ACE® Hemoglobin A1c (HbA1c) Reagent is intended for the quantitative determination of hemoglobin A1c (umol/L) and total hemoglobin (g/dL) in human EDTA whole blood for the calculation of percent hemoglobin A 1 c using the ACE clinical chemistry system. This test is intended for use in clinical laboratories or physician office laboratories to monitor long term blood glucose control in individuals with diabetes mellitus. For in vitro diagnostic use only.

The umol HbA1c and total hemoglobin (THb) values generated are intended for use in the calculation of the HbA1c/THb ratio and cannot be used individually for diagnostic purposes.

Hemoglobin A1c Calibrators are intended for use in the performance of both a multi-point calibration for hemoglobin A1c and a single-point calibration of total hemoglobin on the ACE® clinical chemistry system, for the quantitative determination of percent (%) hemoglobin A1c in whole blood. For in vitro diagnostic use only.

Hemoglobin Alc Controls are intended to reliably monitor the accuracy and precision of quantitative determinations of hemoglobin A1c on the ACE® clinical chemistry system. For in vitro diagnostic use only,

Device Description

ACE® Hemoglobin A1c (HbA1c) Reagent is provided as a single kit and consists of four bottles containing a hemoglobin denaturant, a total hemoglobin reagent, a HbAlc agglutinator reagent and a HbA1c antibody reagent.

The ACE Hemoglobin Alc Calibrators are provided as a single kit and contain ready-to-use liquid calibrators, one of each of six levels.

The ACE Hemoglobin A1c Controls are provided as a single kit and contain lyophilized controls with normal and elevated levels of HbA1c and a reconstitution fluid. The controls are prepared from human whole blood, which has been tested and found negative for antibody to Human Immunodeficiency Virus (anti-HIV) Types 1 and 2, antibody to Hepatitis C (anti-HCV) and for Hepatitis B Surface Antigen (HBsAg) by FDA recommended (approved/licensed) tests.

AI/ML Overview

The document, K063306, describes the Alfa Wassermann, Inc. ACE® Hemoglobin A1c Reagent, Hemoglobin A1c Calibrators and Hemoglobin A1c Controls device. This device is intended for the quantitative determination of hemoglobin A1c and total hemoglobin in human EDTA whole blood for the calculation of percent hemoglobin A1c using the ACE clinical chemistry system.

Here's an analysis of the acceptance criteria and the study that proves the device meets them:

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state "acceptance criteria" in a table format with pass/fail thresholds. However, it presents performance data against which the device's efficacy can be judged, particularly in comparison to a predicate device. The implied acceptance is that the device's performance metrics are comparable to or within acceptable limits relative to established methods, as evidenced by successful correlation and precision studies.

Metric	Implied Acceptance Criteria (Based on Study Design and Predicate Comparison)	Reported Device Performance
Precision	Within-run CV: Low (e.g., < 5%). Total CV: Low (e.g., < 10%). (These are general industry expectations for diagnostic assays, not explicitly defined thresholds in the document, but the study results are presented as meeting these expectations.)	Clinical Chemistry System: - Normal Patient: Within-run CV 2.9%, Total CV 7.0%. - Elevated Patient: Within-run CV 3.0%, Total CV 5.9%. POL Sites (3 sites): - All within-run and total CVs were less than 5.4%.
Measuring Range (Total Hemoglobin)	Statistically acceptable range determined by 95% confidence intervals from linear regression analysis with ±10% limits.	Acceptable range for total hemoglobin: 10-21 g/dL.
Accuracy (Correlation with Predicate)	High correlation coefficient (e.g., > 0.95), slope close to 1, intercept close to 0, and reasonable standard error.	Overall Study (n=unspecified, presumably larger):- Regression Equation: y = 1.052x + 0.23- Correlation Coefficient: 0.975- Standard Error of the Estimate: 0.49- Confidence Interval Slope: 1.000 to 1.104- Confidence Interval Intercept: -0.20 to 0.67POL Labs (n=20 samples per lab):- Lab A: y = 0.940x + 0.41, R=0.9762, SEE=0.46, CI Slp=0.871-1.009, CI Int=-0.16-0.99- Lab B: y = 1.102x - 0.04, R=0.9818, SEE=0.47, CI Slp=1.032-1.172, CI Int=-0.63-0.55- Lab C: y = 1.035x + 0.38, R=0.9844, SEE=0.41, CI Slp=0.974-1.095, CI Int=-0.89-0.13

