Atellica® CH Diazo Total Bilirubin (D_TBil)

K222104 · Siemens Healthcare Diagnostics, Inc. · CIG · Mar 20, 2023 · Clinical Chemistry

Device Facts

• Record ID: K222104
• Device Name: Atellica® CH Diazo Total Bilirubin (D_TBil)
• Applicant: Siemens Healthcare Diagnostics, Inc.
• Product Code: CIG · Clinical Chemistry
• Decision Date: Mar 20, 2023
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1110
• Device Class: Class 2
• Attributes: Pediatric

Indications for Use

The Atellica® CH Diazo Total Bilirubin (D_TBil) assay is for in vitro diagnostic use in the quantitative determination of total bilirubin in adults and children (non-neonates) in human serum and plasma using the Atellica® CH Analyzer. Measurement of total bilirubin, an organic compound formed during the normal and abnormal destruction of red blood cells, is used in the diagnosis and treatment of liver, hemolytic, hematological, and metabolic disorders, including hepatitis and gall bladder block.

Device Story

In vitro diagnostic assay for quantitative total bilirubin measurement in human serum/plasma; utilizes Atellica CH Analyzer. Principle: Diazo chemistry using 2,4-dichloroaniline (DCA) in acidic solution; forms red-colored azocompound measured photometrically at 545nm. Requires 6.0 µL sample. Used in clinical laboratory settings by trained personnel. Output: total bilirubin concentration (mg/dL). Assists clinicians in diagnosing/treating liver, hemolytic, hematological, and metabolic disorders. Performance validated via precision, linearity, and method comparison studies against predicate.

Clinical Evidence

Bench testing only. Precision/reproducibility evaluated per CLSI EP05-A3 (n=80/sample for precision; n=225/sample for reproducibility). Linearity (0.04-27.91 mg/dL) per CLSI EP06. Interference testing per CLSI EP07. Method comparison (n=103) vs. predicate showed Deming regression slope 1.02, intercept 0.08, r=0.997. Matrix comparison (n=57) confirmed equivalency across serum and heparin/EDTA plasma. LoB/LoD/LoQ validated per CLSI EP17-A2.

Technological Characteristics

Photometric diazo colorimetry assay. Reagents: 2,4-dichloroaniline, HCl, Na-Nitrite, phosphate buffer, NaCl, detergents. Measuring interval: 0.10-25.0 mg/dL. Traceable to NIST SRM 916. Analyzed on Atellica CH Analyzer. Software-controlled automated photometric measurement.

Indications for Use

Indicated for quantitative determination of total bilirubin in human serum and plasma for adults and children (non-neonates). Used in diagnosis and treatment of liver, hemolytic hematological, and metabolic disorders (e.g., hepatitis, gall bladder block).

Regulatory Classification

Identification

A bilirubin (total or direct) test system is a device intended to measure the levels of bilirubin (total or direct) in plasma or serum. Measurements of the levels of bilirubin, an organic compound formed during the normal and abnormal distruction of red blood cells, if used in the diagnosis and treatment of liver, hemolytic hematological, and metabolic disorders, including hepatitis and gall bladder block.

Predicate Devices

• Dimension TBI Flex reagent cartridge (K060628)

Submission Summary (Full Text)

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
ASSAY ONLY

## I Background Information:

A 510(k) Number
K222104

B Applicant
Siemens Healthcare Diagnostics Inc.

C Proprietary and Established Names
Atellica® CH Diazo Total Bilirubin (D_TBil)

D Regulatory Information
| Product Code(s) | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| CIG | Class II | 21 CFR 862.1110 - Bilirubin (Total Or Direct) Test System | CH - Clinical Chemistry |

## II Submission/Device Overview:

A Purpose for Submission:
New device

B Measurand:
Total bilirubin

C Type of Test:
Quantitative, photometric test

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.govK222104 - Page 1 of 8

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III Intended Use/Indications for Use:

A Intended Use(s):

See Indications for Use below.

B Indication(s) for Use:

The Atellica® CH Diazo Total Bilirubin (D_TBil) assay is for in vitro diagnostic use in the quantitative determination of total bilirubin in adults and children (non-neonates) in human serum and plasma using the Atellica® CH Analyzer. Measurement of total bilirubin, an organic compound formed during the normal and abnormal destruction of red blood cells, is used in the diagnosis and treatment of liver, hemolytic, hematological, and metabolic disorders, including hepatitis and gall bladder block.

