CHOLESTEROL: Reagent kit intended for the quantitative determination of Cholesterol in human serum. Cholesterol measurements are used in the diagnosis and treatment of disorders involving excess cholesterol in the blood, of lipid and lipoprotein metabolism disorders.

HDL-Cholesterol: Reagent kit intended for the quantitative determination of high-density lipoprotein in human serum. Measurements are used in the diagnosis and treatment of lipid disorders mellitus), atherosclerosis, and various liver and renal diseases.

LDL-Cholesterol: Reagent kit intended for the quantitative determination of low-density lipoprotein in human serum. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.

TRIGLYCERIDES: Reagent kit intended for the quantitative determination of triglycerides (neutral fat) in human serum. 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.

Device Description

CHOLESTEROL: The Cholesterol Oxidase peroxidase (CHOD-PAP) enzymatic method is used. The cholesterol esterase enzyme catalyzes the hydrolysis of cholesterol and free fatty and free fatty acids. Free cholesterol, including that originally present in the sample, is then oxidized by the enzyme cholesterol oxidase (CHOD) to cholest-4-en-3-one, by using molecular oxygen as the electron acceptor and concurrently producing hydrogen peroxide (H2O2). The H2O2 produced is then used in a subsequent chromogenic oxidative coupling reaction, catalyzed by the enzyme peroxidase, in the presence of a redox indicator system, which leads to the formation of a colored compound, absorbing light at 550 nm. The increase in absorbance is directly proportional to the cholesterol concentration in the sample.

HDL-Cholesterol: The Accelerator Selective Detergent method is applied. The determination of HDL-Cholesterol is based on the following reactions: LDL, VLDL, and chylomicrons are neutralized by the combined action of the enzymes Cholesterol Oxidase, Peroxidase, accelerators and N,N-bis-(4-sulfobutyl)-m-toluidine-disodium (DSBmT). HDL remaining in the sample is disrupted by the action of a selective detergent and cholesterol is converted to △4 Cholestenone by the enzymatic action of Cholesterol Esterase and Cholesterol Oxidase, with the subsequent production of H2O2, which reacts with DSBmT and 4-aminoantipyrine in the presence of Peroxidase to a colored complex that absorbs light at 590 nm. The absorbance measured is proportional to the concentration of HDL-Cholesterol in the sample.

LDL-Cholesterol: The Selective Detergent method is applied. The method is in a two-reagent format and depends on the properties of a unique detergent. The first detergent solubilizes only the non-LDL lipoprotein particles. The cholesterol released is consumed by cholesterol esterase and cholesterol oxidase in a non-color forming reaction. The second detergent solubilizes the remaining LDL particles, and a chromogenic coupler allows for color formation. The enzyme reaction with LDL-Cholesterol in the presence of the coupler at 590 nm produces color that is proportional to the amount of LDL cholesterol present in the sample.

TRIGLYCERIDES: The enzymatic glycerol-3-phosphate-peroxidase (GPO-POD) method is used. The method enzymatically hydrolyzes by lipase to free fatty acids and glycerol is phosphorylated by adenosine triphosphate (ATP) with glycerokinase (GK) to produce glycerol-3-phosphate and adenosine diphosphate (ADP). Glycerol-3-phosphate-oxidase oxidizes glycerol-3-phosphate to dihydroxyacetone phosphate and H2O2. The catalytic action of peroxidase (POD) forms quinoneimine from H202, aminoantipyrine, and Dihydrate (N-Ethyl-N-(2hydroxy-3-sulfopropyl)-m-toluidine (TOOS). The absorption change at 550 nm is proportional to the triglycerides concentration in the sample.

AI/ML Overview

Here's a breakdown of the acceptance criteria and the study information for the Medicon Hellas Cholesterol, HDL-Cholesterol, LDL-Cholesterol, and Triglycerides test systems, based on the provided document:

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are generally established by comparison to legally marketed predicate devices and alignment with clinical laboratory guidelines (CLSI). The document presents a clear comparison in the "Device Comparison Table" sections. For this summary, I'll focus on the key performance indicators for each analyte.

CHOLESTEROL

Acceptance Criteria (Predicate: OLYMPUS CHOLESTEROL REAGENT (K925603))	Reported Device Performance (Medicon Hellas CHOLESTEROL)
Method comparison (correlation to comparator): 1.000	Method comparison (correlation to comparator): 0.9980
Reportable range: 20 to 700 mg/dL	Reportable range: 20 to 700 mg/dL
Sensitivity LoD: 1 mg/dL (Predicate LoQ not defined)	Sensitivity LoD / LoQ: LoD 4.4 / LoQ 4.6 (mg/dL)
Precision (within run & total for all LVs): <= 3%	Precision (within run & total for all LVs): <= 4%
Endogenous Interferences: Hemoglobin: up to 500 mg/dL	Endogenous Interferences: Hemoglobin: up to 500 mg/dL
Endogenous Interferences: Triglycerides: up to 1,000 mg/dL	Endogenous Interferences: Triglycerides: up to 1,500 mg/dL
Calibration frequency: 30 days	Calibration frequency: 14 days
On-board stability: Not defined	On-board stability: 28 days
Specimen Type: Human serum, plasma and urine	Specimen Type: Human serum

HDL-Cholesterol

Acceptance Criteria (Predicate: DIRECT HDL (K981224))	Reported Device Performance (Medicon Hellas HDL-Cholesterol)
Method comparison (correlation to comparator): 1.999 (Typo in document, likely 0.999)	Method comparison (correlation to comparator): 0.997
Reportable range: 5.0 to 221 mg/dL	Reportable range: 6.0 to 200 mg/dL
Sensitivity LoD / LoQ: LoD 2.5 / LoQ 5.0 (mg/dL)	Sensitivity LoD / LoQ: LoD 3.0 / LoQ 5.8 (mg/dL)
Precision (within run & total for all LVs): <= 4.0%	Precision (within run & total for all LVs): <=4.0%
Endogenous Interferences: Hemoglobin: up to 2,000 mg/dL	Endogenous Interferences: Hemoglobin: up to 1,000 mg/dL
Endogenous Interferences: Triglycerides: MDL1 1,000mg/dL & MDL2 2,000mg/dL	Endogenous Interferences: Triglycerides: up to 1,500 mg/dL
Calibration frequency: 28 days	Calibration frequency: 28 days
On-board stability: Not defined	On-board stability: 28 days
Specimen Type: Human serum & plasma	Specimen Type: Human serum