2. Sample Size Used for the Test Set and Data Provenance

Precision Studies:
- Main Study: For the ACE clinical chemistry system, two patient samples (one normal, one elevated) were used for determining within-run and total CVs. The number of replicates or days for this specific part is not explicitly stated, but it followed CLSI/NCCLS EP5-A2 which outlines such protocols.
- POL Sites: Three EDTA whole blood samples with varying HbA1c levels were run in triplicate on five different days at each of the three POL sites.
Measuring Range Study: One whole blood sample was used, which was serially diluted.
Accuracy (Correlation) Studies:
- Main Correlation Study: The document does not specify the exact number of samples for the main correlation study against the Tosoh Bioscience's A1c 2.2 Plus Automated Glycohemoglobin Assay. It refers to "patient samples" but no count.
- POL Labs: Each of the three POL labs ran 20 EDTA whole blood samples in duplicate in four different runs (total of 80 measurements per lab, 240 measurements across all POL labs for accuracy comparison).
Data Provenance: The document does not explicitly state the country of origin for the data. Given the submitter's address (West Caldwell, NJ, USA) and FDA submission context, it is highly probable the data was gathered in the United States. The studies appear to be prospective in nature, designed specifically for this submission.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications

For this type of in-vitro diagnostic device (quantitative measurement of a biomarker), "experts" in the sense of human readers (like radiologists) are not typically used to establish ground truth.
The ground truth is established by a predicate device or reference method, which is considered the gold standard. In this case, the Tosoh Bioscience's A1c 2.2 Plus Automated Glycohemoglobin Assay was used as the predicate device against which the accuracy of the ACE Hemoglobin A1c kit was evaluated. The performance characteristics of the predicate device are the "ground truth" here.
There is no mention of a specific number of human experts or their qualifications for establishing ground truth, as the ground truth itself is an established laboratory method.

4. Adjudication Method for the Test Set

Adjudication methods (like 2+1, 3+1) are common in studies where human interpretation or subjective assessment plays a role in establishing ground truth (e.g., image-based diagnostics).
For a quantitative in-vitro diagnostic device like the ACE Hemoglobin A1c kit, which relies on chemical reactions and automated measurement, adjudication by human experts is not applicable or performed. The "adjudication" is inherent in the direct comparison of the device's quantitative output against the quantitative output of the predicate device. Discrepancies are handled through statistical analysis (e.g., linear regression, correlation).

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, an MRMC comparative effectiveness study was not done.
MRMC studies are typically used to assess the impact of an AI system (or new diagnostic tool) on human reader performance, especially in fields like radiology where human interpretation of images is critical.
This device is an automated, in-vitro diagnostic assay for a chemical biomarker (HbA1c). Its primary function is to provide a quantitative measurement, not to assist human interpretation in a comparative effectiveness study setting.

6. Standalone Performance Study (Algorithm Only Without Human-in-the-Loop Performance)

Yes, this entire study is essentially a standalone performance study.
The ACE Hemoglobin A1c kit, when used with the ACE clinical chemistry system, operates as an automated system. The performance data presented (precision, measuring range, accuracy against a predicate) represents the performance of the algorithm/reagent system on its own, without human intervention influencing the measurement outcome once the process is initiated. Personnel (even those without formal medical technology education in POL sites) are involved in sample preparation and loading, but the actual analytical measurement and calculation are automated by the device.

7. Type of Ground Truth Used

The ground truth used for the accuracy studies was the concurrent measurement obtained from a legally marketed and established predicate device: the Tosoh Bioscience's A1c 2.2 Plus Automated Glycohemoglobin Assay. This is a common and accepted method for establishing accuracy for new in-vitro diagnostic devices. It operates as a "reference method" in this context.

