ACE CHOLESTEROL REAGENT, ACE HDL-C REAGENT, ACE LDL-C REAGENT, ACE TRIGLYCERIDES REAGENT
Device Facts
| Record ID | K112538 |
|---|---|
| Device Name | ACE CHOLESTEROL REAGENT, ACE HDL-C REAGENT, ACE LDL-C REAGENT, ACE TRIGLYCERIDES REAGENT |
| Applicant | Alfa Wassermann Diagnostic Technologies, Inc. |
| Product Code | CHH · Clinical Chemistry |
| Decision Date | Mar 29, 2012 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.1175 |
| Device Class | Class 1 |
Indications for Use
ACE Cholesterol Reagent is intended for the quantitative determination of cholesterol in serum and lithium heparin plasma using the ACE and ACE Alera Clinical Chemistry Systems. Cholesterol measurements are used in the diagnosis and treatment of disorders involving excess cholesterol in the blood and lipid and lipoprotein metabolism disorders. This test is intended for use in clinical laboratories or physician office laboratories. For in vitro diagnostic use only. ACE HDL-C Reagent is intended for the homogeneous, quantitative determination of HDL cholesterol (HDL-C) in serum and lithium heparin plasma using the ACE and ACE Alera Clinical Chemistry Systems. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases. This test is intended for use in clinical laboratories or physician office laboratories. For in vitro diagnostic use only. ACE LDL-C Reagent is intended for the quantitative determination of low density lipoprotein cholesterol (LDL-C) in serum and lithium heparin plasma using the ACE and ACE Alera Clinical Chemistry Systems. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases. This test is intended for use in clinical laboratories or physician office laboratories. For in vitro diagnostic use only. ACE Triglycerides Reagent is intended for the quantitative determination of triglycerides in serum and lithium heparin plasma using the ACE and ACE Alera Clinical Chemistry Systems. Triglyceride measurements are used in the diagnosis and treatment of patients with diabetes mellitus, nephrosis, liver obstruction, other diseases involving lipid metabolism or various endocrine disorders. This test is intended for use in clinical laboratories or physician office laboratories. For in vitro diagnostic use only.
Device Story
Reagents for quantitative photometric analysis of cholesterol, HDL-C, LDL-C, and triglycerides in serum and lithium heparin plasma; used on ACE and ACE Alera Clinical Chemistry Systems. Enzymatic reactions (cholesterol esterase, oxidase, peroxidase, lipase, glycerol kinase) produce color via Trinder reaction or specific detergent-based solubilization for lipoproteins. Detergents selectively solubilize HDL or LDL particles to enable specific enzymatic measurement. Systems operate in clinical chemistry labs; results used by physicians for diagnosis/treatment of lipid/metabolic disorders. Output is concentration value (mg/dL) derived from absorbance change at specific wavelengths (505nm, 544nm, 592nm).
Clinical Evidence
Bench testing only. Precision/reproducibility studies performed on ACE and ACE Alera systems using serum and lithium heparin plasma (n=20 per sample). Matrix comparison studies (n=99-103 per analyte) showed high correlation (r=0.9874-0.9977) and slope near 1.0 between serum and lithium heparin plasma. Method comparison studies at three physician office laboratory (POL) sites confirmed performance consistency.
Technological Characteristics
Photometric enzymatic assays. Reagents include cholesterol esterase, cholesterol oxidase, peroxidase, lipase, glycerol kinase, and specific detergents for lipoprotein fractionation. Analysis temperature 37°C. Bichromatic detection at 505/647nm, 592/692nm, 544/692nm, or 505/692nm. Systems are automated clinical chemistry analyzers. Calibration via reference serum/calibrators. 30-day on-board stability.
Indications for Use
Indicated for quantitative determination of cholesterol, HDL-C, LDL-C, and triglycerides in serum and lithium heparin plasma. Used for diagnosis/treatment of lipid/lipoprotein metabolism disorders, diabetes mellitus, atherosclerosis, liver/renal diseases, nephrosis, and endocrine disorders. Intended for clinical or physician office laboratories.
Regulatory Classification
Identification
A cholesterol (total) test system is a device intended to measure cholesterol in plasma and serum. Cholesterol measurements are used in the diagnosis and treatment of disorders involving excess cholesterol in the blood and lipid and lipoprotein metabolism disorders.
Predicate Devices
- Alfa Wassermann ACE plus ISE/Clinical Chemistry System (K931786)
- ACE Reagents (K971526)
- ACE Reagents (K991733)
Related Devices
- K122757 — ACE CHOLESTEROL REAGENT, ACE HDL-C REAGENT, ACE LDL-C REAGENT, ACE TRIGLYCERIDES REAGENT · Alfa Wassermann Diagnostic Technologies, Inc. · Oct 5, 2012
Submission Summary (Full Text)
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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
ASSAY ONLY TEMPLATE
A. 510(k) Number:
k112538
B. Purpose for Submission:
Additional or Expanded Indications: adding lithium heparin plasma as a new claim
C. Measurand:
Cholesterol, High Density Lipoprotein, Low Density Lipoprotein, and Triglycerides
D. Type of Test:
Quantitative, colorimetric
E. Applicant:
Alfa Wassermann Diagnostic Technologies, LLC.
F. Proprietary and Established Names:
ACE Cholesterol Reagent
ACE HDL-C Reagent
ACE LDL-C Reagent
ACE Triglycerides Reagent
G. Regulatory Information:
| Name | Regulation | Product Code | Classification |
| --- | --- | --- | --- |
| ACE Cholesterol Reagent | 21 C.F.R. § 862.1175 | CHH | I, meets limitations per 21 CFR 862.9(c)(4) |
| ACE HDL-C Reagent | 21 C.F.R. § 862.1475 | LBS | I, meets limitations per 21 CFR 862.9(c)(4) |
| ACE LDL-C Reagent | 21 C.F.R. § 862.1475 | MRR | I, meets limitations per 21 CFR 862.9(c)(4) |
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| ACE Triglycerides Reagent | 21 C.F.R. § 862.1705 | CDT | I, meets limitations per 21 CFR 862.9(c)(4) |
| --- | --- | --- | --- |
Panel: Clinical Chemistry (75)
## H. Intended Use:
1. Intended use(s):
See indications for use below.
2. Indications(s) for use:
ACE Cholesterol Reagent is intended for the quantitative determination of cholesterol in serum and lithium heparin plasma using the ACE and ACE Alera Clinical Chemistry Systems. Cholesterol measurements are used in the diagnosis and treatment of disorders involving excess cholesterol in the blood and lipid and lipoprotein metabolism disorders. For in vitro diagnostic use only.