LDL-Cholesterol

Acceptance Criteria (Predicate: DIRECT LDL (K981303))	Reported Device Performance (Medicon Hellas LDL-Cholesterol)
Method comparison (correlation to comparator): 0.960	Method comparison (correlation to comparator): 0.999
Reportable range: 1 to 800 mg/dL	Reportable range: 3 to 800mg/dL
Sensitivity LoD / LoQ: < 10mg/dL	Sensitivity LoD / LoQ: LoD 2 / LoQ 3 mg/dL
Precision (within run & total for all LVs): < 4.0%	Precision (within run & total for all LVs): < 4.0%
Endogenous Interferences: Hemoglobin: up to 500mg/dL	Endogenous Interferences: Hemoglobin: up to 1,000mg/dL
Endogenous Interferences: Triglycerides: up to 1,293 mg/dL	Endogenous Interferences: Triglycerides: up to 1,500 mg/dL
Calibration frequency: 28 days	Calibration frequency: At new lot
On-board stability: Not defined	On-board stability: 28 days
Specimen Type: Human serum & plasma	Specimen Type: Human serum

TRIGLYCERIDES

Acceptance Criteria (Predicate: OLYMPUS TRIGLYCERIDE TEST SYSTEM (K063804))	Reported Device Performance (Medicon Hellas TRIGLYCERIDES)
Method comparison (correlation to comparator): 0.999	Method comparison (correlation to comparator): 0.999
Reportable range: 10 to 1,000mg/dL	Reportable range: 10 to 1,000mg/dL
Sensitivity LoD / LoQ: < 0.31 / 5.0 mg/dL	Sensitivity LoD / LoQ: LoD 5.5 / LoQ 9.7 mg/dL
Precision (within run & total for all LVs): < 5.0%	Precision (within run & total for all LVs): < 4.0%
Endogenous Interferences: Hemoglobin: up to 500mg/dL	Endogenous Interferences: Hemoglobin: up to 400mg/dL
Calibration frequency: 30 days	Calibration frequency: 28 days
On-board stability: 30 days	On-board stability: 28 days
Specimen Type: Human serum, plasma & urine	Specimen Type: Human serum

2. Sample size used for the test set and the data provenance

Accuracy (Method Comparisons):
- A minimum of 75 leftover specimens.
- For the specific analytes:
  - CHOLESTEROL: 93 human serum samples
  - HDL-Cholesterol: 141 human serum samples
  - LDL-Cholesterol: 107 human serum samples
  - TRIGLYCERIDES: 163 human serum samples
- Data Provenance: The document states "left-over specimens," implying retrospective use of clinical samples. The country of origin is not explicitly stated, but the company is Medicon Hellas, S.A. based in Greece, and testing was performed at the company premises.
Precision/Reproducibility:
- Three human serum pools for Cholesterol and Triglycerides.
- Two pools for HDL-Cholesterol.
- Four pools for LDL-Cholesterol.
- Each sample was tested for 20 testing days, two different runs, and two replicate measurements per run (morning and afternoon run), for a total of 80 results per level concentration (e.g., for Cholesterol, 3 pools * 80 results/pool = 240 results).
- Data Provenance: Human serum pools, likely prepared in-house or acquired for the study.
Linearity:
- 11 to 13 levels per analyte, prepared by dilution of a human serum pool.
- Each level was tested in 4 replicates.
- Data Provenance: Human serum pool.
Analytical Specificity / Interference:
- Serum pools at low and high levels of each analyte.
- Each measurement of the blank and the sample containing the interferent was repeated at least 5 times.
- Data Provenance: Serum pools.
Detection Limit:
- LoB: 5 blank serum samples measured in 4 replicates for 3 days (total of 60 measurements).
- LoD: 5 low-level samples measured in 4 replicates for 3 days (total of 60 measurements).
- LoQ: 10 samples that span the low end of linearity, measured 5 times each day for 3 days (total of 150 measurements).
- Data Provenance: Serum samples.
Stability and Calibration Frequency:
- Two fresh serum pools and two serum-based commercial controls.
- Measurements were repeated in triplicate at regular time points.
- Data Provenance: Serum pools and commercial controls.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts

This information is not provided in the document. The ground truth for performance studies like those described (method comparison, precision, linearity, interference, detection limits) for in vitro diagnostic (IVD) devices like these typically involves established reference methods or highly accurate comparative analyzers, rather than expert human interpretation of results. The document states that the performance of the Medicon Hellas reagents was compared with "comparator methods" (Beckman Coulter reagents on AU400, Abbott Diagnostics reagents on Architect c8000), which serve as the reference for ground truth in these types of analytical performance studies. The qualifications of the operators performing these studies are not specified.

4. Adjudication method for the test set

This information is not applicable and therefore not provided. Adjudication methods (e.g., 2+1, 3+1) are typically used in studies where human interpretation of complex data (like medical images) is involved and a consensus is needed to establish ground truth. For quantitative chemical assays, the ground truth is established by the highly precise and accurate measurement of reference methods or predicate devices.

5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

This information is not applicable and therefore not provided. MRMC studies are specific to evaluating diagnostic devices where human readers interpret medical cases, often with and without AI assistance (e.g., radiology AI). The Medicon Hellas devices are in vitro diagnostic reagents for quantitative chemical measurements in serum, not image-based diagnostic tools that require human "readers" in the context of an MRMC study.

6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done

This concept is not applicable in the traditional sense for these in vitro diagnostic reagents. These devices are chemical assays that produce a quantitative numerical output (e.g., cholesterol level in mg/dL). There isn't an "algorithm" making a diagnostic interpretation independently in the way AI software would. The device is the standalone measurement system. Its performance is evaluated independently through analytical studies (precision, linearity, accuracy against reference methods, etc.). The results are then read and interpreted by a human clinician.

