DIAZYME HDL-CHOLESTEROL REAGENT,CALIBRATOR, CONTROLS
Device Facts
| Record ID | K072395 |
|---|---|
| Device Name | DIAZYME HDL-CHOLESTEROL REAGENT,CALIBRATOR, CONTROLS |
| Applicant | Diazyme Laboratories |
| Product Code | LBS · Clinical Chemistry |
| Decision Date | Jan 3, 2008 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.1475 |
| Device Class | Class 1 |
Intended Use
The Diazyme HDL-Cholesterol reagent is intended for the in vitro quantitative determination of High Density Lipoprotein Cholesterol in human serum or plasma. The reagents can assist in the diagnosis and treatment of patients at risk of developing coronary heart disease. Low HDL cholesterol is related to the high risk of coronary heart disease.
Device Story
Diazyme HDL-Cholesterol Reagent is an in vitro diagnostic assay for quantitative HDL cholesterol measurement in human serum or plasma. The assay utilizes a modified polyvinyl sulfonic acid and polyethylene-glycol-methyl ether precipitation method to mask LDL, VLDL, and chylomicrons, preventing their reaction with cholesterol oxidase and cholesterol esterase. HDL cholesterol reacts via a colorimetric Trinder reaction. The device is intended for use in clinical laboratory settings on automated chemistry analyzers (e.g., Roche Hitachi 917). Healthcare providers use the resulting quantitative HDL values to assess patient risk for coronary heart disease, with values <40 mg/dL indicating risk and ≥60 mg/dL considered protective. The kit includes two reagents, calibrators, and separately sold lyophilized human serum-based controls.
Clinical Evidence
Bench testing only. Precision evaluated per CLSI EP5-A (Total CV 2.2-3.7%). Linearity established from 1.06-184.8 mg/dL (r²=0.9999). Method comparison with predicate (n=84) yielded Y=1.0484x-4.6906, r²=0.9871. Interference testing showed <10% deviation for common serum substances (ascorbic acid, bilirubin, hemoglobin, triglycerides). Matrix comparison confirmed suitability for serum, EDTA, citrate, and heparin plasma.
Technological Characteristics
Quantitative colorimetric Trinder reaction. Reagents: modified polyvinyl sulfonic acid and polyethylene-glycol-methyl ether precipitation. Instrumentation: Roche Hitachi 917. Calibrators/Controls: Lyophilized human serum-based. Stability: 12 months at 2-8°C. Standards: CLSI EP05-A2, EP06-A, EP09-A2, EP21-A.
Indications for Use
Indicated for in vitro quantitative determination of HDL cholesterol in human serum or plasma to assist in diagnosis and treatment of patients at risk of coronary heart disease.
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
- Ultra N-Geneous HDL Cholesterol Reagent and calibrator (k021316)
- Bio-Lipid Lipid Controls (k951749)
Related Devices
- K972041 — EZ HDL CHOLESTEROL REAGENT(PROCEDURE 354)/EZ HDL CALIBRATOR (354-5) · Sigma Diagnostics, Inc. · Jul 22, 1997
- K963847 — WAKO DIRECT HDL-C · Wako Chemicals USA, Inc. · Dec 24, 1996
- K971526 — ACE HDL-C REAGENT · Schiapparelli Biosystems, Inc. · May 21, 1997
- K072523 — DIAZYME LDL-CHOLESTEROL REAGENT, CALIBRATOR AND CONTROLS · General Atomics · Jan 22, 2008
Submission Summary (Full Text)
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE
A. 510(k) Number:
k072395
B. Purpose for Submission:
New device
C. Measurand:
High Density Lipoprotein Cholesterol
D. Type of Test:
Quantitative, colorimetric Trinder reaction
E. Applicant:
Diazyme Laboratories
F. Proprietary and Established Names:
Proprietary Name – Diazyme HDL-Cholesterol Reagent
Established Name – HDL Cholesterol
G. Regulatory Information:
1. Regulation section:
21 CFR 862.1475: Lipoprotein Test System
21 CFR 862.1660: Quality control material (assayed and unassayed)
2. Classification:
Class I (meets limitations of exemptions, 21 CFR 862.9 (c) (4)) - Diazyme HDL-Cholesterol Reagent
Class I – Diazyme HDL Controls
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3. Product code:
LBS - Diazyme HDL-Cholesterol
JJX - Diazyme HDL-Cholesterol Control
4. Panel:
75 – Clinical Chemistry
H. Intended Use:
1. Intended use(s):
See Indications for use below.
2. Indication(s) for use:
The Diazyme HDL-Cholesterol reagent is intended for the in vitro quantitative determination of High Density Lipoprotein Cholesterol in human serum or plasma. The reagents can assist in the diagnosis and treatment of patients at risk of developing coronary heart disease.
Calibrator – For calibration of the Diazyme HDL-Cholesterol Reagent Assay in serum or plasma. For In Vitro Diagnostic Use
Control – To monitor the performance of the Diazyme HDL-Cholesterol Reagent. For In Vitro Diagnostic Use
3. Special conditions for use statement(s):
For prescription use only.
