ACE CHOLESTEROL REAGENT, ACE HDL-C REAGENT, ACE LDL-C REAGENT-C, ACE-TRISLYCERIDES REAGENT

{0} K113262 # SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k113262 B. Purpose for Submission: New device C. Measurand: Cholesterol HDL- Cholesterol LDL- Cholesterol Triglycerides D. Type of Test: Quantitative, colorimetric assay 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: 1. Regulation section: 21 CFR 862.1175: Cholesterol (total) test system 21 CFR 862.1475: Lipoprotein test system 21 CFR 862.1475: Lipoprotein test system 21 CFR 862.1705: Triglyceride test system 2. Classification: Class I (for all the four measurands), all meet limitations of exemption per 21 CFR 1 {1} 862.9(c)(4) and 21 CFR 862.9(c)(9) 3. Product code: CHH: Enzymatic Esterase-Oxidase, Cholesterol LBS: LDL & VLDL Precipitation, Cholesterol Via Esterase-Oxidase, HDL MRR: System, Test, Low Density, Lipoprotein CDT: Lipase Hydrolysis/Glycerol Kinase Enzyme, Triglycerides 4. Panel: Clinical Chemistry (75) H. Intended Use: 1. Intended use(s): See indication for use below 2. Indication(s) for use: The ACE Cholesterol Reagent is intended for the quantitative determination of cholesterol concentration in serum using the ACE Axcel Clinical Chemistry System. 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. The ACE HDL-C Reagent is intended for the quantitative determination of high density lipoprotein cholesterol (HDL-C) concentration in serum using the ACE Axcel Clinical Chemistry System. 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. The ACE LDL-C Reagent is intended for the quantitative determination of low density lipoprotein cholesterol (LDL-C) concentration in serum using the ACE Axcel Clinical Chemistry System. 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. The ACE Triglycerides Reagent is intended for the quantitative determination of triglyceride concentration in serum using the ACE Axcel Clinical Chemistry System. Triglyceride measurements are used in the diagnosis and treatment of patients with diabetes mellitus, nephrosis, liver obstruction, other diseases involving lipid metabolism K113262 2 {2} or various endocrine disorders. This test is intended for use in clinical laboratories or physician office laboratories. For in vitro diagnostic use only. 3. Special conditions for use statement(s): For in vitro diagnostic use only. For prescription use. 4. Special instrument requirements: ACE Axcel Clinical Chemistry System I. Device Description: The ACE Cholesterol Reagent is composed of a single reagent bottle containing 0.5 mmol/L 4-Aminoantipyrine (AAP), 25 mmol/L p-Hydroxybenzoic acid, >150 U/L Cholesterol oxidase (Nocardia), >240 U/L Cholesterol esterase (Porcine pancreas and Pseudomonas) >1600 U/L Peroxidase (Horseradish), stabilizers, preservatives, and fillers. The ACE HDL-C Reagent is composed of two reagent bottles (Buffer and Color Reagent). The Buffer (R1) contains: Good's Buffer, <1000 U/L Cholesterol oxidase (E. coli), <1300 U/L Peroxidase (Horseradish), <1 mM N,N-bis(4-sulphobutyl)-m-toluidine-disodium salt (DSBmT), <1 mM Accelerator, <0.06% Preservative and <3000 U/L Ascorbic Oxidase (Curcubita sp.). The Color Reagent (R2) contains: <1500 U/L Cholesterol esterase (Pseudomonas sp.), <1 mM 4-Aminoantipyrine (4-AAP), <2% detergent and preservative. The ACE LDL-C Reagent is composed of two reagent bottles (Buffer and Color Reagent). The Buffer (R1) contains: MES Buffer (pH 6.3), <1.0% Detergent 1, <1500 U/L Cholesterol oxidase (Pseudomonas sp.), <1300 ppg U/L Peroxidase (Horseradish), <0.1% 4-Aminoantipyrine (4-AAP), <3000 U/L Ascorbic acid oxidase (Curcubita sp.), <0.1% Preservative. The Color Reagent (R2) contains: MES Buffer (pH 6.3), <1.0% Detergent 2, <1.0 mmol/L N,N-bis(4-sulphobutyl)-m-toluidine-disodium salt (DSBmT) and <0.1% preservative. The ACE Triglycerides