ACE HDL-C Reagent is intended for the homogeneous, quantitative determination of HDL cholesterol (HDL-C) in serum and lithium heparin plasma using the ACE and ACE Alera Clinical Chemistry Systems. 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.
ACE LDL-C Reagent is intended for the quantitative determination of low density lipoprotein cholesterol (LDL-C) in serum and lithium heparin plasma using the ACE and ACE Alera Clinical Chemistry Systems. 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.
ACE Triglycerides Reagent is intended for the quantitative determination of triglycerides in serum and lithium heparin plasma using the ACE and ACE Alera Clinical Chemistry Systems. Triglyceride measurements are used in the diagnosis and treatment of patients with diabetes mellitus, nephrosis, liver obstruction, other diseases involving lipid metabolism or various endocrine disorders. For in vitro diagnostic use only.
3. Special conditions for use statement(s):
For prescription use.
4. Special instrument requirements:
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# ACE and ACE Alera Clinical Chemistry Systems
## I. Device Description:
### ACE Cholesterol Reagent
The ACE Cholesterol Reagent, used with the ACE and ACE Alera Clinical Chemistry System, is intended for quantitative *in vitro* diagnostic determination of cholesterol concentration in serum and lithium heparin plasma. It is based on a photometric test using a coupled enzymatic reaction (cholesterol esterase, cholesterol oxidase and peroxidase), which produces color via the Trinder reaction. It is composed of one reagent bottle, which is placed in the reagent compartment of the ACE or ACE Alera. It is intended for *in vitro* diagnostic use.
### ACE HDL-C Reagent
The ACE HDL-C Reagent, used with the ACE and ACE Alera Clinical Chemistry System, is intended for quantitative *in vitro* diagnostic determination of high density lipoprotein cholesterol concentration in serum and lithium heparin plasma. It is based on a photometric test measuring the amount of HDL present utilizing two reagents, the second of which contains a unique detergent. This detergent solubilizes only the HDL lipoprotein particles, thus releasing HDL cholesterol to react with the cholesterol esterase and cholesterol oxidase, in the presence of a chromogen to produce color in the reaction cuvette. The detergent also inhibits the reaction of the cholesterol enzymes with LDL and VLDL lipoproteins and chylomicrons by adsorbing to their surfaces. It is composed of two reagent bottles, which are placed in the reagent compartment of the ACE or ACE Alera. It is intended for *in vitro* diagnostic use.
### ACE LDL-C Reagent
The ACE LDL-C Reagent, used with the ACE and ACE Alera Clinical Chemistry System, is intended for quantitative *in vitro* diagnostic determination of low density lipoprotein cholesterol concentration in serum and lithium heparin plasma. It is based on a photometric test measuring the amount of LDL present after other non-LDL cholesterol is solubilized by a detergent and then consumed by cholesterol esterase and cholesterol oxidase in a noncolor forming reaction. LDL is then solubilized by a second detergent in the presence of a chromogenic peroxidase substrate to form a purple-red color. It is composed of two reagent bottles, which are placed in the reagent compartment of the ACE or ACE Alera. It is intended for *in vitro* diagnostic use.
### ACE Triglycerides Reagent
The ACE Triglycerides Reagent, used with the ACE and ACE Alera Clinical Chemistry System, is intended for quantitative *in vitro* diagnostic determination of triglycerides concentration in serum and lithium heparin plasma. It is based on a photometric test using a coupled enzymatic reaction (lipase, glycerol kinase, glycerol phosphate oxidase and peroxidase), which produces color via the Trinder reaction. It is composed of one reagent bottle, which is placed in the reagent compartment of the ACE or ACE Alera. It is intended for *in vitro* diagnostic use.
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# J. Substantial Equivalence Information:
1. Predicate device name(s):
ACE Cholesterol Reagent
ACE HDL-C Reagent
ACE LDL-C Reagent
ACE Triglycerides Reagent
2. Predicate $510(\mathbf{k})$ number(s):
ACE Cholesterol Reagent: k931786
ACE HDL-C Reagent: k931876
ACE LDL-C Reagent: k991733
ACE Triglycerides Reagent: k931786
3. Comparison with predicate:
Device Comparison Table: ACE Cholesterol Reagent
| | New Device | Predicate Device |
| --- | --- | --- |
| 510(k) # | K112538 | K931786 (ACE) |
| Name | ACE Cholesterol Reagent | ACE plus ISE/Clinical Chemistry System, ACE Cholesterol Reagent |
| Intended Use/Indications for Use | ACE Cholesterol Reagent is intended for the quantitative determination of cholesterol using the ACE and ACE Alera Clinical Chemistry Systems. For in vitro diagnostic use only. | Same |
| Sample Type | Serum or lithium heparin plasma | Serum |
| Instrument Platforms | ACE and ACE Alera Clinical Chemistry Systems | ACE, ACE Alera® and NExCT™ Clinical Chemistry Systems |
| Calibration | Calibrated by referencing the change in absorbance of the unknown samples to the change in absorbance of the calibrator; the use of GEMCAL Reference Serum is recommended. | Same |
| Calibration Stability | 30 Days | Same |
| On-Board Stability | 30 Days | Same |
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| | New Device | Predicate Device |
| --- | --- | --- |
| 510(k) # | K112538 | K931786 (ACE) |
| Name | ACE Cholesterol Reagent | ACE plus ISE/Clinical Chemistry System, ACE Cholesterol Reagent |
| Method Traceability | Abell, L.L., Levy, B.B., Brodie, B.B. and Kendall, F.E., J. Biol. Chem. 195, 357 (1952); Allain, C.C., Poon, L.S., Chan, C.S.G, Richmond, W. and Fu, P.C., Clin. Chem. 20, 470 (1974). | Same |