7. The type of ground truth used

The ground truth for the analytical performance studies (precision, linearity, interference, detection limits, and method comparison) was established against:

Reference Methods/Predicate Devices: For method comparison, the device's performance was compared against established comparator methods (Beckman Coulter reagents on AU400 and Abbott Diagnostics reagents on ABBOTT Architect c8000). The document explicitly states these as the comparators.
A Priori Values/Established Standards: For linearity, precision, and detection limits, the ground truth is based on the known concentrations of prepared samples (e.g., serially diluted pools, spiked samples, blank serum) and statistical analysis according to CLSI guidelines.
Traceability to Reference Methods/Materials: For Cholesterol and Triglycerides, traceability is to Gas-chromatography-isotope dilution mass spectrometry (GC-IDMS). For HDL-Cholesterol and LDL-Cholesterol, traceability is to the Abell-Kendall (AK) reference method.

8. The sample size for the training set

This information is not applicable and therefore not provided. These are chemical reagent kits, not machine learning (AI/ML) models that require a "training set" in the computational sense. The development of such reagents involves chemical and enzymatic research and optimization, often tested with various batches and concentrations of samples during R&D. The studies described in this document are for validation and verification of the final device, not for "training" an algorithm.

9. How the ground truth for the training set was established

As noted above, the concept of a "training set" with ground truth established in the AI/ML sense is not applicable to these chemical reagent devices. The "ground truth" for evaluating the analytical performance of the developed reagent kits is established through the reference methods and standardized protocols described in section 7.

Summary

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

August 9, 2024

Medicon Hellas, S.A. Sotia Mitropoulou Regulatory Affairs Manager Melitona 5-7, 153 44 Gerakas Greece

Re: K232404

Trade/Device Name: CHOLESTEROL: HDL-Cholesterol: LDL-Cholesterol: TRIGLYCERIDES Regulation Number: 21 CFR 862.1175 Regulation Name: Cholesterol (Total) Test System Regulatory Class: Class I, meets the limitations of exemptions 21 CFR 862.9(c)(4) Product Code: CHH, LBS, MRR, CDT Dated: July 9, 2024 Received: July 10, 2024

Dear Sotia Mitropoulou:

We have reviewed your section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (the Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database available at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

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Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review. the OS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Paula V. Caposino -S

Paula Caposino, Ph.D. Deputy Division Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

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DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

Indications for Use

Form Approved: OMB No. 0910-0120 Expiration Date: 07/31/2026 See PRA Statement below.

510(k) Number (if known)

Device Name

CHOLESTEROL, HDL-Cholesterol, LDL-Cholesterol, TRIGL YCERIDES

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)

☑ Prescription Use (Part 21 CFR 801 Subpart D)	☐ Over-The-Counter Use (21 CFR 801 Subpart C)
--------------------------------------------------------------------------	-------------------------------------------------------------------------

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

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Image /page/4/Picture/0 description: The image shows the logo for "medicon MEDICON HELLAS S.A.". The word "medicon" is in red, and there is a blue line underneath it. The words "MEDICON HELLAS S.A." are in a smaller font and are also in red. The logo is simple and modern.

510(k) Summary

510(k) Number: K232404

This 510(k) safety and effectiveness summary information is submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

807.92 (a)(1): Name: MEDICON HELLAS S.A. Address: Melitona 5-7, 153 44 Gerakas, Greece Phone: +302.1.0660.6000 FAX: +302.1.0661.2666 Contact: Ms. Sotia Mitropoulou Email: smitropo@mediconsa.com US Agent: Diatron US, Inc. 1.833.228.7931

Mr. Frank Matuszak

frank.matuszak(@diatron.com

All Supplemental testing was performed at the company premises, mentioned above.

807.92 (a)(2): Device name- trade name and common name, and classification

Trade Name: CHOLESTEROL, HDL-Cholesterol, LDL-Cholesterol, and TRIGLYCERIDES.

Common Name: test systems for Cholesterol, HDL, LDL, and Triglycerides for testing human serum.

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Classification Name(s):

21 CFR § 862.1175 - Cholesterol Test system - Product Code CHH

21 CFR § 862.1475 - HDL-Cholesterol Test system - Product Code LBS

21 CFR § 862.1475 - LDL-Cholesterol Test system - Product Code MRR

21 CFR § 862.1705 - Triglycerides Test system - Product Code CDT

807.92 (a)(3): Identification of the legally marketed predicate devices

Cholesterol - OLYMPUS CHOLESTEROL REAGENT (K925603)

HDL-Cholesterol - DIRECT HDL (K981224)

LDL-Cholesterol - DIRECT LDL (K981303)

Triglycerides - OLYMPUS TRIGLYCERIDE TEST SYSTEM (K063804)

Intended Use/Indications for Use:

A) Intended Use(s):

See Indications for Use below.

B) Indications for Use:

HDL-Cholesterol: Reagent kit intended for the quantitative determination of high-density lipoprotein in human serum. Measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.

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C) Special Conditions for Use Statement(s):

Rx - For Prescription Use Only

D) Special Instrument Requirements:

Diatron Pictus®500 Analyzers

807.92 (a)(4): Device Description

CHOLESTEROL:

BEF 1419-0042 Packaging: 6 x 60 mL

Reagent Composition:
4-Chlorophenol	4.7 mM
4-Aminoantipyrine	1 mM
Cholesterol esterase (CHE)	≥ 500 U/L
Cholesterol oxidase (CHOD)	≥ 500 U/L
Peroxidase (POD)	≥ 1000 U/L

The Cholesterol Oxidase peroxidase (CHOD-PAP) enzymatic method is used. The cholesterol esterase enzyme catalyzes the hydrolysis of cholesterol and free fatty and free fatty acids. Free cholesterol, including that originally present in the sample, is then oxidized by the enzyme cholesterol oxidase (CHOD) to cholest-4-en-3-one, by using molecular oxygen as the electron acceptor and concurrently producing hydrogen peroxide (H2O2). The H2O2 produced is then used in a subsequent chromogenic oxidative coupling reaction, catalyzed by the enzyme peroxidase, in the presence of a redox indicator system, which leads to the formation of a colored compound, absorbing light at 550 nm. The increase in absorbance is directly proportional to the cholesterol concentration in the sample.

