HDL CHOLESTEROL (HDLX) ASSAY

K100010 · Beckman Coulter, Inc. · LBS · May 14, 2010 · Clinical Chemistry

Device Facts

• Record ID: K100010
• Device Name: HDL CHOLESTEROL (HDLX) ASSAY
• Applicant: Beckman Coulter, Inc.
• Product Code: LBS · Clinical Chemistry
• Decision Date: May 14, 2010
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1475
• Device Class: Class 1

Indications for Use

HDLX reagent, when used in conjunction with SYNCHRON LX® System(s), UniCel® DxC 800 System(s) and Genzyme Liquid N-geneous® HDL Cholesterol Calibrator is intended for quantitative determination of HDL Cholesterol in the high density lipoprotein fraction of human serum or plasma. HDL cholesterol measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases, and for the assessment of the risk of developing cardiovascular disease.

Device Story

The SYNCHRON® Systems HDL Cholesterol (HDLX) Reagent is a homogeneous, direct assay for quantitative HDL cholesterol measurement in human serum or plasma. Used with SYNCHRON LX® or UniCel® DxC 800 systems, the device automates sample/reagent proportioning (1:93 ratio) into a cuvette. A unique detergent solubilizes HDL particles while inhibiting reaction with LDL, VLDL, and chylomicrons via surface adsorption; a polyanion further enhances selectivity. Cholesterol esterase and oxidase react with released HDL cholesterol to produce a chromogenic product. The system monitors absorbance changes at 560 nm using a timed-endpoint method. The resulting absorbance change is directly proportional to cholesterol concentration, which the system calculates and reports. Operated by laboratory personnel in clinical settings, the output provides clinicians with quantitative lipid data to assist in diagnosing lipid disorders, atherosclerosis, and renal/liver diseases, and assessing cardiovascular risk.

Clinical Evidence

Bench testing only. Precision study (n=20 per level) showed within-run CVs 0.77%–1.83%. Linearity verified per CLSI EP6-A (r=0.9997–0.9998). LoD verified at 5 mg/dL per CLSI EP17-A. Method comparison with predicate using 100 patient samples showed high correlation (R=0.984–0.991). Matrix comparison (serum vs. plasma) confirmed equivalence across sample types.

Technological Characteristics

Homogeneous, direct enzymatic assay. Reagents: detergent (solubilizes HDL, inhibits LDL/VLDL/chylomicrons), polyanion (selectivity enhancer), cholesterol esterase, cholesterol oxidase, and chromogens. Measurement: spectrophotometric timed-endpoint at 560 nm. System: automated clinical chemistry analyzer (SYNCHRON LX/UniCel DxC 800).

Indications for Use

Indicated for quantitative determination of HDL cholesterol in human serum or plasma to aid in diagnosis/treatment of lipid disorders (e.g., diabetes mellitus), atherosclerosis, liver/renal diseases, and cardiovascular risk assessment.

Regulatory Classification

Identification

A lipoprotein test system is a device intended to measure lipoprotein in serum and plasma. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.

Predicate Devices

• Olympus HDL Cholesterol Assay (k040692)

Submission Summary (Full Text)

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

A. 510(k) Number:
k100010

B. Purpose for Submission:
Modification to a previously cleared device (k040767)

C. Measurand:
High Density Lipoprotein Cholesterol (HDL)

D. Type of Test:
Quantitative and semi-quantitative spectrophotometry

E. Applicant:
Beckman Coulter, Inc.

F. Proprietary and Established Names:
SYNCHRON® Systems HDL Cholesterol (HDLX) Reagent

G. Regulatory Information:
1. Regulation section:
21 CFR § 862.1475; Lipoprotein Test System
2. Classification:
Class I; meets limitations of exemptions under 21 CFR § 862.9 (c)(2)
3. Product code:
LBS; LDL & VLDL Precipitation, Cholesterol Via Esterase-Oxidase, HDL
4. Panel:
Clinical Chemistry (75)

H. Intended Use:
1. Intended use(s):
See Indications for use below.
2. Indication(s) for use:
HDLX reagent, when used in conjunction with SYNCHRON LX® System(s), UniCel® DxC 800 System(s) and Genzyme Liquid N-geneous® HDL Cholesterol Calibrator is intended for quantitative determination of HDL Cholesterol in the high density lipoprotein fraction of human serum or plasma.

HDL cholesterol measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases, and for the assessment of the risk of developing cardiovascular disease.

