The FREND™ Testosterone test is a fluorescent nanoparticle immunoassay designed for in vitro quantitative measurement of total testosterone in human serum and plasma (K3-EDTA and lithium heparin). Measurements of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, impotence in males, hirsutisim (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.

The FREND™ Testosterone microfluidic flow cartridge is designed for use in the FREND™ System fluorescent immunoassay reader. The FREND™ Testosterone Test System is intended for use in clinical laboratories. For in vitro diagnostic use only. The test is not intended for use in point-of-care settings.

Device Description

The FREND™ Testosterone is a rapid fluorescence immunoassay that measures testosterone levels in human serum and plasma using the FREND™ system.

The FREND™ Testosterone test is a single-use rapid "competitive" immunoassay utilizing fluorescent nanoparticle in microfluidic flow to capture and quantify total testosterone levels in human serum and plasma (KgEDTA and lithium-heparin) specimens using the FREND™ system. The FREND™ Testosterone Test is a two-step competitive immunoassay with gold micro-particles labeled with mouse monoclonal antitestosterone antibody, testosterone-biotin labeled with fluorescence nanoparticles and fluorescence detection by the FREND™ System.

The FREND™ Testosterone test utilizes microfluidic technology and detects immunecomplexes bound to testosterone. In Step 1, a 70 µL patient sample is first incubated for 5 minutes at 98.6 °F (37 °C) in the Testosterone Gold Antibody pretreatment tube, where the sample interacts with a proprietary mix of a pretreatment solution. In Step 2, the Test Cartridge is placed on the warming platform of the heating block and 35 µL of the mixture from Step 1 is manually loaded into the inlet of the cartridge. The cartridge remains on the warming platform for 30 seconds, while the sample hydrates the testosterone-biotin fluorescent bead conjugate and migrates along the test strip. During migration the bound testosterone in the sample and the testosterone-biotin fluorescent bead conjugates compete to form antigen-antibody complexes in the test zone. Unbound testosterone-biotin fluorescent conjugates flow through and bind to the antitestosterone antibody that is immobilized on the surface in the reference zone. The cartridge is inserted into the FREND instrument for analysis where fluorescent signals in the test and reference zones are measured, typically within 4 minutes.

Testosterone quantification is based upon the ratio of the intensity of the test and reference zones. The magnitude of the fluorescent ratio is inversely proportional to the amount of testosterone in the sample.

The measuring range of the FREND™ Testosterone Test System is 20 to 1500 ng/dL. Results are determined via a lot-specific calibration curve which is generated by the manufacturer using a six-point calibration determined from values averaged from 5 replicates at each level. The established curve is uploaded to the FREND™ System via the Testosterone Code-chip and is valid until the lot expiration date.

The FREND™ Testosterone test cartridge is a disposable plastic device that houses the reagents and contains a port or opening (inlet) where the sample is applied. Once the sample is applied, it will mix with the reagents and travel towards the detection area via capillary action.

The FREND™ System is a portable, automated FREND™ cartridge reader. The FREND™ System is based on quantitative immunoassay technology capable of quantifying single or multiple analytes by measuring laser-induced fluorescence in a single-use disposable reagent cartridge. The FREND™ cartridge utilizes micro-fluidics lateral flow technology where the analyte of interest in the sample forms immune complexes while moving through the fluidics pathway in the cartridge. The concentration of the analyte of interest in an unknown sample is calculated using the ratio of the fluorescent intensity of the test zone and the reference zone.

The FREND™ System is a bench top fluorescence reader containing a touch screen user interface. The System has a slot that accepts the FREND™ Testosterone Test Cartridge (which contains the reagents and sample), and is programmed to analyze the Test when the sample has fully reacted with the on-board in-cartridge reagents. Results of the test are displayed on the screen and can be printed on an optional printer.

The FREND™ System software controls the graphical user interface, communication with hardware, database manaqement and data analysis. The software also controls the functions of the mechanical components including the motor, laser, printer control and acquisition of data from the sensor. The user can set the time and date and enter patient ID through the graphic user interface. The user cannot make any changes to the software.