HDL-Cholesterol:

1419-0240 Packaging: 6 x 13.5 mL (R1) + 6 x 4.5 mL (R2) REF

REF 1419-0242 Packaging: 6 x 35.1 mL (R1) + 6 x 11.7 mL (R2)

Reagent Composition

Reagent 1 (R1)	Reagent 2 (R2)
Cholesterol oxidase: <1000 U/L	Cholesterol esterase: <1500 U/L
Peroxidase: <1300 U/L	4-aminoantipyrine: <1mM
Ascorbate oxidase: <3000U/L	Non-reactive ingredients, preservative
Accelerator: <1mM
DSBmT: <1mM
Non-reactive ingredients, preservative

The Accelerator Selective Detergent method is applied. The determination of HDL-Cholesterol is based on the following reactions: LDL, VLDL, and chylomicrons are neutralized by the combined action of the enzymes Cholesterol Oxidase, Peroxidase, accelerators and N,N-bis-(4-sulfobutyl)-m-toluidine-disodium (DSBmT). HDL remaining in the sample is disrupted by the action of a selective detergent and cholesterol is converted to △4 Cholestenone by the enzymatic action of Cholesterol Esterase and Cholesterol Oxidase, with the subsequent production of H2O2, which reacts with DSBmT and 4-aminoantipyrine in the presence of Peroxidase to a colored complex that absorbs light at 590 nm. The absorbance measured is proportional to the concentration of HDL-Cholesterol in the sample.

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LDL-Cholesterol

国 1419-0220 Packaging: 6 x 13.5 mL (R1) + 6 x 4.5 mL (R2)
Packaging: 6 x 35.1 mL (R1) + 6 x 11.7 mL (R2) REF] 1419-0222

Reagent Composition:

Reagent 1 (R1)	Reagent 2 (R2)
Detergent 1 <1%	Detergent 2 <1%
4-Aminoantipyrine <0.1%	DSBmT <1 mM
Cholesterol Oxidase (CHO) <1500 U/L	Non-reactive ingredients, preservative
Cholesterol Esterase (CHE) <2500 U/L
Peroxidase (POD) <1300 U/L
Ascorbic oxidase <3000 U/L
Non-reactive ingredients, preservative

The Selective Detergent method is applied. The method is in a two-reagent format and depends on the properties of a unique detergent. The first detergent solubilizes only the non-LDL lipoprotein particles. The cholesterol released is consumed by cholesterol esterase and cholesterol oxidase in a non-color forming reaction. The second detergent solubilizes the remaining LDL particles, and a chromogenic coupler allows for color formation. The enzyme reaction with LDL-Cholesterol in the presence of the coupler at 590 nm produces color that is proportional to the amount of LDL cholesterol present in the sample.

TRIGLYCERIDES:

REF] 1419-0068 Packaging: 6 x 30 mL (R1) + 6 x 4 mL (R2)
Reagent Composition:

Reagent 1 (R1)	Reagent 2 (R2)
Pipes buffer (pH: 6.8): 240 mM	4-Aminoantipyrine: < 15 mM
Peroxidase: > 5000 U/L	GPO: > 55000 U/L
Glycerokinase: > 1000 U/L	Non-reactive ingredients, preservative
Lipoprotein Lipase: > 15000 U/L
ATP: 4.5 mM
TOOS: 4.8 mM
Non-reactive ingredients, preservative

The enzymatic glycerol-3-phosphate-peroxidase (GPO-POD) method is used. The method enzymatically hydrolyzes by lipase to free fatty acids and glycerol is phosphorylated by adenosine triphosphate (ATP) with glycerokinase (GK) to produce glycerol-3-phosphate and adenosine diphosphate (ADP). Glycerol-3-phosphate-oxidase oxidizes glycerol-3-phosphate to dihydroxyacetone phosphate and H2O2. The catalytic action of peroxidase (POD) forms quinoneimine from H202, aminoantipyrine, and Dihydrate (N-Ethyl-N-(2hydroxy-3-sulfopropyl)-m-toluidine (TOOS). The absorption change at 550 nm is proportional to the triglycerides concentration in the sample.

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807.92 (a)(5): Intended Use -

CHOLESTEROL: Reagent kit intended for the quantitative determination of Cholesterol in human serum. Cholesterol measurements are used in the diagnosis and treatment of disorders involving excess cholesterol in the blood, of lipid and lipoprotein metabolism disorders.
HDL-Cholesterol: Reagent kit intended for the quantitative determination of high-density lipoprotein in human serum. Measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.
LDL-Cholesterol: Reagent kit intended for the quantitative determination of low-density lipoprotein in human serum. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.
TRIGLYCERIDES: Reagent kit intended for the quantitative determination of triglycerides (neutral fat) in human serum. 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.

807.92 (a)(6): Technological Similarities and Differences to the Predicates

See Comparison with Predicate(s), pg. 6.

807.92 (b)(1,2): Brief Description of Nonclinical Data - Clinical Studies data does not apply

Medicon Hellas (Medicon) has performed a series of studies to confirm the substantial equivalence of their test systems against the legally marketed predicate devices mentioned above, by demonstrating the candidate devices performance against the following current laboratory methods.

Several candidate-reagent lots were used for the studies. These covered early life, mid-life reagents when available (but always included three lots) as well as "end of on-board life". Studies confirmed equivalent performance of the candidate systems during their noted life cycle stages in order to support total shelf-life assignment.

Studies included accuracy (method comparison), reportable range (linearity), precision (reproducibility, between run and within lab [total]) and interfering substances. The following tables provide summary results.

Accuracy (Method Comparisons) - A minimum of 75 left-over specimens, spanning the dynamic ranges, were assayed in singleton and in a blinded fashion on the candidate and predicate systems. Results were tabulated and evaluated using Analyse-it statistics calculator software to generate Passing-Bablok regression statistics. Accuracy statistics are reported as Passing-Bablok regression statistics.

