The IDS-iSYS Aldosterone assay (IS-3300) is a device intended for use in clinical laboratories for the quantitative determination of Aldosterone in human EDTA plasma on the IDS-iSYS Multi-Discipline Automated System. Aldosterone measurements are used in the diagnosis and treatment of primary aldosteronism (a disorder caused by excessive secretion of Aldosterone by the adrenal gland), hypertension caused by primary aldosteronism, selective hyperaldosteronism, edematous states and other conditions of electrolyte balance.

The IDS-iSYS Aldosterone Control Set (IS-3330) is intended for use as assaved quality control samples to monitor the accuracy of the IDS-iSYS Aldosterone assay on the IDS-iSYS Multi-Discipline Automated System.

The IDS-iSYS Aldosterone Calibration Verifiers (IS-3335) are intended for medical purposes for use in the quantitative verification of calibration of the IDS-iSYS Aldosterone assay on the IDS-iSYS Multi-Discipline Automated System.

Device Description

The IDS-iSYS Aldosterone assay is based on chemiluminescence technology. A biotinylated monoclonal anti-Aldosterone antibody is incubated with the sample, after an incubation step an Aldosterone acridinium conjugate is added and after a further incubation step streptavidin coated magnetic particles are added. Following a third incubation step the particles are "captured" using a magnet. After a washing step and addition of trigger reagents, the light emitted by the acridinium label is inversely proportional to the concentration of Aldosterone in the original sample.

AI/ML Overview

Here's an analysis of the provided text to extract the requested information about acceptance criteria and the supporting study:

The provided document is a 510(k) Summary for the IDS-iSYS Aldosterone assay, control set, and calibration verifiers. It focuses on demonstrating substantial equivalence to a predicate device, as required for FDA clearance. The document details performance characteristics but does not explicitly state acceptance criteria in a pass/fail format typical of formal acceptance criteria documents. Instead, it reports performance values found during validation studies.

Here's the breakdown of the information you requested, based on what's available in the document:

1. Table of Acceptance Criteria and Reported Device Performance

As mentioned, explicit "acceptance criteria" are not listed in a single table. However, the performance characteristics are reported, and implicitly, these values are considered acceptable for demonstrating substantial equivalence. The table below compiles the reported performance data from the document.

Note: The document does not provide a column for "Acceptance Criteria" as a separate, pre-defined target. The "Reported Device Performance" is the outcome of the studies aiming to demonstrate acceptable performance.

Performance Characteristic	Implicit or Explicit Acceptance Threshold (Not explicitly stated as "acceptance criteria" but implied targets from regulatory guidance or industry standards)	Reported Device Performance (IDS-iSYS Aldosterone Assay)	Source of Data
Precision (CV%)	Implied: Generally low CV% indicating good reproducibility (e.g., <15-20% for clinical assays, lower for critical ranges)	Within-Run: 2.2% - 8.4% (at 98.7 ng/dL to 7.5 ng/dL)	Table (p.8)
		Total: 5.2% - 12.8% (at 98.7 ng/dL to 7.5 ng/dL)	Table (p.8)
Linearity (R²)	Implied: High R² value (e.g., >0.98 or >0.99) indicating a strong linear relationship.	R² = 1.00	Text (p.9)
Regression Equation (y=mx+b)	Implied: Slope (m) close to 1, intercept (b) close to 0.	y = 1.00x - 1.24 (slope = 1.00, intercept = -1.24)	Text (p.9)
Limit of Blank (LoB)	Implied: Low value to correctly identify absence of analyte.	2.0 ng/dL	Table (p.11)
Limit of Detection (LoD)	Implied: Low value to correctly detect presence of analyte.	3.2 ng/dL	Table (p.11)
Limit of Quantitation (LoQ)	Implied: Low value with acceptable precision (e.g., typically ≤20% CV).	3.9 ng/dL (at 20% CV)	Table (p.11)
Interference (Concentration Bias)	Explicitly stated criterion in study: ≤10% concentration bias to the unspiked sample.	All tested interferents met this criterion.	Text (p.11)
Cross-Reactivity	Implied: Low percentage for non-target analytes, 100% for target analyte.	Aldosterone: 100%. Other analytes: 0.0003% to 3.1% (3α, 5β-Tetrahydroaldosterone: 3.1%)	Table (p.13)
Method Comparison (Slope vs. Predicate)	Implied for substantial equivalence: Slope close to 1, intercept close to 0, high R².	Linear Regression: Slope = 1.053, Intercept = 0.09 ng/dL, R² = 0.980	Text (p.14)
		Passing-Bablok: Slope = 1.070, Intercept = -0.29	Text (p.14)