The FREND™ Testosterone Test System includes the following in the kit:

. 20 FREND™ Testosterone cartridges
20 Testosterone Gold Antibody Pretreatment Tubes .
Disposable pipette tips .
. 1 FREND™ Testosterone Code Chip
1 FREND™ Testosterone Package Insert ●

The FREND™ System (previously cleared in K124056 (FREND™ PSA Plus) K131928 (FREND™ TSH), and K152422 (FREND™ Free T4)) is not provided with the kit but is required for the use of the FREND™ Testosterone test cartridge.

AI/ML Overview

Here's a breakdown of the acceptance criteria and the study that proves the device meets them, based on the provided text:

1. Table of Acceptance Criteria and Reported Device Performance

The document doesn't explicitly state "acceptance criteria" for each performance characteristic as clear pass/fail thresholds. Instead, it provides the results of various analytical performance studies. However, the implicit acceptance criterion for most of these studies is typically that the device performs within expected and acceptable analytical limits for a diagnostic test of its type, and that its performance is comparable to the predicate device.

For the purpose of this table, I will infer the performance goals based on industry standards and the presented data.

Performance Characteristic	Implicit Acceptance Criteria (Inferred)	Reported Device Performance
Precision/Reproducibility	Coefficients of Variation (CV%) should be within acceptable limits for quantitative immunoassays, indicating reliable and consistent results. Often, lower CVs are desirable for higher concentrations.	Repeatability (Intra-assay): - 39.723 ng/dL: 11.2% CV - 202.965 ng/dL: 8.2% CV - 1012.208 ng/dL: 5.4% CV Within-laboratory (Inter-assay): - 39.723 ng/dL: 11.8% CV - 202.965 ng/dL: 8.5% CV - 1012.208 ng/dL: 5.7% CV
Linearity/Reportable Range	The device should demonstrate linearity across its claimed measuring range, meaning results accurately reflect the concentration of the analyte over that range.	Linearity demonstrated across 17 ~ 1650 ng/dL, supporting a reportable range of 20 ng/dL ~ 1500 ng/dL.
Detection Limit (LoD)	The device should be able to reliably detect testosterone at its claimed analytical sensitivity.	LoD established at 14.3 ng/dL. Functional sensitivity established at 19.66 ng/dL. Analytical sensitivity claimed at 20 ng/dL.
Analytical Specificity (Interference)	Key endogenous substances and common pharmaceuticals should not significantly interfere with the test results (e.g., recoveries within 90-110% of expected values).	All tested endogenous substances (Hemoglobin, Bilirubin, Triglyceride, Total protein, Biotin, SHBG) and pharmaceuticals (e.g., Acetylcysteine, Ampicillin-Na, Ascorbic acid, Cyclosporine, Cefoxitin, Heparin, Levodopa, Methyldopa, Metronidazole, Doxycycline, Acetylsalicylic Acid, Rifampin, Acetaminophen, Ibuprofen, Theophylline) showed %Recovery of Testosterone between 90% and 110%. Heterophilic Antibodies (RF, HAMA) also showed recoveries within this range.
Analytical Specificity (Cross-Reactivity)	Structurally similar compounds or other hormones should not lead to significant false-positive readings or inaccurate quantification of testosterone.	No significant cross-reactivity was found for all tested substances (e.g., Androstenedione, Androsterone, Cortisone, Danazol, Estradiol, Estrone, 17a-Ethinyl estradiol, Progesterone, Dexamethasone, Ethisterone, D(-) Norgestrel, Prednisolone, Prednisone, Spironolactone, Cortisol, DHEA, DHEAS, Dihydrotestosterone, Epitestosterone, Ethynodiol diacetate). The % Cross-reactivity was generally very low, with the highest at 0.5160% for Estradiol and 1.0463% for DHEA, and 0.4134% for Dihydrotestosterone.
Method Comparison with Predicate Device	The device's results should correlate well with a legally marketed predicate device (Abbott ARCHITECT 2nd Generation Testosterone), demonstrated by a slope close to 1 and an intercept close to 0 in Passing-Bablok regression analysis.	Passing-Bablok regression analysis comparing FREND™ Testosterone (y) with Abbott ARCHITECT 2nd Generation Testosterone (x) yielded a slope of 0.983 and an intercept of -2.353. This indicates good agreement with the predicate.
Matrix Comparison	The device should produce equivalent results across different sample matrices (serum, K3EDTA plasma, lithium heparin plasma) as specified in its intended use, indicating that the anticoagulant used does not interfere with the test.	Testosterone concentrations measured in serum, lithium heparin, and K3EDTA plasma from 40 individuals using the FREND™ Testosterone Test System gave equivalent results.
Stability	The reagent kit should maintain its performance characteristics for a specified period when stored appropriately.	Reagent stability studies showed the cartridges will meet performance acceptance criteria for one year from the date of manufacture if stored refrigerated as directed.

