Direct LDL Cholesterol (LDL)

K161691 · Randox Laboratories Limited · MRR · Mar 20, 2017 · Clinical Chemistry

Device Facts

• Record ID: K161691
• Device Name: Direct LDL Cholesterol (LDL)
• Applicant: Randox Laboratories Limited
• Product Code: MRR · Clinical Chemistry
• Decision Date: Mar 20, 2017
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1475
• Device Class: Class 1

Intended Use

For the quantitative in vitro determination of LDL-cholesterol concentration in human plasma and serum. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis and various liver and renal diseases. This in vitro diagnostic device is intended for prescription use only.

Device Story

Direct LDL Cholesterol (LDL) is an in vitro diagnostic reagent kit for quantitative measurement of LDL-cholesterol in human plasma and serum. The assay uses a two-step enzymatic clearance method on the RX Daytona Plus analyzer. Reagent 1 eliminates chylomicrons, VLDL, and HDL via cholesterol esterase, cholesterol oxidase, and catalase. Reagent 2 releases LDL-cholesterol, which is then measured via a peroxidase-catalyzed reaction producing a quinoneimine dye proportional to cholesterol concentration (measured at 600 nm). Used in clinical laboratories by trained personnel. Results assist clinicians in diagnosing and managing lipid disorders, atherosclerosis, and liver/renal diseases. Benefits include accurate lipid profiling for cardiovascular risk assessment.

Clinical Evidence

Bench testing only. Precision evaluated per CLSI EP5-A2 (n=80 per sample). Linearity confirmed per CLSI EP6-A (21-740 mg/dL, r=0.997). Detection limits (LoB 1.94, LoD 3.19, LoQ 16.1 mg/dL) determined per CLSI EP17-A2. Interference testing per CLSI EP7-A2 showed no significant interference (<10%) for hemoglobin, bilirubin, triglycerides, Intralipid, or ascorbic acid. Method comparison against predicate (n=139) yielded y=1.01x-1.45, r=0.998. Matrix comparison (n=70) confirmed lithium heparin plasma equivalence to serum (y=1.01x-2.81, r=0.998).

Technological Characteristics

Quantitative colorimetric assay. Reagents: R1 (PIPES buffer, HDAOS, cholesterol esterase/oxidase, catalase, ascorbate oxidase, surfactant); R2 (PIPES buffer, 4-amino antipyrine, peroxidase, surfactant, sodium azide). Sensing principle: enzymatic colorimetric reaction measured at 600 nm. Form factor: liquid ready-to-use reagents. Platform: RX Daytona Plus Chemistry analyzer. Sterilization: not applicable (in vitro diagnostic reagent).

Indications for Use

Indicated for quantitative in vitro determination of LDL-cholesterol in human plasma and serum for patients requiring assessment of lipid disorders, atherosclerosis, or liver/renal disease.

Regulatory Classification

Identification

A lipoprotein test system is a device intended to measure lipoprotein in serum and plasma. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.

Predicate Devices

• Randox Laboratories Ltd., Direct LDL Cholesterol reagent (K982529)

Reference Devices

• Randox Direct HDL-C/LDL-C Calibrator (K122126)
• Randox Lipid Control Sera; Level 1, Level 2 and Level 3 (K022591)

Submission Summary (Full Text)

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#### DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized symbol resembling a caduceus, with three abstract human profiles facing right. The symbol is encircled by the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" in a circular arrangement.

Public Health Service

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002 March 20, 2017

RANDOX LABORATORIES LIMITED PAULINE ARMSTRONG OA/RA MANAGER 55 DIAMOND ROAD CRUMLIN CRUMLIN, GB BT29 40Y ANTRIM UNITED KINGDOM

Re: K161691

Trade/Device Name: Direct LDL Cholesterol (LDL) Regulation Number: 21 CFR 862.1475 Regulation Name: Lipoprotein test system Regulatory Class: I, meets the limitations of exemption 21 CFR 862.9(c)(4) Product Code: MRR Dated: February 6, 2017 Received: February 9, 2017

Dear Dr. Pauline Armstrong:

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 (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. The general controls provisions of the Act include requirements for annual registration. Isting 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.

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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements

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as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely vours.

# Kellie B. Kelm -S

for Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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# Indications for Use

510(k) Number (if known) k161691

Device Name Direct LDL Cholesterol (LDL)

Indications for Use (Describe)

For the quantitative in vitro determination of LDL-cholesterol concentration in human plasma and serum. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders mellitus), atherosclerosis and various liver and renal diseases.