Reagent is composed of a single reagent bottle. The reagent contains: 0.4 mmol/L 4-Aminoantipyrine (AAP), 2.6 mmol/L Adenosine 5'-triphosphate (ATP), 3.0 mmol/L p-Chlorophenol, >2400 U/L Glycerol phosphate oxidase (GPO) (Microorganism), >1000 U/L Lipase (Pseudomonas), >540 U/L Peroxidase (Horseradish), >400 U/L Glycerol kinase (Cellulomonas), buffer, stabilizers and preservatives. J. Substantial Equivalence Information: 1. Predicate device name(s): ACE Cholesterol Reagent ACE HDL-C Reagent ACE LDL-C Reagent ACE Triglycerides Reagent K113262 3 {3} 2. Predicate 510(k) number(s): | ACE Reagent Kit | Reagent 510(k) | | --- | --- | | Cholesterol | k931786 | | HDL-C | k971526 | | LDL-C | k991733 | | Triglycerides | k931786 | 3. Comparison with predicate: | Characteristics | ACE Cholesterol Agent, (Candidate Device) | ACE Cholesterol Reagent (Predicate, k931786) | | --- | --- | --- | | Indications for Use | The ACE Cholesterol Reagent is intended for the quantitative determination of cholesterol concentration in serum. 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. | Same | | Instrument/ Platform | ACE Axcel Clinical Chemistry System | ACE Clinical Chemistry System | | Sample Type | Serum | Same | | Measuring Range | 4 - 600 mg/dL | 2 - 600 mg/dL | | Calibration | GEMCAL | Same | | Calibration Stability | 30 Days | Same | | On-Board Stability | 30 Days | Same | | Characteristics | ACE HDL-C Agent, (Candidate Device) | ACE HDL-C Reagent (Predicate, k971526) | | --- | --- | --- | | Indications for Use | The ACE HDL-C Reagent is intended for the quantitative determination of high density lipoprotein cholesterol (HDL-C) concentration in serum. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases. | Same | K113262 {4} | Instrument/Platform | ACE Axcel Clinical Chemistry System | ACE Clinical Chemistry System | | --- | --- | --- | | Sample Type | Serum | Same | | Reportable | 8 -125 mg/dL | Same | | Calibration | GEMCAL | Same | | Calibration Stability | 30 Days | Same | | On-Board Stability | 30 Days | Same | | Characteristics | ACE LDL-C Agent (Candidate Device) | ACE LDL-C Reagent (Predicate, k991733) | | --- | --- | --- | | Indications for Use | The ACE LDL-C Reagent is intended for the quantitative determination of low density lipoprotein cholesterol (LDL-C) concentration in serum. 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. | Same | | Instrument/Platforms | ACE Axcel Clinical Chemistry System | ACE Clinical Chemistry Systems | | Sample Type | Serum | Same | | Reportable Range | 4 to 430 mg/dL | 3 -500 mg/dL | | Calibration | ACE LDL-C Calibrators | Same | | Calibration Stability | 30 Days | Same | | On-Board Stability | 30 Days | Same | | Characteristics | ACE Triglycerides Agent (Candidate Device) | ACE Triglycerides Reagent (Predicate, k931786) | | --- | --- | --- | K113262 {5} | Indications for Use | The ACE Triglycerides Reagent is intended for quantitative determination of triglycerides concentration in serum. It is intended for in vitro diagnostic use only. | Same | | --- | --- | --- | | Instrument/ Platforms | ACE Axcel Clinical Chemistry System | ACE Clinical Chemistry Systems | | Sample Type | Serum | Same | | Reportable Range | 12 -1000 mg/dL | 6 -1000 mg/dL | | Calibration | GEMCAL | Same | | Calibration Stability | 30 Days | Same | | On-Board Stability | 30 Days | Same | # K. Standard/Guidance Document Referenced: CLSI EP6-A Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach (2003), CLSI EP5-A2 Evaluation of Precision Performance of Quantitative Measurement Methods; Second Edition (2004), CLSI EP7-A2 Interference Testing in Clinical Chemistry; Approved Guideline; Second Edition (2005), CLSI EP9-A2-IR Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline- Second Edition (2002), CLSI EP17-A Protocol for Determination of Limits of Detection; Approved