2. Sample Size Used for the Test Set and Data Provenance

Precision/Reproducibility:
- Sample Size: Six EDTA plasma samples (9 "samples" were run over 20 days, generating 80 replicates per sample concentration).
- Data Provenance: Not explicitly stated, but clinical laboratory setting is implied. The phrase "two sites using three analyzers" could suggest multi-center testing, potentially from different locations/countries, but this is not specified. The studies were performed for the manufacturer for regulatory submission.
Linearity/Assay Reportable Range:
- Sample Size: One high plasma sample diluted with a low sample to create 11 dilution samples. Run in quadruplicate.
- Data Provenance: Not specified, likely internal laboratory data for the manufacturer.
Detection Limit (LoB, LoD, LoQ):
- Sample Size:
  - LoB (Lot 1): One zero-aldosterone plasma sample, 10 replicates for 5 days (total 50 replicates).
  - LoD/LoQ (Lot 1): 7 samples, 2 replicates per sample, once per day for 8 days.
  - LoB (Lot 2): One zero-aldosterone plasma sample, 6 replicates for 5 days (total 30 replicates).
  - LoD (Lot 2): 8 samples, 2 replicates per sample, once per day for 5 days.
  - LoQ (Lot 2): 7 samples, 2 replicates per sample, once per day for 5 days.
- Data Provenance: Not specified, likely internal laboratory data for the manufacturer. Two different sites and two different analyzers were used.
Analytical Specificity (Interference and Cross-Reactivity):
- Sample Size:
  - Interference: Two base plasma samples ("Low" and "High" Aldosterone concentrations), spiked with various potential interferents. For each interferent, 26 replicates for blank and spiked samples were compared.
  - Cross-reactivity: Stock solutions of various compounds diluted serially to create 7-point standard curves for each substance.
- Data Provenance: Not specified, likely internal laboratory data for the manufacturer.
Method Comparison:
- Sample Size: 161 samples (including 12 altered samples).
- Data Provenance: Not specified, implied to be clinical or patient samples. Whether these were retrospective or prospective, or their country of origin, is not mentioned.
Reference Range Study (Expected Values):
- Sample Size: 228 Caucasian adult samples.
- Data Provenance: Collected in the US, prospective (samples collected under specific conditions: 7-10 am after overnight fasting, upright/supine positions).

3. Number of Experts Used to Establish the Ground Truth for the Test Set and the Qualifications of Those Experts

This type of information is generally NOT applicable to in vitro diagnostic (IVD) devices like the Aldosterone assay described. For IVD assays, "ground truth" is typically established by:

Reference methods/predicate devices (e.g., Diasorin Liaison Aldosterone assay for method comparison).
Gravimetric preparation of standards and calibrators for traceability.
Defined sample characteristics (e.g., "zero aldosterone plasma," "high plasma sample").

There were no human experts assessing images or making diagnoses that would require adjudication.

4. Adjudication Method for the Test Set

Not applicable, as this is an IVD assay, not a device requiring human interpretation of results in a diagnostic context that would call for adjudication of different interpretations.

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

Not applicable. This is an IVD assay for quantitative determination of a biomarker; there are no "human readers" interpreting results in the way an MRMC study would be designed for an imaging AI device.

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

Yes, the studies conducted (Precision, Linearity, LoD/LoQ, Analytical Specificity, Method Comparison) represent the standalone performance of the IDS-iSYS Aldosterone assay system. The device quantifies aldosterone in plasma samples directly, without human interpretation of results being a variable in its core analytical performance.

7. The Type of Ground Truth Used

The ground truth used for various studies includes:

Reference materials: Aldosterone (≥95% HPLC; A9477Sigma-Aldrich) dissolved in Dioxane, with concentration calculated by UV quantitation using molar extinction coefficient (for calibrators and traceability).
Predicate device results: The Diasorin Liaison Aldosterone assay (K130321) for method comparison.
Spiked samples: For linearity, interference, and cross-reactivity studies, known concentrations of analyte or interferents were added to base samples to create "true" concentrations.
Patient samples: For method comparison and reference range studies, patient samples were used. Their "truth" for method comparison was established by the predicate device. For reference ranges, the "truth" was the measured value distributed among a healthy population.

8. The Sample Size for the Training Set

The document describes performance studies (validation) but does not explicitly mention a "training set" in the context of machine learning. For an IVD assay, the development process involves reagent optimization, calibration curve fitting, etc., which conceptually use data, but this is not typically termed a "training set" like in AI/ML.