This in vitro diagnostic device is intended for prescription use only.

| Type of Use (Select one or both, as applicable) |
|-----------------------------------------------------------------|
| <span> Prescription Use (Part 21 CFR 801 Subpart D) </span> |
| <span> Over-The-Counter Use (21 CFR 801 Subpart C) </span> |

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## 510(K) SUMMARY, DIRECT LDL CHOLESTEROL (LDL)

## 1. SAFETY AND EFFECTIVENESS AS REQUIRED BY 21 CFR 807.92 STATEMENT

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

## 2. SUBMITTER NAME AND ADDRESS

Name: Dr Pauline Armstrong

Address: Randox Laboratories Limited 55 Diamond Road, Crumlin, County Antrim, BT29 4QY, United Kingdom.

Telephone: [phone] Fax: [phone] E-mail: [e-mail]

Date of Summary Preparation: 16 March 2017

#### 3. 510k NUMBER, DEVICE PROPRIETARY NAME, COMMON NAME, PURPOSE FOR SUBMISSION, REGULATORY CLASSIFCATION, PANEL, PRODUCT CODE AND 21 CFR NUMBER

510k No: k161691

Device Proprietary Name: Direct LDL Cholesterol (LDL)

Common Name: Direct LDL Cholesterol (LDL)

Purpose for Submission: New Device

| Product Code | Regulation Name | Classification | Regulation Section | Panel |
|--------------|-------------------------|----------------------------------------------------------------|------------------------------------------|-------------------------|
| MRR | Lipoprotein test system | Class I, meets the limitation of exemption 21 CFR §862.9(c)(4) | 21 CFR §862.1475 Lipoprotein Test System | Clinical Chemistry (75) |

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## 4. PREDICATE DEVICE PROPRIETARY NAMES AND 510 (k) NUMBERS

#### Predicate Device Proprietary Name:

Randox Laboratories Ltd., Direct LDL Cholesterol Reagent

#### 510 (k) Number: K982529

#### 5. INTENDED USE

For the quantitative in vitro determination of LDL-cholesterol concentration in human plasma and serum. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis and various liver and renal diseases. This in vitro diagnostic device is intended for prescription use only.

## 6. DEVICE DESCRIPTION

The LDL Cholesterol kit assay consists of ready to use reagent solutions. CATALOGUE NUMBER: CH8312

R1. Enzyme Reagent 1 4 x 20 mL R2. Enzyme Reagent 2 4 x 9 mL

#### REAGENT COMPOSITION

| Contents | Initial Concentration of Solution |
|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------|
| R1. Enzyme Reagent 1<br>PIPES Buffer<br>Piperazine-1, 4-bis (2-ethanesulfonic acid)<br>HDAOS<br>N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxylaniline, sodium salt<br>Cholesterol Esterase<br>[E.C.3.1.1.13. Microorganism, 37°C]<br>Cholesterol Oxidase<br>[E.C.1.1.3.6. Streptomyces sp, 37°C]<br>Catalase<br>[E.C.1.11.1.6. Microbial]<br>Ascorbate Oxidase<br>[E.C.1.10.3.3. Acremonium sp.]<br>Surfactant | 50 mmol/l, pH 7.0<br>2.0 mmol/l<br>≥600U/I<br>≥500U/I<br>≥600KU/I<br>≥300U/I<br>0.3% (w/v) |
| R2. Enzyme Reagent 2<br>PIPES Buffer<br>Piperazine-1, 4-bis (2-ethanesulfonic acid)<br>4-Amino antipyrine<br>Peroxidase<br>[E.C.1.11.1.7, Horse Radish, 25°C]<br>Surfactant<br>Sodium Azide | 50 mmol/l, pH 7.0<br>4 mmol/l<br>≥4KU/I<br>1.00% (w/v)<br>0.05% (w/v) |

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## Materials required but not provided.

Direct HDL-C/LDL-C Calibrator, CH2673 (K122126). Randox Lipid Controls (K022591):-LE 2661 or LE 2668 Level 1 Level 2 LE 2662 or LE 2669 Level 3 LE 2663 or LE 2670 RX series Saline (Cat. No. SA8396)

# 7. PREDICATE DEVICE COMPARISON TABLE

Table 1 - Comparison of LDL Cholesterol test system for the RX Daytona Plus to

Predicate device

| Characteristics | LDL Cholesterol Assay for<br>RX daytona plus<br>(New Device) | Randox LDL cholesterol (K982529)<br>(Predicate Device) |
|-----------------------|----------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------|
| | | Similarities |
| Intended Use | For the quantitative <i>in vitro</i> determination<br>of LDL-cholesterol concentration in<br>human plasma and serum. | Same |
| Assay Protocol | Clearance Method | Same |
| Storage<br>(Unopened) | Reagents are stable up to the expiry date<br>when stored unopened at +2 to +8°C | Same |
| Sample Type | Serum, Heparinized Plasma are the<br>recommended samples. | Same |
| | | Differences |
| Measuring Range | 21 - 740 mg/dl | 7.3 to 859 mg/dl |