C Special Conditions for Use Statement(s):

Rx - For Prescription Use Only

D Special Instrument Requirements:

Atellica CH Analyzer

IV Device/System Characteristics:

A Device Description:

The Atellica® CH Diazo Total Bilirubin (D_TBil) assay consists of the following 2 packs:

Pack 1 (P1) 23.5 mL Phosphate buffer (50mmol/L);
NaCl (150mmol/L)

Pack 2 (P2) 8.8ml 2,4-Dicholoroaniline (5mmol/L);
HCl (130mmol/L);
Na-Nitrite (0.5mmol/L)

Materials required but not provided: Atellica CH Bilirubin Calibrator (BILI CAL).

B Principle of Operation:

The Atellica® CH Diazo Total Bilirubin (D_TBil) is an assay that uses 2,4-dichloroaniline (DCA). Direct bilirubin in the presence of diazotized DCA forms a red-colored azocompound in acidic solution that can be measured. A specific mixture of detergents enables the determination of total bilirubin. The detection wavelength is 545nm.

This assay requires 6.0 µL of sample for a single determination.

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.govK222104 - Page 2 of 8

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V Substantial Equivalence Information:

A Predicate Device Name(s):

Dimension TBI Flex and DBI Flex Reagent Cartridges and TBI/DBI Calibrator

B Predicate 510(k) Number(s):

K060628

C Comparison with Predicate(s):

| Device & Predicate Device(s): | K222104 | K060628 |
| --- | --- | --- |
| Device Trade Name | Atellica® CH Diazo Total Bilirubin (D_TBil) | Dimension TBI Flex and DBI Flex Reagent Cartridges and TBI/DBI Calibrator |
| General Device Characteristic Similarities | | |
| Intended Use/Indications For Use | Quantitative determination of total bilirubin in human serum and plasma. Measurement of total bilirubin is used in the diagnosis and treatment of liver, hemolytic, hematological, and metabolic disorders, including hepatitis and gall bladder block. | Same |
| Mode of Detection | Photometric (Diazo chemistry) | Same |
| Measuring Range | 0.10 mg/dL – 25.0 mg/dL | Same |
| General Device Characteristic Differences | | |
| Instrument Platform | Atellica CH Analyzer | Dimension® clinical chemistry system |
| Sample Matrix | Human serum and plasma (lithium heparin, sodium heparin, dipotassium EDTA) | Human serum and plasma (lithium heparin, EDTA) |

Food and Drug Administration
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Silver Spring, MD 20993-0002
www.fda.govK222104 - Page 3 of 8

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VI Standards/Guidance Documents Referenced:

- Clinical and Laboratory Standards Institute (CLSI) EP05-A3 – Evaluation of Precision of Quantitative Measurement Procedures
- CLSI EP09c 3rd Edition – Measurement Procedure Comparison and Bias Estimation Using Patient Samples
- CLSI EP17-A2 – Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline – Second Edition
- CLSI EP06-2nd Edition – Evaluation of Linearity of Quantitative Measurement Procedures
- CLSI EP07-3rd Edition – Interference Testing in Clinical Chemistry
- CLSI EP37 1st Edition – Supplemental Tables for Interference Testing in Clinical Chemistry
- CLSI EP34 1st Edition – Establishing and Verifying an Extended Measuring Interval Through Specimen Dilution and Spiking
- CLSI EP28-A3c (Formerly C28-A3c) – Defining Establishing and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline – Third Edition
- CLSI EP32-R (Formerly X05-R) – Metrological Traceability and Its Implementation; A Report
- CLSI EP25-A (Replaces EP25-P) – Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline.