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3. Special conditions for use statement(s):
For Prescription Use Only

4. Special instrument requirements:
For use with the Unicel DxC 800 and LX20 SYNCHRON Clinical Systems.

I. Device Description:
The device consists of the SYNCHRON® Systems HDLD Reagent (BCI PN 650207). This reagent was cleared under k040767. The new device uses the cleared reagent calibrated using the Genzyme Diagnostics N-geneous Cholesterol Calibrator (PN 5272, cleared under k971162) which is not supplied with the kit.

J. Substantial Equivalence Information:
1. Predicate device name(s):
Olympus HDL Cholesterol Assay
2. Predicate K number(s):
k040692
3. Comparison with predicate:
| Item | Device | Predicate (k040692) |
| --- | --- | --- |
| Similarities | | |
| Intended Use | HDLX reagent, when used in conjunction with SYNCHRON LX® System(s), UniCel® DxC 800 System(s) and Genzyme Liquid N-geneous® HDL Cholesterol Calibrator is intended for quantitative determination of HDL Cholesterol in the high density lipoprotein fraction of human serum or plasma. | same |
| Fundamental technology | Chromogenic colored change measured by spectrophotometer | same |
| Sample types | Serum, heparin and EDTA plasma | same |
| Reagents | LIQUID stable reagents (ready to use) | same |
| Differences | | |
| Range | 5 – 135 mg/dL | 2.5 – 200 mg/dL |
| Sensitivity | 5 mg/dL | 1 mg/dL |

K. Standard/Guidance Document Referenced (if applicable):
- Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline (CLSI EP6-A).
- Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline – Second Edition (CLSI EP9-A2).
- Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline (CLSI EP17-A).

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L. Test Principle:

The SYNCHRON® Systems HDL Cholesterol (HDLX) Reagent is a direct HDL Cholesterol method. It is a homogeneous assay without the need for any offline pretreatment or centrifugation steps. The method depends on a unique detergent which solubilizes only the HDL lipoprotein particles and releases HDL cholesterol to react with cholesterol esterase and cholesterol oxidase in the presence of chromogens, to produce a color product. The same detergent also inhibits the reaction of the cholesterol enzymes with LDL, VLDL, and chylomicrons lipoproteins by adsorbing to their surfaces. A polyanion contained in the reagent enhances the selectivity for HDL cholesterol assay by complexing LDL, VLDL, and chylomicrons lipoproteins.

This HDLX reagent is used to measure the cholesterol concentration by a timed-endpoint method. The change in absorbance at 560 nanometers is monitored. This change in absorbance is directly proportional to the concentration of cholesterol in the sample and is used to calculate the HDL-cholesterol concentration.

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

The sponsor performed a precision study designed to verify that the within-run precision has not changed as a result of changing to the new calibrator. Commercially available multi-level control materials were used. Each control level was aliquoted into 4 samples and each sample was run in 5 replicates to generate a sample number (n) of 20. The study was conducted on one UniCel DxC 800 and one LX20 SYNCHRON Clinical System. The data is summarized below:

| Mean (mg/dL) | Within-run variation | | Mean (mg/dL) | Within-run variation | |
| --- | --- | --- | --- | --- | --- |
| | S.D. | %CV | | S.D. | %CV |
| Unicel DxC 800 | | | LX20 SYNCHRON Clinical System | | |
| 31.82 | 0.53 | 1.66 | 33.13 | 0.35 | 1.05 |
| 46.56 | 0.39 | 0.85 | 47.56 | 0.37 | 0.77 |
| 62.33 | 0.51 | 0.82 | 62.61 | 0.55 | 0.87 |
| 20.20 | 0.37 | 1.83 | 23.11 | 0.38 | 1.64 |
| 57.63 | 0.65 | 1.13 | 58.94 | 0.78 | 1.33 |
| 98.75 | 0.84 | 0.85 | 99.79 | 1.39 | 1.39 |

b. Linearity/assay reportable range:

The study was performed following CLSI protocol EP6-A. Commercially available linearity standards each containing 5 levels (10 different concentration levels in total)) of HDL spanning the reportable range of the assay were used. Each level was run in 3 replicates. The study was conducted on one UniCel DxC 800 and one LX20 SYNCHRON Clinical System. The recovered HDL values were plotted against the expected values and an appropriate line was fitted by standard linear regression. The results of the data analysis are summarized below:

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| Instrument | Range | Linear Regression Analysis |
| --- | --- | --- |
| DxC 800 | 5.0 – 135 mg/dL | Y = 1.004x + 1.00 r = 0.9997 |
| LX20 | 5.0 – 135 mg/dL | Y = 1.004x – 3.3 r = 0.9998 |

The claimed reportable range of the assay is 5.0 to 135 mg/dL.

c. Traceability, Stability, Expected values (controls, calibrators, or methods): The calibrators to be used with this assay are manufactured by Genzyme Corporation and have been previously cleared under k971162.