Guideline (2004) # L. Test Principle: The ACE Cholesterol Reagent: 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 oxidized by cholesterol oxidase to yield hydrogen peroxide $(\mathrm{H}_2\mathrm{O}_2)$ , which then reacts oxidatively with p-hydroxybenzoic acid and 4-aminoantipyrine (AAP) in a reaction catalyzed by peroxidase, producing a red colored quinoneimine complex. The amount of chromogen formed, determined by quantitatively measuring the increase in absorbance at $505~\mathrm{nm} / 647~\mathrm{nm}$ , is directly proportional to the cholesterol concentration in the sample. The ACE HDL-C Reagent is composed of two bottles: Buffer Reagent (R1) and Color Reagent (R2). R2 contains a unique detergent, which solubilizes only the HDL lipoprotein particles; the detergent also inhibits the reaction of the cholesterol enzymes with LDL and VLDL lipoproteins and chylomicrons by adsorbing to their surfaces. The solubilized HDL cholesterol is released and reacts with cholesterol esterase and cholesterol oxidase and a chromogen 4-Aminoantipyrine to produce a color product measurable by the increase of K113262 {6} absorbance at 505/647 nm, which is directly proportional to the cholesterol concentration in the sample. The ACE LDL-C Reagent: There are two detergents, Detergent 1 and Detergent 2, in the ACE LDL-C Reagents R1 and R2 respectively. In the first reaction, 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 product measurable by the increase of absorbance at 505/647 nm, which is directly proportional to the cholesterol concentration in the sample. The ACE Triglycerides Reagent: The ACE Triglycerides Reagent is composed of a single reagent bottle. 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-I-phosphate (G-I-P) and the ATP to adenosine diphosphate (ADP). Glycerol-I-phosphate is oxidized by glycerol phosphate oxidase (GPO) to yield hydrogen peroxide (H₂O₂). 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 nm. ## M. Performance Characteristics (if/when applicable): ### 1. Analytical performance: #### a. Precision/Reproducibility: **In-House: ACE Cholesterol Reagent** Three serum based pools and one normal human serum sample, were tested on one ACE Axcel Clinical Chemistry System two times per run, two runs per day, for a total of 20 or more days. The results are summarized in the table below: | Sample 1 Mean 87.7 mg/dL Cholesterol | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 1.4 | 0.5 | 0.5 | 1.6 | | Coefficient of Variation | 1.6% | 0.5% | 0.6% | 1.8% | | Sample 2 Mean 261.5 mg/dL Cholesterol | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 4.2 | 0.0 | 0.8 | 4.3 | | Coefficient of Variation | 1.6% | 0.0% | 0.3% | 1.6% | K113262 {7} | Sample 3 Mean 430.4 mg/dL Cholesterol | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 8.7 | 0.0 | 3.7 | 9.4 | | Coefficient of Variation | 2.0% | 0.0% | 0.9% | 2.2% | | Sample 4 Mean 176.4 mg/dL Cholesterol | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 2.2 | 2.2 | 0.0 | 3.1 | | Coefficient of Variation | 1.3% | 1.2% | 0.0% | 1.8% | ## Point of Care Laboratory: ACE Cholesterol Reagent Three serum based pools were tested on three ACE Axcel Clinical Chemistry Systems (one at each POL site) at least three times per run, one run per day, for a total of 5 days. The mean, standard deviations and % coefficients of variation (CV) were calculated for each sample. | Cholesterol | | | Within Run | | Total | | | --- | --- | --- | --- | --- | --- | --- | | Lab | Sample | Mean mg/dL | SD | %CV | SD | %CV | | POL 1 | 1 | 84.6 | 1.2 | 1.4 | 1.4 | 1.7 | | POL 2 | 1 | 89.3 | 1.0 | 1.1 | 1.1 | 1.3 | | POL 3 | 1 | 85.3 | 1.3 | 1.5 | 1.5 | 1.7 | | | | | | | | | | POL 1 | 2 | 255.6 | 2.5 | 1.0 | 4.1 | 1.6 | | POL 2 | 2 | 270.3 | 2.1 | 0.8 | 3.3 | 1.2 | | POL 3 | 2 | 256.4 | 2.4 | 0.9 | 2.6 | 1.0 | | | | | | | | | | POL 