CAROLINA LIQUID CHEMISTRIES HDL CHOLESTEROL REAGENT

K132711 · Carolina Liquid Chemistries Corp. · LBS · May 8, 2014 · Clinical Chemistry

Device Facts

• Record ID: K132711
• Device Name: CAROLINA LIQUID CHEMISTRIES HDL CHOLESTEROL REAGENT
• Applicant: Carolina Liquid Chemistries Corp.
• Product Code: LBS · Clinical Chemistry
• Decision Date: May 8, 2014
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1475
• Device Class: Class 1

Indications for Use

For the quantitative determination of high-density lipprotein cholesterol (HDL-C) in serum using the Carolina Liquid Chemistries CLC720® Clinical Chemistry Analyzer. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases. For in vitro diagnostic use only.

Device Story

The Carolina Liquid Chemistries HDL Cholesterol Reagent is an in vitro diagnostic reagent used for the quantitative measurement of HDL-C in human serum. The device is designed for use on the Carolina Liquid Chemistries CLC720® Clinical Chemistry Analyzer. It functions by facilitating a chemical reaction within the analyzer to measure cholesterol levels. The output is a quantitative concentration value of HDL-C, which is provided to clinicians to assist in the diagnosis and management of lipid-related conditions, such as atherosclerosis, diabetes mellitus, and liver or renal diseases. The device is intended for professional use in clinical laboratory settings.

Clinical Evidence

No clinical data provided; bench testing only.

Technological Characteristics

Two-reagent colorimetric enzymatic assay. Reactive ingredients: Cholesterol esterase, Cholesterol oxidase, Peroxidase, DSBmT, 4-aminoantipyrine. Sensing principle: detergent-mediated selective HDL solubilization followed by enzymatic color development. Form factor: liquid reagent for automated clinical chemistry analyzer (CLC720). Storage: 2-8°C. Not CRMLN certified.

Indications for Use

Indicated for quantitative determination of HDL-C in serum for patients requiring assessment of lipid disorders, including diabetes mellitus, atherosclerosis, and liver or renal diseases. For prescription use only.

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.

Submission Summary (Full Text)

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

A. 510(k) Number:
k132711

B. Purpose for Submission:
New device

C. Measurand:
HDL Cholesterol assay

D. Type of Test:
Quantitative, colorimetric assay

E. Applicant:
Carolina Liquid Chemistries Corp.

F. Proprietary and Established Names:
Carolina Liquid Chemistries HDL Cholesterol Reagent

G. Regulatory Information:
1. Regulation section:
CFR 21 section 862.1475 - Lipoprotein test system
2. Classification:
Class I, meets limitations per exemption 21 CFR 862.9(c)(4)
3. Product code:
LBS
4. Panel:
Clinical Chemistry (75)

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H. Intended Use:

1. Intended use(s):

See Indication(s) for use.

2. Indication(s) for use:

For the quantitative determination of high-density lipoprotein cholesterol (HDL-C) in serum using the Carolina Liquid Chemistries CLC720® Clinical Chemistry Analyzer. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases. For in vitro diagnostic use only.

3. Special conditions for use statement(s):

For prescription use only

4. Special instrument requirements:

Carolina Liquid Chemistries CLC720® Clinical Chemistry Analyzer (Cleared in k112377)

I. Device Description:

The Carolina Liquid Chemistries HDL Cholesterol assay is a homogeneous method for directly measuring serum HDL-C levels without the need for any off-line pretreatment or centrifugation steps. The method is in a two-reagent format. The first reagent contains a polyanion and a chromagen. The second reagent contains standard cholesterol enzymes along with a special detergent that has selective affinity for HDL versus other lipoproteins (LDL, VLDL, Chylomicrons).