However, the "Master curve" and "two-point calibration" system is mentioned. The "logistic parameters of the Master calibration curve" are generated using "data of 20 runs Internal Reference Calibrators" (p.9). This could be considered analogous to a training process for establishing the core measurement algorithm, but it's not a training set for an AI inference model.

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

If we consider the generation of the "Master calibration curve" as a "training" process:

Ground Truth: The "Internal Reference Calibrators" referenced (p.9) would serve as the ground truth. These calibrators are traceable to gravimetrically prepared Aldosterone standards using high-purity Aldosterone and UV quantitation (p.9).
Establishment Method: The "new kit calibrator sets are run as 'unknowns' in duplicate in at least 20 assays on one analyser" (p.9). "The data of 20 runs Internal Reference Calibrators are used to generate the logistic parameters of the Master calibration curve by using Prism software package" (p.9).

Summary

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Image /page/0/Picture/1 description: The image shows the seal of the Department of Health & Human Services (HHS) of the United States. The seal features the department's name encircling a symbol. The symbol is a stylized representation of three human profiles facing right, with flowing lines suggesting movement or connection. The text is arranged in a circular pattern around the symbol.

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

April 21, 2015

IMMUNODIAGNOSTIC SYSTEMS LTD. MICK FENTON, REGULATORY AFFAIRS OFFICER 10 DIDCOT WAY, BOLDEN BUSINESS PARK BOLDON, TYNE & WEAR NE35 9PD UNITED KINGDOM

Re: K142994 Trade/Device Name: IDS-iSYS Aldosterone, IDS-iSYS Aldosterone Control Set, IDS-iSYS Aldosterone Calibration Verifiers Regulation Number: 21 CFR 862.1045 Regulation Name: Aldosterone test system Regulatory Class: II Product Code: CJM, JJX Dated: February 6, 2015 Received: February 12, 2015

Dear Mr. Mick Fenton:

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

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

http://www.fda.gov/MedicalDevices/ResourcesforYou/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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DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

Indications for Use

Form Approved: OMB No. 0910-0120 Expiration Date: January 31, 2017 See PRA Statement below.

510(k) Number (if known) K142994

Device Name

IDS-iSYS Aldosterone IDS-iSYS Aldosterone Control Set IDS-iSYS Aldosterone Calibration Verifiers

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.

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.

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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."

FORM FDA 3881 (8/14)

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K142994

510(k) SUMMARY

Introduction	According to the requirements of 21CFR807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.
Submitter	Immunodiagnostic Systems Ltd10 Didcot WayBoldon Business ParkBoldonTyne and WearNE35 9PDUnited KingdomContact Person: Mick FentonPhone: +44 191 5190660Fax: +44 191 5190760Email: michael.fenton@idsplc.comSecondary Contact: Roma YoungPhone: +44 191 5190660Fax: +44 191 5190760Email: roma.young@idsplc.comDate prepared: 04 April 2015
Device Name	Proprietary names:	IDS-iSYS AldosteroneIDS-iSYS Aldosterone Control SetIDS-iSYS Aldosterone Calibration Verifiers
	Common names:	As above
	Classification:	21CFR862.1045 (Class II)21CFR862.1660 (Class I, Reserved)
	Product Code:	CJMJJX
Predicate Device	The IDS-iSYS Aldosterone assay is substantially equivalent to other products in commercial distribution intended for similar use. We claim equivalency to the currently marketed Siemens Healthcare Diagnostics Ltd. Coat-A-Count® Aldosterone; TKAL 6615154 (K831178).The IDS-iSYS Aldosterone Calibration Verifiers are substantially equivalent to other products in commercial distribution intended for similar use. We claim equivalency to the currently marketed

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IDS-iSYS 25-Hydroxy Vitamin DS Calibration Verifiers (K11650). The IDS-iSYS Aldosterone Control Set is substantially equivalent to other products in commercial distribution intended for similar use. We claim equivalency to the currently marketed IDS-iSYS

Device Description The IDS-iSYS Aldosterone assay is based on chemiluminescence technology. A biotinylated monoclonal anti-Aldosterone antibody is incubated with the sample, after an incubation step an Aldosterone acridinium conjugate is added and after a further incubation step streptavidin coated magnetic particles are added. Following a third incubation step the particles are "captured" using a magnet. After a washing step and addition of trigger reagents, the light emitted by the acridinium label is inversely proportional to the concentration of Aldosterone in the original sample.

25-Hydroxy Vitamin DS Control Set (K091849).