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Substantial Equivalence Information:

A. Predicate Device Name(s):

OLYMPUS CHOLESTEROL REAGENT (K925603) DIRECT HDL (K981224) DIRECT LDL (K981303) OLYMPUS TRIGLYCERIDE TEST SYSTEM (K063804)

B. Predicate 510(k) Numbers:

K925603, K981224, K981303, and K063804

C. Comparison with Predicate(s):

Device Comparison Table: CHOLESTEROL

Reagent	CHOLESTEROL	OLYMPUSCHOLESTEROLREAGENT (K925603)
Classification	21 CFR § 862.1175, CHH	Same
Method comparison (correlation to comparator)	0.9980	1.000
Reportable range	20 to 700 mg/dL	20 to 700 mg/dL
Sensitivity LoD / LoQ	LoD 4.4 / LoQ 4.6 (mg/dL)	LoD 1mg/dL / Not defined
Precision (within run & total for all LVs)	<= 4%	<= 3%
Endogenous Interferences - insignificant up to noted concentrations
Ascorbate	up to 5.25 mg/dL	up to 3 mg/dL
Conjugated Bilirubin	up to 40 mg/dL	Not listed / Not defined
Unconj. Bilirubin	up to 40 mg/dL	up to 8 mg/dL
Hemoglobin	up to 500 mg/dL	up to 500 mg/dL
Triglycerides	up to 1,500 mg/dL	up to 1,000 mg/dL
Exogenous Interferences	See pg. 12	Not listed / Not defined
Calibration frequency / On-board stability	14 days / 28 days	30 days / Not defined days
Intended Use	Quantitative determinationof Cholesterol	Quantitative measurement ofcholesterol
Testing Environment	Clinical labs	Same
Method Principle	Photometry	Same
Specimen Type	Human serum	Human serum, plasma andurine

{10}------------------------------------------------

Device Comparison Table: HDL-Cholesterol

Reagent	HDL-Cholesterol	DIRECT HDL(K981224)
Classification	21 CFR § 862.1475, LBS	Same
Method comparison (correlation to comparator)	0.997	1.999
Reportable range	6.0 to 200 mg/dL	5.0 to 221 mg/dL
Sensitivity LoD / LoQ	LoD 3.0 / LoQ 5.8 (mg/dL)	LoD 2.5 / LoQ 5.0 (mg/dL)
Precision (within run & total for all LVs)	<=4.0%	<=4.0%
Endogenous Interferences - insignificant up to noted concentrations
Ascorbate	up to 5.25 mg/dL	up to 3.9 mg/dL
Conjugated Bilirubin	up to 40 mg/dL	MDL1 32.4mg/dL & MDL267.1mg/dL
Unconj. Bilirubin	up to 40 mg/dL	up to 8 mg/dL
Hemoglobin	up to 1,000mg/dL	up to 2,000mg/dL
Triglycerides	up to 1,500 mg/dL	MDL1 1,000mg/dL &MDL2 2,000mg/dL
Exogenous Interferences	See pg. 13	Not listed / Not defined
Calibration frequency / On-board stability	14 days / 28 days	28 days / Not defined days
Intended Use	Quantitative determinationof HDL-Cholesterol	Quantitative measurement ofHDL-Cholesterol
Testing Environment	Clinical labs	Same
Method Principle	Photometry	Same
Specimen Type	Human serum	Human serum & plasma

{11}------------------------------------------------

Device Comparison Table: LDL-Cholesterol

Reagent	LDL-Cholesterol	DIRECT LDL(K981303)
Classification	21 CFR § 862.1475, MRR	Same
Method comparison (correlation to comparator)	0.999	0.960
Reportable range	3 to 800mg/dL	1 to 800mg/dL
Sensitivity LoD / LoQ	LoD 2 / LoQ 3 mg/dL	< 10mg/dL
Precision (within run & total for all LVs	< 4.0%	< 4.0%
Endogenous Interferences - insignificant up to noted concentrations
Conjugated Bilirubin	up to 40 mg/dL	up to 20mg/dL
Unconj. Bilirubin	up to 40 mg/dL	up to 20mg/dL
Hemoglobin	up to 1,000mg/dL	up to 500mg/dL
Triglycerides	up to 1,500 mg/dL	up to 1,293 mg/dL
Exogenous Interferences	See pg. 14	Not listed / Not defined
Acetaminophen	Tested up to 15.6mg/dL	20.0mg/dL
Dipyrone	Tested up to 3.3mg/dL	10mg/dL
N-Acetyl-L-Cysteine	Tested up to 15mg/dL	160mg/dL
Calibration frequency / On-board stability	new lot / 28 days	28 days / not defined
Intended Use	Quantitative determination of LDL-Cholesterol	Quantitative measurement of LDL-Cholesterol
Testing Environment	Clinical labs	Same
Method Principle	Photometry	Same
Specimen Type	Human serum	Human serum & plasma

{12}------------------------------------------------

Device Comparison Table: TRIGLYCERIDES

Reagent	TRIGLYCERIDES	OLYMPUSTRIGLYCERIDE TESTSYSTEM (K063804)
Classification	21 CFR § 862.1705, CDT	Same
Method comparison (correlation to comparator)	0.999	0.999
Reportable range	10 to 1,000mg/dL	10 to 1,000mg/dL
Sensitivity LoD / LoQ	LoD 5.5 / LoQ 9.7 mg/dL	< 0.31 / 5.0 mg/dL
Precision (within run & total for all LVs	< 4.0%	< 5.0%
Endogenous Interferences - insignificant up to noted concentrations
Conjugated Bilirubin	up to 40 mg/dL	Not listed / Not defined
Unconj. Bilirubin	up to 40 mg/dL	up to 40mg/dL
Hemoglobin	up to 400mg/dL	up to 500mg/dL
Exogenous Interferences	See pg. 15	Not listed / Not defined
Calibration frequency / On-board stability	28 days / 28 days	30 days / 30 days
Intended Use	Quantitative determinationof triglycerides	Quantitative measurement oftriglycerides
Testing Environment	Clinical labs	Same
Method Principle	Photometry	Same
Specimen Type	Human serum	Human serum, plasma &urine

Standards/Guidance Documents Referenced:

CLSI EP05-A3 - Evaluation of Precision Performance of Quantitative Measurement Procedures, 3rd ed. 2014

CLSI EP06-Ed02 - Evaluation of Linearity of Quantitative Measurement Procedures, 2nd ed., Nov.2020.

CLSI EP07 - Interference Testing in Clinical Chemistry, 3rd ed. 2018.

CLSI EP09c - Measurement Procedure Comparison and Bias Estimation Using Patient Samples. 3rd ed. 2018.

CLSI EP15-A3 – User Verification of Precision and Estimation of Bias. 3rd ed. 2014.

CLSI EP17-A2 - Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures, Approved guideline, 2nd ed. 2012.

CLSI EP37 - Supplemental Tables for Interference Testing in Clinical Chemistry, 1st ed., 2018

{13}------------------------------------------------

1. Analytical Performance:

Precision/Reproducibility: a)

Precision studies were conducted according to the CLSI EP05-A3 guideline.

