 | 29 | 6.3 | 11 | 2.3 | 10 | 2.1 | 33 | 6.9 |
| 5 | 957 | 50 | 5.1 | 55 | 5.7 | 27 | 3.0 | 16 | 1.6 | 64 | 6.7 |
Gln\Lys
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 43 | 2.8 | 5.9 | 3.7 | 7.8 | 1.0 | 2.5 | 2.9 | 7.2 | 4.8 | 11 |
| 2 | 487 | 24 | 4.6 | 30 | 5.7 | 15 | 3.3 | 0.02 | <0.01 | 34 | 6.9 |
| 3 | 675 | 35 | 4.9 | 43 | 6.0 | 3.3 | 0.52 | <0.01 | <0.01 | 43 | 6.4 |
| 4 | 1064 | 52 | 4.6 | 65 | 5.7 | 37 | 3.7 | 13 | 1.2 | 75 | 7.1 |
| 5 | 2274 | 94 | 3.9 | 136 | 5.7 | 20 | 0.90 | 0.13 | 0.01 | 138 | 6.1 |
Gly
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 247 | 16 | 6.6 | 24 | 9.8 | 7.9 | 3.0 | 3.9 | 1.6 | 25 | 10 |
| 2 | 324 | 19 | 6.3 | 23 | 7.6 | 6.6 | 1.9 | 4.7 | 1.5 | 25 | 7.6 |
| 3 | 524 | 32 | 6.3 | 41 | 8.1 | 6.1 | 1.1 | 0.01 | <0.01 | 42 | 8.0 |
| 4 | 930 | 52 | 5.7 | 82 | 8.9 | 26 | 2.8 | 19 | 2.1 | 88 | 9.5 |
Leu\lle\Pro-OH
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 58 | 2.5 | 4.3 | 3.2 | 5.5 | 0.54 | 0.87 | <0.01 | <0.01 | 3.2 | 5.6 |
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Image /page/12/Picture/0 description: The image shows the logo for PerkinElmer. The logo consists of a blue stylized "P" above the name "PerkinElmer" in black. Below the name is the tagline "For the Better" in a smaller, italicized font.
| 2 | 202 | 9.0 | 4.5 | 11 | 5.7 | 4.7 | 2.3 | <0.01 | <0.01 | 12 | 6.2 |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 3 | 350 | 16 | 4.4 | 16 | 4.4 | 2.7 | 0.80 | <0.01 | <0.01 | 16 | 4.6 |
| 4 | 656 | 31 | 4.7 | 33 | 4.9 | 17 | 2.6 | 0.03 | 0.01 | 37 | 5.6 |
| 5 | 1121 | 45 | 3.9 | 54 | 4.7 | 27 | 2.5 | <0.01 | <0.01 | 60 | 5.4 |
Met
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 2.2 | 0.28 | 18 | 0.28 | 18 | 0.26 | 8.3 | 0.21 | 11 | 0.43 | 20 |
| 2 | 51 | 2.6 | 5.1 | 3.1 | 6.2 | 1.9 | 3.7 | 0.58 | 1.1 | 3.7 | 7.3 |
| 3 | 155 | 8.2 | 5.4 | 9.1 | 5.9 | 2.3 | 1.5 | <0.01 | <0.01 | 9.4 | 6.1 |
| 4 | 369 | 19 | 5.0 | 23 | 6.3 | 9.8 | 2.7 | 4.2 | 1.1 | 26 | 6.9 |
| 5 | 696 | 26 | 3.7 | 37 | 5.2 | 14 | 2.0 | 0.01 | <0.01 | 40 | 5.7 |
Orn
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 29 | 1.9 | 6.7 | 2.9 | 10 | 0.50 | 1.7 | 0.89 | 2.9 | 3.1 | 11 |
| 2 | 109 | 4.1 | 3.8 | 7.0 | 6.4 | 1.5 | 1.4 | 2.0 | 1.8 | 7.4 | 6.8 |
| 3 | 204 | 10 | 4.9 | 12 | 5.7 | 2.3 | 1.2 | <0.01 | <0.01 | 12 | 5.8 |
| 4 | 382 | 14 | 3.8 | 17 | 4.5 | 9.9 | 2.7 | 11 | 2.7 | 23 | 5.9 |
Phe
| Sample | Total meanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | Total Variation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 22 | 1.1 | 5.1 | 1.2 | 5.8 | 0.43 | 1.7 | <0.01 | <0.01 | 1.3 | 5.8 |
| 2 | 127 | 4.3 | 3.4 | 5.6 | 4.5 | 3.5 | 2.7 | 1.2 | 0.94 | 6.7 | 5.3 |
| 3 | 340 | 15 | 4.5 | 16 | 4.8 | 4.7 | 1.4 | <0.01 | <0.01 | 17 | 5.0 |
| 4 | 778 | 35 | 4.5 | 43 | 5.5 | 29 | 3.8 | 7.3 | 0.91 | 52 | 6.7 |
| 5 | 1436 | 39 | 2.6 | 65 | 4.4 | 17 | 1.2 | 23 | 1.6 | 71 | 4.9 |
Pro
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 40 | 1.8 | 4.6 | 2.2 | 5.6 | 0.57 | 1.4 | <0.01 | <0.01 | 2.3 | 5.7 |
| 2 | 178 | 7.0 | 3.9 | 8.1 | 4.5 | 4.6 | 2.6 | 2.0 | 1.1 | 9.5 | 5.3 |
| 3 | 316 | 14 | 4.4 | 15 | 4.7 | 2.6 | 0.86 | <0.01 | <0.01 | 16 | 4.9 |
| 4 | 596 | 28 | 4.5 | 30 | 4.8 | 16 | 2.8 | 6.9 | 1.1 | 34 | 5.7 |
{13}------------------------------------------------
Image /page/13/Picture/1 description: The image shows the logo for PerkinElmer. The logo consists of the company name in a bold, sans-serif font, with a blue graphic above it. The graphic is a stylized "P" shape with an arrow pointing to the right. Below the company name is the tagline "For the Better" in a smaller, italicized font.
| Tyr | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample | Totalmean | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
| μmol/L | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | |
| 1 | 20 | 1.2 | 6.3 | 1.5 | 7.5 | 0.52 | 2.6 | <0.01 | <0.01 | 1.5 | 7.7 |
| 2 | 109 | 4.1 | 3.9 | 4.7 | 4.5 | 3.6 | 3.3 | 1.4 | 1.3 | 6.1 | 5.6 |
| 3 | 264 | 9.2 | 3.6 | 9.2 | 3.6 | 4.1 | 1.6 | <0.01 | <0.01 | 10 | 3.8 |
| 4 | 586 | 21 | 3.6 | 26 | 4.6 | 15 | 2.6 | 2.4 | 0.39 | 30 | 5.1 |
Val
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 55 | 2.4 | 4.5 | 2.8 | 5.2 | 0.81 | 1.3 | <0.01 | <0.01 | 2.9 | 5.2 |
| 2 | 205 | 8.7 | 4.2 | 11 | 5.4 | 5.6 | 2.8 | 3.6 | 1.8 | 13 | 6.4 |
| 3 | 314 | 15 | 4.8 | 15 | 4.8 | 4.2 | 1.4 | <0.01 | <0.01 | 16 | 5.1 |
| 4 | 540 | 27 | 4.9 | 29 | 5.2 | 15 | 3.0 | 10 | 1.9 | 34 | 6.4 |
C0
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 8.4 | 0.46 | 5.6 | 0.54 | 6.5 | 0.04 | 0.47 | <0.01 | 0.01 | 0.55 | 6.5 |
| 2 | 42 | 1.8 | 4.4 | 2.1 | 5.0 | 1.5 | 3.7 | 1.1 | 2.6 | 2.8 | 6.7 |
| 3 | 89 | 4.9 | 5.4 | 5.4 | 5.9 | 1.1 | 1.2 | 0.01 | 0.02 | 5.5 | 6.1 |
| 4 | 186 | 8.3 | 4.3 | 11 | 5.5 | 6.7 | 3.7 | 3.0 | 1.6 | 13 | 6.9 |
C2
| Sample | Totalmean | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| μmol/L | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | |
| 1 | 3.6 | 0.17 | 4.8 | 0.21 | 5.7 | 0.09 | 2.5 | <0.01 | <0.01 | 0.22 | 6.3 |
| 2 | 12 | 0.58 | 4.7 | 0.72 | 5.9 | 0.47 | 3.9 | 0.06 | 0.47 | 0.87 | 7.1 |
| 3 | 18 | 0.87 | 4.7 | 0.91 | 4.9 | 0.09 | 0.49 | <0.01 | <0.01 | 0.91 | 5.0 |
| 4 | 30 | 1.3 | 4.3 | 1.5 | 4.9 | 0.66 | 2.2 | <0.01 | <0.01 | 1.7 | 5.5 |
СЗ
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.44 | 0.03 | 6.7 | 0.04 | 8.1 | 0.01 | 2.9 | <0.01 | 0.01 | 0.04 | 9.1 |
| 2 | 4.5 | 0.18 | 3.8 | 0.27 | 5.5 | 0.14 | 3.2 | 0.03 | 0.76 | 0.30 | 6.6 |
| 3 | 13 | 0.73 | 5.1 | 0.95 | 6.6 | 0.16 | 1.2 | <0.01 | <0.01 | 0.96 | 7.2 |
| 4 | 32 | 1.6 | 4.8 | 2.3 | 6.8 | 1.2 | 4.0 | 0.36 | 1.2 | 2.6 | 8.3 |
{14}------------------------------------------------
Image /page/14/Picture/1 description: The image shows the logo for PerkinElmer. The logo consists of a blue abstract shape resembling a stylized "P" with a forward-pointing arrow, positioned above the company name "PerkinElmer" in a bold, sans-serif font. Below the company name, the tagline "For the Better" is written in a smaller, italicized font.