| Reactive Ingredients | 4-Aminoantipyrene p-Hydroxybenzoic acid Cholesterol oxidase (Nocardia) Cholesterol esterase (porcine pancreas and Pseudomonas) Peroxidase (Horseradish) | Same |
| Analysis Temperature | 37°C | Same |
| Sample Volume | 3 μL | Same |
| Precision (mg/dL) | **Within run:** Sample A: Mean 139, SD 3.0, CV 2.2% Sample B: Mean 212, SD 2.4, CV 1.1% Sample C: Mean 290, SD 5.7, CV 2.0%
**Total:** Sample A: Mean 139, SD 3.2, CV 2.3% Sample B: Mean 212, SD 3.3, CV 1.6% Sample C: Mean 290, SD 6.0, CV 2.1% | Same |
| Matrix Comparison | x = serum, y = lithium heparin plasma
ACE: Regression Equation: y = 0.985x - 1.7
Correlation Coefficient: 0.9947
Sample Range: 40-568
ACE Alera: Regression Equation: y = 0.994x - 2.5
Correlation Coefficient: 0.9934
Sample Range: 42-577 | Serum only |
| Detection Wavelength | 505/647 nm | Same |
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Device Comparison Table: ACE HDL-C Reagent
| | New Device | Predicate Device |
| --- | --- | --- |
| 510(k) # | K112538 | K931786 (ACE) |
| Name | ACE HDL-C Reagent | ACE plus ISE/Clinical Chemistry System, ACE HDL-C Reagent |
| Intended Use/Indication for Use | ACE HDL-C Reagent is intended for the homogeneous, quantitative determination of HDL cholesterol (HDL-C) using the ACE and ACE Alera Clinical Chemistry Systems. For in vitro diagnostic use only. | Same |
| Sample Type | Serum or lithium heparin plasma | Serum |
| Instrument Platforms | ACE and ACE Alera Clinical Chemistry Systems | ACE and ACE Alera® Clinical Chemistry System |
| Calibration | Calibrated by referencing the change in absorbance of the unknown samples to the change in absorbance of the calibrator; the use of GEMCAL Reference Serum is recommended. | Same |
| Calibration Stability | 30 Days | Same |
| On-Board Stability | 30 Days | Same |
| Method Traceability | Gotto, A.M., Lipoprotein metabolism and the etiology of hyperlipidemia, Hospital Practice, 23:Suppl. 1, 4 (1988). | Same |
| Reactive Ingredients | Buffer Cholesterol oxidase (E. coli) Peroxidase (Horseradish) N, N-bis(4-sulphobutyl)-m-toluidine-disodium salt Accelerator Ascorbic oxidase (Curcurbita sp.) | Same |
| Analysis Temperature | 37°C | Same |
| Sample Volume | 3 μL | Same |
| Precision (mg/dL) | Within run: Sample A: Mean 32, SD 0.7, CV 2.0% Sample B: Mean 61, SD 1.2, CV 2.0% | Same |
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| | New Device | Predicate Device |
| --- | --- | --- |
| 510(k) # | K112538 | K931786 (ACE) |
| Name | ACE HDL-C Reagent | ACE plus ISE/Clinical Chemistry System, ACE HDL-C Reagent |
| | Sample C: Mean 85, SD 1.5, CV 1.8%
**Total:**
Sample A: Mean 32, SD 1.3, CV 4.2%
Sample B: Mean 61, SD 2.6, CV 4.3%
Sample C: Mean 85, SD 3.3, CV 3.9% | |
| Comparative Analysis Regression Evaluation | Regression Equation: y = 0.954x + 1.9
Correlation Coefficient: 0.988
Sample Range: 15-120 mg/dL | Same |
| Expected Values | 40 to 60 mg/dL | Same |
| Matrix Comparison | x = serum, y = lithium heparin plasma
ACE:
Regression Equation: y = 1.015x - 0.6
Correlation Coefficient: 0.9884
Sample Range: 6-120
ACE Alera:
Regression Equation: y = 0.989x + 0.4
Correlation Coefficient: 0.9874
Sample Range: 7-123 | Serum only |
| Detection Wavelength | 592/692 nm | Same |
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Device Comparison Table: ACE LDL-C Reagent
| | New Device | Predicate Device |
| --- | --- | --- |
| 510(k) # | K112538 | K931786 (ACE) |
| Company | Alfa Wassermann Diagnostic Technologies, LLC | Alfa Wassermann Diagnostic Technologies, LLC |
| Name | ACE LDL-C Reagent | ACE plus ISE/Clinical Chemistry System, ACE LDL-C Reagent |
| Intended Use/Indications for Use | ACE LDL-C Reagent is intended for the quantitative determination of low density lipoprotein cholesterol (LDL-C) using the ACE and ACE Alera Clinical Chemistry Systems. For in vitro diagnostic use only. | Same |
| Sample Type | Serum or lithium heparin plasma | Serum |
| Instrument Platforms | ACE and ACE Alera Clinical Chemistry Systems | ACE and the NExCT Clinical Chemistry System |
| Calibration | Calibrated by referencing the change in absorbance of the unknown samples to the change in absorbance of the calibrator; the use of LDL-C Calibrator is recommended. | Same |
| Calibration Stability | 30 days | Same |
| On-Board Stability | 30 days | Same |
| Method Traceability | Gotto, A.M., Lipoprotein metabolism and the etiology of hyperlipidemia, Hospital Practice, 23: Suppl. 1, 4 (1998); Crouse, J.R. et al. Studies of low density lipoprotein molecular weight in human beings with coronary artery disease J. Lipids Res., 26, 566 (1985). | Same |
| Reactive Ingredients | Cholesterol esterase
Cholesterol oxidase
Peroxidase
4-Aminoantipyrine
Ascorbic acid oxidase
Buffer
N,N-bis (4-sulfobutyl)-m-toluidine, disodium salt | Same |
| Analysis Temperature | 37°C | Same |
| Sample Volume | 3 μL | Same |
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| | New Device | Predicate Device |
| --- | --- | --- |
| 510(k) # | K112538 | K931786 (ACE) |
| Company | Alfa Wassermann Diagnostic Technologies, LLC | Alfa Wassermann Diagnostic Technologies, LLC |
| Name | ACE LDL-C Reagent | ACE plus ISE/Clinical Chemistry System, ACE LDL-C Reagent |
| Reaction Volume (total) | 418 μL | Same |
| Precision (mg/dL) | **Within-Run:**
Sample A: Mean 95, SD 2.1, CV 2.2%
Sample B: Mean 146, SD 3.8, CV 2.6%
Sample C: Mean 196, SD 2.8, CV 1.4%
**Total:**
Sample A: Mean 95, SD 2.5, CV 2.6%
Sample B: Mean 146, SD 4.7, CV 3.2%
Sample C: Mean 196, SD 6.3, CV 3.2% | Same |
| Comparative Analysis Regression Evaluation | Regression Equation: y = 1.111x - 15.5
Correlation Coefficient: 0.9747
Sample Range: 41-226 mg/dL | Same |
| Expected Values | <130 mg/dL | Same |
| Matrix Comparison | x = serum, y = lithium heparin plasma
ACE:
Regression Equation: y = 1.008x - 2.6
Correlation Coefficient: 0.9954
Sample Range: 9-460