4. Special instrument requirements:
All testing was performed on the Roche Diagnostics Hitachi 917.
I. Device Description:
The Diazyme HDL-Cholesterol kit contains two reagents: Reagent 1 4X70ml bottles and Reagent 2 2X50ml bottles and 2X1ml calibrator materials. The control materials consist of three lyophilized human serum based materials which are sold separately. All donor pools for the serum pool used to make the control and calibrator materials have been tested by FDA licensed methods and found non reactive for hepatitis B surface antigen, hepatitis C and HIV I and II. The assay is based upon a modified polyvinyl sulfonic acid and polyethylene-glycol-methyl ether coupled precipitation to
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LDL, VLDL and chylomicrons making them inaccessible to Cholesterol oxidase and cholesterol esterase. These enzymes react with HDL via a Trinder reaction.
This device has not been certified or tested by the Cholesterol Reference Method Laboratory Network.
## J. Substantial Equivalence Information:
1. Predicate device name(s):
Ultra N-Geneous HDL Cholesterol Reagent and calibrator. Bio-Lipid Lipid Controls
2. Predicate K number(s):
k021316, k951749
3. Comparison with predicate:
| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Indication for Use | The Diazyme HDL-Cholesterol reagent is intended for the in vitro quantitative determination of High Density Lipoprotein Cholesterol in human serum or plasma. The reagents can assist in the diagnosis and treatment of patients at risk of developing coronary heart disease. | For the quantitative determination of high density lipoprotein cholesterol (HDL-C) in human serum or plasma |
| Type of Test | Quantitative | Quantitative |
| Specimen Type | Serum or Plasma | Serum or Plasma |
| Calibrator | Lyophilized | Lyophilized |
| Control | 3 levels, lyophilized | 3 levels, lyophilized |
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K. Standard/Guidance Document Referenced (if applicable):
The following standards have been used to support this submission:
EP05-A2 – Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline-Second Edition
EP06-A - Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline
EP09-A2 – Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline -Second Edition
EP21-A - Estimation of Total Analytical Error for Clinical Laboratory Methods; Approved Guideline
L. Test Principle:
Colorimetric Trinder reaction (Cholesterol Oxidase and Cholesterol Esterase)
M. Performance Characteristics (if/when applicable):
1. Analytical performance:
a. Precision/Reproducibility:
Precision of the Diazyme HDL- Cholesterol Reagent was evaluated using the CLSI EP5-A guideline. In the study three levels of serum samples were tested with 2 runs per day in duplicate over 20 days. Testing was performed on the Roche Diagnostics Hitachi 917. The results are presented in the table below.
Within Run Precision
| | Level 1 | Level 2 | Level 3 |
| --- | --- | --- | --- |
| N | 80 | 80 | 80 |
| Mean | 29 | 53.07 | 90.56 |
| SD | 0.3 | 0.41 | 0.84 |
| CV% | 1.0 | 0.8 | 0.9 |
Total Precision
| | Level 1 | Level 2 | Level 3 |
| --- | --- | --- | --- |
| N | 80 | 80 | 80 |
| Mean | 29 | 53.07 | 90.56 |
| SD | 0.65 | 1.36 | 2.02 |
| CV% | 2.3 | 2.6 | 2.2 |
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Additional precision studies were performed at or near the clinical significant ranges of HDL. These precision studies of the Diazyme HDL- Cholesterol Reagent were evaluated using the CLSI EP5-A guideline. In the study three levels of serum samples were tested with 2 runs per day in duplicate over 5 days. Testing was performed on the Roche Diagnostics Hitachi 917.
Within Run Precision
| | Level 1 | Level 2 | Level 3 |
| --- | --- | --- | --- |
| N | 20 | 20 | 20 |
| Mean | 21.63 | 44.28 | 159.59 |
| SD | 0.18 | 0.30 | 1.77 |
| CV% | 0.9 | 0.7 | 1.10 |
Total Precision
| | Level 1 | Level 2 | Level 3 |
| --- | --- | --- | --- |
| N | 20 | 20 | 20 |
| Mean | 21.63 | 44.28 | 159.59 |
| SD | 0.61 | 0.79 | 5.90 |
| CV% | 2.8 | 1.8 | 3.7 |
# b. Linearity/assay reportable range:
Linearity determination was based upon NCCLS (CLSI) EP6-A and was performed on the Roche Diagnostics Hitachi 917. Each level was measured in triplicate. Samples were serially diluted from a level of 184.18 to $0\mathrm{mg / dL}$ . The linearity for this method is $1.06 - 184.8\mathrm{mg / dL}$ , $\mathrm{Y} = 0.998\mathrm{x} + 0.7275$ , $\mathrm{r}^2 = 0.9999$ . The sponsor determined an acceptable $\%$ error was $\leq 10\%$ . Results of the study support the claimed linearity range.