| C4 | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.06 | 0.01 | 11 | 0.01 | 12 | <0.01 | 3.5 | <0.01 | 0.01 | 0.01 | 3 |
| 2 | 0.57 | 0.03 | 4.9 | 0.03 | 5.5 | 0.02 | 2.9 | <0.01 | 0.78 | 0.04 | 6 |
| 3 | 1.8 | 0.09 | 4.8 | 0.09 | 4.8 | 0.01 | 0.31 | <0.01 | <0.01 | 0.09 | 4 |
| 4 | 4.2 | 0.18 | 4.1 | 0.20 | 4.6 | 0.08 | 2.0 | 0.04 | 0.93 | 0.22 | 5 |
CE
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.04 | 0.01 | 19 | 0.01 | 19 | <0.01 | 2.6 | <0.01 | 0.01 | 0.01 | 17 |
| 2 | 1.0 | 0.04 | 4.2 | 0.06 | 6.3 | 0.03 | 2.8 | <0.01 | <0.01 | 0.07 | 6.7 |
| 3 | 3.6 | 0.17 | 4.9 | 0.18 | 5.2 | 0.04 | 1.1 | <0.01 | <0.01 | 0.19 | 5.2 |
| 4 | 8.9 | 0.37 | 4.2 | 0.43 | 4.9 | 0.31 | 3.6 | <0.01 | <0.01 | 0.53 | 6.0 |
C5DC\C6OH
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.03 | 0.01 | 17 | 0.01 | 23 | <0.01 | 9.2 | <0.01 | 5.5 | 0.01 | 25 |
| 2 | 0.44 | 0.03 | 7.5 | 0.03 | 7.9 | 0.01 | 3.1 | 0.01 | 1.5 | 0.04 | 8.6 |
| 3 | 1.5 | 0.09 | 5.9 | 0.10 | 6.2 | 0.02 | 1.1 | <0.01 | <0.01 | 0.10 | 6.3 |
| 4 | 3.8 | 0.22 | 5.7 | 0.24 | 6.2 | 0.07 | 1.9 | 0.07 | 1.7 | 0.26 | 6.7 |
CE
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.38 | 0.02 | 4.4 | 0.02 | 6.0 | 0.01 | 2.4 | <0.01 | <0.01 | 0.02 | 6.4 |
| 2 | 1.4 | 0.07 | 5.0 | 0.07 | 5.0 | 0.03 | 1.8 | 0.01 | 0.59 | 0.07 | 5.3 |
| 3 | 3.4 | 0.16 | 4.7 | 0.17 | 4.9 | 0.10 | 3.1 | <0.01 | <0.01 | 0.20 | 5.8 |
C8
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 2.0 | 0.10 | 4.8 | 0.12 | 6.0 | 0.08 | 3.8 | <0.01 | <0.01 | 0.14 | 7.1 |
| 2 | 7.5 | 0.35 | 4.7 | 0.37 | 4.9 | 0.12 | 1.6 | <0.01 | <0.01 | 0.39 | 5.2 |
| 3 | 18 | 0.86 | 4.6 | 0.99 | 5.3 | 0.64 | 3.6 | 0.10 | 0.55 | 1.2 | 6.4 |
| 4 | 35 | 1.3 | 3.5 | 1.6 | 4.4 | 0.44 | 1.3 | 0.56 | 1.6 | 1.7 | 5.0 |
CV% 11 6.2 4.8 5.2
{15}------------------------------------------------
Image /page/15/Picture/1 description: The image shows the logo for PerkinElmer. The logo consists of a blue stylized "P" above the name "PerkinElmer" in black font. Below the name is the tagline "For the Better" in a smaller, italicized blue font.
C10
| Sample | Totalmean | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| μmol/L | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | |
| 1 | 0.46 | 0.02 | 5.5 | 0.03 | 6.8 | 0.01 | 2.4 | <0.01 | <0.01 | 0.03 | 7.2 |
| 2 | 1.6 | 0.09 | 5.8 | 0.11 | 7.1 | 0.02 | 1.2 | <0.01 | <0.01 | 0.11 | 7.1 |
| 3 | 3.9 | 0.24 | 6.2 | 0.29 | 7.7 | 0.11 | 2.9 | 0.04 | 1.1 | 0.32 | 8.2 |
C12
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.49 | 0.02 | 5.1 | 0.03 | 6.4 | 0.02 | 3.4 | <0.01 | <0.01 | 0.04 | 7.2 |
| 2 | 1.8 | 0.10 | 5.8 | 0.10 | 5.8 | 0.01 | 0.76 | <0.01 | <0.01 | 0.10 | 5.8 |
| 3 | 4.3 | 0.25 | 5.8 | 0.26 | 6.0 | 0.16 | 3.7 | <0.01 | <0.01 | 0.30 | 7.0 |
C14
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | 0.05 | <0.01 | 9.0 | 0.01 | 10 | <0.01 | 3.5 | <0.01 | 0.02 | 0.01 | 11 |
| 2 | 0.56 | 0.03 | 5.1 | 0.03 | 6.0 | 0.01 | 1.6 | 0.01 | 1.9 | 0.04 | 6.4 |
| 3 | 1.8 | 0.09 | 5.0 | 0.09 | 5.0 | 0.03 | 1.6 | <0.01 | <0.01 | 0.09 | 5.3 |
| 4 | 4.3 | 0.24 | 5.7 | 0.25 | 5.8 | 0.12 | 2.8 | 0.09 | 1.9 | 0.29 | 6.8 |
C16
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.99 | 0.05 | 5.1 | 0.06 | 5.8 | 0.02 | 2.3 | <0.01 | <0.01 | 0.06 | 6.1 |
| 2 | 3.6 | 0.16 | 4.4 | 0.20 | 5.7 | 0.13 | 3.6 | <0.01 | <0.01 | 0.24 | 6.6 |
| 3 | 11 | 0.52 | 5.0 | 0.66 | 6.4 | 0.07 | 0.62 | <0.01 | <0.01 | 0.67 | 6.3 |
| 4 | 24 | 1.2 | 5.2 | 1.5 | 6.3 | 0.35 | 1.5 | <0.01 | <0.01 | 1.5 | 6.4 |
C18
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.49 | 0.03 | 5.5 | 0.03 | 5.9 | 0.01 | 2.0 | <0.01 | 0.01 | 0.03 | 6.2 |
| 2 | 1.2 | 0.06 | 4.5 | 0.08 | 6.1 | 0.03 | 2.7 | 0.01 | 1.1 | 0.09 | 6.9 |
| 3 | 2.9 | 0.15 | 4.9 | 0.15 | 5.1 | 0.03 | 1.0 | <0.01 | <0.01 | 0.16 | 5.4 |
| 4 | 6.2 | 0.28 | 4.4 | 0.34 | 5.3 | 0.03 | 0.58 | <0.01 | <0.01 | 0.34 | 5.5 |
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| SA | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
| 1 | 0.39 | 0.04 | 17 | 0.05 | 19 | 0.01 | 2.1 | <0.01 | 0.02 | 0.05 | 13 |
| 2 | 3.5 | 0.20 | 6.3 | 0.25 | 7.9 | 0.09 | 2.4 | <0.01 | 0.01 | 0.27 | 7.7 |
| 3 | 13 | 0.62 | 5.4 | 1.0 | 8.6 | 0.22 | 1.6 | <0.01 | <0.01 | 1.0 | 8.1 |
| 4 | 35 | 1.3 | 4.0 | 2.6 | 7.8 | 0.77 | 2.2 | 0.01 | 0.04 | 2.7 | 7.8 |
| 5 | 85 | 5.4 | 5.6 | 8.4 | 8.7 | 1.7 | 3.1 | <0.01 | <0.01 | 8.6 | 10 |
ADO
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.13 | 0.02 | 19 | 0.02 | 20 | <0.01 | 2.1 | <0.01 | 0.02 | 0.02 | 17 |
| 2 | 1.4 | 0.07 | 5.1 | 0.10 | 6.8 | 0.02 | 1.3 | <0.01 | <0.01 | 0.10 | 7.1 |
| 3 | 5.7 | 0.20 | 3.5 | 0.22 | 3.8 | 0.07 | 1.2 | <0.01 | <0.01 | 0.23 | 4.0 |
| 4 | 15 | 0.50 | 3.2 | 0.58 | 3.7 | 0.23 | 1.6 | 0.04 | 0.28 | 0.62 | 4.1 |
| 5 | 30 | 0.81 | 2.6 | 1.2 | 3.8 | 0.43 | 1.5 | 0.35 | 1.2 | 1.3 | 4.3 |
C26:0-LPC
| Sample | Totalmeanμmol/L | Repeatability | Within-Lab | Between-lot | Between-instrument | TotalVariation | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.26 | 0.04 | 16 | 0.06 | 23 | 0.01 | 4.5 | <0.01 | 0.01 | 0.06 | 23 |
| 2 | 0.78 | 0.08 | 10 | 0.10 | 12 | 0.01 | 1.1 | <0.01 | <0.01 | 0.10 | 12 |
