BIO-RAD D-10 DUAL PROGRAM

K041444 · Bio-Rad Laboratories, Inc. · LCP · Jun 9, 2004 · Hematology

Device Facts

• Record ID: K041444
• Device Name: BIO-RAD D-10 DUAL PROGRAM
• Applicant: Bio-Rad Laboratories, Inc.
• Product Code: LCP · Hematology
• Decision Date: Jun 9, 2004
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.7470
• Device Class: Class 2
• Attributes: 3rd-Party Reviewed

Intended Use

The Bio-Rad D-10™ Dual Program is intended for the percent determination of hemoglobin A₁c, A₂ and F, and for the detection of abnormal hemoglobins in human whole blood using ion-exchange high performance liquid chromatography (HPLC). The measurement of the percent hemoglobin A₁c is effective in monitoring long-term glucose control in individuals with diabetes mellitus, and measurement of the percent HbA₂ and HbF are effective in long-term monitoring of β-thalassemias (i.e., hereditary hemolytic anemias characterized by decreased synthesis of one or more types of abnormal hemoglobin polypeptide chains). Detection of hemoglobin thalassemia variants such as hemoglobins S, C, D and E by HPLC is effective in presumptive identification of these variants. The Bio-Rad D-10™ Dual Program is intended for Professional Use Only. For in vitro diagnostic use.

Device Story

The Bio-Rad D-10 Dual Program is an HPLC-based diagnostic system for human whole blood analysis. It utilizes ion-exchange chromatography to separate and quantify hemoglobin fractions (HbA1c, HbA2, HbF) and identify abnormal hemoglobin variants. The system operates via two selectable programs: a 3-minute short program for HbA1c and a 6.5-minute extended program for HbA1c, HbA2, and HbF. Used in clinical laboratories by professional staff, the system processes EDTA-anticoagulated whole blood samples. The device provides quantitative area percentage results for hemoglobin fractions, which clinicians use to monitor long-term glycemic control in diabetic patients and manage beta-thalassemia. By automating the separation and identification of hemoglobin variants, the system aids in the presumptive diagnosis of hemoglobinopathies, facilitating timely clinical intervention and patient management.

Clinical Evidence

Bench testing only. Precision evaluated over 20 days (n=80); total CVs ranged 0.9-5.3%. Linearity confirmed across reportable ranges for HbA1c, HbA2, and HbF. Interference studies showed no significant impact from bilirubin (20 mg/dL), lipids (5680 mg/dL), or high EDTA. Method comparison against predicates using 40-148 patient samples showed strong correlation (r² > 0.98 for all analytes). Reference ranges established via non-parametric analysis of 53 healthy subjects.

Technological Characteristics

Ion-exchange high performance liquid chromatography (HPLC) system. Uses cation exchange cartridges. Visible detection at 415 nm. Sample type: EDTA-anticoagulated human whole blood. Connectivity: Standalone system with software-driven program parameters (via floppy diskette). Standardization: HbA1c traceable to DCCT/IFCC and NGSP certified. Software-controlled automated analysis.

Indications for Use

Indicated for individuals with diabetes mellitus for long-term glucose monitoring and individuals with β-thalassemias for long-term monitoring of hemoglobin levels. Used for presumptive identification of hemoglobin variants (S, C, D, E) in human whole blood. For professional use in clinical chemistry laboratories.

Regulatory Classification

Identification

A glycosylated hemoglobin assay is a device used to measure the glycosylated hemoglobins (A1a , A1b , and A1c ) in a patient's blood by a column chromatographic procedure. Measurement of glycosylated hemoglobin is used to assess the level of control of a patient's diabetes and to determine the proper insulin dosage for a patient. Elevated levels of glycosylated hemoglobin indicate uncontrolled diabetes in a patient.

Predicate Devices

• VARIANT™ II Hemoglobin A1c Program (K984268)
• VARIANT™ II β-thalassemia Short Program (K991127)
• Bio-Rad D-10™ Hemoglobin Testing System (K031043)

Submission Summary (Full Text)

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K0414444

## JUN - 9 2004

# Summary of Safety and Effectiveness

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is: K______________________________________________________________________________________________________________________________________________

| Submitter: | Bio-Rad Laboratories<br>Diagnostics Group<br>4000 Alfred Nobel Drive,<br>Hercules, California 94547<br>Phone: [phone]<br>FAX: [phone] |
|------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------|
| Contact Person: | Jackie Buckley<br>Regulatory Affairs Representative |
| Date of Summary Preparation: | April 16, 2004 |
| Device Name: | Bio-Rad D-10™ Dual Program |
| Classification Name: | HbA₁c: Assay, Glycosylated Hemoglobin<br>[21CFR 864.7470 / Prod. Code LCP] and<br>HbA₂: Hemoglobin A₂ Quantitation<br>[21CFR 864.7400 / Prod. Code: JPD] |
| Predicate Devices: | HbA₁c: VARIANT™ II Hemoglobin A₁c Program<br>Bio-Rad Laboratories<br>[K984268; 12/17/98] |
| | HbA₂/F: VARIANT™ II β-thalassemia Short Program<br>Bio-Rad Laboratories<br>[K991127; 06/10/99] |
| | Presumptive Hemoglobin Identification:<br>VARIANT™ II β-thalassemia Short Program<br>Bio-Rad Laboratories<br>[K991127; 06/10/99] |
| | Special Instrument Requirement:<br>Bio-Rad D-10™ Hemoglobin Testing System<br>[K031043; 08/27/03] |

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Image /page/1/Figure/0 description: The image is a predicate map for the D-10 Dual Program. It shows two assays, one for 3 minutes (short) and one for 6.5 minutes (extended). The 3-minute assay analyzes HbA1c, while the 6.5-minute assay analyzes HbF, HbA2, and HbA1c. The predicate methods are Variant II B-Thal Short (K991127), Variant II B-Thal Short (K991127), and Variant II Hemoglobin A1c (K984268), which is the same as D-10 Hemoglobin A1c (K031043).