A precision evaluation study procedure was applied for one site and one analyzer in a standard 20x2x2 experiment. Three human serum pools were tested for CHOLESTEROL and TRIGLYCERIDES, two pools for HDL-Cholesterol and four pools for LDL-Cholesterol, near the Medical Decision Limits, for every analyte. The protocol uses measurements of each sample for 20 testing days, two different runs, and two replicate measurements per run (morning and afternoon run) for a total of 80 results per level concentration. When results were collected, analysis of the data was done using the 2-way factor nested ANOVA model. From the above analysis were calculated the mean, the standard deviation, the coefficient of variation (%CV) for repeatability (within-run), between run precision and total precision for each reagent Lot of Cholesterol reagent.

The Within-run precision, Between Day and Total precision results for a representative reagent lot for each analyte are presented in the tables below.

CHOLESTEROL		Mean	Repeatability	Between Run	Between Day	Total precision
Sample	N	(mg/dL)	%CV	%CV	%CV	%CV
Serum	80	92	1.6	0.0	1.6	2.2
Pool	80	241	1.4	2.0	1.5	2.8
	80	345	1.4	0.8	2.5	3.0

HDL-Cholesterol		Mean	Repeatability	Between Run	Between Day	Total precision
Sample	N	(mg/dL)	%CV	%CV	%CV	%CV
Serum	80	31	2.3	2.0	2.3	3.8
Pool	80	57	1.2	0.8	2.2	2.6

	LDL-Cholesterol	Mean	Repeatability	Between Run	Between Day	Total precision
Sample	N	(mg/dL)	%CV	%CV	%CV	%CV
SerumPool	80	98	1.0	1.2	3.6	3.9
SerumPool	80	126	1.8	2.3	1.9	3.5
SerumPool	80	156	1.1	1.5	3.1	3.6
SerumPool	80	193	1.1	1.6	2.5	3.2

TRIGLYCERIDES		Mean	Repeatability	Between Run	Between Day	Total precision
Sample	N	(mg/dL)	%CV	%CV	%CV	%CV
Serum	80	41	1.0	1.6	0.9	2.1
Pool	80	148	1.3	2.1	1.8	3.0
	80	399	0.7	1.3	1.0	1.8

{14}------------------------------------------------

b) Linearity:

Linearity studies were conducted according to the CLSI EP06-EdA2 guideline.

For each analyte 11 to 13 levels were prepared by dilution of a human serum pool with a concentration of analyte higher than the claimed upper limit of linearity. The human serum high pool was prepared from a human base pool after spiking with a purified concentrated analyte. The diluent it was a delipidized commercial preparation free of each analyte. The samples were assigned their reference values arithmetically from serial dilutions of the high-level sample. Each level was tested in 4 replicates using 1 instrument, and 3 Lots of reagents. Sample included target MDLs within the range tested. To validate the linearity a Least Square Linear Regression analysis was performed with intercept in the model. Statistics were calculated using Analyse-it standard algorithms. Levels below and above the defined reportable range limits were included. Results for a representative reagent lot for are presented in the tables below.

Device	SampleMatrix	ClaimedMeasurementRange	Slope	Intercept	R2
CHOLESTEROL	Serum	20 – 700 mg/dL	1.001	0.409	0.9997
HDL-Cholesterol	Serum	6 – 200 mg/dL	1.033	-0.668	0.9984
LDL-Cholesterol	Serum	3 – 800 mg/dL	1.099	-0.40	0.9996
TRIGLYCERIDES	Serum	10 – 1000 mg/dL	0.978	-0.236	0.9994

c) Analytical Specificity / Interference:

Interference studies were conducted according to the CLSI EP07-A3 guideline.

The study measured the effect of endogenous substances to act as interferents at recommended test level (CLSI EP37. Table 2) on two concentration levels of analyte (CLSI EP07. Table A1). The study measured also the effect of a number of exogenous substances (CLSI EP37, Table 1) which when present might interfere the final test reported by Medicon candidate reagents. Again, as previously noted the effect on 2 concentration levels was tested as per indicated in CLSI EP07 (Table A1). Serum pools at low and high levels of each analyte were prepared, then spiked with the interference compounds. The results define the level at which interferences of the noted compounds will affect results by more than +/- 10%.

Endogenous interference procedure

Low and high levels of each analyte were spiked with a recommended concentration of endogenous substance. The recommended concentration for endogenous substance which was withdrawn from CLSI EP37 procedure.

Interferent	Highest recommended conc.
Hemoglobin	1000 mg/dL
Conjugated Bilirubin	40 mg/dL
Unconjugated Bilirubin	40 mg/dL
Triglycerides	1500 mg/dL

In each case, the mean value of each spiked sample was compared to its neat (zero interferent) mean and % recovered values were calculated. Each measurement of the blank and the relative sample containing the interferent was repeated at least 5 times and means are compared. When interference was found for any tested compound(s), dose response studies were conducted to define the highest concentration which showed a deviation from control value lower than 10%.

{15}------------------------------------------------

Exogenous interference procedure

To assess the effect observed by the exogenous substance on two levels concentrations of the analyte in question (low and high), two different drug concentrations were studied. These are the highest drug concentration under the therapeutic treatment and the recommended test concentration in the sample as proposed by CLSI EP-37 protocol or from relative bibliographic data. Each measurement of the blank and the relative sample containing the interferent was repeated at least 5 times and means are compared. No interference was confirmed when the observed mean in the presence of interferent was within 10% of the blank mean. When interference was found for any tested compound(s), dose response studies were conducted to define interference limits for the drugs.

The following provides endogenous and exogenous study results for each reagent. In the following tables were summarized the highest concentrations for each interferent that that did not cause significant interference.