| 3 | 1.5 | 0.11 | 7.1 | 0.11 | 7.7 | 0.01 | 0.47 | <0.01 | <0.01 | 0.11 | 7.7 |
| 4 | 3.0 | 0.18 | 5.8 | 0.19 | 6.4 | 0.01 | 0.55 | 0.10 | 3.0 | 0.22 | 7.3 |
| 5 | 5.5 | 0.29 | 5.0 | 0.37 | 6.5 | 0.10 | 2.1 | 0.11 | 1.9 | 0.40 | 7.3 |
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Analytical Sensitivity:
The analytical sensitivity (the lowest measurable analyte concentrations) has been demonstrated using two QSights. Results are summarized in the table below.
Analytical sensitivity limits for the NeoBase 2 Non-derivatized MSMS kit using QSight:
| Analyte | QSightAnalytical sensitivity limit (μmol/L) |
|---|---|
| Ala | 3.66 |
| Arg | 0.64 |
| Asa 1 | 0.16 |
| Cit | 2.63 |
| Gln\Lys | 6.31 |
| Gly | 8.61 |
| Leu\Ile\Pro-OH | 0.40 |
| Met | 1.56 |
| Orn | 1.83 |
| Phe | 0.29 |
| Pro | 0.34 |
| Tyr | 1.84 |
| Val | 0.84 |
| C0 | 0.18 |
| C2 | 0.04 |
| C3 | 0.02 |
| C4 | 0.01 |
| C5 | 0.01 |
| C5DC\C6OH | 0.04 |
| C6 | 0.03 |
| C8 | 0.10 |
| C10 | 0.04 |
| C12 | 0.10 |
| C14 | 0.01 |
| C16 | 0.02 |
| C18 | 0.01 |
| SA | 0.24 |
| ADO | 0.07 |
¹ Asa is measured as a total concentration of Asa and its anhydrides.
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Linearity:
The linearity was determined in accordance with CLSI document EP06-A using one QSight. The assay is demonstrated to be linear as presented in the table below.
| Analyte | Linear range lowerlimit (µmol/L) | Linear range upperlimit (µmol/L) |
|---|---|---|
| Ala | 163 | 1450 |
| Arg | 1.84 | 359 |
| Asa 1 | 0.22 | 67.2 |
| Cit | 9.18 | 1040 |
| Gln\Lys | 42.2 | 2450 |
| Gly | 268 | 2070 |
| Leu\lle\Pro-OH | 57.8 | 1430 |
| Met | 1.66 | 802 |
| Orn | 25.6 | 807 |
| Phe | 20.9 | 1500 |
| Pro | 37.3 | 1240 |
| Tyr | 19.7 | 1270 |
| Val | 51.0 | 1130 |
| C0 | 7.80 | 407 |
| C2 | 3.53 | 147 |
| C3 | 0.41 | 64.2 |
| C4 | 0.05 | 11.3 |
| C5 | 0.04 | 17.8 |
| C5DC\C6OH | 0.04 | 7.30 |
| C6 | 0.03 | 7.99 |
| C8 | 0.10 | 41.2 |
| C10 | 0.04 | 7.24 |
| C12 | 0.10 | 10.3 |
| C14 | 0.05 | 9.45 |
| C16 | 0.84 | 46.7 |
| C18 | 0.48 | 12.8 |
| SA | 0.24 | 88.2 |
| ADO | 0.17 | 41.3 |
| C26:0-LPC | 0.22 | 7.08 |
Linear ranges determined for the NeoBase 2 Non-derivatized MSMS kit using QSight:
4 Asa is measured as a total concentration of Asa and its anhydrides.
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Interference:
The NeoBase 2 Non-derivatized MSMS kit was evaluated for interference in accordance with CLSI document EP07 using QSight. The substances potentially interfering with the assay and additional 9 substances that are abundant in the DBS sample matrix and have potential for unspecific effects on the test results were further tested. The substances potentially interfering with the assay were added into whole blood. The interference samples included NeoBase 2 Control analytes at two concentrations (below and above typical cut-off range). The NeoBase 2 surrogate analytes, which are not included in NeoBase 2 Controls, were studied at endogenous concentration level. The following substances were found not to interfere with the assay on QSight at the concentration indicated:
| Tested substance | Added concentration of tested substance in blood |
|---|---|
| Formiminoglutamic acid (Figlu) | 37.1 µmol/L |
| O-Acetyl-L-serine | 1000 µmol/L |
| 6-Aminocaproic Acid | 6.07 µmol/L |
| DL-Malic acid | 3000 µmol/L |
| 4-Aminoantipyrine | 500 µmol/L |
| Propranolol | 7.74 µmol/L |
| 2,5-dihydroxybenzoic acid | 127 µmol/L |
| Bilirubin conjugated | 15 mg/dL |
| Bilirubin unconjugated | 10 mg/dL |
| Calcifediol | 250 nmol/L |
| Acetaminophen | 5.5 mg/dL |
| Lidocaine | 51.2 µmol/L |
In the study, the following interferents were identified:
Sarcosine:
Amino acid derivative Sarcosine was found to interfere with the assay by increasing the measured Ala concentration. Sarcosine concentrations above 31.3 umol/L may theoretically cause a false positive screening result. Sarcosine concentration in plasma ranges from 0-625 umol/L in newborns aged 0-1 months. However, Sarcosine does not exist in healthy newborns; it is only found at detectable amount when a newborn is affected by hypersarcosinemia. Therefore, Sarcosine is unlikely to interfere with Ala in routine testing.
Creatine:
Non-essential amino acid Creatine was found to interfere with the assay by increasing the measured Ala, Glu and Leu concentrations. Creatine concentrations above 450 umol/L with Ala, above 900 umol/L with Glu or above 1500 umol/L with Leu may theoretically cause a false positive screening result. The normal expected Creatine level in newborns aged 0-1 months is 107-640 umol/L in whole blood. Therefore, Creatine is unlikely to interfere with Glu and Leu in routine testing. The proposed cut-off level measured with NeoBase 2 assay for Ala is approximately 600 umol/L (e.g. 99th percentile, 627 umol/L). An additional 2619 µmol/L dose of Creatine would be needed to raise the Ala concentration from endogenous level (351
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umol/L) to the cut-off level (600 umol/L).