2. Sample Sizes and Data Provenance (for test set/comparison studies)

Precision/Reproducibility: Three serum pools were assayed, each in duplicate, twice per day for 20 days (80 total measurements per level).
Linearity/Assay Reportable Range: Seven levels of serum testosterone were tested in quadruplicate using three reagent lots.
Detection Limit (LoD): Not specified in terms of sample number, but established according to CLSI EP17-A2 protocol.
Analytical Specificity (Interference): Not explicitly stated how many samples were used, but testing involved samples spiked with various interferents.
Analytical Specificity (Cross-Reactivity): Not explicitly stated how many samples were used, but testing involved samples spiked with various cross-reactants.
Method Comparison with Predicate Device: 157 de-identified leftover samples were used. An additional 2 samples were outside the range of both methods.
- Data Provenance: The method comparison study was performed in a CLIA-certified laboratory testing facility. The document does not specify the country of origin for the samples directly, but the applicant company is located in Korea, and the US FDA is reviewing the device for the US market. The samples were "de-identified leftover samples," indicating they were retrospective.
Matrix Comparison: 40 individuals' samples (serum, lithium heparin, and K3EDTA plasma) were used.
Expected Values/Reference Range: 488 normal, apparently healthy adult male and female individuals.

3. Number of Experts and Qualifications for Ground Truth

This document describes a diagnostic in vitro medical device (an assay for testosterone levels) and does not involve image interpretation or clinical diagnosis by human experts for establishing ground truth as typically seen in AI/CADe devices.

Instead, the "ground truth" for the comparison studies is established by:

Reference methods/predicate devices (e.g., Abbott ARCHITECT 2nd Generation Testosterone assay).
Gravimetric preparation for standards/calibrators.
Protocols and standards like CLSI guidelines for analytical performance.

Therefore, the concept of "number of experts" and "qualifications of those experts" in the context of clinical interpretation of a test set is not directly applicable here. The experts involved would be laboratory scientists and method developers ensuring adherence to CLSI guidelines and proper validation of the reference method.

4. Adjudication Method for the Test Set

Not applicable. For an in vitro diagnostic device measuring a quantitative analyte, adjudication methods like 2+1 or 3+1 consensus are not generally used. The ground truth for method comparison is the result obtained from the predicate device or a recognized reference method.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, a multi-reader multi-case (MRMC) comparative effectiveness study was not done. This type of study is relevant for imaging devices or AI tools that assist human readers in making diagnoses or interpretations. The FREND™ Testosterone Test System is a laboratory assay that provides a quantitative value, not an assistive AI tool for human interpretation.

6. Standalone Performance

Yes, a standalone performance was done for the device itself. All the analytical performance studies (Precision, Linearity, LoD, Analytical Specificity, Stability) and matrix comparison were conducted to assess the performance of the FREND™ Testosterone Test System independently. The method comparison study also shows the device's standalone performance compared to another established device. The device generates a quantitative result directly, rather than assisting a human in an interpretative task.