Indications for Use Statement and Intended Uses:

The Bio-Rad D-10TM Dual Program system is intended for the percent determination of hemoglobins A1c, A2 and F, and for the detection of abnormal hemoglobins in human whole blood using ion-exchange high performance liquid chromatography (HPLC).

Measurement of the percent hemoglobin Aie is effective in monitoring long-term glucose control in individuals with diabetes mellitus, and measurement of the percent HbA 2and HbF are effective in long-term monitoring of ß-thalassemias (i.e., hereditary hemolytic anemias characterized by decreased synthesis of one or more types of abnormal hemoglobin polypeptide chains).

Detection of hemoglobin thalassemia variants such as hemoglobins S, C, D and E by HPLC is effective in presumptive identification of these variants. The Bio-Rad D-10TM Dual Program is intended for Professional Use Only. For in vitro diagnostic use.

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#### Description of Device

The Bio-Rad D-10™ Dual Program is a new device system that utilizes the principles of high performance liquid chromatography (HPLC), by which chromatographic separation of performance nquild chromatography (THLC), of willen exchange cartridge. The Bio-Rad D-10™ nemogrooms is a new program system that combines the determination of percent hemoglobin A = 1 = d for diabetes monitoring with percent hemoglobins A2 and F used for evaluation of (3 = thalassemia. The D-10™ Dual Program system consists of two different reagent programs with two intended uses. The D-10TM Dual Program reagent kit has a short program (3 minutes) for the determination of hemoglobin Are in which the components exactly the same as the D-10™ Hemoglobin Atc Program (K031043) reagents. The second program includes an extended program (6.5 minutes) that can be used for the determination of HbA2, HbF as well as HbA1c The components are exactly the same as the D-10™ Hemoglobin A12 Program (K031043) system and reagents with an additional HbA2/F/A1e Calibrator/Diluent Set and floppy diskette for the new program parameters.

#### Technical Characteristics Compared to Predicate

The Bio-Rad D-10™ Dual Program [identified herein as the: "D-10″ Dual" or "D-10™ Dual Program [6.5 minute)" system] and its 2 cleared predicates, the VARIANT™ II Hemoglobin A (K984268) and VARIANT™ II B-thalassemia (K991127) Programs, have the same technical HPLC and general program characteristics that are summarized in the following tables:

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## HbA1c

| Characteristics | Bio-Rad D-10™ Dual Program<br>(6.5 Minutes) | VARIANT II Hemoglobin A1c Program<br>[Cleared: / K984268; 12/17/98] |
|---------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use(s) | The Bio-Rad D-10 Dual Program system is<br>intended for the percent determination of<br>hemoglobins A1c, A2 and F, and for the<br>detection of abnormal hemoglobins in human<br>whole blood using ion-exchange high<br>performance liquid chromatography (HPLC).<br>The Bio-Rad D-10 Dual Program is intended<br>for in Professional Use only. For in vitro<br>diagnostic Use. | The VARIANT II Hemoglobin A1c Program is<br>intended for the determination of hemoglobin<br>A1c in human whole blood using ion-exchange<br>high performance liquid chromatography<br>(HPLC).<br>The VARIANT II Hemoglobin A1c Program is<br>intended for use only with the Bio-Rad<br>VARIANT II Hemoglobin Testing System.<br><br>For in vitro diagnostic use. |
| Indication(s) for Use | Measurement of the percent hemoglobin A1c is<br>effective in monitoring long-term glucose<br>control in individuals with diabetes mellitus,<br>and measurement of the percent HbA2 and HbF<br>are effective in monitoring of β-thalassemia<br>(i.e., hereditary hemolytic anemias<br>characterized by decreased synthesis of one or<br>more types of abnormal hemoglobin<br>polypeptide chains).<br><br>Detection of hemoglobin thalassemia variants<br>such as hemoglobins S, C, D and E by HPLC is<br>effective in presumptive identification of these<br>variants. | Measurement of the percent hemoglobin A1c is<br>effective in monitoring long-term glucose control<br>in individuals with diabetes mellitus. |
| Assay Principle | Cation exchange high performance liquid<br>chromatography | Cation exchange high performance liquid<br>chromatography |
| Sample Type | Human anticoagulated whole blood (EDTA) | Human anticoagulated whole blood (EDTA) |
| Visible Detection | 415 nm | 415 nm |
| Standardization | Traceable to the Diabetes Control and<br>Complications Trial (DCCT) reference method<br>and IFCC. Certified via the National<br>Glycohemoglobin Standardization Program<br>(NGSP) for HbA1c. | Traceable to the Diabetes Control and<br>Complications Trial (DCCT) reference method<br>and IFCC. Certified via the National<br>Glycohemoglobin Standardization Program<br>(NGSP) for HbA1c. |
| Results | Quantitative Area % HbA1c | Quantitative Area % HbA1c |
| Time to process<br>sample | 6.5 minutes | 3.0 minutes |
| Expected Value<br>Range | 4.27 – 6.07 % HbA1c | 4.27 – 6.07 % HbA1c |
| Linearity | 3.7 – 18.4 % HbA1c | 1.3 – 18.9% HbA1c |