L-Asparagine:
Non-essential amino acid L-Asparagine was found to interfere with the assay by increasing the measured Orn concentration. L-Asparagine concentrations above 750 umol/L may theoretically cause a false positive screening result on Orn. The reference plasma level of L-Asparagine in newborns aged 0-1 months is 29-132 umol/L [4]. Therefore, L-Asparagine is unlikely to interfere with Orn in routine testing.
L-Lysine:
L-Lysine was found to interfere with the assay by decreasing the measured Arg concentrations and by increasing the measured Gln and Glu concentrations above 1000 µmol/L caused a decrease in the measured Arg by 19%. Newborn screening samples with arginine levels close to the cutoff and with suspected hyperlysinemia should be tested using a method that shows no interference by lysine. L-Lysine is an essential amino acid and is isobaric to NeoBase 2 analyte Gln. NeoBase 2 assay cannot separate the compounds, and the result of Gln is a sum of Gln and L-Lysine (Gln\Lys). The reference plasma level of L-Lysine, Gln and Glu in newborns aged 0-1 months are 92-325 umol/L, 376-709 umol/L and 62-620 umol/L, respectively. L-Lysine may theoretically cause a false positive screening result to Gln. The proposed cut-off area measured with NeoBase 2 assay for Gln is generally high (e.g. 99th percentile, 1200 μmol/L). An additional 1059 µmol/L dose of L-Lysine would be needed to raise the Gln concentration from endogenous level (435 µmol/L) to the cut-off area (1200 µmol/L). For Glu, L-Lysine concentrations from 1500 umol/L may theoretically cause a false positive screening result. Since the whole blood used in the interference samples contains also endogenous L-Lysine, the sum of spiked and endogenous L-Lysine levels are in the upper part of newborn physiological L-Lysine range. When considering the positive rate in neonatal screening, it can be expected that any possible effect of endogenous L-lysine would be incorporated in the users established cut-offs for Gln and Glu (upper population percentiles), and would not therefore cause false positives in routine screening. In hyperlysinemia, the concentration of L-Lysine in blood plasma is relatively high. Plasma L-Lysine levels have been reported to exceed 600 umol/L and can reach up to 2000 µmol/L. When blood specimen is taken from newborn with such a condition, L-Lysine may cause false positive screening results to Gln and/or Glu.
L-Glutamic acid:
Non-essential amino acid and NeoBase 2 analyte L-Glutamic acid (Glu) was found to interfere with the assay by increasing the measured Met concentrations above 2250 µmol/L may theoretically cause a false positive screening result on Met. The reference plasma level of Glu in newborns aged 0-1 months is 62-620 µmol/L. Therefore, Glu is very unlikely to interfere with Met in routine testing.
L-Methionine sulfone:
Amino acid derivative L-Methionine sulfone was found to interfere with the assay, thus increasing the measured Tyr concentration. L-Methionine sulfone concentrations above 31.3 µmol/L may theoretically cause a false positive screening result on Tyr. No reference concentration in newborns was found for methionine sulfone. However, because methionine sulfone is an oxidation product of methionine sulfoxide, and methionine sulfoxide is a product of methionine, the highest concentration of methionine sulfone should not exceed the normal level of methionine in infants aged 0–1 months (reference plasma level is 10–60 µmol/L). L-Methionine sulfone dose of 62.5 µmol/L increased the endogenous Tyr concentration (56 umol/L) to 69 umol/L. Since the proposed cut-off area measured with NeoBase 2 assay for Tyr is higher (e.g. 99.0% percentile, 192 µmol/L), L-Methionine sulfone is unlikely to interfere routine testing. In addition, any possible effect of endogenous L-Methionine sulfone would be incorporated in the users established cut-offs (upper population percentiles), and would not therefore cause false in routine
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screening.
Verapamil metabolite D617:
D617 is a metabolite of calcium channel blocker Verapamil. D617 was found to interfere with the assay by increasing the measured ASA concentration. D617 concentrations from 0.72 umol/L may theoretically cause a false positive screening result on ASA. Therapeutic concentration range in plasma for Verapamil is 0.11–1.32 umol/L. D617 has been found to present approximately 20% of the given oral dose excreted in to urine, i.e. the D617 level in blood is very unlikely to exceed the concentration of 0.72 µmol/L in whole blood. Therefore, Verapamil metabolite D617 is unlikely to interfere with ASA in routine testing. Nevertheless, newborns given Verapamil or exposed to the compound during pregnancy or breastfeeding could screen positive for ASA.
L-Ornithine (Orn):
NeoBase 2 analyte L-Ornithine (Orn) was found to interfere with the assay by increasing the measured Pro concentration. The interference is caused by mass transition overlap between a fragment of Orn formed in the ion source and Pro. Orn concentrations above 93.8 umol/L may theoretically cause a false positive screening result on Pro. The proposed cut-off area measured with NeoBase 2 assay for Pro is above 200 umol/L (e.g. 99th percentile, 243 umol/L). An additional 297 umol/L dose of Orn would be needed to raise Pro concentration from endogenous level (180 umol/L) to the cut-off area (240 umol/L). The proposed normal population for Orn measured with NeoBase 2 assay (e.g. 99th percentile, 152 µmol/L) is on much lower level than the needed additional dose. It is unlikely that Orn interferes with Pro in routine testing. In addition, any possible effect of endogenous L-Ornithine would be incorporated in the users established cutoffs for Pro (upper population percentiles), and would not therefore cause false positives in routine screening.
Albumin:
High albumin concentrations were found to interfere with the assay. When total albumin is above 3.16 g/dL, the interference caused an increase in the measured ADO concentrations. The reference range for albumin in infant plasma/serum aged 0–12 months is 2.8–4.7 g/dL corresponding to albumin concentration of 1.4–2.4 g/dL in whole blood. Therefore it is unlikely that albumin interferes with ADO in routine testing.
Intralipid (Triglycerides):
Intralipid was found to interfere with the assay. When more than 0.25 g/dL of intralipid was added to blood containing 0.07 g/dL of endogenous triglycerides (i.e tested total triglycerides above 0.32 g/dL) the interference caused an increase in the measured C24:0-LPC concentration. The reference range for triglycerides in newborns aged 0–7 days has been reported to be from 0.02 to 0.18 g/dL in serum corresponding to triglyceride concentration in whole blood of 0.01 to 0.09 g/dL. Therefore it is unlikely that triglycerides interfere with C24:0-LPC in routine testing.
Chlorhexidine digluconate:
Chlorhexidine digluconate was found to interfere with the assay by increasing the measured C24:0-LPC and C26:0-LPC concentrations. Chlorhexidine digluconate is a cationic broad-spectrum antimicrobial agent belonging to the bis(biguanide) family. Its mechanism of action involves destabilization of the outer bacterial membrane. Chlorhexidine digluconate amounts above 0.03% may theoretically cause false positive screening results on C24:0-LPC and C26:0-LPC. If disinfectant pads containing chlorhexidine digluconate are used to wipe off the heels of newborns in preparation for sample collection, there is potential for chlorhexidine digluconate to be carried into the sample. It has been estimated that in the worst case, 1 µL of the 3% skin disinfection solution might contaminate a 75 µL blood droplet, which
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corresponds to an amount of 0.04% Chlorhexidine digluconate in the sample. Therefore, it is unlikely that Chlorhexidine digluconate will interfere with C24:0-LPC and C26:0-LPC in routine testing.
Hemoglobin:
High hemoglobin concentrations were found to interfere with the assay. Total hemoglobin above 21.7 g/dL caused a decrease in the measured SA by 20% and an elevation in Val by 16%. Hemoglobin interference was observed at an SA concentration of 8.7 µmol/L, but not at 0.32 µmol/L. Total hemoglobin above 20.4 g/dL caused a decrease in the measured C24:0-LPC by 18-20%. Total hemoglobin above 19.2 g/dL caused an elevation in the measured ADO by 21-36%. The effect of hematocrit on Arg was also evaluated. Hematocrit values below 43% increased Arg results by 36%, and hematocrit values above 63% decreased Arg results by 28%. Although interference by hemoglobin and hematocrit was observed with concentrations within the newborn reference ranges (12.0-22.0 g/dL for hemoglobin, 35-65% for hematocrit), it is concluded based on the external study results that the interferences are not pronounced enough to impair the separation of the affected and unaffected cases. Interference by hemoglobin could cause false negative screening results only if C24:0-LPC would be used as the sole marker for X-ALD. Therefore, C24:0-LPC should always be used together with the primary marker C26:0-LPC.