Intended Use The IDS-iSYS Aldosterone assay (IS-3300) is a device intended for use in clinical laboratories for the quantitative determination of Aldosterone in human EDTA plasma on the IDS-iSYS Multi-Discipline Automated System. Aldosterone measurements are used in the diagnosis and treatment of primary aldosteronism (a disorder caused by excessive secretion of Aldosterone by the adrenal gland), hypertension caused by primary aldosteronism, selective hyperaldosteronism, edematous states and other conditions of electrolyte balance.
The IDS-iSYS Aldosterone Control Set (IS-3330) is intended for use as assayed quality control samples to monitor the accuracy of the IDS-iSYS Aldosterone assay on the IDS-iSYS Multi-Discipline Automated System.

The IDS-iSYS Aldosterone Calibration Verifiers (IS-3335) is a device intended for medical purposes for use in the quantitative verification of calibration of the IDS-iSYS Aldosterone assay when performed on the IDS-iSYS Multi-Discipline Automated System.

Comparison Table Table 1 Similarities and differences

Performance	Predicate	New device
Intended Use	IVD; For the quantitativemeasurement of aldosterone inserum (or heparin/EDTA plasma)	IVD; For the quantitativemeasurement of aldosterone inEDTA plasma
Analyte	Aldosterone	same
Sample matrix(primary tube type)	EDTA plasma	same
Calibrators -matrix	Lyophilised human serum		same
Reagent storage	2-8°C		same
Sample volume	200μl		same
Range of assay	2.5 to120 ng/dL (reported as 25 –1200 pg/mL)		3.9 to 120 ng/dL
Spiking recovery	Average 96.1% [range 86-111%between 232-847 pg/mL (23.2 -84.7 ng/dL)]		Average 94.8% [range 77-107.9%between 27.2-71.3ng/dL]
Analytical specificity	Bilirubin: Presence of bilirubin inconcentrations of up to 200 mg/Lhas no effect on results, within theprecision of the assay.	PotentiallyInterferingAgent	ThresholdConcentration
	Hemolysis: Presence of packed redblood cells in concentrations up to30 µL/mL has no effect on results,within the precision of the assay.	Triglyceride	500 mg/dL
	Lipemia: the 600 pg/mL calibratorwas serially diluted with each oftwo lipemic serum pools, resultsshowed excellent recoveries, evenin presence of severe lipemia.	Haemoglobin	200 mg/dL
		Bilirubin	15 mg/dL
		Albumin	8 g/dL
		Red BloodCells	0.2%
		Biotin	22 nM
		RheumatoidFactor	1000 IU/mL
		Human anti-mouseAntibodies(HAMA)	30 ng/mL
Antibody	Antiserum against aldosterone(species origin not stated)		Monoclonal mouse-anti-aldosteroneantibody
Capture	Antibody-coated polypropylenetubes		Streptavidin covalently coupled toparamagnetic particles
Method of detection	Radioactivity using 125I-labelledaldosterone		Chemiluminescence using anacridinium-ester derivative
Analytical sensitivity	11 pg/mL (1.1 ng/dL)		3.2 ng/dL
Linearity	The assay has a tendency to over-recover upon dilution. Patientsamples having values greater thanthe highest calibrator (1200pg/mL)can be reported as greater than1200 pg/mL or diluted using aserum pool. Dilution with the zerocalibrator is not recommended. Theuncorrected value observed afterdilution should be greater than 600pg/mL.		y=1.00x – 1.24ng/dL; r² = 1.00
Precision	Intra-assay: 5.4 - 2.3% at 65-813pg/mL (6.5 - 81.3 ng/dL)		Within-Run: 8.4-2.2% at 7.5 - 98.7ng/dL
	Inter-assay: 15.7- 3.8% at 58-548pg/mL (5.8 – 54.8 ng/dL)		Total: 12.8-5.2% at 7.5 – 98.7 ng/dL
Reference range	Standing: 4 - 31 ng/dL		Female Supine: 3.9- 33.6 ng/dL
	Recumbent: 1 – 16 ng/dL		Female Upright:3.9 – 50.1 ng/dLMale Supine: 3.9 – 19.5 ng/dLMale Upright: 3.9 - 23.2 ng/dL