Sample Matrix	Interferent	Highest concentration tested that did notcause significant interference
Serum	Hemoglobin	500 mg/dL
	Conjugated Bilirubin	40 mg/dL
	Unconjugated Bilirubin	40 mg/dL
	Triglycerides	1500 mg/dL
	Acetaminophen	15.6 mg/dL
	Acetylsalicylic Acid	3 mg/dL
	Ampicillin	7.5 mg/dL
	Ascorbic Acid	5.25 mg/dL
	Atorvastatin	0.075 mg/dL
	Cefotaxime	52.8 mg/dL
	Cefoxitin	660 mg/dL
	Cyclosporine	0.18 mg/dL
	Dipyrone	3.3 mg/dL
	Dobesilate Calcium	6 mg/dL
	Dobutamine	0.121 mg/dL
	Doxycycline	1.8 mg/dL
	Fenofibrate	4.5 mg/dL
	Heparin	330 U/dL
	Ibuprofen	21.9 mg/dL
	Intralipid	2000 mg/dL
	Levodopa	0.75 mg/dL
	Methotrexate	136 mg/dL
	Methyldopa	2.25 mg/dL
	Metronidazole	12.3 mg/dL
	N-Acetyl-Cysteine	15 mg/dL
	Phenylbutazone	32.1 mg/dL
	Pravastatin	0.0207 mg/dL
	Rifampicin	4.8 mg/dL
	Rosuvastatin	0.0111 mg/dL
	Simvastatin	0.168 mg/dL
	Theophylline	6 mg/dL

CHOLESTEROL:

{16}------------------------------------------------

HDL-Cholesterol:

Sample Matrix	Interferent	Highest concentration tested that did notcause significant interference
Serum	Hemoglobin	1000 mg/dL
	Conjugated Bilirubin	40 mg/dL
	Unconjugated Bilirubin	40 mg/dL
	Triglycerides	1500 mg/dL
	Acetaminophen	15.6 mg/dL
	Acetylsalicylic Acid	3 mg/dL
	Ampicillin	7.5 mg/dL
	Ascorbic Acid	5.25 mg/dL
	Atorvastatin	0.075 mg/dL
	Cefotaxime	52.8 mg/dL
	Cefoxitin	660 mg/dL
	Cyclosporine	0.18 mg/dL
	Dipyrone	3.3 mg/dL
	Dobesilate Calcium	6 mg/dL
	Dobutamine	0.121 mg/dL
	Doxycycline	1.8 mg/dL
	Fenofibrate	4.5 mg/dL
	Heparin	330 U/dL
	Ibuprofen	21.9 mg/dL
	Intralipid	2000 mg/dL
	Levodopa	0.75 mg/dL
	Methotrexate	136 mg/dL
	Methyldopa	1.35 mg/dL
	Metronidazole	12.3 mg/dL
	N-Acetyl-Cysteine	15 mg/dL
	Phenylbutazone	32.1 mg/dL
	Pravastatin	0.0207 mg/dL
	Rifampicin	4.8 mg/dL
	Rosuvastatin	0.0111 mg/dL
	Simvastatin	0.168 mg/dL
	Theophylline	6 mg/dL

{17}------------------------------------------------

LDL-Cholesterol:

Sample Matrix	Interferent	Highest concentration tested that did notcause significant interference
Serum	Hemoglobin	1000 mg/dL
	Conjugated Bilirubin	40 mg/dL
	Unconjugated Bilirubin	40 mg/dL
	Triglycerides	1500 mg/dL
	Acetaminophen	15.6 mg/dL
	Acetylsalicylic Acid	3 mg/dL
	Ampicillin	7.5 mg/dL
	Ascorbic Acid	5.25 mg/dL
	Atorvastatin	0.075 mg/dL
	Cefotaxime	52.8 mg/dL
	Cefoxitin	660 mg/dL
	Cyclosporine	0.18 mg/dL
	Dipyrone	3.3 mg/dL
	Dobesilate Calcium	6 mg/dL
	Dobutamine	0.121 mg/dL
	Doxycycline	1.8 mg/dL
	Fenofibrate	4.5 mg/dL
	Heparin	330 U/dL
	Ibuprofen	21.9 mg/dL
	Intralipid	2000 mg/dL
	Levodopa	0.75 mg/dL
	Methotrexate	136 mg/dL
	Methyldopa	2.25 mg/dL
	Metronidazole	12.3 mg/dL
	N-Acetyl-Cysteine	15 mg/dL
	Phenylbutazone	32.1 mg/dL
	Pravastatin	0.0207 mg/dL
	Rifampicin	4.8 mg/dL
	Rosuvastatin	0.0111 mg/dL
	Simvastatin	0.168 mg/dL
	Theophylline	6 mg/dL

{18}------------------------------------------------

TRIGLYCERIDES:

Sample Matrix	Interferent	Highest concentration tested that did not cause significant interference
Serum	Hemoglobin	400 mg/dL
	Conjugated Bilirubin	40 mg/dL
	Unconjugated Bilirubin	40 mg/dL
	Acetaminophen	15.6 mg/dL
	Acetylsalicylic Acid	3 mg/dL
	Ampicillin	7.5 mg/dL
	Ascorbic Acid	5.25 mg/dL
	Atorvastatin	0.075 mg/dL
	Cefotaxime	52.8 mg/dL
	Cefoxitin	660 mg/dL
	Cyclosporine	0.18 mg/dL
	Dipyrone	3.3 mg/dL
	Dobesilate Calcium	6 mg/dL
	Dobutamine	0.121 mg/dL
	Doxycycline	1.8 mg/dL
	Fenofibrate	4.5 mg/dL
	Heparin	330 U/dL
	Ibuprofen	21.9 mg/dL
	Intralipid	2000 mg/dL
	Levodopa	0.75 mg/dL
	Methotrexate	136 mg/dL
	Methyldopa	2.25 mg/dL
	Metronidazole	12.3 mg/dL
	N-Acetyl-Cysteine	15 mg/dL
	Phenylbutazone	32.1 mg/dL
	Pravastatin	0.0207 mg/dL
	Rifampicin	4.8 mg/dL
	Rosuvastatin	0.0111 mg/dL
	Simvastatin	0.168 mg/dL
	Theophylline	6 mg/dL

d) Assay Reportable Range:

Device	SampleMatrix	ClaimedMeasurementRange
CHOLESTEROL	Serum	20 - 700 mg/dL
HDL -Cholesterol	Serum	6 - 200 mg/dL
LDL - Cholesterol	Serum	3 - 800 mg/dL
TRIGLYCERIDES	Serum	10 - 1000 mg/dL

{19}------------------------------------------------

e) Traceability, Stability, Expected Values (Controls, Calibrators or Methods):

Device	Reference Method/Reference Materials
CHOLESTEROL	Gas-chromatography-isotope dilution mass spectrometry (GC-IDMS) referencemethod
HDL-Cholesterol	Abell-Kendall (AK) reference method
LDL-Cholesterol	Abell-Kendall (AK) reference method
TRIGLYCERIDES	Gas-chromatography-isotope dilution mass spectrometry (GC-IDMS) referencemethod

The Medicon reagents are traceable to the following reference materials:

f) Detection Limit:

The Limit of Blank (LoB), Limit of Detection (LoD) and Limit of Quantitation studies were performed according to CLSI EP17-A2 guideline.