J. Substantial Equivalence Information:

1. Predicate device name(s):

Olympus HDL Cholesterol Reagent

2. Predicate 510(k) number(s):

k040692

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3. Comparison with predicate:

| Reagent Similarities | | |
| --- | --- | --- |
| Item | Candidate Device: Carolina Liquid Chemistries HDL Cholesterol Reagent (k132711) | Predicate Device: Olympus HDL Cholesterol Reagent (k040692) |
| Intended Use | System reagent for the quantitative determination of HDL-Cholesterol. For in vitro diagnostic use | Same |
| Measurand Analyte | HDL-Cholesterol | Same |
| Test Principle | Detergent solubilization of HDL to selectively measure HDL cholesterol using an enzymatic method | Same |
| Reagent Composition | Reactive ingredients: Cholesterol esterase, Cholesterol oxidase, Peroxidase, DSBmT (N,N-bis(4-sulphobutyl)-m-toluidine-disodium salt), 4-aminoantipyrine | Same |
| Reagent Storage and Stability | Unopened reagent is stable until the expiration date stated on the vial label when stored at 2-8°C; opened reagents are stable for 30 days when stored in the refrigerator compartment of the analyzer | Same |
| Reagent Differences | | |
| --- | --- | --- |
| Item | Candidate Device: Carolina Liquid Chemistries HDL Cholesterol Reagent (k132711) | Predicate Device: Olympus HDL Cholesterol Reagent (k040692) |
| Sample Type | Serum | Serum and Plasma |

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K. Standard/Guidance Document Referenced (if applicable):

CLSI EP5-A2, Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline-Second Edition.

CLSI EP6-A, Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline.

CLSI EP17-A, Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline.

CLSI EP07-A2, Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition.

CLSI EP9-A2, Method Comparison and Bias Estimation Using Patient Samples, 2002

L. Test Principle:

The method is in a two-reagent format. The first reagent contains a polyanion and a chromagen. The second reagent contains standard cholesterol enzymes along with a special detergent that has selective affinity for HDL verses the other lipoproteins (LDL, VLDL, Chylomicrons).

The reaction can be described as follows:

Detergent
LDL, VLDL, Chylomicrons → Stable Complexes
Polyanion

Detergent
HDL → HDL Disrupted

Cholesterol Esterase
HDL Cholesterol → Cholest-4-en-3-one
Cholesterol Oxidase

H2O2 + DSBmT + 4-AA → Color Development
Peroxidase

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M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

Precision Studies were conducted utilizing the Carolina Liquid Chemistries HDL Cholesterol Reagent on the Carolina Liquid Chemistries CLC 720 Chemistry Analyzer following the recommendations in the CLSI guideline EP5-A2. A total of three serum-based control materials were analyzed 40 times for the within-run precision, and were analyzed in triplicate two times per day for 20 non-consecutive days for the total precision analysis. A single reagent lot and instrument was used for the analysis. The results are summarized below:

| Precision | | | | | | |
| --- | --- | --- | --- | --- | --- | --- |
| | WITHIN RUN | | | TOTAL PRECISION | | |
| | Level 1 | Level 2 | Level 3 | Level 1 | Level 2 | Level 3 |
| N | 40 | 40 | 40 | 120 | 120 | 120 |
| MEAN (mg/dL) | 65.3 | 45.0 | 26.9 | 65.3 | 45.0 | 26.9 |
| SD (mg/dL) | 1.04 | 0.92 | 0.66 | 1.60 | 1.32 | 1.13 |
| CV (%) | 1.6 | 2.0 | 2.5 | 2.4 | 2.9 | 4.2 |

The sponsor performed the following additional precision studies on the Carolina Liquid Chemistries CLC 720 Chemistry Analyzer following recommendations in the CLSI guideline EP5-A2. Two clinical sample pools were analyzed 20 times for the within-run precision, and were analyzed two times per day, for 30 non-consecutive days, for the total precision determination. A single reagent lot (different lot from above described study) and instrument (same as above study) was used for the analysis. The results are summarized below:

| Precision | | | | | |
| --- | --- | --- | --- | --- | --- |
| | WITHIN RUN | | | TOTAL PRECISION | |
| | Level 1 | Level 2 | | Level 1 | Level 2 |
| N | 20 | 20 | | 60 | 60 |
| MEAN (mg/dL) | 43.2 | 83.6 | | 41.2 | 81.4 |
| SD (mg/dL) | 0.796 | 0.582 | | 1.80 | 2.01 |
| CV | 1.8 | 0.7 | | 4.4 | 2.5 |

b. Linearity/assay reportable range:

Linearity studies were carried out using six commercially available HDL Linearity

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standards. All samples were tested five times. The sample range tested was 8.6 mg/dL to 165.2 mg/dL.

| Range tested | Intercept | Slope | r² |
| --- | --- | --- | --- |
| 8.6 to 165.2 mg/dL | 0.83 | 1.002 | 1.00 |

The sponsor demonstrated acceptable linearity from 8.6 to 165.2 mg/dL.

The claimed measuring range of the device is from 9 to 150 mg/dL.

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

The sponsor recommends the use of a commercially available calibrator (the JAS Diagnostics HDL Calibrator traceable to NIST SRM 911b) which was previously cleared in k021671.

Stability testing was performed by the manufacturer. The reagents are stable for 12 months when stored between 2-8 °C.

On board stability was performed on the CLC 720 Chemistry Analyzer demonstrated that the reagents are stable for 30 days when stored on board the CLC 720 Chemistry Analyzer when stored at 2-8 °C.

The reagent system has not been tested or certified by the Cholesterol Reference Method Laboratory Network (CRMLN). The labeling contains language that the device has not been certified by the CRMLN.

d. Detection limit:

A limit of detection study was conducted based on procedures described in CLSI EP17-A. The limit of the blank (LoB) of 0.16 mg/dL was calculated from data using a pooled serum sample diluted with 0.85 % sodium chloride assayed 8 times and repeated for a total of ten individually calibrated runs to produce 80 results. The LoD of 0.53 mg/dL was calculated as the LoB plus the one-tailed 95% confidence interval of the test results. The LoQ of 6.48 mg/dL was determined with a performance goal of ≤20% CV and is lower than the lower limit of the measuring range which is set at 9 mg/dL.

e. Analytical specificity:

To determine the level of interference from substances normally present in human serum, the Carolina Liquid Chemistries HDL Cholesterol Reagent was tested with low level HDL serum samples containing about 32-38 mg/dL HDL-C and high level HDL serum samples containing about 62-72 mg/dL HDL-C. Both levels were spiked with various concentrations of potentially interfering substances following the recommendations in the

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CLSI guideline EP7-A2 on the CLC 720 Chemistry Analyzer. A total of four or six levels were tested for interferents and all samples were tested in six runs with eight replicates. The following substances normally present in serum produced less than 10% deviation when tested at levels equal to the following concentrations:

| Interference Substance | Concentration |
| --- | --- |
| Bilirubin
(conjugated and unconjugated) | 20 mg/dL |
| Hemolysis | 200 mg/dL |
| Lipemia | 800 mg/dL |
| Ascorbic Acid | 30 mg/dL |
| γ-Globulin | 5 g/dL |

f. Assay cut-off:

Not applicable

2. Comparison studies:

a. Method comparison with predicate device:

Method comparison studies were conducted following the recommendations in the CLSI EP9-A2 guideline.

80 serum samples were assayed in parallel by singlicate runs on both the Carolina Liquid Chemistries HDL Cholesterol Reagent on the Carolina Liquid Chemistries CLC 720 Chemistry Analyzer and a predicate method. The results were analyzed by using Deming regression. The range tested was 11.2 mg/dL to 150.2 mg/dL. Altered samples were included in the study (16%).

The comparison by Deming regression resulted in a slope of 0.958 (95%CI = 0.939 to 0.977), an intercept of 1.06 (95%CI = -0.32 to 2.45), correlation coefficient = 0.9961, and a standard error of 2.43.

b. Matrix comparison:

Not applicable

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 expected values for serum HDL cholesterol are as follows:

Low: <40 mg/dL

High: ≥60 mg/dL

* National Cholesterol Education Program. Third report of the expert panel on detection, evaluation, and treatment of high blood cholesterol in adults. NIH Pub No. 02-5215. Bethesda, MD: National Heart, Lung, and Blood Institute (2002).

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.