1 | 3 | 423.6 | 6.2 | 1.5 | 6.4 | 1.5 | | POL 2 | 3 | 441.6 | 5.1 | 1.2 | 6.1 | 1.4 | | POL 3 | 3 | 423.2 | 3.0 | 0.7 | 6.6 | 1.6 | ## In-House: ACE HDL-C Reagent Three serum based pools and one normal human serum sample, were tested on one ACE Axcel Clinical Chemistry System two times per run, two runs per day, for a total of 20 or more days. The results are summarized in the table below: K113262 {8} | Sample 1 Mean 27.8 mg/dL HDL-C | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 0.7 | 0.6 | 0.9 | 1.3 | | Coefficient of Variation | 2.7% | 2.3% | 3.4% | 4.8% | | Sample 2 Mean 63.0 mg/dL HDL-C | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 1.1 | 0.0 | 1.9 | 2.2 | | Coefficient of Variation | 1.7% | 0.0% | 3.0% | 3.4% | | Sample 3 Mean 98.3 mg/dL HDL-C | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 1.6 | 1.3 | 2.3 | 3.1 | | Coefficient of Variation | 1.7% | 1.3% | 2.4% | 3.2% | | Sample 4 Mean 73.9 mg/dL HDL-C | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 1.1 | 0.9 | 1.9 | 2.4 | | Coefficient of Variation | 1.4% | 1.2% | 2.6% | 3.2% | Point of Care Laboratory: ACE HDL-C Reagent Three serum based pools were tested on three ACE Axcel Clinical Chemistry Systems (one at each POL site) at least three times per run, one run per day, for a total of 5 days. The mean, standard deviations and % coefficients of variation (CV) were calculated for each sample. | HDL-C | | | Within Run | | Total | | | --- | --- | --- | --- | --- | --- | --- | | Lab | Sample | Mean mg/dL | SD | %CV | SD | %CV | | POL 1 | 1 | 25.8 | 0.4 | 1.6 | 0.6 | 2.3 | | POL 2 | 1 | 27.0 | 0.7 | 2.6 | 1.0 | 3.5 | | POL 3 | 1 | 26.8 | 0.6 | 2.2 | 0.8 | 3.0 | | | | | | | | | | POL 1 | 2 | 59.4 | 0.6 | 1.1 | 0.6 | 1.1 | | POL 2 | 2 | 61.5 | 1.0 | 1.6 | 1.3 | 2.2 | K113262 {9} K113262 10 | POL 3 | 2 | 31.3 | 0.6 | 1.0 | 1.1 | 1.8 | | --- | --- | --- | --- | --- | --- | --- | | | | | | | | | | POL 1 | 3 | 93.5 | 1.0 | 1.1 | 1.6 | 1.7 | | POL 2 | 3 | 97.6 | 1.6 | 1.6 | 1.9 | 1.9 | | POL 3 | 3 | 97.0 | 0.7 | 0.7 | 2.1 | 2.1 | **In-House: ACE LDL-C Reagent** Three serum based pools and one normal human serum sample, were tested on one ACE Axcel Clinical Chemistry System two times per run, two runs per day, for a total of 20 or more days. The results are summarized in the table below: | **Sample 1** Mean 45.1 mg/dL LDL-C | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 2.1 | 0.5 | 0.5 | 2.2 | | Coefficient of Variation | 4.6% | 1.2% | 1.0% | 4.9% | | **Sample 2** Mean 181.7 mg/dL LDL-C | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 4.7 | 3.2 | 3.4 | 6.6 | | Coefficient of Variation | 2.6% | 1.8% | 1.9% | 3.6% | | **Sample 3** Mean 328.5 mg/dL LDL-C | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 8.7 | 0.0 | 6.1 | 10.6 | | Coefficient of Variation | 2.7% | 0.0% | 1.8% | 3.2% | | **Sample 4** Mean 86.3 mg/dL LDL-C | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 2.2 | 1.9 | 0.9 | 3.0 | | Coefficient of Variation | 2.5% | 2.2% | 1.0% | 3.5% | **Point of Care Laboratory: ACE LDL-C Reagent** Three serum-based pools were tested on three ACE Axcel Clinical Chemistry Systems (one at each POL site) at least three times per run, one run per day, for a total of 5 days. The mean, standard deviations and % coefficients of variation (CV) were calculated for each sample. {10} | LDL-C | | | Within Run | | Total | | | --- | --- | --- | --- | --- | --- | --- | | Lab | Sample | Mean mg/dL | SD | %CV | SD | %CV | | POL 1 | 1 | 45.9 | 2.0 | 4.4 | 2.1 | 4.6 | | POL 2 | 1 | 41.9 | 1.3 | 3.1 | 2.5 | 5.9 | | POL 3 | 1 | 46.0 | 1.9 | 4.1 | 2.4 | 5.2 | | | | | | | | | | POL 1 | 2 | 185.9 | 4.9 | 2.6 | 7.0 | 3.8 | | POL 2 | 2 | 164.0 | 6.2 | 3.8 | 8.2 | 5.0 | | POL 3 | 2 | 180.9 | 3.7 | 2.0 | 4.9 | 2.7 | | | | | | | | | | POL 1 | 3 | 336.7 | 8.0 | 2.4 | 8.0 | 2.4 | | POL 2 | 3 | 307.6 | 6.9 | 2.2 | 10.5 | 