Calibration Frequency: The calibration is valid for 14 days on the UniCel DxC 800 and the LX20 SYNCHRON Clinical System.

The method has not been certified or tested by the Cholesterol Reference Method Laboratory Network. The package insert includes the following statement: “This method has not been certified or tested by the Cholesterol Reference Method Laboratory Network.”

d. Detection limit:

The sponsor performed a study designed to verify that the limit of detection (LoD) has not changed as a result of changing to the new calibrator. The study was performed in accordance with the recommendations of CLSI EP17-A. The limit of the blank (LoB) was calculated from data using a blank sample (5% human serum albumin solution) run in replicates of 20. The LoD was calculated using a low level sample (a diluted commercially available plasma-based standard) run in replicated of 20 on one UniCel DxC 800 and one LX20 SYNCHRON Clinical System. The sponsor stated that 100% of the replicates of the low level sample exceeded the calculated LoB.

The claimed LoD of the assay is 5 mg/dL.

e. Analytical specificity:

Established in k040767

f. Assay cut-off:

Not applicable

2. Comparison studies:

a. Method comparison with predicate device:

Studies were performed to compare the performance of the proposed device to the predicate device using 100 serum patient samples. The samples were run in singlet on the proposed device using the UniCel DxC 800 and the LX20 SYNCHRON Clinical System and in replicates of 2 on the predicate device. The results of the linear regression including the 95% confidence interval (CI) are summarized below:

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| Instrument | Range (mg/dL) | Slope (95% CI) | Intercept (95% CI) | R |
| --- | --- | --- | --- | --- |
| DxC 800 | 6.3 to 130 | 1.057 (1.030 to 1.085) | -4.927 (-6.696 to -3.159) | 0.991 |
| LX20 | 5 to 137 | 1.057 (1.020 to 1.095) | -5.465 (-7.870 to -3.060) | 0.984 |

b. Matrix comparison:
Forty eight paired samples (serum, lithium heparin plasma, sodium heparin plasma and EDTA plasma) were drawn from healthy volunteers. The samples were run in singlet on the proposed device using the UniCel DxC 800 and the LX20 SYNCHRON Clinical System. The plasma recoveries were compared to the corresponding serum recoveries for the same donor. Results of the linear regression including the 95% confidence interval (CI) are summarized below:

Lithium heparin plasma vs. serum

| Instrument | Range (mg/dL) | Slope (95% CI) | Intercept (95% CI) | R |
| --- | --- | --- | --- | --- |
| DxC 800 | 7 to 137 | 0.991 (0.977 to 1.004) | 1.011 (0.018 to 2.005) | 0.999 |
| LX20 | 9 to 125 | 0.999 (0.986 to 1.013) | -0.876 (-1.864 to 0.112) | 0.999 |

Sodium heparin plasma vs. serum

| Instrument | Range (mg/dL) | Slope (95% CI) | Intercept (95% CI) | R |
| --- | --- | --- | --- | --- |
| DxC 800 | 7 to 137 | 0.988 (0.972 to 1.004) | 1.198 (0.033 to 2.363) | 0.998 |
| LX20 | 9 to 125 | 1.000 (0.986 to 1.015) | -0.867 (-1.911 to 0.176) | 0.999 |

EDTA plasma vs. serum

| Instrument | Range (mg/dL) | Slope (95% CI) | Intercept (95% CI) | R |
| --- | --- | --- | --- | --- |
| DxC 800 | 7 to 137 | 0.977 (0.963 to 0.992) | 0.264 (-0.779 to 1.307) | 0.999 |
| LX20 | 9 to 125 | 0.994 (0.977 to 1.010) | -1.724 (-2.914 to -0.535) | 0.998 |

3. Clinical studies:
a. Clinical Sensitivity: Not applicable
b. Clinical specificity:

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Not applicable

c. Other clinical supportive data (when a. and b. are not applicable):

Not applicable

4. Clinical cut-off:

Not applicable

5. Expected values/Reference range:

The reference intervals listed were taken from literature*.

| Cardiovascular risk | Conventional units | S.I. units |
| --- | --- | --- |
| Low | ≥ 60 mg/dL | ≥ 1.55 mmol/L |
| High | < 40 mg/dL | < 1.03 mmol/L |

*NIH Publication No. 01 3305, ATP III Guidelines At-A-Glance, Quick Desk Reference, May (2001).
NIH Publication No. 01 3670, Third Report of National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation and Treatment of High Cholesterol in Adults (Adult Treatment Panel III), May (2001).

The package insert includes precautionary language that each laboratory should establish its own reference intervals based upon its patient population.

N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:

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