7. Type of Ground Truth Used

The ground truth used for various performance evaluations includes:

Reference Method: For method comparison, the Abbott ARCHITECT 2nd Generation Testosterone assay (K120009) was used as the reference method.
Gravimetric Preparation: For standards/calibrators, the ground truth is established through gravimetric preparation and confirmed by the reference method.
Expected Values/Spiked Samples: For interference and cross-reactivity studies, known concentrations of analytes and interferents are spiked into samples, creating a known "ground truth" to measure recovery and specificity.
CLSI Protocols: Adherence to CLSI (Clinical and Laboratory Standards Institute) protocols for various analytical studies (e.g., EP5-A3 for precision, EP6-A for linearity, EP17-A2 for LoD, EP07-A2 for interference/cross-reactivity, EP14-A3 for matrix comparison, C28-A3 for reference intervals) implies that the ground truth is established based on recognized industry standards and best practices for in vitro diagnostic device validation.

8. Sample Size for the Training Set

The document does not explicitly mention a "training set" in the context of machine learning or AI algorithm development because the FREND™ Testosterone Test System is a fluorescent nanoparticle immunoassay. While it uses the "FREND™ System" which includes software and data analysis, it's a fixed algorithm for quantifying a chemical analyte based on fluorescent ratio, not a learning algorithm that requires a traditional training set like a deep learning model.

The "calibration curve" is "generated by the manufacturer using a six-point calibration determined from values averaged from 5 replicates at each level." This is a form of calibration rather than algorithm training.

9. How the Ground Truth for the Training Set Was Established

As there is no traditional "training set" in the AI/ML sense, the concept of establishing ground truth for it is not applicable. The device's operational parameters (like the calibration curve) are established by the manufacturer through rigorous calibration using control materials with known testosterone concentrations derived from a six-point calibration curve, averaged from 5 replicates at each level. The standards/calibrators themselves are prepared gravimetrically and confirmed by measurement on the predicate device.

Summary

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Image /page/0/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo is circular, with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. In the center of the circle is an emblem featuring a stylized image of three human profiles facing to the right, with a wing-like shape extending above them. The emblem is rendered in a single color, creating a silhouette effect.

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

NANOENTEK USA INC. C/O MAŬREEN GARNER NEW WORLD REGULATORY SOLUTIONS, INC 1983 HAZELWOOD ROAD TOMS RIVER NJ 08753

October 14, 2016

Re: K153577

Trade/Device Name: FREND ™ Testosterone Test System Regulation Number: 21 CFR 862.1680 Regulation Name: Testosterone test system Regulatory Class: I, reserved Product Code: CDZ Dated: September 29, 2016 Received: October 3, 2016

Dear Maureen Garner:

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, 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.

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 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.

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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 (800) 638 2041 or (301) 796-7100 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 (800) 638-2041 or (301) 796-7100 or at its Internet address

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

Sincerely yours.

Katherine Serrano -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) K153577

Device Name FREND™ Testosterone Test System

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)

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510(k) Summary

As required by the Safe Medical Devices Act (SMDA) of 1990 and in accordance with 21 CFR §807.92, a 510(k) summary is provided.

Applicant A.

Company Name:	NanoEnTek, Inc.
Address:	12F 5 Digital-ro, 26-gil, Guro-guSeoul 152-740, KOREA
Contact Person:	Sunmi Han
Phone Number:	+82-2-6220-7940
Facsimile Number:	+82-2-6220-7721

B. 510(k) Preparer Information (Contact Person)

Company Name:	New World Regulatory Solutions, Inc.
Address:	P.O. Box 5374Toms River, New Jersey 08754, USA
Contact Person:	Maureen Garner
Phone Number:	(732) 779-7422
Facsimile Number:	(732) 270-4829
Email:	NWRSinc@gmail.com

C. Purpose for Submission:

New Analyte on FREND™ System

D. Measurand:

Testosterone

E. Type of Test:

Quantitative, Fluorescence Immunoassay

E. Proprietary and Established Device Name:

FREND™ Testosterone Test System

G. Regulatory Information:

	Assay
Proprietary Name	FREND™ Testosterone (reagent cartridge)
Generic Name	Competitive Immunoassay, Testosterone
Regulation Number:	21 CFR §862.1680
Product Code	CDZ
Classification	Class I, Reserved
Classification Name	Testosterone Test System
Panel	Chemistry (75)

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H. Intended Use:

1. Intended use(s): See indications for use below:

1. Indication(s) for use:
  The FREND™ Testosterone test is a fluorescent nanoparticle immunoassay designed for in vitro quantitative measurement of total testosterone in human serum and plasma (KsEDTA and lithium heparin). Measurements of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, impotence in males and, in females, hirsutisim (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.