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## HbA2

| Characteristics | Bio-Rad D-10TM Dual Program<br>(6.5 Minutes) | VARIANT II β-thalassemia Short<br>[Cleared: / K991127; 06/10/1999] |
|------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Uses | The Bio-Rad D-10 Dual Program system is<br>intended for the percent determination of<br>hemoglobins A1c, A2 and F, and for the<br>detection of abnormal hemoglobins in human<br>whole blood using ion-exchange high<br>performance liquid chromatography (HPLC).<br><br>The Bio-Rad D-10 Dual Program is intended<br>for Professional Use Only. For in vitro<br>diagnostic use. | The VARIANT II β-thalassemia Short Program<br>is intended for the separation and area percent<br>determinations of hemoglobins A2 and F, and<br>as an aid in the identification of abnormal<br>hemoglobins in whole blood using ion-<br>exchange high performance liquid<br>chromatography.<br><br>The VARIANT II β-thalassemia Short Program<br>is intended for use only with the Bio-Rad<br>VARIANT II Hemoglobin Testing System.<br><br>For in vitro diagnostic use. |
| Indication(s) for Use | Measurement of the percent hemoglobin A1c is<br>effective in monitoring long-term glucose<br>control in individuals with diabetes mellitus,<br>and measurement of the percent HbA2 and<br>HbF are effective in monitoring of β-<br>thalassemia (i.e., hereditary hemolytic anemias<br>characterized by decreased synthesis of one or<br>more types of abnormal hemoglobin<br>polypeptide chains).<br><br>Detection of hemoglobin thalassemia variants<br>such as hemoglobins S, C, D and E by HPLC<br>is effective in presumptive identification of<br>these variants. | Measurement of the percent HbA2 and HbF are<br>effective in monitoring of β-thalassemia (i.e.,<br>hereditary hemolytic anemias characterized by<br>decreased synthesis of one or more types of<br>abnormal hemoglobin polypeptide chains).<br><br>Identification of hemoglobin thalassemia<br>variants such as hemoglobins S, C, D and E by<br>HPLC is effective in presumptive identification<br>of these variants. |
| Assay Principle | Cation exchange high performance liquid<br>chromatography | Cation exchange high performance liquid<br>chromatography |
| Sample Type | Human anticoagulated whole blood (EDTA) | Human anticoagulated whole blood (EDTA) |
| Visible Detection | 415 nm | 415 nm |
| Standardization | The Joint Committee on Traceability in<br>Laboratory Medicine has not identified a<br>higher order reference method or reference<br>material for the quantitation of HbA2 and HbF | The Joint Committee on Traceability in<br>Laboratory Medicine has not identified a higher<br>order reference method or reference material for<br>the quantitation of HbA2 and HbF |
| Results | Quantitative Area % HbA2 | Quantitative Area % HbA2 |
| Time to process<br>sample | 6.5 minutes | 6.5 minutes |
| Expected Value<br>Range | 2.2 - 3.7 % HbA2 | 2.3 - 3.3% HbA2 |
| Linearity | 1.5 - 11.4 % HbA2 | 1.6 - 18.7% HbA2 |
| Characteristics | Bio-Rad D-10TM Dual Program<br>(6.5 Minutes) | VARIANTTM II β-thalassemia Short<br>[Cleared: / K991127; 06/10/1999] |
| Intended Uses | The Bio-Rad D-10 Dual Program system is<br>intended for the percent determination of<br>hemoglobins A1c, A2 and F, and for the<br>detection of abnormal hemoglobins in human<br>whole blood using ion-exchange high<br>performance liquid chromatography (HPLC).<br><br>The Bio-Rad D-10 Dual Program is intended<br>for Professional Use Only. For <i>in vitro</i><br>diagnostic use. | The VARIANT II β-thalassemia Short Program<br>is intended for the separation and area percent<br>determinations of hemoglobins A2 and F, and<br>as an aid in the identification of abnormal<br>hemoglobins in whole blood using ion-<br>exchange high performance liquid<br>chromatography.<br><br>The VARIANT II β-thalassemia Short Program<br>is intended for use only with the Bio-Rad<br>VARIANT II Hemoglobin Testing System.<br><br>For <i>in vitro</i> diagnostic use. |
| Indication(s) for Use | Measurement of the percent hemoglobin A1c is<br>effective in monitoring long-term glucose<br>control in individuals with diabetes mellitus,<br>and measurement of the percent HbA2 and<br>HbF are effective in monitoring of β-<br>thalassemia (i.e., hereditary hemolytic anemias<br>characterized by decreased synthesis of one or<br>more types of abnormal hemoglobin<br>polypeptide chains).<br><br>Detection of hemoglobin thalassemia variants<br>such as hemoglobins S, C, D and E by HPLC<br>is effective in presumptive identification of<br>these variants. | Measurement of the percent HbA2 and HbF are<br>effective in monitoring of β-thalassemia (i.e.,<br>hereditary hemolytic anemias characterized by<br>decreased synthesis of one or more types of<br>abnormal hemoglobin polypeptide chains).<br><br>Identification of hemoglobin thalassemia<br>variants such as hemoglobins S, C, D and E by<br>HPLC is effective in presumptive identification<br>of these variants. |
| Assay Principle | Cation exchange high performance liquid<br>chromatography | Cation exchange high performance liquid<br>chromatography |
| Sample Type | Human anticoagulated whole blood (EDTA) | Human anticoagulated whole blood (EDTA) |
| Visible Detection | 415 nm | 415 nm |
| Standardization | The Joint Committee on Traceability in<br>Laboratory Medicine has not identified a<br>higher order reference method or reference<br>material for the quantitation of HbA2 and HbF | The Joint Committee on Traceability in<br>Laboratory Medicine has not identified a higher<br>order reference method or reference material for<br>the quantitation of HbA2 and HbF |
| Results | Quantitative Area % HbF | Quantitative Area % HbF |
| Time to process<br>sample | 6.5 minutes | 6.5 minutes |
| Expected Value<br>Range | 0 - 0.8% HbF | <1.0% HbF |
| Linearity | 0.8 – 16.5 % HbF | 1.3 – 44.3% HbF |
| Characteristics | Bio-Rad D-10TM Dual Program<br>(6.5 Minutes) | VARIANT II β-thalassemia Short [Cleared: /<br>K991127; 06/10/1999] |
| Intended Uses | The Bio-Rad D-10 Dual Program system is<br>intended for the percent determination of<br>hemoglobins A1c, A2 and F, and for the<br>detection of abnormal hemoglobins in human<br>whole blood using ion-exchange high<br>performance liquid chromatography (HPLC).<br><br>The Bio-Rad D-10 Dual Program is intended<br>for Professional Use Only. For in vitro<br>diagnostic use. | The VARIANT II β-thalassemia Short Program<br>is intended for the separation and area percent<br>determinations of hemoglobins A2 and F, and<br>as an aid in the identification of abnormal<br>hemoglobins in whole blood using ion-<br>exchange high performance liquid<br>chromatography.<br><br>The VARIANT II β-thalassemia Short Program<br>is intended for use only with the Bio-Rad<br>VARIANT II Hemoglobin Testing System.<br><br>For in vitro diagnostic use. |
| Indication(s) for Use | Measurement of the percent hemoglobin A1c is<br>effective in monitoring long-term glucose<br>control in individuals with diabetes mellitus,<br>and measurement of the percent HbA2 and<br>HbF are effective in monitoring of β-<br>thalassemia (i.e., hereditary hemolytic anemias<br>characterized by decreased synthesis of one or<br>more types of abnormal hemoglobin<br>polypeptide chains).<br><br>Detection of hemoglobin thalassemia variants<br>such as hemoglobins S, C, D and E by HPLC<br>is effective in presumptive identification of<br>these variants. | Measurement of the percent HbA2 and HbF are<br>effective in monitoring of β-thalassemia (i.e.,<br>hereditary hemolytic anemias characterized by<br>decreased synthesis of one or more types of<br>abnormal hemoglobin polypeptide chains).<br><br>Identification of hemoglobin thalassemia<br>variants such as hemoglobins S, C, D and E by<br>HPLC is effective in presumptive identification<br>of these variants. |
| Assay Principle | Cation exchange high performance liquid<br>chromatography | Cation exchange high performance liquid<br>chromatography |
| Sample Type | Human anticoagulated whole blood (EDTA) | Human anticoagulated whole blood (EDTA) |
| Visible Detection | 415 nm | 415 nm |
| Results | Variant windows for S, C and any additional<br>unknown peaks will be detected as "unknown" | Variant windows for S, C, D and any additional<br>unknown peaks will be detected as "unknown" |
| Normal Reference<br>Interval | Compared to "normal" pattern of<br>chromatography | Compared to "normal" pattern of<br>chromatography |
| Time to process<br>sample | 6.5 minutes | 6.5 minutes |