In addition to above findings, following potential interferences have been reported:
Benzocaine:
Disinfectants such as alcohol swabs with a topical anesthetic benzocaine should not be used to wipe off the heel of a newborn. Benzocaine and Phe are isomers having the same mass to charge ratio of 166.1. Therefore, benzocaine may cause falsely elevated Phe concentration and a false positive phenylketonuria (PKU) screening result.
C5 isomer pivalylcarnitine:
Pivalic acid may cause false positive screening result for Isovaleric acidemia (IVA), whose marker is acylcarnitine C5. Pivalic acid can be liberated from a prodrug containing esterified pivalic acid (such as pivalic-ester containing antibiotics, corticosteroids or other pharmaceuticals) administered to mothers or newborns. Pivalic acid is further metabolized to pivalylcarnitine which is an isomer of C5, and therefore pivalylcarnitine can cause falsely elevated C5 results. Falsely elevated C5 concentrations have been measured in newborns due to pivaly carnitine interference. Administration of pivalic acid containing prodrugs can lead to carnitine depletion, falsely elevated C5 have been connected to cases where pivalylcarnitine originated from neopentanoate esters present in nipple-fissure unguent used by the breastfeeding mothers.
C8 isomer valproylcarnitine:
Medication valproic acid administered to mothers or newborns may interfere with the screening of Medium-chain acyl-CoA dehydrogenase deficiency (MCAD) or Medium-chain ketoacyl-CoA thiolase deficiency (MCKAT), whose marker is acylcarnitine C8. Valproic acid is metabolized to valproylcarnitine, which is an isomer of C8, and can cause falsely elevated C8 concentration. False positive MCAD results have been measured in newborns due to valproylcarnitine interference.
C3DC\C4OH, C4DC\C5OH and C5DC\C6OH:
Analytes in the pairs C3DC C40H; C4DC C50H; and C5DC (C60H; are all natural acycarnitines that can be present in dried blood spots and cannot be separated in the NeoBase 2 assay due to mass transition overlap. The mass to charge ratios of the precursor ions are 248 (for C3DC, C4OH), 262 (for C4DC, C5OH), and 276 (for C5DC, C6OH) and they all have the same identifying product ion (m/z 85). As a result, the
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NeoBase 2 Non-derivatized MSMS kit reports the results for these analytes in the pair together as a sum, which is very much the same as the case for Leu\lle\Hydroxyproline, and Gln\Lys. Because of this overlap, the results for these analytes should be reported as the cumulative concentration of the two analytes in the pair.
C16:10H\C17
C16:10H is a marker among other hydroxylated long chain acylcarnitines for Long-chain L-3-hydroxyacyl-CoA dehydrogenase deficiency (LCHAD) and Trifunctional protein deficiency (TFP). Heptadecanoylcarnitine (C17) has been identified as a marker specific for propionic acidemia (PROP) and Methylmalonic acidemia (MUT). C17 and C16:10H are isomers, having the same mass to charge ratio of m/z 414, and are always measured in NeoBase 2 assay as a sum of both analytes. In screening of LCHAD, TFP, PROP and MUT, the cumulative sum concentration of C16:10H and C17 increases. It is recommended to confirm positive screening result with 2nd tier analysis, which is capable of separating C16:10H and C17 and identify specifically the disorder.
C26:0-LPC
Elevated C26:0-LPC concentrations have been measured in newborn blood spots and post-natal blood samples taken from children diagnosed by Aicardi Goutières Syndrome (AGS) leading to false positive results in first tier X-ALD screening.
Interference from M+2 Isotopic Peaks:
The analytes and internal standards measured in NeoBase 2 assay not only produce an M peak (the monoisotopic peak that is used in the measurement), but also M+1, M+2, and M+3 peaks. These additional M+n peaks are due to the naturally occurring heavier stable isotopes such as 13C, 15N or 180. In tandem mass spectra of complex samples where many analytes are analyzed simultaneously (as is the case of the NeoBase 2 assay) the M+n peaks of one compound have the potential to overlap with the peaks generated by other compounds of neighboring m/z and cause falsely elevated peaks. Potential M+2 peak interferences are as follows: M+2 peak of C5 overlaps with C3DC\C4OH; M+2 peak of C6 overlaps with C4DC\C5OH; and M+2 peak of C8 overlaps with C6DC. However, the effect is only significant when C5, C6, and C8 are present in high concentrations. At the endogenous concentrations the risk for false positive result with C3DC\C4OH, C4DC\C5OH and C6DC is negligible. When elevated level of C5, which is a marker for Isovaleric acidemia (IVA) and 2-methylbutyry|g|ycinuria (2MBG), is observed, concentration of C3DC\C40H must be evaluated. When elevated level of C6, which is a marker for Medium-chain acyl-CoA dehydrogenase deficiency (MCAD), is observed, concentration of C4DC)C50H must be evaluated. When elevated level of C8, which is a marker for MCAD and Medium-chain ketoacyl-CoA thiolase deficiency (MCKAT), is observed, concentration of C6DC must be evaluated. Conversely, when elevations are detected on C3DC\C40H; C4DC\C50H; or C6DC measurements; it is recommended to evaluate the concentrations of C5, C6, and C8 to ensure these observations are not due to the M+2 effect described here.
Plasticizers and contaminants from other consumables:
Plasticizers or other additives may leach from the plastic material used in the sample preparation, storage packages and medical equipment and interfere with the newborn screening results. For example, slip agent Oleamide (m/z 282) is known to interfere with C5DC IS having the same mass to charge ratio. Elevated C5DC IS intensity falsely decreases acycarnitine C5DC concentration and may cause false negative screening result on C5DC, which is a marker for Glutaric acidemia type I (GAI). Similarly, a common antistatic agent Lauric acid diethanolamide (LDEA, m/z 288) has been found to interfere with acycarnitine C8 having the same mass to charge ratio. LDEA can lead to false positive C8 result, which is a marker for Medium-chain acyl-CoA dehydrogenase deficiency (MCAD) and Medium-chain ketoacyl-CoA thiolase
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deficiency (MCKAT) disorders. In addition, falsely elevated C8 levels in two neonates treated with extracorporeal membrane oxygenation (ECMO) has been identified. The C8 interference was traced to PVC tubing used in ECMO and a plasticizer Di-ethylhexyl phthalate (DEHP) used commonly in the manufacturing of PVC. Interference originated from a DEHP metabolite, 2-Ethylhexanoic acid, which was further metabolized in the exposed neonates to C8 isomer, 2-ethylhexacosanoylcarnitine, can lead to false positive C8 test result. In the other neonate sample, also increased level of acylcarnitine C6DC was detected. Interference was likely because of another plasticizer, di-(2-ethylhexyl) adipate (DEHA) metabolite adipic acid, which was metabolized to C6DC, which is a marker for 3-hydroxy-3-methylglutaric acidemia (HMG), the false positive C6DC test result may occur. The NeoBase 2 assay is validated with the specific microplates and plate covers provided with the kit and any other items should not be used to avoid plasticizer or other additive contamination. Diethylethanolamine (DEAE), which is used e.g. in cleaning agents, is known to interfere with amino acid Val and can cause falsely elevated results as observed during the external study. Similarly, dimethylethanolamine and ethylaminoethanol are known to interfere with amino acid Ala and can cause falsely elevated results as observed during the external study.
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Reproducibility:
Reproducibility of the NeoBase 2 Non-derivatized MSMS assay was determined on QSight across 2 external sites and one internal site. The reproducibility is based on 75 determinations: in each laboratory 5 plates measured over 5 working days using one kit lot and each plate having 5 replicates per sample. The results of reproducibility, between- and within-laboratory precisions are presented in the table below.