ACE Alera:
Regression Equation: y = 0.995x - 1.3
Correlation Coefficient: 0.9954
Sample Range: 9-464 | Serum only |
| Detection Wavelength | 544/692 nm | Same |
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Device Comparison Table: ACE Triglycerides Reagent
| | New Device | Predicate Device |
| --- | --- | --- |
| 510(k) # | K112538 | K931786 (ACE) |
| Name | ACE Triglycerides Reagent | ACE plus ISE/Clinical Chemistry System, ACE Triglycerides Reagent |
| Intended Use/Indications for Use | ACE Triglycerides Reagent is intended for the quantitative determination of triglycerides using the ACE and ACE Alera Clinical Chemistry Systems. For in vitro diagnostic use only. | Same |
| Sample Type | Serum or lithium heparin plasma | Serum |
| Instrument Platforms | ACE and ACE Alera Clinical Chemistry Systems | ACE, ACE Alera® and the NExCT™ Clinical Chemistry Systems |
| Calibration | Calibrated by referencing the change in absorbance of the unknown samples to the change in absorbance of the calibrator; the use of GEMCAL Reference Serum is recommended. | Same |
| Calibration Stability: | 30 Days | Same |
| On-Board Stability | 30 Days | Same |
| Method Traceability | Frederickson, D.S., et al., New Engl. J. Med. 276, 32 (1976); Tietz, N.W. (Ed.), Textbook of Clinical Chemistry, W.B. Saunders Co., Philadelphia, PA (1986). | Same |
| Reactive Ingredients | 4-Aminoantipyrine adenosine 5'-triphosphate p-Chlorophenol Glycerol phosphate oxidase (Microorganism) Lipase (Pseudomonas) Peroxidase (Horseradish) Glycerol kinase (Cellulomonas) Buffer | Same |
| Analysis Temperature | 37°C | Same |
| Sample Volume | 3 μL | Same |
| Reaction Volume (total) | 243 μL | Same |
| Precision | Within run: | Same |
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| | New Device | Predicate Device |
| --- | --- | --- |
| 510(k) # | K112538 | K931786 (ACE) |
| Name | ACE Triglycerides Reagent | ACE plus ISE/Clinical Chemistry System, ACE Triglycerides Reagent |
| (mg/dL) | Sample A: Mean 73 SD 1.3, CV 1.7%
Sample B: Mean 142, SD 1.6, CV 1.2%
Sample C: Mean 203, SD 2.4, CV 1.2%
Total:
Sample A: Mean 73, SD 1.4, CV 1.9%
Sample B: Mean 142, SD 2.6, CV 1.8%
Sample C: Mean 203, SD 4.0, CV 2.0% | |
| Comparative Analysis Regression Evaluation | Regression Equation: y = 0.993x + 1.2
Correlation Coefficient: 0.997
Sample Range: 27-531 mg/dL | Same |
| Expected Values | <150 mg/dL | Same |
| Matrix Comparison | x = serum, y = lithium heparin plasma
ACE:
Regression Equation: y = 1.005x - 7.9
Correlation Coefficient: 0.9977
Sample Range: 39-887
ACE Alera:
Regression Equation: y = 1.007x - 7.4
Correlation Coefficient: 0.9973
Sample Range: 38-884 | Serum only |
| Detection Wavelength | 505/692 nm | Same |
K. Standard/ Guidance Document Referenced (if applicable):
EP9-A2 Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline- Second Edition (2002).
L. Test Principle:
ACE Cholesterol Reagent
The ACE Cholesterol Reagent is composed of a single reagent bottle.
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The reagent contains:
| Component | Quantity |
| --- | --- |
| ACE Cholesterol Reagent | |
| 4-Aminoantipyrine (AAP) | 0.5 mmol/L |
| p-Hydroxybenzoic acid | 25 mmol/L |
| Cholesterol oxidase (Nocardia) | >150 U/L |
| Cholesterol esterase (Porcine pancreas and Pseudomonas) | >240 U/L |
| Peroxidase (Horseradish) | >1600 U/L |
| Stabilizers, Preservatives, and Fillers | |
Cholesterol esters in serum are completely hydrolyzed by cholesterol esterase to free cholesterol and free fatty acids. The cholesterol liberated by the esterase, plus any endogenous free cholesterol, are both oxidized by cholesterol oxidase to yield hydrogen peroxide $(\mathrm{H}_2\mathrm{O}_2)$ . The hydrogen peroxide then acts to oxidatively couple p-hydroxybenzoic acid and 4-aminoantipyrine (AAP) in a reaction catalyzed by peroxidase, producing a red colored quinoneimine complex which absorbs strongly at $505~\mathrm{nm}$ .
Cholesterol esters $+\mathrm{H}_2\mathrm{O}$ cholesteryl + fatty acids
Cholesterol $+\mathrm{O}_2$ cholesteryl +O2 cholest-4-ene-3-one $+\mathrm{H}_2\mathrm{O}_2$
$\mathrm{H}_2\mathrm{O}_2 + \mathrm{p}$ -hydroxybenzoic acid $+\mathrm{AAP}$ quinoneimine $+\mathrm{H}_2\mathrm{O}$ (red color)
The amount of chromogen formed, determined by measuring the increase in absorbance, bichromatically at $505~\mathrm{nm} / 647~\mathrm{nm}$ , is directly proportional to the cholesterol concentration in the sample.
# ACE HDL-C Reagent
The ACE HDL-C Reagent is composed of two reagent bottles (Buffer and Color Reagent). The Buffer (R1) contains:
| Component | Quantity |
| --- | --- |
| ACE HDL-C Buffer (R1) | |
| Good's Buffer | |
| Cholesterol oxidase (E. coli) | <1000 U/L |
| Peroxidase (Horseradish) | <1300 U/L |
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| N,N-bis(4-sulphobutyl)-m-toluidine-disodium salt (DSBmT) | <1 mM |
| --- | --- |
| Accelerator | <1 mM |
| Preservative | <0.06% |
| Ascorbic Oxidase (Curcubita sp.) | <3000 U/L |
The Color Reagent (R2) contains:
| Component | Quantity |
| --- | --- |
| ACE HDL-C Color Reagent (R2) | |
| Cholesterol esterase (Pseudomonas sp.) | <1500 U/L |
| 4-Aminoantipyrine (4-AAP) | <1 mM |
| Detergent | <2% |
| Preservative | |
The HDL-C Assay utilizes two reagents, the second containing a unique detergent. This detergent solubilizes only the HDL lipoprotein particles, thus releasing HDL cholesterol to react with the cholesterol esterase and cholesterol oxidase, in the presence of a chromogen to produce color. The detergent also inhibits the reaction of the cholesterol enzymes with LDL, VLDL and chylomicron lipoproteins by adsorbing to their surfaces. The amount of chromogen formed, determined by measuring the increase in absorbance bichromatically at $592 / 692\mathrm{nm}$ , is directly proportional to the HDL cholesterol concentration in the sample.