3.4 | | POL 3 | 3 | 328.7 | 5.7 | 1.7 | 8.6 | 2.6 | In-House: ACE Triglycerides Reagent Three serum-based pools and one normal human serum sample were tested on one ACE Axcel Clinical Chemistry System two times per run, two runs per day, for a total of 20 or more days. The results are summarized in the table below: | Sample 1 Mean 73.3 mg/dL Triglycerides | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 1.3 | 0.8 | 0.4 | 1.5 | | Coefficient of Variation | 1.8% | 1.0% | 0.5% | 2.1% | | Sample 2 Mean 138.9 mg/dL Triglycerides | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 2.5 | 1.1 | 0.8 | 2.9 | | Coefficient of Variation | 1.8% | 0.8% | 0.6% | 2.1% | | Sample 3 Mean 950.4 mg/dL Triglycerides | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 11.5 | 7.4 | 9.6 | 16.7 | | Coefficient of Variation | 1.2% | 0.8% | 1.0% | 1.8% | K113262 {11} | Sample 4 Mean 58.6 mg/dL Triglycerides | Within Run | Between Run | Between Day | Total | | --- | --- | --- | --- | --- | | Standard Deviation, mg/dL | 1.7 | 0.7 | 0.0 | 1.8 | | Coefficient of Variation | 2.9% | 1.3% | 0.0% | 3.2% | Point of Care Laboratory: ACE Triglycerides Reagent Four serum-based pools were tested on three ACE Axcel Clinical Chemistry Systems (one at each POL site) at least three times per run, one run per day, for a total of 5 days. The mean, standard deviations and % coefficients of variation (CV) were calculated for each sample. | Triglycerides | | | Within Run | | Total | | | --- | --- | --- | --- | --- | --- | --- | | Lab | Sample | Mean mg/dL | SD | %CV | SD | %CV | | POL 1 | 1 | 70.5 | 1.7 | 2.3 | 1.7 | 2.3 | | POL 2 | 1 | 73.0 | 1.1 | 1.5 | 1.1 | 1.5 | | POL 3 | 1 | 72.7 | 1.2 | 1.7 | 3.0 | 4.1 | | | | | | | | | | POL 1 | 2 | 136.0 | 2.1 | 1.5 | 2.3 | 1.7 | | POL 2 | 2 | 137.9 | 2.1 | 1.5 | 2.4 | 1.8 | | POL 3 | 2 | 136.5 | 1.6 | 1.2 | 1.8 | 1.3 | | | | | | | | | | POL 1 | 3 | 182.1 | 3.9 | 2.1 | 4.6 | 2.5 | | POL 2 | 3 | 185.4 | 2.8 | 1.5 | 2.8 | 1.5 | | POL 3 | 3 | 182.1 | 1.1 | 0.6 | 1.1 | 0.6 | | | | | | | | | | POL 1 | 4 | 933.4 | 4.3 | 0.5 | 6.4 | 0.7 | | POL 2 | 4 | 933.4 | 8.4 | 0.9 | 9.0 | 1.0 | | POL 3 | 4 | 944.0 | 7.8 | 0.8 | 16.6 | 1.8 | b. Linearity/assay reportable range: ACE Cholesterol Reagent Linearity studies were carried out using dilutions of a spiked serum samples. Eleven concentrations were prepared by mixing spiked serum samples in known proportion K113262 {12} with blank samples. All samples were measured in triplicate. The sample range tested was 3 to 966 mg/dL. | Claimed Measuring Range | Intercept | Slope | r² | | --- | --- | --- | --- | | 4 to 600 mg/dL | -1.1 | 1.016 | 0.9993 | Based on the linearity data, the measuring range claimed from 4 to 600 mg/dL was supported. ## ACE HDL-C Reagent Linearity studies were carried out using dilutions of a spiked serum samples. Eleven concentrations were prepared by mixing spiked serum samples in known proportion with blank samples. All samples were measured in triplicate. The sample range tested was 7.6 to 137.7 mg/dL. | Claimed Measuring Range | Intercept | Slope | r² | | --- | --- | --- | --- | | 8 to 125 mg/dL | 0.1 | 1.011 | 0.9986 | Based on the linearity data, the measuring range claimed from 8 to 125 mg/dL was supported. ## ACE LDL-C Reagent Linearity studies were carried out using dilutions of a spiked serum samples. Eleven concentrations were prepared by mixing spiked serum samples in known proportion with blank samples. All samples were measured in triplicate. The sample range tested was 0.3 to 430 mg/dL. | Claimed Measuring Range | Intercept | Slope | r² | | --- | --- | --- | --- | | 4 to 430 mg/dL | -4.5 | 0.985 | 0.9981 | Based on the linearity data, the measuring range claimed from 4 to 430 mg/dL was supported. ## ACE Triglycerides Reagent Linearity studies were carried out using dilutions of a spiked serum samples. Eleven