Reproducibility determined for the NeoBase 2 Non-derivatized MSMS kit using QSight across 2 external sites and one internal site:
Ala
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 313 | 17 | 5.4 | 11 | 3.5 | 20 | 6.5 |
| 2 | 426 | 25 | 5.9 | 4.4 | 1.0 | 25 | 6.0 |
| 3 | 755 | 38 | 5.0 | 10 | 1.4 | 39 | 5.2 |
Arg
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 8.6 | 0.52 | 6.0 | 0.10 | 1.2 | 0.53 | 6.2 |
| 2 | 45 | 2.3 | 5.0 | 1.7 | 3.8 | 2.8 | 6.3 |
| 3 | 154 | 6.5 | 4.2 | 4.7 | 3.0 | 8.0 | 5.2 |
Asa 1
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.73 | 0.17 | 24 | 0.11 | 15 | 0.21 | 28 |
| 2 | 9.9 | 0.79 | 8.0 | 1.3 | 14 | 1.6 | 16 |
| 3 | 39 | 2.3 | 5.9 | 4.6 | 12 | 5.1 | 13 |
4 Asa is measured as a total concentration of Asa and its anhydrides.
Cit
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 27 | 3.1 | 12 | 0.29 | 1.1 | 3.1 | 12 |
| 2 | 141 | 7.4 | 5.3 | 6.0 | 4.2 | 9.5 | 6.8 |
| 3 | 463 | 25 | 5.4 | 8.7 | 1.9 | 27 | 5.8 |
{26}------------------------------------------------
Image /page/26/Picture/1 description: The image shows the logo for PerkinElmer. The logo features a blue stylized "P" with an arrow-like shape pointing to the right. Below the symbol, the text "PerkinElmer" is written in a bold, sans-serif font. Underneath "PerkinElmer", the tagline "For the Better" is written in a smaller, italicized font.
Gln\Lys
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 520 | 29 | 5.6 | 12 | 2.4 | 31 | 6.0 |
| 2 | 741 | 40 | 5.4 | 40 | 5.4 | 57 | 7.6 |
| 3 | 1402 | 82 | 5.9 | 54 | 3.8 | 98 | 7.0 |
Gly
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 316 | 20 | 6.4 | 6.0 | 1.9 | 21 | 6.7 |
| 2 | 515 | 35 | 6.7 | 10 | 2.0 | 36 | 7.0 |
| 3 | 1112 | 70 | 6.3 | 14 | 1.3 | 71 | 6.4 |
Leu\lle\Pro-OH
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 189 | 9.0 | 4.8 | 2.2 | 1.2 | 9.2 | 4.9 |
| 2 | 325 | 16 | 5.1 | 13 | 4.1 | 21 | 6.5 |
| 3 | 725 | 38 | 5.2 | 12 | 1.7 | 40 | 5.5 |
Met
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 16 | 1.1 | 6.7 | 0.73 | 4.5 | 1.3 | 8.1 |
| 2 | 108 | 5.9 | 5.5 | 4.2 | 3.9 | 7.2 | 6.7 |
| 3 | 372 | 20 | 5.4 | 7.9 | 2.1 | 22 | 5.8 |
Orn
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 112 | 4.4 | 4.0 | 5.0 | 4.5 | 6.7 | 6.0 |
| 2 | 179 | 9.5 | 5.3 | 5.3 | 2.9 | 11 | 6.1 |
| 3 | 374 | 20 | 5.2 | 4.3 | 1.2 | 20 | 5.3 |
{27}------------------------------------------------
Image /page/27/Picture/1 description: The image shows the logo for PerkinElmer. The logo features a blue stylized "P" above the company name in black, sans-serif font. Below the company name is the tagline "For the Better" in a smaller, blue, italicized font.
Phe
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 60 | 2.7 | 4.6 | 0.52 | 0.86 | 2.8 | 4.6 |
| 2 | 215 | 12 | 5.4 | 6.4 | 3.0 | 13 | 6.1 |
| 3 | 663 | 31 | 4.7 | 4.2 | 0.63 | 32 | 4.8 |
Pro
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| 1 | 138 | 6.0 | 4.3 | 2.8 | 2.0 | 6.6 | 4.8 |
| 2 | 242 | 13 | 5.4 | 3.8 | 1.6 | 14 | 5.6 |
| 3 | 543 | 29 | 5.4 | 3.2 | 0.59 | 30 | 5.4 |
Tyr
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 60 | 2.4 | 4.1 | 0.04 | 0.06 | 2.4 | 4.1 |
| 2 | 197 | 9.4 | 4.8 | 6.2 | 3.2 | 11 | 5.7 |
| 3 | 601 | 29 | 4.8 | 7.3 | 1.2 | 30 | 4.9 |
Val
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 207 | 9.5 | 4.6 | 2.1 | 1.0 | 9.7 | 4.7 |
| 2 | 315 | 18 | 5.8 | 14 | 4.6 | 23 | 7.3 |
| 3 | 637 | 36 | 5.7 | 15 | 2.4 | 39 | 6.2 |
CO
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 25 | 1.3 | 5.0 | 0.35 | 1.4 | 1.3 | 5.1 |
| 2 | 73 | 4.1 | 5.7 | 1.7 | 2.3 | 4.5 | 6.1 |
| 3 | 210 | 11 | 5.3 | 1.4 | 0.66 | 11 | 5.3 |
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Image /page/28/Picture/1 description: The image shows the logo for PerkinElmer. The logo consists of the company name in bold, black font, with the tagline "For the Better" in a smaller, blue font underneath. To the left of the company name is a blue graphic that resembles a stylized "P" or an arrow pointing to the right.
| C2 | |||||||
|---|---|---|---|---|---|---|---|
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 12 | 0.53 | 4.5 | 0.14 | 1.2 | 0.55 | 4.7 |
| 2 | 28 | 1.4 | 4.9 | 1.1 | 3.8 | 1.8 | 6.2 |
| 3 | 75 | 3.8 | 5.1 | 0.92 | 1.2 | 3.9 | 5.2 |
C3
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 1.1 | 0.07 | 6.2 | 0.03 | 2.3 | 0.08 | 6.6 |
| 2 | 9.7 | 0.52 | 5.3 | 0.30 | 3.1 | 0.60 | 6.2 |
| 3 | 34 | 1.8 | 5.2 | 0.35 | 1.0 | 1.8 | 5.3 |
C4
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.15 | 0.01 | 7.1 | <0.01 | 2.1 | 0.01 | 7.4 |
| 2 | 1.4 | 0.08 | 5.5 | 0.06 | 4.6 | 0.10 | 7.2 |
| 3 | 4.9 | 0.27 | 5.5 | 0.09 | 1.9 | 0.29 | 5.9 |
CE
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| 1 | 0.084 | 0.01 | 8.7 | <0.01 | 2.4 | 0.01 | 9.0 |
| 2 | 2.2 | 0.13 | 6.1 | 0.07 | 3.2 | 0.15 | 6.9 |
| 3 | 8.2 | 0.42 | 5.2 | 0.11 | 1.3 | 0.44 | 5.3 |
C5DC/C6OH
| C5DC\C6OH | |||||||
|---|---|---|---|---|---|---|---|
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.057 | 0.01 | 14 | 0.01 | 16 | 0.01 | 21 |
| 2 | 0.78 | 0.05 | 6.5 | 0.06 | 7.2 | 0.08 | 9.7 |
| 3 | 2.9 | 0.17 | 6.1 | 0.13 | 4.4 | 0.22 | 7.5 |
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Image /page/29/Picture/1 description: The image shows the logo for PerkinElmer. The logo consists of the company name in a bold, sans-serif font, with the "PerkinElmer" text in black. Above the name is a blue graphic element that resembles a stylized "P" or an arrow pointing to the right. Below the company name is the tagline "For the Better" in a smaller, italicized font.