HDL, LDL, VLDL, Non-Reactive LDL, VLDL, chylomicrons + DSBmT chylomicrons
HDL
HDL Cholesterol
$\mathrm{H}_2\mathrm{O}_2 + \mathrm{DSBmT} + \mathrm{AAP}$
$\xrightarrow{\text{Stable}}$
Disrupted HDL
$\Delta^4$ cholestenone $+\mathrm{H}_2\mathrm{O}_2$
Color Development
# ACE LDL-C Reagent
The ACE LDL-C Reagent is composed of two reagent bottles (Buffer and Color Reagent).
The Buffer (R1) contains:
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| Component | Quantity |
| --- | --- |
| ACE LDL-C Buffer (R1) | |
| MES Buffer (pH 6.3) | |
| Detergent 1 | <1.0% |
| Cholesterol esterase (Cellulomonas sp.) | <1500 U/L |
| Cholesterol oxidase (Pseudomonas sp.) | <1500 U/L |
| Peroxidase (Horseradish) | <1300 ppg U/L |
| 4-Aminoantipyrine (4-AAP) | <0.1% |
| Ascorbic acid oxidase (Curcubita sp.) | <3000 U/L |
| Preservative | <0.1% |
The Color Reagent (R2) contains:
| Component | Quantity |
| --- | --- |
| ACE LDL-C Color Reagent (R2) | |
| MES Buffer (pH 6.3) | |
| Detergent 2 | <1.0% |
| N,N-bis(4-sulphobutyl)-m-toluidine-disodium salt (DSBmT) | <1.0 mmol/L |
| Preservative | <0.1% |
Detergent 1 solubilizes non-LDL lipoprotein particles (HDL, VLDL and chylomicrons) and releases cholesterol. The cholesterol is consumed by cholesterol esterase and cholesterol oxidase in a non-color forming reaction. In a second reaction, detergent 2 solubilizes the remaining LDL particles and forms peroxide, via the enzymes cholesterol esterase and cholesterol oxidase. The peroxide, in the presence of peroxidase and two peroxidase substrates, 4-aminoantipyrine and DSBmT, results in a purple-red color.
The first reaction:
HDL, VLDL
Micellar Cholesterol
H2O2 +AAP
Colorless
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The second reaction:
LDL
Micellar cholesterol
Micellar Cholesterol
$\mathrm{H}_2\mathrm{O}_2$
$\mathrm{H}_2\mathrm{O}_2 + \mathrm{AAP} + \mathrm{DSMbT}$
Purple-red pigment
The amount of color formed, determined by measuring the increase in absorbance bichromatically at $544 / 692\mathrm{nm}$ , is directly proportional to the LDL cholesterol concentration in the sample.
# ACE Triglycerides Reagent
The ACE Triglycerides Reagent is composed of a single reagent bottle.
The reagent contains:
| Component | Quantity |
| --- | --- |
| ACE Triglycerides Reagent | |
| 4-Aminoantipyrine (AAP) | 0.4 mmol/L |
| Adenosine 5'-triphosphate (ATP) | 2.6 mmol/L |
| p-Chlorophenol | 3.0 mmol/L |
| Glycerol phosphate oxidase (GPO) (Microorganism) | >2400 U/L |
| Lipase (Pseudomonas) | >1000 U/L |
| Peroxidase (Horseradish) | >540 U/L |
| Glycerol kinase (Cellulomonas) | >400 U/L |
| Buffer, Stabilizers and Preservatives | |
Triglycerides in serum are hydrolyzed by lipase to form glycerol and free fatty acids. In the presence of adenosine triphosphate (ATP) and glycerol kinase (GK), the glycerol is converted to glycerol-1-phosphate (G-1-P) and the ATP to adenosine diphosphate (ADP). Glycerol-1-phosphate is oxidized by glycerol phosphate oxidase (GPO) to yield hydrogen peroxide $(\mathrm{H}_2\mathrm{O}_2)$ . The hydrogen peroxide then acts to oxidatively couple p-chlorophenol and 4-aminoantipyrine (AAP) in a reaction catalyzed by peroxidase, producing a red colored quinoneimine complex which absorbs strongly at $505~\mathrm{nm}$ .
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Triglycerides → Glycerol + fatty acids
Glycerol +ATP → G-GP → G-1-ADP
G-1-P+O₂ → G-P+O₂ → H₂O₂ + dihydroxyacetone phosphate
H₂O₂ + p-chlorophenol +AAP → Quinomeimine (red color +H₂O)
The amount of chromogen formed, determined by measuring the increase in absorbance bichromatically at 505 nm/692 nm, is directly proportional to the triglycerides concentration in the sample.
## M. Performance Characteristics (if/when applicable):
### 1. Analytical performance:
#### a. Precision/Reproducibility:
Precision studies were performed on one lot of the ACE Cholesterol, HDL, LDL, and Triglyceride Reagents using three serum and three lithium heparin plasma samples on one ACE and one ACE Alera Clinical Chemistry Systems. The low level samples were from a normal human patient, the high levels samples were spiked with analyte, and the mid-level samples were prepared by mixing the high and low level samples. The samples were run two times per run, with two runs being performed per day, with each run consisting of each sample being run on the ACE and the ACE Alera Clinical Chemistry Systems (total n for each sample is 20). This was done for three days (HDL) or five days (Cholesterol, LDL, Triglycerides). The results are presented below.