VII Performance Characteristics (if/when applicable):

A Analytical Performance:

1. Precision/Reproducibility:

Precision

Precision studies were conducted according to CLSI EP05-A3. Total precision was assessed in 2 runs (minimum 2 hours between each run), 2 replicates per run per day, over 20 days, for a total of 80 results per sample. Three spiked serum samples were tested using 3 reagent lots on one Atellica CH Analyzer. Repeatability and within-lab imprecision were calculated. Results were similar between reagent lots. The precision data from a representative reagent lot are summarized below.

| Sample | Mean (mg/dL) | Repeatability | | Within-lab (total) | |
| --- | --- | --- | --- | --- | --- |
| | | SD (mg/dL) | %CV | SD (mg/dL) | %CV |
| Serum 1 | 1.02 | 0.015 | 1.5 | 0.034 | 3.3 |
| Serum 2 | 13.40 | 0.053 | 0.4 | 0.140 | 1.0 |
| Serum 3 | 22.39 | 0.067 | 0.3 | 0.189 | 0.8 |

Reproducibility

Testing was conducted according to CLSI EP05-A3. Reproducibility was assessed using 3 serum pool samples measured in 5 replicates, using 3 reagent lots, over 5 days, on 3 Atellica CH Analyzers, for a total of 225 replicates per sample. Repeatability, between-day, between-lot,

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between-instrument, and reproducibility were calculated. The data for reproducibility studies are summarized below.

| Sample | Mean (mg/dL) | Repeatability | | Between-Day | | Between-Lot | | Between-Instrument | | Reproducibility | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD (mg/dL) | % CV | SD (mg/dL) | % CV | SD (mg/dL) | % CV | SD (mg/dL) | % CV | SD (mg/dL) | % CV |
| Serum 1 | 1.03 | 0.012 | 1.2 | 0.009 | 0.9 | 0.013 | 1.3 | 0.011 | 1.1 | 0.023 | 2.2 |
| Serum 2 | 13.24 | 0.037 | 0.3 | 0.062 | 0.5 | 0.028 | 0.2 | 0.047 | 0.4 | 0.091 | 0.7 |
| Serum 3 | 22.14 | 0.057 | 0.3 | 0.106 | 0.5 | 0.053 | 0.2 | 0.063 | 0.3 | 0.146 | 0.7 |

# 2. Linearity:

Linearity studies were conducted according to CLSI-EP06 $2^{\text{nd}}$ Edition. A dilution series composed of 9 levels (0.04 - 27.91 mg/dL) was prepared by mixing different proportions of the high serum pool sample and the low serum pool sample. Four replicates for each sample were measured on the Atellica CH Analyzer using 3 reagent lots. The data was analyzed using a linear regression model as described in the guideline. The results support the claimed measuring range of 0.10 mg/dL - 25.0 mg/dL.

# 3. Analytical Specificity/Interference:

Interference studies were conducted according to the CLSI EP07 $3^{\mathrm{rd}}$ Edition guideline. The effect of endogenous and exogenous substances on the D_TBil assay was evaluated using two human serum pools spiked to obtain bilirubin concentrations of $\sim 1.0\mathrm{mg / dL}$ or $\sim 14.0$ mg/dL. Each sample was tested in 5 replicates. Testing used the paired-difference method of comparing a sample spiked with interferent to a sample spiked with control material (diluent used in the test sample). The sponsor considered interference to be non-significant if the difference between the samples with and without interferent are within $10.0\%$ . The results of the interference studies are summarized below.

| Interferent | Highest interferent concentration tested that showed no significant interference |
| --- | --- |
| Hemoglobin | 1000 mg/dL |
| Triglyceride | 1000 mg/dL |
| Acetaminophen | 20 mg/dL |
| Acetylsalicylic acid | 100 mg/dL |
| Ascorbic acid | 5 mg/dL |
| Carbenicillin | 3 mg/dL |
| Cholesterol | 500 mg/dL |
| Cyanokit (Hydroxocobalamin) | 40 μg/ml |
| Diazepam | 20 μg/mL |
| Eltrombopag | 25 μg/mL |
| Ethanol | 800 mg/dL |

Food and Drug Administration

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www.fda.govK222104 - Page 5 of 8

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| Interferent | Highest interferent concentration tested that showed no significant interference |
| --- | --- |
| Ibuprofen | 50 mg/dL |
| Immunoglobin G | 5 g/dL |
| Levodopa | 225 μg/mL |
| Phenazopyridine | 80 μg/mL |
| Phloroglucinol | 250 ng/mL |
| Rifampicin | 6 mg/dL |
| Indican | 1.5 mg/dL |

4. Assay Reportable Range:

The assay measuring interval is 0.10 to 25.00 mg/dL.