| C6 | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||||
| SD | CV% | SD | CV% | SD | CV% | ||||
| 1 | 0.037 | <0.01 | 12 | <0.01 | 6.2 | 0.01 | 13 | ||
| 2 | 1.1 | 0.06 | 5.5 | 0.04 | 3.5 | 0.07 | 6.5 | ||
| 3 | 4.0 | 0.22 | 5.6 | 0.07 | 1.7 | 0.23 | 5.8 |
C8
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.078 | 0.01 | 12 | <0.01 | 2.2 | 0.01 | 12 |
| 2 | 4.9 | 0.26 | 5.3 | 0.17 | 3.5 | 0.31 | 6.4 |
| 3 | 19 | 1.0 | 5.4 | 0.17 | 0.93 | 1.0 | 5.5 |
C10
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.11 | 0.01 | 7.9 | <0.01 | 2.0 | 0.01 | 8.2 |
| 2 | 0.97 | 0.06 | 6.3 | 0.05 | 5.1 | 0.08 | 8.1 |
| 3 | 3.5 | 0.23 | 6.4 | 0.08 | 2.4 | 0.24 | 6.9 |
C12
| Sample | Total meanμmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.045 | 0.01 | 11 | <0.01 | 2.3 | 0.01 | 11 |
| 2 | 1.1 | 0.07 | 6.2 | 0.04 | 3.6 | 0.08 | 7.2 |
| 3 | 4.3 | 0.26 | 6.1 | 0.06 | 1.3 | 0.27 | 6.3 |
C14
| C14 | |||||||
|---|---|---|---|---|---|---|---|
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.10 | 0.01 | 6.5 | <0.01 | 1.9 | 0.01 | 6.7 |
| 2 | 1.1 | 0.07 | 5.7 | 0.04 | 3.4 | 0.08 | 6.7 |
| 3 | 4.1 | 0.22 | 5.4 | 0.04 | 0.88 | 0.22 | 5.5 |
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Image /page/30/Picture/1 description: The image shows the logo for PerkinElmer. The logo consists of the company name in a bold, sans-serif font, with the "PerkinElmer" text in black. Above the name is a blue graphic element that resembles a stylized "P" or an arrow pointing to the right. Below the company name is the tagline "For the Better" in a smaller, italicized font.
| C16 | |||||||
|---|---|---|---|---|---|---|---|
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 1.0 | 0.05 | 4.8 | 0.03 | 2.9 | 0.06 | 5.6 |
| 2 | 7.6 | 0.43 | 5.6 | 0.30 | 3.9 | 0.52 | 6.8 |
| 3 | 26 | 1.4 | 5.3 | 0.38 | 1.5 | 1.4 | 5.5 |
C18
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.70 | 0.04 | 5.1 | 0.02 | 2.3 | 0.04 | 5.6 |
| 2 | 2.1 | 0.10 | 5.0 | 0.08 | 3.7 | 0.13 | 6.3 |
| 3 | 6.0 | 0.27 | 4.5 | 0.08 | 1.4 | 0.28 | 4.7 |
SA
| Sample | Total mean μmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.23 | 0.04 | 19 | 0.03 | 12 | 0.05 | 22 |
| 2 | 15 | 1.3 | 8.5 | 0.43 | 2.9 | 1.3 | 9.0 |
| 3 | 59 | 4.5 | 7.7 | 1.5 | 2.5 | 4.8 | 8.1 |
ADO
| Sample | Totalmeanμmol/L | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 0.11 | 0.01 | 12 | 0.01 | 9.5 | 0.02 | 16 |
| 2 | 3.8 | 0.23 | 5.9 | 0.21 | 5.5 | 0.31 | 8.1 |
| 3 | 17 | 0.90 | 5.4 | 0.70 | 4.2 | 1.1 | 6.8 |
C26:0-LPC
| Sample | Total | Within-lab | Between-lab | Reproducibility | |||
|---|---|---|---|---|---|---|---|
| meanumol/L | SD | CV% | SD | CV% | SD | CV% | |
| 1 | 0.27 | 0.07 | 24 | 0.13 | 48 | 0.15 | 54 |
| 2 | 0.69 | 0.08 | 11 | 0.16 | 23 | 0.18 | 25 |
| 3 | 2.0 | 0.16 | 7.9 | 0.21 | 11 | 0.26 | 14 |
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Image /page/31/Picture/1 description: The image shows the logo for PerkinElmer. The logo features a blue stylized "P" shape, with the company name "PerkinElmer" written in black, bold font below it. Underneath the company name, the tagline "For the Better" is written in a smaller, blue, italicized font.
Screening Performance:
The screening performance of the NeoBase 2 Non-derivatized MSMS kit on QSight and TQD platforms was compared in a clinical study at one routine screening laboratory in the United States. Newborn population distributions were determined by measuring the analyte concentrations. Using data from routine newborn screening specimens, the cut-offs of the 3044-001U NeoBase 2 Non-derivatized MSMS kit analytes were determined by calculating analyte concentrations corresponding to the 1st, 10th, 99th and 99.5th percentiles. The percentiles and descriptive values calculated from results obtained from 2530 routine screening specimens are presented in the table below.
| Analyte | Mean(μmol/L) | Median(μmol/L) | 1st | 10th | 99th | 99.5th |
|---|---|---|---|---|---|---|
| Ala | 374 | 362 | 226 | 281 | 627 | 672 |
| Arg | 8.17 | <LOQ | <LOQ | <LOQ | 25.8 | 32.7 |
| Asa | 0.39 | 0.37 | 0.19 | 0.26 | 0.75 | 0.82 |
| Cit | 14.7 | 14.1 | 7.85 | 10.3 | 26.9 | 29.7 |
| Gln\Lys | 739 | 724 | 439 | 556 | 1200 | 1300 |
| Glu | 254 | 245 | 147 | 183 | 443 | 471 |
| Gly | 534 | 519 | 314 | 394 | 924 | 997 |
| Leu\lle\Pro-OH | 98.0 | 94.9 | 57.6 | 71.8 | 172 | 207 |
| Met | 21.0 | 20.2 | 11.6 | 15.4 | 38.0 | 44.1 |
| Orn | 74.8 | 71.2 | 36.4 | 49.4 | 152 | 166 |
| Phe | 54.3 | 53.0 | 35.2 | 42.1 | 91.1 | 97.3 |
| Pro | 141 | 137 | 82.2 | 103 | 243 | 267 |
| Tyr | 89.4 | 83.7 | 40.5 | 57.0 | 192 | 206 |
| Val | 95.7 | 92.3 | 55.7 | 69.3 | 174 | 187 |
| C0 | 20.5 | 19.3 | 8.87 | 12.4 | 43.1 | 48.2 |
| C2 | 21.8 | 20.8 | 9.73 | 13.6 | 44.4 | 48.4 |
| C3 | 2.20 | 2.05 | 0.86 | 1.27 | 4.89 | 5.29 |
| C3DC\C4OH | 0.19 | 0.18 | 0.07 | 0.11 | 0.39 | 0.41 |
| C4 | 0.23 | 0.21 | 0.10 | 0.14 | 0.65 | 0.73 |
| C4DC\C5OH | 0.25 | 0.24 | 0.12 | 0.16 | 0.51 | 0.56 |
| C5 | 0.10 | 0.09 | 0.05 | 0.06 | 0.24 | 0.27 |
| C5:1 | 0.01 | 0.01 | <LOQ | 0.01 | 0.02 | 0.02 |
| C5DC\C6OH | 0.11 | 0.10 | 0.05 | 0.07 | 0.23 | 0.26 |
| C6 | 0.05 | 0.05 | 0.03 | 0.03 | 0.12 | 0.13 |
| C6DC | 0.07 | 0.06 | <LOQ | 0.04 | 0.13 | 0.14 |
| C8 | <LOQ | <LOQ | <LOQ | <LOQ | 0.14 | 0.17 |
| C8:1 | 0.10 | <LOQ | <LOQ | <LOQ | 0.23 | 0.25 |
| C10 | 0.10 | 0.10 | 0.04 | 0.06 | 0.22 | 0.25 |
| C10:1 | 0.04 | 0.04 | <LOQ | <LOQ | 0.07 | 0.08 |
| C10:2 | <LOQ | <LOQ | <LOQ | <LOQ | <LOQ | <LOQ |
| C12 | 0.12 | 0.11 | <LOQ | <LOQ | 0.28 | 0.30 |
| Mean(μmol/L) | Median(μmol/L) | Percentiles (μmol/L) | ||||
| Analyte | 1st | 10th | 99th | 99.5th | ||
| C12:1 | <LOQ | <LOQ | <LOQ | <LOQ | 0.25 | 0.28 |
| C14 | 0.25 | 0.24 | 0.12 | 0.17 | 0.46 | 0.50 |
| C14:1 | 0.14 | 0.13 | 0.05 | 0.08 | 0.33 | 0.37 |
| C14:2 | 0.03 | 0.03 | 0.01 | 0.02 | 0.06 | 0.06 |
| C14OH | 0.03 | 0.03 | 0.01 | 0.02 | 0.06 | 0.06 |
| C16 | 3.72 | 3.62 | 1.75 | 2.47 | 6.69 | 6.94 |
| C16:1 | 0.24 | 0.24 | 0.10 | 0.15 | 0.42 | 0.46 |
| C16OH | 0.04 | 0.04 | 0.02 | 0.03 | 0.09 | 0.09 |
| C16:1OH\C17 | 0.06 | 0.06 | 0.03 | 0.04 | 0.11 | 0.12 |
| C18 | 0.92 | 0.88 | 0.44 | 0.60 | 1.75 | 1.91 |
| C18:1 | 1.22 | 1.19 | 0.62 | 0.84 | 2.15 | 2.32 |
| C18:2 | 0.17 | 0.16 | 0.08 | 0.11 | 0.38 | 0.43 |
| C18OH | 0.02 | 0.02 | 0.01 | 0.01 | 0.04 | 0.04 |
| C18:1OH | 0.05 | 0.05 | 0.02 | 0.03 | 0.13 | 0.14 |
| C18:2OH | 0.02 | 0.01 | 0.01 | 0.01 | 0.03 | 0.04 |
| SA | 0.25 | 0.24 | <LOQ | <LOQ | 0.40 | 0.44 |
| ADO | 0.77 | 0.74 | 0.38 | 0.53 | 1.40 | 1.47 |
| D-ADO | <LOQ | <LOQ | <LOQ | <LOQ | <LOQ | <LOQ |
| C24:0-LPC | 0.50 | 0.47 | 0.29 | 0.36 | 0.95 | 1.02 |
| C26:0-LPC | 0.35 | 0.33 | 0.18 | 0.24 | 0.71 | 0.78 |
Descriptive statistics of the study using QSight (n=2530). The values below the measuring range that are not recommended to be considered as accurate are indicated by <LOQ (lower limit of quantitation):
{32}------------------------------------------------
Image /page/32/Picture/1 description: The image shows the logo for PerkinElmer, a company focused on diagnostics, life sciences, and environmental solutions. The logo features the company name in a bold, sans-serif font, with "Perkin" in black and "Elmer" in a lighter gray. Above the name is a stylized graphic consisting of a blue vertical bar and a blue triangle pointing to the right. Below the company name is the tagline "For the Better" in a smaller, italicized font.