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Cross-reactivity	Analyte	Cross-Reactivity	Analyte	Cross-Reactivity
	Aldosterone	100%	Aldosterone	100%
	Androstenedione	Not detectable	Androstenedione	<0.001%
	Androsterone	0.0005%	Androsterone	0.0003%
	Corticosterone	0.002%	Cortisol	<0.001%
	18-OH-corticosterone	0.033%	11-Deoxycortisol	<0.001%
	Cortisol	Not detectable	Cortisone	<0.001%
	Cortisone	0.0003%	Corticosterone	<0.001%
	11-Deoxycorticosterone	0.006%	11-Deoxy-corticosterone	<0.001%
	11-Deoxycortisol	0.0004%	18-OH-corticosterone	0.2%
	Dexamethasone	0.00005%	Dexamethasone	<0.001%
	DHEA	0.0005%	DHEA	<0.001%
	Estradiol	Not detectable	Estradiol	<0.001%
	Estriol	Not detectable	Estrone	<0.001%
	Estrone	Not detectable	Prednisone	<0.001%
	Fludrocortisone	Not detectable	Prednisolone	<0.001%
	Prazosin	Not detectable	Progesterone	<0.001%
	Prednisolone	Not detectable	Spironolactone	<0.001%
	Prednisone	0.00003%	Testosterone	<0.001%
	Pregnenolone	Not detectable	Prazosin HCl	<0.001%
	Progesterone	0.007%	Verapamil HCl	<0.001%
	17α-OH-Progesterone	Not detectable	Doxazosin mesylate	<0.001%
	Spironolactone	0.06%	Fludrocortisone acetate	<0.001%
	Testosterone	Not detectable	Cholesterol	<0.001%
			3α, 5β-Tetrahydoaldosterone	3.1%
Automation	Manual assay		Automated assay
Onboard reagent stability	N/A		35 days
Calibration	Full standard curve to be run with each assay run		Uses a Master Curve and a two-point, user-initiated calibration to calibrate the assay. The system stores the calibration for the interval specified in the kit IFU

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Calibrators – numberof vials	7 levels including the zero	2 levels
Calibration interval	Per assay run	4 days
Assay duration	Minimum 18hours (overnightincubation)	43 minutes to first result
Quality Control	A tri-level, human serum-basedimmunoassay control, containingaldosterone as one of over 25constituents, is available fromSiemens Healthcare Diagnostics,not included in the kit (cataloguenumber: CON6)	Requires three serum-based qualitycontrol samples to verify thecalibration. The control samples aresupplied in a separate Kit.
Kit components ofReagent Kit	- Aldosterone Antibody-coatedtubes (TAL1)- 125I-Aldosterone tracer (TAL2)- Aldosterone Calibrators Athrough G (ALC3-9)	- Reagent cartridge of:Streptavidin-Magnetic particles,Aldosterone-Acridinium conjugatetracer, Biotinylated anti-aldosteroneantibody, Assay buffer- Two levels of Calibrators (A&B)- Mini CD
Kit components ofControls Set	N/A	- Three levels of assay controls (6vials per level, 1mL per vial)- Mini CD
Method Comparison	N/A	IDS-iSYS = 1.070 (x) – 0.29,R2=0.98, n=161x= Diasorin Liaison Aldosterone
IDS-iSYSAldosteroneCalibration Verifiers	IDS-iSYS 25OHD® CalibrationVerifiers	New Device
Similarities
Intended use/Indications for use	Intended for medical purposes foruse in the quantitative verificationof calibration and assay measurablerange of the IDS-iSYS	The same
Levels	Four	Four
Unopened Stability	Six months	Six months
Differences
Format	Ready to use	Lyophilized
Analyte	25-Hydroxy Vitamin D	Aldosterone
Matrix	Horse serum	Human serum
ReconstitutedStability	2.5 hours	4 hours

IDS-iSYS	IDS-iSYS 250HD Control Set
Aldosterone Control		New Device
Set
Similarities
Intended use/	Intended for medical purposes for
Indications for use	used for quality control of the assay	The same

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	on the IDS-iSYS Multi-DisciplineAutomated System.
Levels	Three	Three
Unopened Stability	Six months	Six months
Differences
Format	Ready to use	Lyophilized
Analyte	25-Hydroxy Vitamin D	Aldosterone
Matrix	Horse serum	Human serum
ReconstitutedStability	2.5 hours	4 hours

Performance Characteristics

1. Analytical performance:

a. Precision/Reproducibility:

Precision was determined in accordance with CLSI EP5-A2, "Evaluation of Precision Performance of Quantitative Measurement Methods". Assessment was made for the following variables:

within run precision;
between run precision
total precision

Precision studies were performed on two sites using three analyzers and three manufacturing batches (cartridge reagents and calibrators) by four operators.

To assess the variables of reproducibility, three lots of IDS-iSYS Aldosterone assay reagents were used to assay nine samples over a minimum of 20 assay days. Samples were assayed in duplicate, twice a day, to provide 80 replicates over 40 runs. Three instruments were used with each reagent batch and operated by a total of four personnel during the study. The sample concentrations were interpolated by using two-point calibration (Kit Cal A and B) as intended for use by the end user.