concentrations were prepared by mixing spiked serum samples in known proportion with blank samples. All samples were measured in triplicate. The sample range tested was 12.3 to 1249.3 mg/dL. | Claimed Measuring Range | Intercept | Slope | r² | | --- | --- | --- | --- | | 12 to 1000 mg/dL | 8.6 | 0.9915 | 0.9995 | Based on the linearity data, the measuring range claimed from 12 to 850 mg/dL was supported. K113262 {13} c. Traceability, Stability, Expected values (controls, calibrators, or methods): ## Traceability ACE Cholesterol Reagent is traceable to NIST SRM 1951. The calibrator (GEMCAL) was previously cleared under k931786. ACE HDL-C Reagent is traceable to a CDC certified method. The calibrator (GEMCAL) was previously cleared under k931786. ACE LDL-C Reagent is traceable to the CDC LDL cholesterol reference method. The ACE LDL-C Calibrators were previously cleared under k931733. ACE Triglycerides Reagents is traceable to NIST SRM 1951. The calibrator (GEMCAL) was previously cleared under k931786 ## On-Board stability: The stability protocols and acceptance criteria were reviewed and determined to be adequate. All ACE reagent in this submission demonstrated on-board stability of 30 days. The real-time close-vial stability study protocols and acceptance criteria were reviewed and determined to be adequate. The stability for each reagent is shown in the table below: | Reagent | Closed-Vial Stability (months) | | --- | --- | | Cholesterol | 15 | | HDL-C | 24 | | LDL-C | 24 | | Triglycerides | 15 | ## d. Detection limit: Protocols for the determination of the limit of the blank (LoB) and the limit of detection (LoD) were performed in accordance with the recommendations in the CLSI Guideline EP17-A. Testing was carried out using true blanks and low level samples (total 60 each) over three days on two ACE Axcel Clinical Chemistry Systems. The resulting LoB and LoD for the four measurands are summarized below: | Reagent | Limit of Blank (LoB) | Limit of Detection (LoD) | | --- | --- | --- | | Cholesterol | 2.9 mg/dL | 3.6 mg/dL | | HDL-C | 1.0 mg/dL | 1.5 mg/dL | | LDL-C | 2.7 mg/dL | 3.9 mg/dL | | Triglycerides | 10.4mg/dL | 11.6 mg/dL | K113262 {14} e. Analytical specificity: ACE Cholesterol Reagent Interference studies were performed by using serum pools containing Cholesterol with individual interferents at a range of concentrations. The sera were assayed for Cholesterol (n = 3 replicates) and the mean result calculated. Interference was considered to be significant by the sponsor if the analyte recovery changed by ± 10%. The results reported were obtained on the ACE Axcel Clinical Chemistry System analyzer. | Interferent | Cholesterol Concentrations Tested (mg/dL) | No Significant Interference At or Below: | | --- | --- | --- | | Unconjugated Bilirubin | Low Pool: 182 | 30 mg/dL | | | High Pool: 405 | | | Hemoglobin | Low Pool: 159 | 500 mg/dL | | | High Pool: 445 | | | Triglycerides | Low Pool: 170 | 2180 mg/dL | | | High Pool: 378 | | | Ascorbic Acid | Low Pool: 185 | 6 mg/dL | | | High Pool: 417 | | ACE HDL-C Reagent: Interference studies were performed by using serum pools containing HDL with individual interferents at a range of concentrations. The sera were assayed for HDL (n = 3 replicates) and the mean result calculated. Interference was considered to be significant by the sponsor if the analyte recovery changed by ± 10%. The results reported were obtained on the ACE Axcel Clinical Chemistry System analyzer. | Interferent | HDL Concentrations Tested (mg/dL) | No Significant Interference At or Below: | | --- | --- | --- | | Unconjugated Bilirubin | Low Pool: 43 | 59 mg/dL | | | High Pool: 98 | | | Hemoglobin | Low Pool: 64 | 1000 mg/dL | | | High Pool: 91 | | K113262 {15} | Triglycerides | Low Pool: 30 | 652 mg/dL | | --- | --- | --- | | | High Pool: 101 | | | Ascorbic