**Cholesterol on the ACE Analyzer**
| Sample | Mean (mg/dL) | Within Run | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- |
| | | SD | CV % | SD | CV % |
| Serum 1 | 145.6 | 2.2 | 1.5 | 3.0 | 2.0 |
| Plasma 1 | 126.0 | 1.4 | 1.2 | 2.1 | 1.7 |
| Serum 2 | 246.8 | 1.0 | 0.4 | 2.0 | 0.8 |
| Plasma 2 | 332.0 | 4.4 | 1.3 | 5.4 | 1.6 |
| Serum 3 | 512.2 | 4.9 | 1.0 | 8.9 | 1.7 |
| Plasma 3 | 539.2 | 4.1 | 0.8 | 6.8 | 1.3 |
**Cholesterol on the ACE Alera Analyzer**
| Sample | Mean (mg/dL) | Within Run | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- |
| | | SD | CV % | SD | CV % |
| Serum 1 | 144.5 | 2.0 | 1.4 | 2.8 | 1.9 |
| Plasma 1 | 124.6 | 1.2 | 1.0 | 2.0 | 1.6 |
| Serum 2 | 244.4 | 2.0 | 0.8 | 2.1 | 0.9 |
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| Plasma 2 | 326.4 | 3.1 | 0.9 | 3.9 | 1.2 |
| --- | --- | --- | --- | --- | --- |
| Serum 3 | 503.8 | 3.0 | 0.6 | 5.5 | 1.1 |
| Plasma 3 | 531.8 | 2.1 | 0.4 | 7.7 | 1.4 |
HDL-C on the ACE Analyzer
| Sample | Mean (mg/dL) | Within Run | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- |
| | | SD | CV % | SD | CV % |
| Serum 1 | 51.3 | 1.7 | 3.3 | 1.9 | 3.7 |
| Plasma 1 | 44.4 | 0.9 | 1.9 | 1.3 | 3.0 |
| Serum 2 | 83.0 | 1.1 | 1.3 | 1.6 | 1.9 |
| Plasma 2 | 76.8 | 0.7 | 0.9 | 0.9 | 1.1 |
| Serum 3 | 115.4 | 1.0 | 0.8 | 2.1 | 1.8 |
| Plasma 3 | 110.8 | 1.1 | 1.0 | 2.2 | 2.0 |
HDL-C on the ACE Alera Analyzer
| Sample | Mean (mg/dL) | Within Run | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- |
| | | SD | CV % | SD | CV % |
| Serum 1 | 49.0 | 0.6 | 1.2 | 1.0 | 2.1 |
| Plasma 1 | 42.5 | 0.6 | 1.4 | 1.0 | 2.4 |
| Serum 2 | 77.9 | 0.8 | 1.0 | 1.2 | 1.6 |
| Plasma 2 | 71.6 | 1.3 | 1.8 | 1.3 | 1.9 |
| Serum 3 | 107.1 | 1.0 | 1.0 | 1.5 | 1.4 |
| Plasma 3 | 102.6 | 1.3 | 1.2 | 1.9 | 1.9 |
LDL-C on the ACE Analyzer
| Sample | Mean (mg/dL) | Within Run | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- |
| | | SD | CV % | SD | CV % |
| Serum 1 | 89.4 | 1.9 | 2.2 | 2.1 | 2.3 |
| Plasma 1 | 75.4 | 1.7 | 2.3 | 2.3 | 3.0 |
| Serum 2 | 154.6 | 2.4 | 1.5 | 3.4 | 2.2 |
| Plasma 2 | 212.4 | 3.5 | 1.6 | 6.5 | 3.1 |
| Serum 3 | 339.8 | 5.4 | 1.6 | 8.3 | 2.4 |
| Plasma 3 | 360.4 | 6.3 | 1.8 | 6.8 | 1.9 |
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LDL-C on the ACE Alera Analyzer
| Sample | Mean (mg/dL) | Within Run | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- |
| | | SD | CV % | SD | CV % |
| Serum 1 | 89.2 | 2.2 | 2.4 | 2.2 | 2.4 |
| Plasma 1 | 76.2 | 1.7 | 2.1 | 2.3 | 2.8 |
| Serum 2 | 156.1 | 2.6 | 1.7 | 3.5 | 2.3 |
| Plasma 2 | 214.2 | 3.2 | 1.5 | 4.3 | 2.0 |
| Serum 3 | 341.4 | 4.3 | 1.3 | 7.8 | 2.3 |
| Plasma 3 | 365.9 | 5.6 | 1.5 | 7.7 | 2.1 |
Triglyceride on the ACE Analyzer
| Sample | Mean (mg/dL) | Within Run | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- |
| | | SD | CV % | SD | CV % |
| Serum 1 | 58.0 | 0.9 | 1.6 | 1.8 | 3.2 |
| Plasma 1 | 51.0 | 1.2 | 2.4 | 2.0 | 3.9 |
| Serum 2 | 329.5 | 2.2 | 0.7 | 4.2 | 1.3 |
| Plasma 2 | 324.3 | 7.3 | 2.2 | 9.4 | 2.9 |
| Serum 3 | 601.8 | 5.1 | 0.8 | 9.5 | 1.6 |
| Plasma 3 | 596.7 | 4.5 | 0.7 | 7.3 | 1.2 |
Triglyceride on the ACE Alera Analyzer
| Sample | Mean (mg/dL) | Within Run | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- |
| | | SD | CV % | SD | CV % |
| Serum 1 | 58.1 | 0.6 | 1.1 | 1.7 | 3.0 |
| Plasma 1 | 51.1 | 0.8 | 1.6 | 2.1 | 4.2 |
| Serum 2 | 327.9 | 2.5 | 0.8 | 4.3 | 1.3 |
| Plasma 2 | 324.6 | 5.4 | 1.7 | 5.4 | 1.7 |
| Serum 3 | 599.8 | 4.1 | 0.7 | 4.4 | 0.7 |
| Plasma 3 | 588.4 | 7.0 | 1.2 | 13.5 | 2.3 |
Precision studies were also conducted in three physician office laboratory (POL) sites on three serum samples at three concentrations over five days. The results are as follows:
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Cholesterol on the ACE Analyzer
| Sample 1 | | | |
| --- | --- | --- | --- |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 111.4 | 110.5 | 109.5 |
| Within-Run SD (% CV) | 1.3 (1.2%) | 0.8 (0.7%) | 1.2 (1.1%) |
| Total SD (%CV) | 1.4 (1.3%) | 0.9 (0.8%) | 1.4 (1.3%) |
| Sample 2 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 239.6 | 230.3 | 236.7 |
| Within-Run SD (% CV) | 2.5 (1.0%) | 1.8 (0.8%) | 2.1 (0.9%) |
| Total SD (%CV) | 3.0 (1.2%) | 3.0 (1.3%) | 2.7 (1.1%) |
| Sample 3 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 364.4 | 347.3 | 356.5 |
| Within-Run SD (% CV) | 2.5 (0.7%) | 3.9 (1.1%) | 2.4 (0.7%) |
| Total SD (%CV) | 4.2 (1.2%) | 4.1 (1.2%) | 3.1 (0.9%) |
Cholesterol on the ACE Alera Analyzer
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| Sample 1 | | | |
| --- | --- | --- | --- |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 110.5 | 111.3 | 111.0 |
| Within-Run SD (% CV) | 2.4 (2.2%) | 2.4 (2.2%) | 1.9 (1.7%) |
| Total SD (%CV) | 2.6 (2.4%) | 2.6 (2.4%) | 2.1 (1.9%) |
| Sample 2 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 238.5 | 239.0 | 237.7 |
| Within-Run SD (% CV) | 4.2 (1.8%) | 3.4 (1.4%) | 2.5 (1.1%) |
| Total SD (%CV) | 4.9 (2.1%) | 4.4 (1.8%) | 2.5 (1.1%) |
| Sample 3 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 356.3 | 359.9 | 358.7 |