The data used to provide the claims to be inserted in the Labelling were obtained from six EDTA plasma samples which were assayed using three lots of reagents. The samples were run in duplicate twice per day for 20 days on 2 instruments and are the following:

Concentrationng/dL (pmol/L)	n	Within-run		Total
		SD	CV%	SD	CV%
7.5 (208)	80	0.6	8.4	1.0	12.8
21.5 (594)	80	1.0	4.8	1.5	6.9
29.3 (812)	80	0.9	3.2	1.6	5.5
49.3 (1366)	80	1.6	3.3	2.6	5.2
65.7 (1820)	80	1.4	2.2	4.1	6.2
98.7 (2734)	80	4.0	4.1	5.9	6.0

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b. Linearity/Assay Reportable Range:

The linear range of the assay was determined following a protocol based on the CLSI guidance EP6-A "Evaluation of the linearity of quantitative measurement procedures: a statistical approach".

A high plasma sample with aldosterone concentrations at 133.6ng/dL was diluted with a low sample at 2.7 ng/dL in order to assess the linearity covering the range below and including the LoQ. The 11 dilution samples were run in quadruplicate with one reagent batch. This one linearity data set was chosen in order to justify the claims to be included in the Instructions for use as it covered the entire claimed measuring range (LoQ 3.9 to 120 ng/dL).

The linear regression equation obtained when the observed results were plotted against the expected results was: y = 1.00x - 1.24, R2 = 1.00

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

Traceability

The IDS-iSYS Aldosterone calibrators, controls and calibration verifiers are traceable to in-house reference calibrators produced by dissolving Aldosterone ( ≥95% HPLC; A9477Sigma-Aldrich) in Dioxane (anhydrous, 99.8%; 296309 Sigma-Aldrich). The concentration was calculated by UV quantitation using the molar extinction coefficient of s =15000 at an absorbance of 240mm. Preparation was gravimetric with no adjustment.

Value Assignment:

Value assignment of the IDS-iSYS Aldosterone Kit Calibrators: for each manufacturing batch the new kit calibrator sets are run as 'unknowns' in duplicate in at least 20 assays on one analyser. The concentrations of the calibrators to be calculated by using Prism software package following issued QC procedures.

The concentration values obtained for the calibrators must fall within specified ranges. The data of 20 runs Internal Reference Calibrators are used to generate the logistic parameters of the Master calibration curve by using Prism software package. The calibrator concentration and master curve parameters are reagent batch specific and linked together. Verification of the calibration is performed by running three assays with On Board 2 point calibration on three different analyzers with controls of known levels.

The Kit Calibrator A and Calibrator B, when assayed by the user, provides an adjustment to trace the user's calibration curve to the master calibration curve for the specific reagents batch of IDS-iSYS Aldosterone kit.

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Value assignment of the IDS-iSYS Aldosterone Kit Controls:

To assign acceptable ranges to the Kit Assay Controls (CTL1, CTL2 and CTL3), the controls are assayed in the IDS-iSYS Aldosterone assay with On Board 2 point calibration. The new Assay Controls are run in triplicate in at least 18 assays on a total of at least three analyzers. The acceptable range is then calculated as the mean ± 3 standard deviations for Kit Controls 1, 2, and 3, respectively.

Value assignment of the IDS-iSYS Aldosterone Calibration Verifiers: The value allocation of calibration verifiers (CV0, 1, 2 and 3) is performed the same way as the Kit Assay controls.

Stability:

The stability of the IDS-iSYS Aldosterone assay Cartridge, Calibrators A and B, Kit Controls 1-3 and Calibration verifiers was derived from assessment by storage at various conditions, following EN 13640:2002 guideline (CLSI EP25-A), Stability Testing of In Vitro Diagnostic Reagents.

Reagentshelf life	Cartridge	Calibrators	Controls	CalibrationVerifiers
Beforeopening at2 - 8 °C	Six months	twelvemonths	Fifteen months
Cartridge,afteropening at2 - 8 °C	35 days	NA	NA	NA
On boardthe System	35 days	4 Hours	4 Hours	4 Hours
Frozen at<-20°C	N/A	5 weeks	5 weeks	5 weeks

d. Detection limit:

Study Protocol

The limit of blank (LoB) and limit of detection (LoD) and limit of quantitation (LoO) were determined based on guidance from CLSI EP17-A "Protocols for the determination of limits of detection and limits of quantitation"

LoB, LoD and LoQ for two reagent lots were performed on 2 different analyzers, at two different sites. For the first reagent lot the LoB was determined in a plasma sample with zero aldosterone levels, in 10 replicates for 5 separate days for a total of 50 replicates. The resulting LoB level was 1.71 ng/dL. The LoD and LoQ for the same reagent batch were determined in 7 samples (concentrations ranging from 2.1 to 8.8ng/dL), run in 2 replicates, once per day

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for 8 days. The resulting parametric LoD was 3.05 ng/dL and the LoQ at 20% CV was 3.9 ng/dL.