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## HbF

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## Hemoglobin Variants

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## Testing To Establish Substantial Equivalence:

#### Accuracy:

#### HbAıc

Method correlation between Bio-Rad D-10™ Dual Program (6.5 minutes) and VARIANT™ II Hemoglobin At Program was evaluated using 40 EDTA whole blood samples ranging from 4.7% to 11.2% HbA1. The results are presented in the following table:

| ﻟﻠﻘﻀﺎﺀ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮ | THESE T FORT SET ( Dir TATTERS ( COLL CRECCRON FOR BERT FL | | | |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------|--------|--------|-----------|
| Regression Method | | | Slope | Intercept |
| Least Squares | 40 | 0.9843 | 0.9906 | 0.4310 |

#### D. 10TM Dual Program (6.5 Minutes) Correlation for HbA.

#### HbAz

Method correlation between Bio-Rad D-10TM Dual Program (6.5 minutes) and VARIANT™ II 8thalassemia Short Program was evaluated with 40 EDTA whole blood samples ranging from 1.9% to 8.9% HbA2. The results are presented in the following table:

| D-10™ Dual Program (6.5 Minutes) Correlation for HbA₂ | | | | |
|-------------------------------------------------------|----|--------|--------|-----------|
| Regression Method | n | r² | Slope | Intercept |
| Least Squares | 40 | 0.9832 | 1.0898 | -0.2407 |