1. Special conditions for use statement(s): For prescription use only.
1. Special instrument requirements: NanoEnTek FREND™ System

l. Device Description

The FREND™ Testosterone is a rapid fluorescence immunoassay that measures testosterone levels in human serum and plasma using the FREND™ system.

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The FREND™ Testosterone Test System includes the following in the kit:

. 20 FREND™ Testosterone cartridges
20 Testosterone Gold Antibody Pretreatment Tubes .
Disposable pipette tips .
. 1 FREND™ Testosterone Code Chip
1 FREND™ Testosterone Package Insert ●

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J. Substantial Equivalence Information:

An overview of the similarities and differences between the FREND™ Testosterone Test System and the predicate is provided in the table below:

Similarities
Item	FREND™ Testosterone Test System	Abbott ARCHITECT 2nd Generation Testosterone (K120009)
Intended Use	The FREND™ Testosterone test is a fluorescent nanoparticle immunoassay designed for in vitro quantitative measurement of total testosterone in human serum and plasma (K3EDTA and lithium heparin). Measurements of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, impotence in males and, in females, hirsutisim (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.The FREND™ Testosterone microfluidic flow cartridge is designed for use in the FREND™ System fluorescent immunoassay reader. The FREND™ Testosterone Test System is intended for use in clinical laboratories. The FREND™ Testosterone Test is for in vitro diagnostic use only. The test is not intended for use in point-of-care settings.	The ARCHITECT 2nd Generation Testosterone is a Chemiluminescent Microparticle Immunoassay (CMIA) for the quantitative determination of testosterone in human serum and plasma. Measurements of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delayed or precocious puberty, impotence in males and, in females, hirsutisim (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.The ARCHITECT 2nd Generation Testosterone Calibrators are for the calibration of the ARCHITECT i System when used for the quantitative determination of testosterone in human serum and plasma.The ARCHITECT 2nd Generation Testosterone Controls are for the verification of the accuracy and precision of the ARCHITECT i System when used for the quantitative determination of testosterone in human serum and plasma.For in vitro diagnostic use only.
Sample Type	Human serum and K3EDTA and lithium heparin plasma	Same
Analyte	Testosterone	Same
Type of Test	Fluorescent immunoassay determination of testosterone	Chemiluminescent immunoassay determination of testosterone
Quality Control	Internal procedural/instrument quality controls; commercially-available external positive and negative assay controls	Internal procedural/instrument quality controls; external positive and negative assay controls cleared with test kit
Interpretation of Results	Interpolation from a lot-specific calibration curve	Same

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Differences
Item	FREND™ Testosterone Test System	Abbott ARCHITECT Testosterone(K120009)
Sample Size	70 µL for the incubation step and 35 µLfor running the test	150 µL for the first Testosterone testplus 100 µL for each additionalTestosterone from the same test cup
Test Cartridge	Disposable single-use cartridge	No single-use cartridge
RandomAccess /Degree ofAutomation	No random access, manualmanipulation	Random access, semi-automated

K. Performance Characteristics (if/when applicable)

1. Analytical performance

a. Precision/Reproducibility

A single lot imprecision study was performed at the NanoEnTek laboratory as described in the CLSI protocol EP5-A3. Three serum pools with low, intermediate and high testosterone levels were assayed in duplicate twice per day for 20 days (80 total measurements per level). The results are summarized below:

Sample	MeanTestosterone	Repeatability		Within-laboratory
Pool	Level, (ng/dL)	SD	CV%	SD	CV%
1	39.723	4.451	11.2%	4.692	11.8%
2	202.965	16.670	8.2%	17.296	8.5%
3	1012.208	54.748	5.4%	57.480	5.7%

		FREND™ Testosterone Single Site Single Lot Precision

b. Linearity/assay reportable range:

Linearity was established according to CLSI-EP6-A in three reagent lots using seven levels of serum testosterone tested in quadruplicate. Linearity was demonstrated across a measuring interval of 17 ~ 1650 ng/dL, in support of the FREND™ Testosterone reportable range of 20 ng/dL ~ 1500 ng/dL.