For the second reagent lot the LoB was determined in another plasma sample with zero aldosterone levels, in 6 replicates for 5 separate days for a total of 30 replicates. The resulting LoB was 2.03 ng/dL. The LoD in the second reagent lot was determined in 8 samples (with concentrations between 0.31 and 8.5 ng/dL), run in 2 replicates, once per day for 5 days. The LoD was determined in 7 samples (with concentrations between 1.1 and 7.8 ng/dL), run in 2 replicates, once per day for 5 days. The resulting LoQ was 3.8 ng/dL. In each case for the LoB, LoD and LoQ the highest values were taken for the Instruction for use claims.

LoB	2.0 ng/dL(55.4 pmol/L)
LoD	3.2 ng/dL(88.6 pmol/L)
LoQ	3.9 ng/dL(108 pmol/L)

e. Analytical specificity:

The sponsor performed interference and cross-reactivity studies were in accordance with CLSI guidance EP7-A2"Interference testing in clinical chemistry".

Interference Screen testing was performed for the following substances: Triglycerides, Haemoglobin, Bilirubin, protein (albumin), Human anti-mouse antibody (HAMA), Red Blood Cells, Biotin and Rheumatoid Factor. To determine potential interference in the specific detection of Aldosterone, two base plasma samples, one in the range of approximately 10ng/dL Aldosterone ("Low") and another at approximately 40ng/dL ("High"), were spiked with the potential interferent. Control samples (blank) were spiked with a volume of relevant diluent equal to that of the spiked interferent. The mean of 26 replicates, for both spiked and control samples (Blank), were then compared. One reagent batch was used for interference testing. The differences observed between the mean spiked and control sample values were examined and assessed according to acceptance criteria of ≤10% concentration bias to the unspiked sample.

All interferent spiking was performed just prior the assay.

In the case of protein interference, the native content was first measured by the Bradford test before the albumin spiking was performed.

% Interference was calculated using the formula % Interference = (mean spiked value - mean control value) / mean control value x 100

Interference was tested on two base plasma samples at two different clinically relevant Aldosterone concentrations. One Aldosterone level sample was used

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with approx 10-15 ng/dL concentration. Where a neat sample at the desired concentration was not available Aldosterone was spiked into plasma pools created from various low Aldosterone content native plasma samples. Similarly for the high sample Aldosterone was either spiked into plasma pools (at approx. 40-60ng/dL) or native high sample pools were created to assess the interference at the higher part of the assay range. The interference materials were spiked into both samples. The baseline concentrations (Blank) were established in 26 replicates during the interference measurement. The control and interferent spiked samples (test) were measured in alternating order.

In the case of protein interference testing, the native protein content in the sample was first measured by the Bradford test to ascertain if the native content was at the level required in the specifications (6g/dL). If the native protein content was lower then human serum albumin (HSA) was spiked to bring the concentration up to 6g/dL and this was used to compare the interference in Aldosterone levels with the same sample spiked to 8g/dL HSA. The Bradford test was therefore carried out at least twice following the spiking of HSA and the sample was then put to the iSYS analyzer for determination of Aldosterone levels.

Stock solutions of each interferent were prepared in a diluent appropriate for the particular substance to be tested for interference (in most cases Scantibodies charcoal stripped lipid stripped serum). Samples were then spiked with the stock solution to a defined concentration of the potential interferent. Un-spiked control (Blank) samples were prepared using an equivalent volume of the diluent used to prepare the potential interfering substance.

Percent interference was calculated using the formula below:

% Interference = (mean spiked concentration - mean un-spiked concentration) x 100 mean un-spiked concentration

For assessing RF interference, recovery and linearity studies were performed. One plasma RF sample was used to compare the observed versus expected recovery by using ~10ng/dL and ~40ng/dL spiking with aldosterone antigen. The percent recoveries were calculated using the following calculation:

Recovery = Obs mean spiked value - Obs mean unspiked value

% Recovery = (Observed Recovery value / Expected Recovery value (Analyte added)) x 100

Cross-reactivity experiments were performed from stock materials prepared gravimetrically to a top dose (up to 20000x the Aldosterone assay top dose). Cross-reactivity was defined as the point where the reduction in signal corresponds to 50% of the signal achieved in the absence of analyte (B/Bo of 50%), as a percentage of the analyte concentration giving the same fall in signal

% Cross-reactivity = ED50 (ng/dL) Aldosterone ED50 (ng/dL) compound

Stock concentrations of the substances to be checked for cross-reactivity were prepared initially in an organic solvent. This stock solution of the cross-reactant

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was then spiked into Scantibodies serum with zero aldosterone concentration and subsequently diluted down serially to create a 7-point standard curve for each substance. Curves were run in the same experiment for comparison of ED-50 values of the potential cross-reactant against the ED50 of the Aldosterone assay displacement curve.