# c. Traceability, Stability, Expected values (controls, calibrators, or methods):
This device has not been certified or tested by the Cholesterol Reference Method Laboratory Network. The NIST SRM 1915b material was tested with the Diazyme HDL-Cholesterol reagent and found to have a deviation from the NIST target value of $1.9\%$ for level I $(48.7\mathrm{mg / dL})$ and $-3.4\%$ for level II $(51.9\mathrm{mg / dL})$ .
Values are assigned to the control materials by repeat testing on the Hitachi 917 using the Diazyme HDL-cholesterol reagent and calibrator. The assigned ranges which are lot specific are as follows: Level I $23 - 35\mathrm{mg / dL}$ ; Level II $44 - 66\mathrm{mg / dL}$ and Level III $74 - 110\mathrm{mg / dL}$ .
Reagent Stability: Reagents from two lots were stored in incubators at $37^{\circ}\mathrm{C}$ , $25^{\circ}\mathrm{C}$ and $4^{\circ}\mathrm{C}$ . Reagents were tested at 0 day, 2 days, 6 days, 9 days and 12 days. Accelerated study results show that the reagent is stable for up to 12 months when stored at $2 - 8^{\circ}\mathrm{C}$ . Real time studies are being conducted and
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adjustments to shelf life will be made as needed. Stability on board the Hitachi 917 was studied by storing the two reagents (R1 and R2) in the 2-8°C storage chamber of the Hitachi 917. Results indicate the reagents are suitable for on board storage for up to 30 days.
Calibrator Stability: Calibrators from two lots were stored in incubators at 37°C, 25°C and 4°C. The calibrators were removed from storage at predetermined times and tested with the Diazyme HDL-Cholesterol reagent. The accuracy of the Calibrators was determined by testing HDL-C samples of known concentrations determined by the predicate kit. The shelf life of the calibrators was determined to be 12 months at 2-8°C. Real time studies are being conducted and adjustments to shelf life will be made as needed.
Control Stability: Controls from two lots were stored in incubators at 37°C, 25°C and 4°C. The controls were removed from storage at predetermined times and tested in duplicate with the Diazyme HDL-Cholesterol reagent. The accuracy of the controls was determined by testing HDL-C samples of known concentrations determined by the predicate kit. The shelf life of the control materials was determined to be 12 months at 2-8°C. Open vial stability for controls was determined to be one month. Real time studies are being conducted and adjustments to shelf life will be made as needed.
d. Detection limit:
To demonstrate the limit of blank of Diazyme HDL-Cholesterol, HDL zero calibrator was tested in 12 replicates on the Hitachi 917. The Limit of blank is defined as the mean + 3SD. N = 12, Mean = 0.37, SD = 0.27 LOD = 1.06. To determine the limit of quantitation (LOQ), a serum sample was diluted into 5 other concentrations and assayed over 3 days with 2 runs per day for a total of 150 replicates on the Hitachi 917. The five concentrations were 0.30, 0.75, 1.50, 3.00 and 5.00 mg/dL. In the study, the estimated LOQ where the inter-assay CV is 20% was 1.39 mg/dL.
e. Analytical specificity:
To determine the level of interference from substance normally present in human serum, the Diazyme HDL-Cholesterol Assay was tested with normal serum samples containing about 34 mg/dL HDL-C spiked with various concentrations of interference substances following CLSI guideline EP7-A and was performed on the Roche Diagnostics Hitachi 917. The following substance normally present in serum produced less than 10% deviation when tested at levels equal to the following concentrations:
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| Interference Substance | Concentration |
| --- | --- |
| Ascorbic Acid | 10mM |
| Bilirubin | 40 mg/dL |
| Bilirubin Conjugated | 30 mg/dL |
| Hemoglobin | 1000 mg/dL |
| Triglycerides | 1000 mg/dL |
f. Assay cut-off:
Not applicable
2. Comparison studies:
a. Method comparison with predicate device:
A total of 84 samples (68 non-altered, 16 altered) were compared with the Equal HDL Ultra Cholesterol and calibrator on the Hitachi 917.
Y = 1.0484x-4.6906, r² = 0.9871. Samples ranged from 7.17 to 181.84 mg/dL were tested based upon recommendations in NCCLS (CLSI) EP-9A2 guidance.
b. Matrix comparison:
Samples were compared between serum and EDTA, citrate and heparin plasma on the Hitachi 917. Samples were compared across the measuring range, from 14.03 to 184.4 mg/dL some samples were spiked or diluted to cover the entire range.
Serum vs EDTA plasma Y = 1.0264X + 1.1334, R² = 0.9959
Serum vs Citrate plasma Y = 1.0661X + 2.0933, R² = 0.996
Serum vs Heparin plasma Y = 1.05X + 0.3388, R² = 0.994
3. Clinical studies:
a. Clinical Sensitivity:
Not applicable
b. Clinical specificity:
Not applicable
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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:
Less than 40 mg/dL – A major risk factor for heart disease
60 mg/dL and above – Considered protective against heart disease
* Third report of the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and treatment of High Blood Cholesterol in Adults (NCEP ATPIII)
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.