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

The standards/calibrators are prepared gravimetrically and confirmed bv measurement on the ARCHITECT i 2nd Generation Testosterone assay (K120009). There is no need for calibration by the operator as the calibration information is coded in the individual cartridge.

d. Stability
Real-time stability testing for the Testosterone reagent kit was performed according to CLSI EP25-A, Evaluation of Stability of In Vitro Diagnostic Reagents. Reagent stability studies based on procedures and criteria in the NanoEnTek quality system showed that the cartridges for FREND™ Testosterone will meet performance acceptance criteria for one year from the date of manufacturer if stored refrigerated appropriately as directed.

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e. Detection Limit:
The Limit of Detection (LoD) for the FREND™ Testosterone was established at 14.3 ng/dL according to the CLSI EP17-A2 protocol. The functional sensitivity was established at 19.66 ng/dL. The analytical sensitivity of the FREND™ Testosterone is claimed at 20 ng/dL.
f. Analytical specificity:

Interference Studies

Interference was determined according to CLSI EP07-A2. No interference was found if recoveries were between 90% to 110% of the expected testosterone value. Results are summarized in the table below.

Class	Interferent (Concentration Tested)	%RecoveryTestosteroneLow	%RecoveryTestosteroneHigh
Endogenoussubstances	Hemoglobin (500 mg/dL)	97.2	96.5
	Bilirubin, conjugated (30 mg/dL)	106.6	100.4
	Bilirubin, unconjugated (30 mg/dL)	99.6	98.9
	Triglyceride (3 g/dL)	99.3	94.4
	Total protein (12 g/dL)	96.5	100.8
	Biotin (30 ng/mL)	101.8	104.3
	Sex Hormone Binding Globulin (100 nmol/L)	99.9	100.2
Pharmaceuticals	Acetylcysteine (415 $ μ $ g/mL)	97.9	99.4
	Ampicillin-Na (50.3 $ μ $ g/mL)	98.7	100.6
	Ascorbic acid (60 $ μ $ g/mL)	96.3	101.2
	Ca-Dobesilate (40 $ μ $ g/mL)	101.3	101.5
	Cyclosporine (3 $ μ $ g/mL)	97.4	96.8
	Cefoxitin (66 $ μ $ g/mL)	101.1	97.9
	Heparin (3,000 U/L)	102.6	99.7
	Levodopa (4 mg/mL)	96.7	100.9
	Methyldopa (15 $ μ $ g/mL)	101.0	97.1
	Metronidazole (120 $ μ $ g/L)	104.7	104.8
	Doxycycline (30 $ μ $ g/mL)	100.5	97.9
	Acetylsalicylic Acid (250 $ μ $ /g/mL)	94.7	98.4
	Rifampin (640 $ μ $ g/mL)	97.1	99.7
	Acetaminophen (200 $ μ $ g/mL)	99.6	100.3
	Ibuprofen (250 $ μ $ g/mL)	95.7	97.4
	Theophylline (400 $ μ $ g/mL)	102.1	99.5
HeterophilicAntibodies	RF (1075 IU/mL)	104.3	98.2
	HAMA (70 ng/mL)	103.8	100.2

Cross Reactivity

The following substances were evaluated for potential cross-reactivity with the FREND™ Testosterone at two concentrations. Testing was done according to the CLSI protocol EP07-A2. No significant cross-reactivity was found.