Acid | Low Pool: 45 | 6 mg/dL | | | High Pool: 105 | | ## ACE LDL-C Reagent: Interference studies were performed by using serum pools containing LDL with individual interferents at a range of concentrations. The sera were assayed for LDL (n = 3 replicates) and the mean result calculated. Interference was considered to be significant by the sponsor if the analyte recovery changed by ± 10%. The results reported were obtained on the ACE Axcel Clinical Chemistry System analyzer. | Interferent | LDL Concentrations Tested (mg/dL) | No Significant Interference At or Below: | | --- | --- | --- | | Unconjugated Bilirubin | Low Pool: 116 | 59 mg/dL | | | High Pool: 374 | | | Hemoglobin | Low Pool: 74 | 1000 mg/dL | | | High Pool: 273 | | | Triglycerides | Low Pool: 113 | 1233 mg/dL | | | High Pool: 352 | | | Ascorbic Acid | Low Pool: 122 | 6 mg/dL | | | High Pool: 396 | | ## ACE Triglycerides Reagent: Interference studies were performed by using serum pools containing Triglycerides with individual interferents at a range of concentrations. The sera were assayed for Triglycerides (n = 3 replicates) and the mean result calculated. Interference was considered to be significant by the sponsor if the analyte recovery changed by ± 10%. The results reported were obtained on the ACE Axcel Clinical Chemistry System analyzer. K113262 {16} | Interferent | Triglycerides Concentrations Tested (mg/dL) | No Significant Interference At or Below: | | --- | --- | --- | | Unconjugated Bilirubin | Low Pool: 78 | 7 mg/dL | | | High Pool: 726 | | | Hemoglobin | Low Pool: 50 | 125 mg/dL | | | High Pool: 270 | | | Ascorbic Acid | Low Pool: 75 | 0.75 mg/dL | | | High Pool: 716 | | The following limitations are stated in the labeling for ACE Triglyceride Reagent: “Use clear, unhemolyzed serum. Hemolyzed and icteric samples cannot be used.” “Advice patients not to take any alcohol or vitamin supplements 24 hours prior to fasting blood work.” f. Assay cut-off: Not applicable g. Cleaning and Disinfection Not applicable. 2. Comparison studies: Studies were carried out according to CLSI EP09-A2-IR. In house: ACE Cholesterol Reagent One hundred ten serum samples were assayed in parallel by both the test and predicate methods. The results were analyzed by using Deming regression. The range tested was 7 to 527 ng/dL. Altered samples were included in the study (no more than 10%). The comparison by Deming regression resulted in a slope of 1.012 (95%CI = 0.999 to 1.026), an intercept of -3.3 (95%CI = -6.2 to -0.5), correlation coefficient (R²) = 0.9977, and a standard error of 5.0. Point of Care Laboratory: ACE Cholesterol Reagent Serum samples were assayed in parallel by both the test and predicate methods at three POC sites. The results were analyzed by using Deming regression. (Some altered samples were used among all three sites.) K113262 {17} | POL | n | Range | Regression Equation | Correlation Coefficient | Standard Error | Confidence Interval Slope | Confidence Interval Intercept | | --- | --- | --- | --- | --- | --- | --- | --- | | 1 | 57 | 22-571 | Y = 1.003x - 2.7 | 0.9995 | 3.0 | 0.995 to 1.012 | -4.6 to -0.7 | | 2 | 60 | 17-515 | Y = 1.017x - 1.9 | 0.9978 | 6.9 | 0.999 to 1.034 | -6.1 to 2.3 | | 3 | 46 | 62-538 | y = 0.996x - 0.3 | 0.9945 | 8.4 | 0.964 to 1.028 | -7.3 to 6.7 | In house: ACE HDL-C Reagent One hundred eight serum samples were assayed in parallel by both the test and predicate methods. The results were analyzed by using Deming regression. The range tested was 8 to 122 ng/dL. Altered samples were included in the study (no more than 10%). The comparison by Deming regression resulted in a slope of 0.972 (95%CI = 0.9564 to 0.989), an intercept of 0.5 (95%CI = -0.4 to 1.5), correlation coefficient (R²) = 0.9957, and a standard error of 1.7. Point of Care Laboratory: ACE HDL-C Reagent Serum samples were