## TM Duel Program (6.5 Minutes) Correlation for HbA

#### HbF

Method correlation between Bio-Rad D-10'™ Dual Program (6.5 minutes) and VARIANT™ II ß-thalassemia Short Program was evaluated with 40 EDTA whole blood samples ranging from 0% to 12.91% HbF. The results are presented in the following table:

| Regression Method | n | r2 | Slope | Intercept |
|-------------------|----|--------|--------|-----------|
| Least Squares | 40 | 0.9959 | 0.9497 | -0.1785 |

## D-10TM Dual Program (6,5 Minutes) Correlation for HbF

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### Precision:

#### HbAjc

The following precision table provides comparison data on the precision between D-101™ Dual Program (6.5 Minutes) and VARIANT™ II Hemoglobin Air Program, each utilizing EDTA I rogum (0.5 Minites) and both tested against samples with normal (5.4-5.9) and diabetic (13.1-13.7) % Alc content.

Method precision was performed using a protocol based on the NCCLS Evaluation protocol, Vol.12, No. 4, EP5-A (Feb. 1999) for the D-10TM Dual Program (6.5 Minutes) and NCCLS Fealuation protocol, Vol.12, No. 4, EP5-T2 (Mar. 1992) for the VARIANT II Hemoglobin Are Program. The protocols for both the D-10TM Dual Program (6.5 Minutes) and VARIANT II Hemoglobin Aic Programs are similar. Using these protocols, 40 runs (2 per day) were remoglobin 1 ( 1 1 0 g 1 1 1 (or V ARIANT ™ II) Hemoglobin Testing System over 20 working days. In each duplicate daily run, duplicate aliquots of normal HbAretic HbAr, patient samples were each analyzed per run. Although the precision samples are different, since they were run at different time periods, the precision results between the D-101™ Dual Program (6.5 Minutes) and the VARIANT ™ II Hemoglobin Aic (HbA1) Program are equivalent. A summary of combined comparative precision results is presented in the following precision table.

| D-10TM Dual (6.5 Minutes)<br>HbA1c Program | | VARIANTTM II Hemoglobin A1c<br>(HbA1c) Program | | |
|--------------------------------------------|-----------------|------------------------------------------------|-----------------|------|
| Normal Sample | Diabetic Sample | Normal Sample | Diabetic Sample | |
| n= (number of samples) | 80 | 80 | 80 | 80 |
| Mean (%HbA1c) | 5.9 | 13.1 | 5.4 | 13.7 |
| Within run (%CV) | 0.8 | 0.3 | 1.8 | 0.7 |
| Total Precision (%CV) | 1.8 | 0.9 | 2.1 | 1.7 |

D-10™ Dual Program (6.5 Minutes) HbAj¿ vs. VARIANT™ II HbAi¿ Program - Precision

### HbAz

The following precision table provides comparison data on the precision between D-1014 Dual Program (6.5 minutes) and VARIANT™ II ß -thalassemia Short Programs, each utilizing EDTA whole blood patient samples. The HbA2 tested samples had moderate (2.2-2.8) and high (4.6-5.4) % HbA2 content.

Method precision was performed using a protocol based on the NCCLS Evaluation protocol, Vol.12, No. 4, EP5-A (Feb. 1999) for the D-10TM Dual Program (6.5 minutes) and NCCLS Evaluation protocol, Vol.12, No. 4, EP5-T2 (Mar. 1992) for the VARIANT II ß-thalassemia Short Program. The protocols for both the D-10™ Dual Program and VARIANT II ßthalassemia Short Program are similar. Using these protocols, 40 runs (2 per day) were performed on one D-10™ (or VARIANT II) Hemoglobin Testing System over 20 working days. In each duplicate daily run, duplicate aliquots of low HbA2 and of high HbA2 patient samples were each analyzed in run. Although the precision samples are different, since they were run at different time periods, the precision results between the D-10™ Dual Program (6.5 minutes) and the VARIANT II ß-thalassemia Short Program are equivalent. A summary of combined comparative precision results is presented in the following precision table.

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#### Precision: (continued)

| | D-10 Dual Program (6.5 Minutes) | VARIANT II β-thalassemia Short | | |
|------------------------|---------------------------------|--------------------------------|------------|-------------|
| | HbA₂ | HbA₂ | | |
| | Low Sample | High Sample | Low Sample | High Sample |
| n= (number of samples) | 80 | 80 | 80 | 80 |
| Mean (%HbA₂) | 2.2 | 5.4 | 2.8 | 4.6 |
| Within run (%CV) | 4.5 | 1.7 | 1.6 | 0.9 |
| Total Precision (%CV) | 5.3 | 3.1 | 2.0 | 2.1 |

D-10™ Dual Program (6.5 minute HbA2) vs. VARIANT II (8-thalassemia Short(HbA2)-Precision

#### HbF

The following precision table provides comparison data on the precision between D-10™ Dual Program (6.5 minutes) and VARIANT™ II ß -thalassemia Short Programs, each utilizing EDTA whole blood patient samples. The HbF tested samples had moderate (1.6-2.1) and high (8.2-8.7) % HbF content.