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	Conc. of cross-	% Cross-reactivity
Cross-reactant	reactant (ng/dL)	Low	High
Androstenedione (1,000 nmol/L)	28641	0.0032	0.0337
Androsterone (1,000 nmol/L)	29044	0.0004	0.0158
Cortisone (1,000 nmol/L)	36044	0.0077	0.0300
Danazol (1,000 nmol/L)	33746	0.0007	0.0496
Estradiol (200 nmol/L)	5447.6	0.0033	0.5160
Estrone (500 nmol/L)	13518.5	0.0144	0.0378
17a-Ethinyl estradiol (1,000 ng/mL)	100000	0.0033	0.0081
Progesterone (2,000 nmol/L)	62892	0.0007	0.0167
Dexamethasone (5 µmol/L)	196230	0.0255	0.0122
Ethisterone (20 nmol/L)	624.9	0.0437	0.3840
D(-) Norgestrel (20 ng/mL)	2000	0.0125	0.1667
Prednisolone (2,000 nmol/L)	61288	0.0001	0.0175
Prednisone (2,000 nmol/L)	71686	0.0031	0.0040
Spironolactone (500 ng/mL)	50000	0.0039	0.0172
Cortisol (10,000 nmol/L)	362460	0.0004	0.0070
DHEA (50 nmol/L)	1442.1	0.0178	1.0463
DHEAS (50 µmol/L)	1842500	0.0002	0.0009
Dihydrotestosterone (40 nmol/L)	1161.68	0.2040	0.4134
Epitestosterone (100 nmol/L)	2884.2	0.0467	0.1502
Ethynodiol diacetate (50 ng/mL)	5000	0.0274	0.1280

% Cross-reactivity = 100 x ((Measured value - true value)/concentration of cross-reactant))

g. Assay cut-off:

Not applicable

2. Comparison studies

a. Method comparison with predicate device:
Comparison studies with 157 de-identified leftover samples were performed in a CLIA-certified laboratory testing facility. The reference method was the Abbott ARCHITECT 200 Generation Testosterone Assay (K120009) run on the Abbott ARCHITECT i System. The samples spanned the measuring range of the FREND Testosterone Test System. An additional 2 samples gave values beyond the range of one or both test methods. All samples were assayed using sera split between the applicant device and the reference method.

Results from the FREND™ Testosterone on the FREND™ System (y) were compared with the reference results (x) by Passing-Bablok regression analysis, giving a slope of 0.983 and intercept of -2.353.

b. Matrix comparison:
The matrix comparison study was performed at the NanoEnTek laboratory according to CLSI EP14-A3. Testosterone concentrations in the serum, lithium heparin and K3EDTA from 40 individuals were measured using the FREND™ Testosterone Test System, giving equivalent results.

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1. Clinical studies:
- a. Clinical Sensitivity Not applicable
- b. Clinical Specificity Not applicable
- c. Other clinical supportive data (when a. and b. are not applicable) Not applicable
- 1. Clinical cut-off: Not applicable
- 1. Expected values/Reference range:

Serum samples from a total of 488 normal, apparently healthy adult male and female individuals were assayed on 3 lots of the FREND™ Testosterone assay using two FREND™ Systems according to CLSI C28-A3 guidelines. The reference interval for the FREND™ Testosterone Test System, stratified by age range and gender, is provided in the table below.

Gender (Age Range)	Testosterone, ng/dL	Number Tested
F (21 - 49)	<20 ~ 107.5	120
F (50 - 90)	<20 ~ 150.3	124
M (21 - 49)	170.1 ~ 1263.6	123
M (50 - 88)	152.4 ~ 1095.2	121

Proposed Labeling L.

The labeling is sufficient and it satisfies the requirements of 21 CFR §809.10.

M. Conclusion

The submitted information in this premarket notification is complete and supports a substantial equivalence determination.

Regulation Number and Section

§ 862.1680 Testosterone test system.

(a)
Identification. A testosterone test system is a device intended to measure testosterone (a male sex hormone) in serum, plasma, and urine. Measurement of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delayed or precocious puberty, impotence in males and, in females hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.(b)
Classification. Class I.