The disorders and amount of confirmed positive specimens included in the study are listed in tables below. In total, there were 19 specimens included in the group of amino acid disorders, 12 in the group of fatty acid oxidation disorders, 16 in the group of organic acid conditions and 5 in the group of other conditions. The results for the screening performance data including confirmed positive specimens tested with QSight and TQD are presented in tables below.
| Amino acid disorders (AA) | TQD | ||||||
|---|---|---|---|---|---|---|---|
| 99th percentile | 99.5th percentile | ||||||
| Screening positive | Screening negative | Total | Screening positive | Screening negative | Total | ||
| QSight | Screening positive | 881 | 12 | 100 | 492 | 9 | 58 |
| Screening negative | 85 | 2414 | 2499 | 443 | 2497 | 2541 | |
| Total | 173 | 2426 | 2599 | 93 | 2506 | 2599 |
4 Includes all 19 specimens confirmed positive for AA detected with both platforms using 99th percentile cut-offs.
2 Includes 18 specimens confirmed positive for AA detected with both platforms using 99.50 percentile cut-offs.
3 Includes 1 specimen confirmed positive for OTCD with secondary marker not detected with QSight using 99.50 percentile cut-off, but detected with primary marker Cit with 1* percentile cutoff presented in the next table.
{33}------------------------------------------------
Image /page/33/Picture/0 description: The image shows the logo for PerkinElmer. The logo features a blue vertical bar with a triangle pointing to the right. Below the symbol, the name "PerkinElmer" is written in a bold, sans-serif font, with "Perkin" in black and "Elmer" in gray. Underneath the name, the tagline "For the Better" is written in a smaller, italicized font.
| Amino acid disorders (AA) | TQD | |||
|---|---|---|---|---|
| 1st percentile | ||||
| Screeningpositive | Screeningnegative | Total | ||
| QSight | Screeningpositive | 201 | 31 | 51 |
| Screeningnegative | 12 | 2519 | 2531 | |
| Total | 32 | 2550 | 2582 |
1 Includes 2 specimens confirmed positive for OTCD detected with both platforms using 15 percentile cut-offs.
| Fatty acid oxidation (FAO) | TQD | |||||||
|---|---|---|---|---|---|---|---|---|
| 99th percentile | 99.5th percentile | |||||||
| Screeningpositive | Screeningnegative | Total | Screeningpositive | Screeningnegative | Total | |||
| QSight | Screeningpositive | 1651 | 52 | 217 | 901 | 33 | 123 | |
| Screeningnegative | 69 | 23042 | 2373 | 70 | 23972 | 2467 | ||
| Total | 234 | 2356 | 2590 | 160 | 2430 | 2590 |
1 Includes 9 specimens confirmed positive for FAO detected with both platforms using upper percentile cut-offs. ² Includes 1 specimen confirmed positive for CPT-II not detected with either platform using upper percentile cutoffs.Specimen detected with secondary marker in table below.
| TQD | ||||
|---|---|---|---|---|
| Low percentile1 | ||||
| Fatty acid oxidation (FAO) | Screening positive | Screening negative | Total | |
| QSight | Screening positive | 2831 | 193 | 476 |
| Screening negative | 5 | 2103 | 2108 | |
| Total | 288 | 2296 | 2584 |
1 "Low Percentile" reflects the positive CO specimens detected with a 10% cut-off with other carnitine-positive samples at the 1% cut-off
2 Includes all 4 specimens confirmed positive for FAO detected with both platforms using low percentile cut-offs.
{34}------------------------------------------------
Image /page/34/Picture/0 description: The image shows the logo for PerkinElmer. The logo features a blue vertical bar and a blue triangle pointing to the right, positioned above the company name. The text "PerkinElmer" is in a bold, sans-serif font, with "Elmer" in a slightly lighter shade of gray. Below the company name is the tagline "For the Better" in a smaller, italicized font.
| Organic acid condition (OA) | TQD | ||||||
|---|---|---|---|---|---|---|---|
| 99th percentile | 99.5th percentile | ||||||
| Screening positive | Screening negative | Total | Screening positive | Screening negative | Total | ||
| QSight | Screening positive | 861 | 11 | 97 | 511 | 8 | 59 |
| Screening negative | 42 | 2457 | 2499 | 33 | 2504 | 2537 | |
| Total | 128 | 2468 | 2596 | 84 | 2512 | 2596 |
4 Includes all 16 specimens confirmed positive for OA detected with both platforms using upper percentile cut-offs.
| ADA-SCID | TQD | ||||||
|---|---|---|---|---|---|---|---|
| 99th percentile | 99.5th percentile | ||||||
| Screeningpositive | Screeningnegative | Total | Screeningpositive | Screeningnegative | Total | ||
| QSight | Screeningpositive | 81 | 6 | 14 | 71 | 0 | 7 |
| Screeningnegative | 40 | 2528 | 2568 | 25 | 2550 | 2575 | |
| Total | 48 | 2534 | 2582 | 32 | 2550 | 2582 |
4 Includes 2 specimens confirmed positive for ADA-SCID detected with both platforms using upper percentile cut-offs.
| X-ALD | TQD | TQD | |||||
|---|---|---|---|---|---|---|---|
| 99th percentile | 99.5th percentile | ||||||
| Screeningpositive | Screeningnegative | Total | Screeningpositive | Screeningnegative | Total | ||
| QSight | Screeningpositive | 71 | 96 | 103 | 31 | 67 | 70 |
| Screeningnegative | 262 | 2454 | 2480 | 15 | 24983 | 2513 | |
| Total | 33 | 2550 | 2583 | 18 | 2565 | 2583 |
4 Includes 2 specimens confirmed positive for X-ALD detected with both platforms using upper percentile cut-offs. 2 Includes one borderline sample with variant of unknown significance (VOUS), not detected with QSight using 99th percentile cut-off and detected with TQD. Primary marker C26-LPC result was 0.45 µmo//l with QSight and 0.42 µmo//l with TQD. Secondary marker C24-LPC result was 0.61 µmol/l with QSight and 0.41 µmol/l with TQD. 3 Includes one borderline sample with variant of unknown significance (VOUS), not detected with either method using
99.5th percentile cut-off.
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 (K173568).
§ 862.1055 Newborn screening test system for amino acids, free carnitine, and acylcarnitines using tandem mass spectrometry.
(a)
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.(b)
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.