Method precision was performed using a protocol based on the NCCLS Evaluation protocol, Vol.12, No. 4, EP5-A (Feb. 1999) for the D-10TM Dual Program (6.5 minutes) and NCCLS Evaluation protocol, Vol.12, No. 4, EP5-T2 (Mar. 1992) for the VARIANT II ß-thalassemia Short Program. The protocols for both the D-10TM Dual Program and VARIANT II ßthalassemia Short Program are similar. Using these protocols, 40 runs (2 per day) were performed on one D-10™ (or VARIANT II) Hemoglobin Testing System over 20 working days. In each duplicate daily run, duplicate aliquots of low HbF and of high HbF patient samples were each analyzed per run. Although the precision samples are different, since they were run at different time periods, the precision results between the D-10™ Dual Program (6.5 minutes) and the VARIANT II ß-thalassemia Short Program are equivalent. A summary of combined comparative precision results is presented in the following precision table.

| D-10 Dual Program (6.5 minutes) HbF | | VARIANT II β-thalassemia Short HbF | | |
|-------------------------------------|------------|------------------------------------|------------|-------------|
| | Low Sample | High Sample | Low Sample | High Sample |
| n= (number of samples) | 80 | 80 | 80 | 80 |
| Mean (%HbF) | 2.1 | 8.7 | 1.6 | 8.2 |
| Within run (%CV) | 1.7 | 1.4 | 2.1 | 0.6 |
| Total Precision (%CV) | 3.3 | 2.0 | 3.9 | 1.4 |

D-10™ Dual Program (6.5 minutes) HbF vs. VARIANT II ß-thalassemia Short (HbF) - Precision

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### Linearity:

#### HbAjc

The following linearity table provides comparison data on the linearity and recovery analyses between D-10 TM Dual Program (6.5 Minutes) and VARIANT II Hemoglobin A1e Programs, each utilizing eight EDTA-based blood standards (n=2 for each standard). This second linearity study was performed to compare the D-10th Dual Program with the VARAINT II Hemoglobin AIc linearity using the same standards. The % Recovery for HbAic by the D-10 ™ Dual Program is essentially the same as the VARIANT II Hemoglobin A16 Program. The results are presented in the following linearity table.

The linear range as stated in the Instruction Manual on the D-10™ Dual Program is 3.7 to 18.4% HbAr which was performed in the first linearity study, each using a total of seven standards (n=2 for each standard) below, at, and substantially above blood levels of typical normal levels of hemoglobin Air and found in normal and diabetic patients.

| % Contribution | | D-10 Dual Program (6.5 Minutes) | | | VARIANT II Hemoglobin A1c | | |
|----------------|----------|---------------------------------|------------------|------------|---------------------------|------------------|------------|
| Normal | Diabetic | Theoretical % HbA1c | Observed % HbA1c | % Recovery | Theoretical % HbA1c | Observed % HbA1c | % Recovery |
| 100 | 0 | 3.8 | 3.8 | 100 | 4.0 | 4.0 | 100 |
| 90 | 10 | 5.3 | 5.3 | 100 | 5.4 | 5.4 | 100 |
| 80 | 20 | 6.8 | 6.7 | 98.5 | 6.8 | 6.7 | 98.5 |
| 67 | 33 | 8.8 | 8.6 | 97.7 | 8.8 | 8.7 | 98.9 |
| 50 | 50 | 11.3 | 11.1 | 98.2 | 11.3 | 11.3 | 100 |
| 33 | 67 | 13.8 | 13.7 | 99.3 | 13.8 | 13.7 | 99.3 |
| 20 | 80 | 15.7 | 15.7 | 100 | 15.8 | 15.9 | 100.6 |
| 0 | 100 | 18.6 | 18.6 | 100 | 19.0 | 19.0 | 100 |

D-10 " Dual Program (6.5 Minutes) vs. VARIANT II Hemoglobin A12 Linearity

#### HbAz

The following linearity table provides comparison data on the linearity and recovery analyses between D-10 ™ Dual Program (6.5 minutes) and VARIANT II B-thalassemia, each utilizing eight EDTA-based blood standards (n=2 for each standard). This second linearity study was performed to compare the D-10 ™ Dual Program with the VARAINT II ß-thalassemia Short Program linearity using the same standards. The % Recovery for HbA2 by the D-10 100 Dual Program is essentially the same as the VARIANT II ß-thalassemia Short Program (HbA2). Results are presented in the linearity table below.

The linear range as stated in the Instruction Manual on the D-10 ™ Dual Program is 1.5 to 11.4% HbA2 which was performed in the first linearity study, each using a total of seven standards (n=2 for each standard) below, at, and substantially above blood levels of typical normal levels of hemoglobin A. and found in normal patients or patients with ß-thalassemia.

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### Linearity: (continued)

| D-10 Dual Program (6.5 Minutes) | | | VARIANT II - thalassemia Short | | | | |
|---------------------------------|------|-----------------------|--------------------------------|---------------|-----------------------|--------------------|---------------|
| % Contribution<br>Low | High | Theoretical<br>% HbA₂ | Observed<br>% HbA₂ | %<br>Recovery | Theoretical<br>% HbA₂ | Observed<br>% HbA₂ | %<br>Recovery |
| 100 | 0 | 1.7 | 3.8 | 100 | 1.8 | 1.8 | 100 |
| 90 | 10 | 2.5 | 2.5 | 100 | 2.6 | 2.6 | 100 |
| 80 | 20 | 3.4 | 3.1 | 91.2 | 3.4 | 3.3 | 97.1 |
| 67 | 33 | 4.5 | 4.2 | 93.3 | 4.4 | 4.3 | 97.1 |
| 50 | 50 | 6.0 | 5.8 | 96.7 | 5.7 | 5.6 | 98.3 |
| 33 | 67 | 7.4 | 7.2 | 97.3 | 7.0 | 6.9 | 98.6 |
| 20 | 80 | 8.6 | 8.5 | 98.8 | 8.1 | 8.1 | 100 |
| 0 | 100 | 10.3 | 10.3 | 100 | 9.7 | 9.7 | 100 |