| Within-Run SD (% CV) | 2.2 (0.6%) | 6.6 (1.8%) | 2.0 (0.5%) |
| Total SD (%CV) | 4.2 (1.2%) | 7.1 (2.0%) | 2.9 (0.8%) |
HDL on the ACE Analyzer
| Sample 1 | | | |
| --- | --- | --- | --- |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 35.7 | 33.4 | 35.6 |
| Within-Run SD (% CV) | 0.9 (2.4%) | 0.6 (1.8%) | 0.7 (2.0%) |
| Total SD (%CV) | 1.1 (3.2%) | 1.5 (4.4%) | 2.1 (6.0%) |
| Sample 2 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 58.6 | 54.4 | 58.5 |
| Within-Run SD (% CV) | 0.9 (1.6%) | 0.5 (1.0%) | 1.1 (1.9%) |
| Total SD (%CV) | 1.2 (2.1%) | 2.0 (3.7%) | 2.9 (5.0%) |
| Sample 3 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 81.2 | 77.3 | 82.0 |
| Within-Run SD (% CV) | 1.4 (1.8%) | 1.0 (1.3%) | 1.1 (1.3%) |
| Total SD (%CV) | 1.8 (2.3%) | 2.9 (3.8%) | 4.1 (5.0%) |
HDL on the ACE Alera Analyzer
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| Sample 1 | | | |
| --- | --- | --- | --- |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 33.2 | 38.3 | 35.4 |
| Within-Run SD (% CV) | 1.1 (3.2%) | 0.9 (2.3%) | 0.6 (1.8%) |
| Total SD (%CV) | 1.2 (3.7%) | 1.0 (2.7%) | 1.8 (5.0%) |
| Sample 2 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 55.0 | 61.2 | 58.0 |
| Within-Run SD (% CV) | 1.5 (2.7%) | 1.2 (1.9%) | 1.1 (1.9%) |
| Total SD (%CV) | 1.9 (3.5%) | 1.5 (2.5%) | 2.1 (3.7%) |
| Sample 3 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 76.0 | 86.5 | 81.2 |
| Within-Run SD (% CV) | 1.1 (1.4%) | 1.6 (1.8%) | 1.5 (1.8%) |
| Total SD (%CV) | 2.2 (2.8%) | 2.2 (2.6%) | 3.2 (4.0%) |
LDL on the ACE Analyzer
| Sample 1 | | | |
| --- | --- | --- | --- |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 67.5 | 67.0 | 67.0 |
| Within-Run SD (% CV) | 2.0 (3.0%) | 0.6 (0.9%) | 2.5 (3.8%) |
| Total SD (%CV) | 2.1 (3.1%) | 2.3 (3.5%) | 2.7 (4.0%) |
| Sample 2 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 152.7 | 138.4 | 150.8 |
| Within-Run SD (% CV) | 4.1 (2.7%) | 4.0 (2.9%) | 3.0 (2.0%) |
| Total SD (%CV) | 5.3 (3.5%) | 4.2 (3.0%) | 4.2 (2.8%) |
| Sample 3 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 237.1 | 217.8 | 240.2 |
| Within-Run SD (% CV) | 3.8 (1.6%) | 3.3 (1.5%) | 4.6 (1.9%) |
| Total SD (%CV) | 5.2 (2.2%) | 6.0 (2.8%) | 6.6 (2.7%) |
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LDL on the ACE Alera Analyzer
| Sample 1 | | | |
| --- | --- | --- | --- |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 70.7 | 68.4 | 67.2 |
| Within-Run SD (% CV) | 3.3 (4.7%) | 3.0 (4.4%) | 1.1 (1.6%) |
| Total SD (%CV) | 3.6 (5.0%) | 3.2 (4.7%) | 1.8 (2.6%) |
| Sample 2 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 154.8 | 153.6 | 150.8 |
| Within-Run SD (% CV) | 2.1 (1.4%) | 3.6 (21.4%) | 3.4 (2.3%) |
| Total SD (%CV) | 4.5 (2.9%) | 4.9 (3.2%) | 3.5 (2.3%) |
| Sample 3 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 244.6 | 244.9 | 241.9 |
| Within-Run SD (% CV) | 3.6 (1.5%) | 6.5 (2.7%) | 4.3 (1.8%) |
| Total SD (%CV) | 6.5 (2.7%) | 7.1 (2.9%) | 5.3 (2.2%) |
Triglyceride on the ACE Analyzer
| Sample 1 | | | |
| --- | --- | --- | --- |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 66.7 | 68.8 | 69.3 |
| Within-Run SD (% CV) | 1.0 (1.5%) | 1.5 (2.1%) | 1.5 (2.1%) |
| Total SD (%CV) | 2.0 (3.0%) | 1.9 (2.8%) | 1.5 (2.1%) |
| Sample 2 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 267.8 | 271.9 | 262.8 |
| Within-Run SD (% CV) | 1.9 (0.7%) | 3.2 (1.2%) | 2.1 (0.8%) |
| Total SD (%CV) | 2.0 (0.7%) | 3.7 (1.3%) | 2.9 (1.1%) |
| Sample 3 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 450.4 | 461.9 | 439.5 |
| Within-Run SD (% CV) | 9.7 (2.1%) | 8.8 (1.9%) | 3.8 (0.9%) |
| Total SD (%CV) | 11.2 (2.5%) | 9.7 (2.1%) | 5.1 (1.2%) |
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Triglyceride on the ACE Alera Analyzer
| Sample 1 | | | |
| --- | --- | --- | --- |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 65.3 | 69.5 | 68.8 |
| Within-Run SD (% CV) | 1.9 (2.9%) | 1.4 (2.1%) | 1.1 (1.7%) |
| Total SD (%CV) | 2.1 3.2%) | 1.5 (2.2%) | 1.8 (2.6%) |
| Sample 2 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 249.4 | 265.9 | 260.1 |
| Within-Run SD (% CV) | 4.8 (1.9%) | 2.2 (0.8%) | 1.1 (0.4%) |
| Total SD (%CV) | 5.9 (2.4%) | 2.3 (0.9%) | 1.5 (0.6%) |
| Sample 3 | | | |
| | POL 1 | POL 2 | POL 3 |
| Mean (mg/dL) | 410.5 | 442.9 | 432.1 |
| Within-Run SD (% CV) | 6.8 (1.7%) | 3.7 (0.8%) | 3.5 (0.8%) |
| Total SD (%CV) | 7.5 (1.8%) | 3.7 (0.8%) | 3.7 (0.9%) |
b. Linearity/assay reportable range:
The linearity remains the same as in predicate devices. The measuring ranges previously established are:
Cholesterol: 6 to 600 mg/dL
HDL: 4-125 mg/dL
LDL: 3-800 mg/dL
Triglycerides 6-1000 mg/dL
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Not applicable, the traceability and stability remains the same as in predicate devices.
d. Detection limit:
Not applicable, the detection limit remains the same as in predicate devices.
e. Analytical specificity:
Not applicable, the specificity remains the same as in predicate devices.
f. Assay cut-off:
Not applicable.