The sponsor provided data to support that the measuring interval can be extended up to 50.00 mg/dL following manual or automated dilution.

5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):

The assay is traceable to SRM916 (NIST) Standard Reference Material.

6. Detection Limit:

Limit of Blank (LoB), Limit of Detection (LoD) and Limit of Quantitation (LoQ) were validated based on the CLSI guideline EP17-A2.

To calculate the LoB, four blank samples in replicates of five were tested using three reagent lots on one Atellica CH Analyzer over three days for a total of 60 measurements per reagent lot. LoB was determined using the 95% nonparametric percentile of the replicates for each of three reagent lots. The claimed LoB is 0.01 mg/dL.

To calculate the LoD, two low native serum samples and two samples prepared from diluted serum pools were tested in replicates of five on one Atellica CH Analyzer using three reagent lots over three days for a total of 60 measurements per lot. The maximum observed LoD across the 3 lots tested was 0.02 mg/dL. The claimed LoD is 0.02 mg/dL.

To calculate the LoQ, five low level total bilirubin sample pools targeted between 0.03 mg/dL and 0.17 mg/dL were tested in replicates of five on one Atellica CH Analyzer using three reagent lots over five days for a total of 125 determinations per lot. The maximum observed LoQ across all reagent lots was determined to be 0.04 mg/dL. The claimed LoQ is 0.1 mg/dL which is the lowest concentration that met the performance goal of less than 20% CV.

7. Assay Cut-Off:

Not applicable.

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www.fda.govK222104 - Page 6 of 8

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B Comparison Studies:

1. Method Comparison with Predicate Device:
A method comparison study was conducted in accordance with CLSI EP09c-A3 using 103 serum samples with 3 reagent lots on one Atellica CH Analyzer. Each serum sample was tested in duplicate. Results from the Atellica CH Diazo Total Bilirubin assay were compared to results using the predicate device. Results were analyzed using Deming regression. Only the first replicate result was used in the statistical analysis.

The table below summarizes the data using serum samples from a representative lot:

| N | Sample Range (mg/dL) | Intercept (mg/dL) | Slope | Correlation Coefficient (r) | Claimed Measuring Range (mg/dL) |
| --- | --- | --- | --- | --- | --- |
| 103 | 0.14 -22.55 | 0.08 | 1.02 | 0.997 | 0.1-25.0 |

2. Matrix Comparison:
A matrix comparison study was conducted according to CLSI EP09c-A3 using 1 reagent lot. Fifty-seven (57) matched serum and plasma (lithium heparin, sodium heparin, dipotassium EDTA) samples were assayed with one Atellica CH Analyzer in singlicate. Results were analyzed using Deming regression.

The table below summarizes the matrix comparison data:

| Analysis | Slope | Intercept (mg/dL) | Sample Range | N | r |
| --- | --- | --- | --- | --- | --- |
| Lithium heparin plasma vs. serum | 0.98 | 0.05 | 0.24-22.61 mg/dL | 57 | 0.997 |
| Sodium heparin plasma vs serum | 1.00 | 0.02 | 0.24-22.61 mg/dL | 57 | 0.998 |
| K2 EDTA plasma vs serum | 0.99 | 0.03 | 0.24-22.61 mg/dL | 57 | 0.998 |

Results demonstrated equivalency between serum, lithium heparin plasma, sodium heparin plasma, and K2 EDTA plasma specimens.

C Clinical Studies:

1. Clinical Sensitivity:
Not applicable.

2. Clinical Specificity:
Not applicable.

3. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):
Not applicable.

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D Clinical Cut-Off:

Not applicable.

E Expected Values/Reference Range:

The reference interval was verified by testing 21 serum samples (age range from 4 to 83) with the total bilirubin assay in accordance with CLSI EP28-A3c. Data support the pre-established reference interval of 0.3 - 1.2mg/dL (5.13 - 20.52 μmol/L)¹.

¹Wu AHB, ed. Tietz Clinical Guide to Laboratory Tests. 4th ed. St. Louis, MO: Saunders; 2006:316.

VIII Proposed Labeling:

The labeling supports the finding of substantial equivalence for this device.

IX Conclusion:

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

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.govK222104 - Page 8 of 8