Result Summary

Potentially InterferingAgent	ThresholdConcentration
Triglyceride	500mg/dL
Haemoglobin	200mg/dL
Bilirubin	15mg/dL
Albumin	8g/dL
Red Blood Cells	0.2%
Biotin	22nM
Rheumatoid Factor	1000IU/mL
Human anti-mouseAntibodies (HAMA)	30ng/mL

Limitations stated in the package inset:

In patients receiving therapy with a high biotin dose (i.e. >5 mg/day), no sample should be taken until at least 8 hours after the last biotin administration.

Hemolyzed samples should not be used with this assay.

Analyte	Cross-Reactivity
Aldosterone	100%
Androstenedione	<0.001%
Androsterone	<0.001%
Cortisol	<0.001%
11-Deoxycortisol	<0.001%
Cortisone	<0.001%
Corticosterone	<0.001%
11-Deoxycorticosterone	<0.001%
18-Hydroxycorticosterone	0.2%
Dexamethasone	<0.001%
DHEA	<0.001%
Estradiol	<0.001%
Estrone	<0.001%
Prednisone	<0.001%
Prednisolone	<0.001%
Pregnenolone	<0.001%
Progesterone	<0.001%
Spironolactone	<0.001%
Testosterone	<0.001%
Prazosin HCl	<0.001%
Verapamil HCl	<0.001%
Doxazosin mesylate	<0.001%
Fludrocortisone acetate	<0.001%
Cholesterol	<0.001%
3α, 5β-Tetrahydroaldosterone	3.1%

2. Comparison Studies:

Correlation studies based on guidance from the CLSI protocol EP9-A2 were performed to compare the IDS-iSYS Aldosterone assay and the Diasorin Liaison Aldosterone assay (previously cleared in K130321). The results of the statistical methods used for comparison of agreement between the IDS-iSYS Aldosterone

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assay and the Liaison Aldosterone assay in 161 samples (including 12 altered samples) were as follows:

The linear regression correlation of the IDS-iSYS to the Diasorin Aldosterone assay in this set of plasma samples gave a slope of 1.053 (95%CI: 1.029 to 1.076) and intercept of 0.09ng/dL (95%CI: -0.86 to 1.05), R2= 0.980. Sample range tested was 3.9 - 110.9 ng/dL on the candidate device and 3.0 - 98.0 ng/dL on the comparative device.

Passing-Bablok analysis of the same data gave an intercept of -0.29 (95%CI: -0.98 to 0.46) and slope of 1.070 (95%CI: 1.043 to 1.096).

3. Reference range study (Expected values):

In order to determine the normal ranges for the IDS-iSYS Aldosterone assay, 228 Caucasian adult samples, collected in the US, 18-65 years of age, with normal blood pressure (systolic/diastolic ≤120/80) and normal BMI (18.5-24.9) were analysed using the IDS-iSYS Aldosterone assay. Samples were taken between 7-10am after overnight fasting, upright (30 minutes standing or walking) and supine (lying down for at least 30 minutes). The results obtained are given in the table below:

	n	Mean	SD	Range(Central 95%)
FemaleSupine	55	9.9 ng/dL(274 pmol/L)	9.4 ng/dL	<3.9 ng/dL-33.6 ng/dL
FemaleUpright	56	13.8 ng/dL(382 pmol/L)	11.9 ng/dL	<3.9 ng/dL-50.1 ng/dL
MaleSupine	56	5.1 ng/dL(141 pmol/L)	4.7 ng/dL	<3.9 ng/dL-19.5 ng/dL
MaleUpright	61	7.8 ng/dL(216 pmol/L)	4.8 ng/dL	<3.9 ng/dL-23.2 ng/dL

Conclusion

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

Regulation Number and Section

§ 862.1045 Aldosterone test system.

(a)
Identification. An aldosterone test system is a device intended to measure the hormone aldosterone in serum and urine. Aldosterone measurements are used in the diagnosis and treatment of primary aldosteronism (a disorder caused by the excessive secretion of aldosterone by the adrenal gland), hypertension caused by primary aldosteronism, selective hypoaldosteronism, edematous states, and other conditions of electrolyte imbalance.(b)
Classification. Class II.