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# 2. Comparison studies:
a. Method comparison with predicate device:
A method comparison study was performed using serum samples at Physician Office Laboratories. Operators assayed serum samples on the ACE and the ACE Alera Clinical Chemistry Systems. The following linear regression data was obtained by comparing samples run in house to the POL sites using Deming analysis:
ACE Analyzer
| Cholesterol | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Site | N | Range | Slope | 95% CI of Slope | Intercept | 95% CI of Intercept | r2 |
| In-house vs. POL 1 | 46 | 41 to 431 | 1.017 | 1.001 to 1.034 | -4.5 | 8.2 to -0.9 | 0.9986 |
| In-house vs. POL 2 | 46 | 42 to 425 | 0.983 | 0.967 to 1.000 | 1.4 | -2.4 to 5.1 | 0.9985 |
| In-house vs. POL 3 | 46 | 40 to 425 | 0.996 | 0.982 to 1.009 | -0.9 | -3.9 to 2.0 | 0.9990 |
| HDL | | | | | | | |
| In-house vs. POL 1 | 45 | 14 to 102 | 0.955 | 0.927 to 0.983 | 0.1 | -1.4 to 1.7 | 0.9954 |
| In-house vs. POL 2 | 45 | 15 to 103 | 0.958 | 0.932 to 0.984 | 0.8 | -0.6 to 2.3 | 0.9960 |
| In-house vs. POL 3 | 45 | 14 to 103 | 1.002 | 0.964 to 1.041 | 0.5 | -1.7 to 2.6 | 0.9921 |
| LDL | | | | | | | |
| In-house vs. POL 1 | 45 | 21 to 329 | 0.979 | 0.949 to 1.009 | 0.0 | -4.2 to 4.1 | 0.9949 |
| In-house vs. POL 2 | 45 | 23 to 329 | 0.955 | 0.924 to | 1.7 | -2.5 to 5.9 | 0.9945 |
| | | | | 0.996 | | | |
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ACE Alera Analyzer
| Cholesterol | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Site | N | Range | Slope | 95% CI of Slope | Intercept | 95% CI of Intercept | r² |
| In-house vs. POL 1 | 46 | 43 to 429 | 1.001 | 0.961 to 1.042 | -5.1 | -14.5 to 4.2 | 0.9911 |
| In-house vs. POL 2 | 46 | 43 to 428 | 1.002 | 0.971 to 1.032 | -6.1 | -13.2 to 1.0 | 0.9948 |
| In-house vs. POL 3 | 46 | 29 to 762 | 1.006 | 0.997 to 1.016 | -0.5 | -3.0 to 2.1 | 0.9995 |
| HDL | | | | | | | |
| In-house vs. POL 1 | 45 | 12 to 101 | 0.968 | 0.918 to 1.017 | 0.5 | -2.3 to 3.2 | 0.9863 |
| In-house vs. POL 2 | 45 | 16 to 102 | 0.959 | 0.910 to 1.008 | 1.6 | -1.1 to 4.3 | 0.9862 |
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| | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| In-house vs. POL 3 | 44 | 14 to 103 | 0.953 | 0.916 to 0.989 | 0.3 | -1.7 to 2.4 | 0.9924 |
| LDL | | | | | | | |
| In-house vs. POL 1 | 46 | 24 to 333 | 0.982 | 0.929 to 1.035 | 2.5 | -4.9 to 9.8 | 0.9840 |
| In-house vs. POL 2 | 46 | 23 to 325 | 0.983 | 0.920 to 1.045 | -0.2 | -8.8 to 8.4 | 0.9782 |
| In-house vs. POL 3 | 46 | 20 to 322 | 1.011 | 0.984 to 1.038 | -4.7 | -8.3 to -1.0 | 0.9962 |
| Triglyceride | | | | | | | |
| In-house vs. POL 1 | 46 | 24 to 333 | 0.977 | 0.956 to 0.999 | -1.0 | -6.9 to 4.9 | 0.9975 |
| In-house vs. POL 2 | 46 | 31 to 755 | 0.997 | 0.970 to 1.023 | 0.5 | -6.9 to 8.0 | 0.9962 |
| In-house vs. POL 3 | 46 | 29 to 750 | 0.977 | 0.957 to 0.997 | -1.0 | -6.6 to 4.7 | 0.9977 |
# b. Matrix comparison:
Matrix comparison was done by using Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline - Second Edition (2002), CLSI EP9-A2 as a guideline. Paired samples were drawn from the same patients in serum and lithium heparin plasma tubes drawn at hospital sites. Some of the samples were spiked with a solution containing a high level of analyte obtained from commercial sources.
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| Reagent | Sample # | # Diluted/# Spiked | Serum Range | Results ACE | Results ACE Alera |
| --- | --- | --- | --- | --- | --- |
| Cholesterol | ACE: 102 | 0/5 ACE | ACE: 40 to 568 mg/dL | Slope: 0.985(0.965, 1.005) | Slope: 0.994 (0.971, 1.016) |
| | Alera: 103 | 0/6 Alera | Alera: 42 to 577 mg/dL | Intercept: -1.7 (-5.7 to 2.3) Correlation: 0.9947 | Intercept: -2.5 (-7.0, 2.1) Correlation: 0.9934 |
| HDL | ACE: 101 | 0/0 | ACE: 6 to 120 mg/dL | Slope: 1.015 (0.984, 1.045) | Slope: 0.989 (0.957, 1.02) |
| | Alera: 100 | | Alera: 7 to 123 mg/dL | Intercept: -0.6 (-2.1, 0.8) Correlation: 0.9884 | Intercept: 0.4 (-1.2, 1.9) Correlation: 0.9874 |
| LDL | 99 | 0/4 | ACE: 9 to 460 mg/dL | Slope: 1.008 (0.989, 1.028) | Slope: 0.995 (0.976, 1.014) |
| | | | Alera: 9 to 464 mg/dL | Intercept: -2.6 (-5.0, -0.2) Correlation: 0.9954 | Intercept: -1.3 (-3.7, 1.0) Correlation: 0.9954 |
| Triglyceride | 101 | 0/5 | ACE: 39 to 887 mg/dL | Slope: 1.005 (0.991, 1.019) | Slope: 1.007 (0.992, 1.021) |
| | | | Alera: 38 to 884 mg/dL | Intercept: -7.9 (-11.1, -4.7) Correlation: 0.9977 | Intercept: -7.4 (-10.8, -4.0) Correlation: 0.9973 |
# 3. Clinical studies:
a. Clinical Sensitivity:
Not Applicable
b. Clinical specificity:
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
Specific ranges for each analyte/methodology are listed in the package insert.
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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.