#### D-10™ Dual Program (6.5 Minutes) vs. VARIANT II Hemoglobin A16 - Linearity

#### HPF

The following linearity table provides comparison data on the linearity and recovery analyses between D-10 TM Dual Program (6.5 minutes) and VARIANT TM II B-thalassemia, each utilizing eight EDTA-based blood standards (n=2 for each standard). This second linearity study was performed to compare the D-10 TM Dual Program with the VARAINT ™ II B-thalassemia Short Program linearity using the same standards. The % Recovery for HbF by the D-10 TM Dual Program is essentially the same as the VARIANT II ß-thalassemia Program. The results are presented in the following linearity table.

The linear range as stated in the Instruction Manual on the D-10 ™ Dual Program is 0.8 to 16.5% HbF which was performed in the first linearity study, each using a total of seven standards (n=2 for each standard) below, at, and substantially above blood levels of typical normal levels of hemoglobin F and found in normal patients or patients with ß-thalassemia.

| D-10 Dual Program (6.5 Minutes HbF) | | | VARIANT II β-thalassemia Short | | | | |
|-------------------------------------|----------------------|-------------------|--------------------------------|----------------------|-------------------|---------------|-------|
| % Contribution | Theoretical<br>% HbF | Observed<br>% HbF | %<br>Recovery | Theoretical<br>% HbF | Observed<br>% HbF | %<br>Recovery | |
| Low | High | 0.4 | 0.4 | 100 | 0.1 | 0.1 | 100 |
| 100 | 0 | | | | | | |
| 95 | 5 | 1.4 | 1.5 | 107.1 | 1.0 | 1.1 | 110.0 |
| 90 | 10 | 2.4 | 2.7 | 112.5 | 1.9 | 1.7 | 89.5 |
| 80 | 20 | 4.5 | 4.8 | 106.7 | 3.7 | 4.0 | 108.1 |
| 67 | 33 | 7.2 | 7.7 | 106.9 | 6.2 | 6.6 | 106.5 |
| 50 | 50 | 10.8 | 11.1 | 102.8 | 9.4 | 9.9 | 105.3 |
| 33 | 67 | 14.4 | 14.6 | 101.4 | 12.6 | 13.0 | 103.2 |
| 20 | 80 | 17.4 | 17.5 | 100.6 | 15.2 | 15.5 | 102.0 |
| 0 | 100 | 22.0 | 22.0 | 100 | 19.3 | 19.3 | 100 |

#### D-10 ™ Dual Program (6.5 Minutes) vs. VARIANT II {-thalassemia Short (HbF) - Linearity

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### Specificity and Interference Testing

#### HbA1c

In evaluating the specificity of the Bio-Rad D-10™ Dual Program for %HbAre in EDTAtreated blood samples, two closely related but chemical derived analogs of HbA1, where evaluated as part of a detailed analytical specificity study. The influence of carbamylated hemoglobin was studied by spiking specimens with sodium cyanate until the carbamylated hemoglobin levels increased to a range of 2.0%. Also, influence of unstable labile hemoglobin Ate was studied by spiking samples with glucose until unstable labile Ate in hemoglobin reached 3.5%. As was the case for the predicate Bio-Rad VARIANT™ II HbA1c system, the results with this new Rad D-10101 Dual Program system demonstrated that the final measurement of %HbAre at normal and diabetic levels was not significantly influenced by either added carbamylated hemoglobin or added glucose-labile hemoglobin Aic at the above indicated limits.

Additional normal and diabetic blood samples were obtained as patient bloods that were anticoagulated with EDTA in the standard manner. In three separate trials of patient pools or individual blood samples: a) concentrated bilirubin was added to a final level of 20 mg/dL: b) concentrated lipids were added to a final level of 5680 mg/dL; and c) additional dipotassium EDTA was added to a concentration of ~1980 mg/dL (11x the normal level) to determine the effect of high concentrations of EDTA that can occur in cases of "short draws." For the final measurement of normal and high diabetic HbA1c in blood samples, neither the Bio-Rad D-10TM Dual Program system nor cleared predicate Bio-Rad VARIANT™ II HbAic Program system were influenced by these excess biochemicals or excess EDTA anticoagulant, as illustrated in the interference evaluation table on the next page.

#### HbA2

In evaluating for specificity of the Bio-Rad D-10™ Dual Program for %HbA2 in EDTAtreated blood samples, additional normal, moderate and high blood %HbA2 samples were obtained as patient bloods that were anticoagulated with EDTA in the standard manner. In three separate trials of patient pools or individual blood samples: a) concentrated bilirubin was added to a final level of 20 mg/dL; b) concentrated lipids were added to a final level between 5680 mg/dL; and c) additional dipotassium EDTA was added to a concentration of ~1980 mg/dL (11x the normal level) to determine the effect of high concentrations of EDTA that can occur in cases of "short draws." For the final measurement of normal, moderate and high HbA2 in blood samples, neither the Bio-Rad D-10130 Dual Program system nor the cleared predicate Bio-Rad VARIANT II 8-thalassemia Program system were influenced significantly by these excess biochemicals or excess EDTA anticoagulant, as illustrated in the interference evaluation table on the next page.