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## 8. TEST PRINCIPLE (1, 2)

The assay consists of 2 distinct reaction steps:

- 1. Elimination of chylomicron, VLDL-Cholesterol and HDL-Cholesterol by cholesterol esterase, cholesterol oxidase and subsequently catalase.
Cholesterol esterase Cholesterol ester -→Cholesterol + fatty acid Cholesterol oxidase Cholesterol + O2 → Cholestenone + H2O2 Catalase 2H2O + O2 2H2O2

- 2. Specific measurement of LDL-Cholesterol after release of LDL-Cholesterol by detergents in Reagent 2.

$$\text{Cholesterol ester} \xrightarrow{\text{Cholesterol esterase}} \text{Cholesterol + fatty acid}$$

$$\text{Cholesterol + O}_{2} \xrightarrow{\text{Cholesterol oxidase}} \text{Cholestenone + H}_{2}\text{O}_{2}$$

$$2\text{H}_{2}\text{O}_{2} + \text{4-AA + HDAOS} \xrightarrow{\text{Peroxidase}} \text{Quinone pigment+4H}_{2}\text{O}$$

The intensity of the quinoneimine dye produced is directly proportional to the cholesterol concentration when measured at 600 nm.

In the second reaction catalase is inhibited by sodium azide in Enzyme Reagent 2.

Key: 4 - AA - 4 - Aminoantipyrine HDAOS = N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxylaniline, sodium salt

- 1. Weiland H. and Seidel D., J Lip Res, 24: 904-909, 1983.
2. Friedewald W.F., et al, Clin Chem, 18: 499-502, 1972.
-

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# 9. PERFORMANCE CHARACTERISTICS

## Analytical performance:

## a. Precision/Reproducibility:

Precision was evaluated consistent with C.L.S.I documents EP5-A2. Precision studies were performed by two operators on two RX Daytona plus svstems using control material and unaltered human serum samples that were spiked with LDL cholesterol concentrations or diluted to achieve concentrations based on established ranges <100mg/dl optimal LDL cholesterol. 130 - 159 mg/dL borderline high LDL Cholesterol, 160 – 189 mg/dL high LDL Cholesterol, >190 mg/dL very high LDL Cholesterol. Testing was conducted for two reagent lots of Cholesterol levels, one lot on each RX Daytona plus system, twice per day for 20 non-consecutive days. Two replicates per run were performed for each sample. The assay was calibrated on the first day of the study and no assay recalibrations were required throughout the duration of the study. The results of Lot 2, which is representative of both lots of reagent, is summarized in the following table.

| Lot 2 | | | Mean | | Within Run | | Among Run | | Among<br>Day | | Total |
|--------|--------------|----|---------|------|------------|------|-----------|-----|--------------|------|-------|
| Method | Product | N | (mg/dl) | SD | CV | SD | CV | SD | CV | SD | CV |
| LDL | QC 1 | 80 | 92.0 | 2.7 | 3.0 | 4.6 | 5.0 | 0.8 | 0.9 | 5.4 | 5.9 |
| LDL | QC 2 | 80 | 135.9 | 3.8 | 2.8 | 4.1 | 3.0 | 2.6 | 1.9 | 6.2 | 4.6 |
| LDL | QC 3 | 80 | 186.7 | 5.5 | 2.9 | 5.9 | 3.2 | 1.4 | 0.7 | 8.2 | 4.4 |
| LDL | Serum pool 1 | 80 | 65.0 | 1.7 | 2.6 | 3.2 | 5.0 | 1.2 | 1.8 | 3.8 | 5.9 |
| LDL | Serum pool 2 | 80 | 154.0 | 4.4 | 2.9 | 6.0 | 3.9 | 2.1 | 1.4 | 7.8 | 5.0 |
| LDL | Serum pool 3 | 80 | 200.1 | 5.8 | 2.9 | 8.2 | 4.1 | 0.0 | 0.0 | 10.0 | 5.0 |
| LDL | Serum pool 4 | 80 | 343.7 | 10.3 | 3.0 | 14.2 | 4.1 | 4.3 | 1.2 | 18.0 | 5.3 |

Table 2 Precision Summary

## b. Linearity/assay reportable range:

Linearity studies have been carried out in accordance with C.L.S.I. standard EP6-A. Linearity studies were performed at 11 levels to determine the analytical range of an assay - that is the range where the reported result is a linear function to the analyte concentration (or where deviation from linearity is less than 10%).