8. Sample Size for the Training Set

The document does not explicitly mention a training set sample size. This device is a reagent/assay kit for a clinical chemistry system, not a machine learning or AI algorithm that typically goes through a distinct "training phase" on a dataset in the same way.
The methods described (latex agglutination inhibition assay, conversion to alkaline hematin, measurement of absorbance) are biochemical principles. While there might have been internal development/optimization data, it's not described as a formal "training set" in the context of this 510(k) summary. The calibration procedure itself (using calibrators) is a form of "training" the system for accurate measurements, but this is distinct from a dataset used for training an AI model.

9. How the Ground Truth for the Training Set Was Established

As concluded in point 8, a formal "training set" in the AI sense is not described.
For the calibration process (which could be considered analogous to a system's "learning" or setting up its operational parameters), the Hemoglobin A1c Calibrators are used. These calibrators are presumably manufactured with known, precise concentrations of HbA1c and total hemoglobin, established through rigorous manufacturing and quality control standards. The ground truth for these calibrators would be established through a combination of gravimetric/volumetric preparation and validation against higher-order reference methods and materials (e.g., IFCC reference methods for HbA1c primary calibration). However, this specific process for the calibrators' ground truth is not detailed in this 510(k) summary.

Summary

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K063306

510(k) SUMMARY

Alfa Wassermann, Inc. ACE® Hemoglobin A1c Reagent, Hemoglobin A1c Calibrators and Hemoglobin A1c Controls

Submitter

Alfa Wassermann, Inc. 4 Henderson Drive West Caldwell, NJ 07006 Phone: 973-852-0120 Facsimile: 973-852-0121

Contact Person

Murray Rosenthal, Ph.D.

Date Prepared

October 31, 2006

Product Trade and Common Name

ACE® Hemoglobin A1c (HbA1c) Reagent Hemoglobin A1c Calibrators Hemoglobin Alc Controls

Classification Name

Class II, Product Code LCP CFR § 864.7470: Glycosylated Hemoglobin Assay

Class II, Product Code JIT CFR § 862.1150: Calibrator, Secondary

Class I, Product Code JJX CFR § 862.1660: Quality Control Material (assayed and unassayed)

Predicate Devices

Tosoh Bioscience, Inc. A1c 2.2 Plus Automated Glycohemoglobin Assay

General Atomics, Inc. Hemoglobin A1c Enzymatic Assay

ALFA WASSERMANN, INC. Henderson Drive st Caldwell, NJ 07006 3) 882-8630 (973) 227-6700 faWassermannUS.com APR 1 7 2007

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Description and Intended Use of the Device

ACE® Hemoglobin A1c (HbA1c) Reagent is intended for the quantitative determination of hemoglobin A1c (umol/L) and total hemoglobin (g/dL) in human EDTA whole blood for the calculation of percent hemoglobin A1c using the ACE clinical chemistry system. This test is intended for use in clinical laboratories or physician office laboratories to monitor long term blood glucose control in individuals with diabetes mellitus. For in vitro diagnostic use only.

The umol HbAlc and total hemoglobin (THb) values generated are intended for use in the calculation of the HbA1c/THb ratio and cannot be used individually for diagnostic purposes.

Hemoglobin A1c Calibrators are intended for use in the performance of both a multi-point calibration for hemoglobin A1c and a single-point calibration of total hemoglobin on the ACE clinical chemistry system for the quantitative determination of percent (%) hemoglobin A1c in whole blood. For in vitro diagnostic use only.

Hemoglobin A1c Controls are intended to reliably monitor the accuracy and precision of quantitative determinations of hemoglobin A1c on the ACE clinical chemistry system. For in vitro diagnostic use only.

Technological Characteristics

The ACE Hemoglobin A1c Reagent is provided as a single kit and consists of four bottles containing a hemoglobin denaturant, a total hemoglobin reagent, a HbAlc agglutinator reagent and a HbA1c antibody reagent.