{13}------------------------------------------------

## Specificity and Interference Testing - continued

#### HbF

The HbF assay was evaluated using the Bio-Rad D-10™ Dual Program system as part of a detailed analytical specificity study. First the influence of an unstable complex of glucose & hemoglobin known as labile Hemoglobin A16 (Which chromatographs in proximity to HbF) was studied by spiking samples with glucose until labile A . in Hemoglobin reached 0-2.6%. Final measurement of HbF in these blood-based human specimens was not influenced significantly by labile Hemoglobin A te at the above-indicated limits, as was the case also for the predicate, the Bio-Rad VARIANT™ II ß-thalassemia Program system.

In evaluating for specificity of this Bio-Rad D-10™ Dual Program system for %HbF in EDTA-treated blood samples, additional normal, moderate and high blood samples were obtained as patient bloods that were anticoagulated with EDTA in the standard manner. In three separate trials of patient pools or individual blood samples: a) concentrated bilirubin was added to a final level of 20 mg/dL; b) concentrated lipids were added to a final level between 5680 and 6000 mg/dL; and c) additional dipotassium EDTA was added to a concentration of ~1980 mg/dL (11x the normal level) to determine the effect of high concentrations of EDTA that can occur in cases of "short draws." For the final measurement of HbF, as well as HbA1c and HbA2, neither the Bio-Rad D-10 Dual Program system, nor cleared predicate Bio-Rad VARIANT II ß-thalassemia Program system were influenced significantly by these excess biochemicals or excess EDTA anticoagulant, as illustrated in the interference evaluation table below.

| Interfering Substance | D-10 TM Dual Program<br>(Extended)<br>(HbA1c & HbA2/F) | VARIANT TM II<br>Hemoglobin A1c<br>(HbA1c) | VARIANT TM II β-<br>thalassemia Short<br>(HbA2/F) |
|-------------------------------------------------------|-----------------------------------------------------------------|--------------------------------------------------------------------|---------------------------------------------------------|
| Potential Labile Hb<br>(glucose + Hb)<br>Interference | No significant<br>interference up to 3.5%<br>Labile Hb on HbA1c | No significant<br>interference up to<br>4.8% Labile Hb on<br>HbA1c | Not Applicable |
| Potential Labile Hb<br>(glucose + Hb)<br>Interference | No significant<br>interference up to 2.6%<br>Labile Hb on HbF | Not Applicable | Not Applicable |
| Bilirubin | No interference up to<br>20 mg/dL | No interference up to<br>20 mg/dL | No interference up to<br>20 mg/dL |
| Lipids<br>(Triglycerides) | No interference up to<br>5680 mg/dL | No interference up to<br>6000 mg/dL | No interference up to<br>4600 mg/dL |
| EDTA | No interference up to<br>11X EDTA | No interference up to<br>11X EDTA | No interference up to<br>11X EDTA |

## Summary of Testing for Interfering Substances:

{14}------------------------------------------------

## Conclusion:

The similarities of the intended use and the general performance characteristics and results of the The smilliarthed and evaluated Bio-Rad D-10™ Dual Program system are nearly identical to or newily desorrocd and Crafacted 210 reacleared program systems [i.e., the Bio-Rad logical extensions of the two oceared program and the Bio-Rad VARIANT T™ II B-thalassemia Short VARINT - In Tremogious: 11] - 10gs sed on the use of the same HPLC technology, and the I tograin. - Theo, one most of the correlation, precision, linearity, and interfering substances tests versus the corresponding results obtained with the two predicate systems that the Substances tests versus the occreepom system is substantially equivalent to these 2 cleared and currently marketed predicate systems.

{15}------------------------------------------------

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

Image /page/15/Picture/2 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized eagle-like symbol with three curved lines forming its body and wings. The symbol is positioned to the right of a circular arrangement of text that reads "DEPARTMENT OF HEALTH & HUMAN SERVICES USA".

Bio-Rad Laboratories, Inc. c/o Alfredo J. Quattrone, Ph.D., D.A.B.T. Third Party 510(k) Review Coordinator California Department of Health Food & Drug Branch 1500 Capitol Avenue Mailstop 7602 Sacramento, CA 95814

JUN - 9 2004

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Re: k041444

Trade/Device Name: Bio-Rad D-10TM Dual Program Regulation Number: 21 CFR 864.7470 Regulation Name: Glycosylated hemoglobin assay Regulatory Class: Class II Product Code: LCP Dated: May 28, 2004 Received: June 1, 2004

Dear Dr. Quattrone

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate for associous 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). Tou may, therefore, market the device, subject to the general controls provisions of the Act. The r ou may, dieres provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device n may be subject to bach adde of Federal Regulations (CFR), Parts 800 to 895. 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 i tease be day nou a carreraination that your device complies with other requirements of the Act that I Dr has Intatutes and regulations administered by other Federal agencies. You must or any I with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

{16}------------------------------------------------

#### Page 2

This letter will allow you to begin marketing your device as described in your Section 510(k) This letter will anow you to oegin mailing of substantial equivalence of your device to a legally prematication: The PDF Intentigssification for your device and thus, permits your device to proceed to the market.

If you desire specific information about the application of labeling requirements to your device, If you destic specific monitiation as advertising of yo…