The linearity samples were prepared at 11 levels. The sponsor set a range from approximately 21 mg/dl analyte concentration up to a high concentration of approximately 850 mg/dl using low and high serum pools. The low and high pools were mixed to make nine intermediate levels. Each level was run in replicates of five on two lots of LDL Cholesterol reagent on one RX Daytona plus

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system. The observed values were compared to the expected values; the linear regression correlation between the expected values and the observed values are summarized in the following table:

Table 3 Linearity Summary including Regression equation and correlation co-efficient.

| Analyte Tested | LDL Cholesterol<br>(mg/dl) |
|----------------------|----------------------------|
| Linear<br>Regression | $Y = 0.99x +6.34$ |
| r | 0.997 |

The reportable range of the assay is 21-740 mg/dl.

Samples with concentrations greater than the linearity limit established are returned > 740 mg/dL and are re-run automatically by the RX daytona plus using the established high re-run conditions.

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

Refer to K122126 Direct HDL/LDL Calibrator and K022591 Lipid Controls.

The Direct HDL-C/LDL-C Calibrator is traceable to an internal master reference material. Calibrators are value assigned using one instrument and multiple repetitions. The mean, standard deviation, and % CV are calculated and evaluated against acceptance criteria.

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

## d. Detection limit:

Sensitivity studies have been carried out in accordance with C.L.S.I. guideline EP17-A2 'Protocols for Determination of Limits of Detection and Limits of Quantification; Approved Guideline'. A Limit of Blank (L.o.B), a Limit of Detection (L.o.D) and a Limit of Quantification (L.o.Q) were performed on two lots of reagents tested by two operators on one RX Daytona Plus system.

The Limit of Detection (LoD) for LDL Cholesterol on the RX Daytona Plus is 3.19 mg/dl based on 240 determinations, with 4 low level samples.

The Limit of Blank (LoB) is 1.94 mg/dl.

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The Limit of Quantitation (LoQ) is 16.1 mg/dl as determined by the lowest concentration detected with ≤20% imprecision.

### e. Analytical Specificity:

Interference studies have been carried out in accordance with C.L.S.I. guideline EP7-A2 'Interference testing in clinical chemistry; Approved Guideline Second Edition'. The effects of potential interferents were determined by calculating the mean value of the spiked interferent with the corresponding control solution. The spiked sample results were compared to control samples prepared without the potential interferents.

#### Acceptance Criteria: % of Control ± 10%

The following analytes were tested up to the levels indicated at LDL Cholesterol concentrations of 96.75 mg/dl and 193.5 mg/dl and found not to interfere:

| Hemoglobin | No significant interference up to 1000mg/dl. |
|---------------------|----------------------------------------------|
| Total Bilirubin | No significant interference up to 60mg/dl. |
| Conjugate Bilirubin | No significant interference up to 60mg/dl. |
| Triglycerides | No significant interference up to 500mg/dl. |
| Intralipid® | No significant interference up to 500mg/dl. |
| Ascorbic Acid | No significant interference up to 6mg/dl. |

#### f. Method comparison with predicate device:

Correlation studies were carried out in accordance with C.L.S.I. guideline EP9-A2 'Method Comparison and Bias Estimation Using Patient Samples: Approved Guideline - Second Edition'.

The candidate device on the RX Daytona Plus analyzer, was tested against the predicate device on the RX Imola analyzer.

139 serum patient samples spanning the range 22.45 to 735.68 mg/dl were tested by two operators on two lots of LDL cholesterol reagent, across 3 working days with each sample tested in singlicate.

The candidate device was compared to the predicate device and the following linear regression equation was obtained:

Y = 1.01x - 1.45 Correlation coefficient of r = 0.998

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## q. Matrix comparison:

Matrix method comparisons for the LDL cholesterol assay was tested by one operator on one RX Daytona plus system and was assessed for two lots of LDL cholesterol reagents. Both serum and lithium heparin plasma were tested to determine whether method accuracy with plasma specimens are equivalent to serum results and that lithium heparin plasma does not interfere with either the method or the system.

#### LDL Cholesterol matrix comparison on the RX Daytona plus (Lithium Heparin)

Patient samples were drawn in matched pairs – one sample serum (x) and the second sample lithium heparin plasma (y). 70 matched patient sample pairs were analyzed spanning the range 25.45 to 665.64 mg/dl and the following linear regression equation was obtained:

Y = 1.01x -2.81 Correlation coefficient of r = 0.998

Expected values/Reference range:

Referenced from literature

Table 4 Reference Ranges

| Analyte | Serum |
|---------------------|--------------------------------------------------------------------------------------------------------|
| LDL Cholesterol (3) | < 100 mg/dL optimal<br>130 – 159 mg/dL borderline high<br>160 – 189 mg/dL high<br>>190 mg/dL very high |

3. Third Report of the National Cholesterol Education Programme (NCEP) Expert Panel on Detection, Evaluation and treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No. 02-5215 September 2002

#### 10. CONCLUSION

Testing results indicate that the proposed device is substantially equivalent to the predicate device.