The ACE Hemoglobin Alc Calibrators are provided as a single kit and contain ready-to-use liquid calibrators, one of each of six levels.

Principles of Operation

Prior to assay, whole blood samples require a pretreatment step. The red blood cells in the sample are lysed by the Hemoglobin Denaturant and the hemoglobin chain hydrolyzed.

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For determination of HbAlc, a latex agglutination inhibition assay is used. In the absence of HbA 1c in the sample, the agglutinator (synthetic polymer containing the immunoreactive portion of HbA1c) in the HbAIc Agglutinator Reagent and the antibody-coated microparticles in the HbAIc Antibody Reagent will agglutinate. The presence of HbA1c in the sample competes for the antibody binding sites and inhibits agglutination. The increase in absorbance, monitored monochromatically at 592 nm on the ACE clinical chemistry system, is inversely proportional to the HbA1c present in the sample.

For the determination of total hemoglobin, all hemoglobin derivatives in the sample are converted to alkaline hematin. The reaction produces a green colored solution, which is measured bichromatically at 573 nm/692 nm on the ACE clinical chemistry system. The intensity of color produced is directly proportional to the total hemoglobin concentration in the sample.

Both the concentration of HbA1c and total hemoglobin are measured; then the ratio is calculated and the result reported as percent HbA1c.

The ACE Hemoglobin A1c Calibrators are used to calibrate the ACE clinical chemistry system for both the Hemoglobin HbA1c and Total Hemoglobin assays.

The ACE Hemoglobin A 1c Controls are run in the same way as patient samples.

Performance Data

The performance of the ACE Hemoglobin A 1 c kit was evaluated for precision, measuring range and accuracy.

The precision studies were done according to CLSI/NCCLS EP5-A2, Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline - Second Edition. Using this protocol, precision was measured on the ACE clinical chemistry system using the ACE Hemoglobin A1c reagents, calibrated with the ACE Hemoglobin A1c Calibrators on a normal and elevated patient. The within-run CV for the normal patient was 2.9% and for the elevated patient was 3.0%. The total CV for the normal patient was 7.0% and for the elevated patient was 5.9%.

Further precision studies were done in three separate Physician Office Laboratory (POL) sites. These studies were conducted by personnel without formal medical technology education. The studies consisted of running three EDTA whole blood samples with varying levels of HbA1c in triplicate on five different days. The within-run and total CVs were all less than 5.4%.

The measuring range for total hemoglobin was determined using a whole blood sample, by serial dilution of the packed cells with plasma from the blood sample, HbAlc levels were measured on the ACE clinical chemistry system using the

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ACE Hemoglobin Alc reagents, calibrated with the ACE Hemoglobin A1c Calibrators. The acceptable range was determined by using 95% confidence intervals from linear regression analysis with ±10% limits. The acceptable range for total hemoglobin was determined to be 10-21 g/dL.

A correlation study, to determine the accuracy of the ACE Hemoglobin A1c kit. calibrated with the ACE Hemoglobin A1c calibrators, was done against the Tosoh Bioscience's A1c 2.2 Plus Automated Glycohemoglobin Assay. The study followed the protocol in CLSI/NCCLS EP9-T, Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline - Second Edition. Linear regression analysis was performed: ACE clinical chemistry system (y) and Tosoh HPLC (x). The analysis yielded the following results:

RegressionEquation	CorrelationCoefficient	Standard Error ofthe Estimate	ConfidenceInterval Slope	ConfidenceIntervalIntercept
y = 1.052x+0.23	0.975	0.49	1.000 to 1.104	-0.20 to 0.67

Further accuracy studies were done in three separate POL labs. These studies were conducted by personnel without formal medical technology education. At each lab, these studies consisted of running the same 20 EDTA whole blood samples with varying levels of HbA1c (determined using the Tosoh Bioscience's A 1 c 2.2 Plus Automated Glycohemoglobin Assay) in duplicate in four different runs of ten samples each. Linear regression analysis was performed: ACE clinical chemistry system (y) and Tosoh HPLC (x). The analysis yielded the following results:

Lab	RegressionEquation	CorrelationCoefficient	Standard Errorof theEstimate	ConfidenceInterval Slope	ConfidenceIntervalIntercept
A	$y = 0.940x+0.41$	0.9762	0.46	0.871 to 1.009	-0.16 to 0.99
B	$y = 1.102x-0.04$	0.9818	0.47	1.032 to 1.172	-0.63 to 0.55
C	$y = 1.035x+0.38$	0.9844	0.41	0.974 to 1.095	-0.89 to 0.13

In all instances, the ACE Hemoglobin A1c kit run on the ACE clinical chemistry analyzer functioned as intended.

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Substantial Equivalence

The evaluations and comparative analysis indicated substantial equivalence between the results obtained for the ACE Hemoglobin A1c Reagent, ACE Hemoglobin Alc Calibrators and ACE Hemoglobin A1c Controls and the predicate devices (Tosoh Bioscience's A1c 2.2 Plus Automated Glycohemoglobin Assay and General Atomics Enzymatic HbA1c Assay). The ACE Hemoglobin Alc Reagent, ACE Hemoglobin A1c Calibrators and ACE Hemoglobin A1c Controls have the same intended use and similar indications, technological characteristics, performance characteristics, and similar principles of operation as its predicate devices. Therefore, the ACE Hemoglobin A1c Reagent, ACE Hemoglobin A1c Calibrators and ACE Hemoglobin A1c Controls are substantially equivalent to commercially available products that measure HbA1c in whole blood samples.

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/5/Picture/1 description: The image shows the seal of the U.S. Department of Health & Human Services. The seal features a stylized eagle with its wings spread, and the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the eagle. The seal is black and white and appears to be a simple graphic representation.

Public Health Service

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Alfa Wassermann Diagnostic Technologies, LLC c/o Dr. Murray Rosenthal Manager, Reagent Technology 4 Henderson Drive West Caldwell, NJ 07006

APR 1 7 2007

Re: K063306

Trade/Device Name: ACE® Hemoglobin A1c (HbA1c) Reagent Hemoglobin Alc Calibrators Hemoglobin Alc Controls

Regulation Number: 21 CFR 864.7470 Regulation Name: Glycosylated Hemoglobin assay. Regulatory Class: Class II Product Code: LCP, JIT, JJX Dated: March 2, 2007 Received: March 5, 2007

Dear Dr. Rosenthal:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

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 can be found in Title 21, Code 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 that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

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Page 2 -

This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification 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, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (240) 276-0490. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its tollil the (800) 638-2041 or (240) 276-3150 or at its Internet address at http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely vours.

Jean M. Cooper, M.S., D.V.M.

Yean M. Cooper, M.S., D.V.M. Director Division of Chemistry and Toxicology Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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Section 4 Section 1.01 Indications for Use Statement

510(k) Number (if known): K063306

Device Name:

ACE® Hemoglobin A1c (HbA1c) Reagent Hemoglobin A1c Calibrators Hemoglobin Alc Controls

Indications for Use:

ACE® Hemoglobin A1c (HbA1c) Reagent is intended for the quantitative determination of hemoglobin Alc (umol/L) and total hemoglobin (g/dL) in human EDTA whole blood for the calculation of percent hemoglobin A 1 c using the ACE clinical chemistry system. This test is intended for use in clinical laboratories or physician office laboratories to monitor long term blood glucose control in individuals with diabetes mellitus. For in vitro diagnostic use only.

The umol HbA1c and total hemoglobin (THb) values generated are intended for use in the calculation of the HbA1c/THb ratio and cannot be used individually for diagnostic purposes.

Prescription Use X (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 801 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (OD

Page of

Carol C. Benson
Division Sign-Off

Division Sign-

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K063306

Regulation Number and Section

§ 864.7470 Glycosylated hemoglobin assay.

(a)
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.(b)
Classification. Class II (performance standards).