LoB: For the Limit of Blank study 5 blank serum were measured in 4 replicates for 3 days for a total of 60 measurements. Each set of measurements was performed with 3 reagent Lots. LoB was estimated using nonparametric analysis of the data.

LoD: For the Limit of Detection study, 5 low level samples were measured in 4 replicates for 3 days for a total of 60 measurements. Each set of measurements was performed with 3 reagent Lots. LoD was calculated from the formula LoD= LoB+1.654*SD.

LoQ: The LoQ was calculated using a precision profile approach according to CLSI EP17-A2 procedure. For the determination of LoQ, 10 samples that span the low end of linearity were measured 5 times each day for a total of 150 measurements in 3 days with 3 Reagent Lots. The LoO level was determined from the concentration that generated < 20% CV.

The results are summarized in the table below:

Device	Sample Matrix	LoB (mg/dL)	LoD(mg/dL)	LoQ(mg/dL)
CHOLESTEROL	Serum	2.5	4.4	4.6
HDL-Cholesterol	Serum	1.0	3.0	5.8
LDL-Cholesterol	Serum	1.0	2.0	3.0
TRIGLYCERIDES	Serum	4.0	5.5	9.7

g) Assay Cut-Off:

Not applicable.

{20}------------------------------------------------

h) Stability and Calibration frequency and shelf-life confirmation;

Calibration frequency / on-board stability -

For the On-Board Stability and Calibration Frequency study, two (2) fresh serum pools with known analyte concentrations, and two (2) serum based commercial controls were analyzed on the Pictus 500 analyzer.

Reagents were loaded on to the Pictus P500 analyzer's cooled reagent was calibrated at the start of each study. During the above stability studies, reagents were calibrated and 2 controls were measured in triplicate. Then the reagents were remained on the analyzer. The measurements were repeated in triplicate at regular time points to cover the claimed shelf life of the reagent (plus a 10% of this).

Reagent is calibrated at fixed time points according to calibration frequency mentioned in the relevant IFU the measured values at a time point should not exceed the 10% of initial value of the calculated mean for any concentration.

On Board stability /Calibration frequency: The un-capped on-board reagent stability claim is defined as the number of days between the start testing date and the last test day that results were within the acceptance limits. Calloration frequency is defined as the required time interval between calibrations to ensure reagent recovers values within the acceptance criteria. Limits are defined in each device's IFU and were followed during all studies.

Results were as follows:

Device	On-Board Stability (days)	Calibration Frequency (days)
CHOLESTEROL	28	14
HDL-Cholesterol	28	28
LDL-Cholesterol	28	At new lot
TRIGLYCERIDES	28	28

{21}------------------------------------------------

2. Comparison Studies:

Method Comparison with Predicate Device: a.

Performance of the CHOLESTEROL and TRIGLYCERIDES reagents for serum assayed on the Pictus P500 analyzer was compared with the comparator methods, Beckman Coulter reagents assayed on the Beckman Coulter AU400 analyzer. Performance of the HDL-Cholesterol and LDL-Cholesterol reagents assayed on the Pictus P500 analyzer was compared with the comparator methods, Abbott Diagnostics reagents on the ABBOTT Architect c8000 analyzer.

A minimum of 75 left over specimens, spanning the dynamic ranges, were assayed in singleton and in a blinded fashion on the candidate and predicate systems. Between 93-162 human serum samples were tested in a single measurement on the candidate and predicate systems using at least three lots of Medicon Hellas reagents. Less than 10% of samples were spiked or diluted to cover the analytical measurement ranges. Results were tabulated and evaluated using Analyse-it statistics calculator software to generate Passing-Bablok regression statistics for Cholesterol, HDL-Cholesterol, LDL-Cholesterol and Triglycerides.

Device	SampleMatrix	N	Sampleconcentrationrange tested	Slope	Intercept	R2
CHOLESTEROL	Serum	93	44 – 666 mg/dL	0.9769	5.098	0.999
HDL-Cholesterol	Serum	141	6 – 177 mg/dL	1.0180	-0.028	0.997
LDL-Cholesterol	Serum	107	5 – 721 mg/dL	0.9821	1.750	0.999
TRIGLYCERIDES	Serum	163	26 – 975 mg/dL	0.9774	2.041	0.999

The summary of the results for a representative lot are provided in the table below:

b. Matrix Comparison:

Not applicable.

{22}------------------------------------------------

3. Expected Values / Reference Range:

The following reference ranges, cited from the scientific literature', were included in the labeling.

Analyte	Sample Matrix	Reference Range
Cholesterol	Serum	Desirable < 200 mg/dLBorderline 200-239 mg/dLAbnormal ≥ 240 mg/dL
HDL-Cholesterol	Serum	Low Risk: HDL-C ≥ 60 mg/dLHigh Risk: HDL-C ≤ 40 mg/dL
LDL-Cholesterol	Serum	Optimal: < 100 mg/dLNear optimal/above optimal: 100 – 129 mg/dLBorderline high risk: 131 – 159 mg/dLHigh risk: 160 – 189 mg/dLVery high risk: ≥ 190 mg/dL
Triglycerides	Serum	Normal: < 150mg/dLBorderline high: 150 - 199 mg/dLHigh: 200 - 499 mg/dLVery high: ≥ 500 mg/dL

CONCLUSION

The summary includes the conclusions drawn from the nonclinical tests (discussed above) that demonstrate that the device is as safe, as effective, and performs as well as or better than the predicate device.

4 National Cholesterol Education Program. Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). National Institutes of Health, National Heart, Lung, and Blood Institute, NIH Publication No. 01-3670 May 2001

Regulation Number and Section

§ 862.1175 Cholesterol (total) test system.

(a)
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.(b)
Classification. Class I (general controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to § 862.9.