assayed in parallel by both the test and predicate methods at three POC sites. The results were analyzed by using Deming regression. Some altered samples were used among all three sites. | Lab | n | Range | Regression Equation | Correlation Coefficient | Standard Error | Confidence Interval Slope | Confidence Interval Intercept | | --- | --- | --- | --- | --- | --- | --- | --- | | POL 1 | 60 | 13-110 | y = 0.959x - 2.6 | 0.9961 | 1.8 | 0.937 to 0.981 | -4.0 to -1.3 | | POL 2 | 56 | 20-110 | y = 0.969x - 1.8 | 0.9964 | 1.7 | 0.947 to 0.991 | -3.1 to -0.4 | | POL 3 | 43 | 21-117 | y = 1.015x - 0.5 | 0.9898 | 2.4 | 0.970 to 1.061 | -3.0 to 2.0 | In house: ACE LDL-C Reagent One hundred eight serum samples were assayed in parallel by both the test and predicate methods and the results were analyzed by using Deming regression. The range tested was 32 to 422 ng/dL. Altered samples were included in the study (no more than 10%). K113262 {18} The comparison by Deming regression resulted in a slope of 0.982 (95%CI = 0.968 to 0.996), an intercept of -1.0 (95%CI = -3.1 to 1.0), correlation coefficient (R²) = 0.9973, and a standard error of 4.7. ## Point of Care Laboratory: ACE LDL-C Reagent Serum samples were assayed in parallel by both the test and predicate methods at three POC sites. The results were analyzed by using Deming regression. Some altered samples were used among all three sites. | POL | N | Range | Regression Equation | Correlation Coefficient | Standard Error | Confidence Interval Slope | Confidence Interval Intercept | | --- | --- | --- | --- | --- | --- | --- | --- | | 1 | 74 | 4-395 | y = 1.012x + 1.4 | 0.9970 | 6.0 | 0.994 to 1.031 | -1.3 to 4.0 | | 2 | 58 | 9-397 | y = 1.010x - 2.5 | 0.9974 | 5.9 | 0.991 to 1.030 | -5.5 to 0.4 | | 3 | 47 | 30-417 | y = 1.037x - 3.2 | 0.9940 | 9.0 | 1.002 to 1.071 | -8.4 to 2.1 | ## In house: ACE Triglycerides Reagent One hundred eleven serum samples were assayed in parallel by both the test and predicate methods and the results compared by Deming regression. The range tested was 24 to 975 µg/dL. Altered samples were included in the study (no more than 10%). The comparison by Deming regression resulted in a slope of 1.031 (95%CI = 1.025 to 1.037), an intercept of -0.8 (95%CI = -2.7 to 1.1), correlation coefficient (R²) = 0.9995, and a standard error of 6.6. ## Point of Care Laboratory: ACE Triglycerides Reagent Serum samples were assayed in parallel by both the test and predicate methods at three POC sites. The results were analyzed by using Deming regression. Some altered samples were used among all three sites. | POL | N | Range | Regression Equation | Correlation Coefficient | Standard Error | Confidence Interval Slope | Confidence Interval Intercept | | --- | --- | --- | --- | --- | --- | --- | --- | | 1 | 54 | 23-976 | y = 1.008x - 1.6 | 0.9996 | 6.6 | 1.000 to 1.016 | -3.9 to 0.7 | | 2 | 60 | 20-910 | y = 0.997x - 4.0 | 0.9996 | 5.9 | 0.989 to 1.004 | -6.3 to -1.7 | | 3 | 47 | 37-978 | y = 1.012x - 2.8 | 0.9992 | 8.4 | 1.000 to 1.024 | -6.3 to 0.6 | K113262 {19} 3. Clinical studies: Not applicable 4. Clinical cut-off: Not applicable 5. Expected values/Reference range: Cholesterol: Desirable: <200 mg/dL, Borderline: 200-239 mg/dL, High: >239 mg/dL) HDL: <40 mg/dL = Low, ≥60 mg/dL = High LDL: Optimal: <100 mg/dL, Near optimal: 100 – 129 mg/dL, Borderline high: 130 – 159 mg/dL, High: 160 – 189, Very high: >189 mg/dL Triglycerides: <150 = normal, 150 – 199 = borderline high, 200-499 = high, ≥500 = very high NCEP, Third Report of National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation and Treatment of High Cholesterol in Adults (Adult Treatment Panel III); Executive Summary. NIH Publication No. 02-5215. National Institutes of Health. Bethesda, Maryland: September 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. K113262 20