The Immunalysis Ethyl Alcohol Enzyme Assay is an in vitro diagnostic device for the quantitative analysis of ethyl alcohol (ethanol) in human urine, serum or plasma with automated clinical chemistry analyzers. The measurement of ethanol is used for the diagnosis and treatment of alcohol intoxication and poisoning. This assay is calibrated against ethyl alcohol. This device is intended for prescription use only.

Device Description

The Immunalysis Ethyl Alcohol Assay is based on the oxidation of ethyl alcohol to acetaldehyde by alcohol dehydrogenase (ADH) and nicotinamide adenine dinucleotide (NAD) reduced to NADH resulting in an absorbance change measured spectrophotometrically at 340nm. The concentration of ethanol in the sample is directly proportional to the ADH activity.

AI/ML Overview

The Immunalysis Ethyl Alcohol Enzyme Assay is an in vitro diagnostic device for the quantitative analysis of ethyl alcohol (ethanol) in human urine, serum, or plasma using automated clinical chemistry analyzers. The measurement of ethanol is used for the diagnosis and treatment of alcohol intoxication and poisoning.

1. Table of Acceptance Criteria and Reported Device Performance

Performance Characteristic	Acceptance Criteria (Implicit from Study Design)	Reported Device Performance
Limit of Blank (LoB)	Calculation from sixty (60) blank measurements of unaltered drug-free negative human urine, serum, and plasma.	Urine: 0.481 mg/dL Serum: 0.668 mg/dL Plasma: 0.652 mg/dL
Limit of Detection (LoD)	Testing of four samples per matrix (urine, serum, plasma) at concentrations in the range LoB to 4xLoB, in replicates of five, on two reagent lots over three days.	Urine: 1.3 mg/dL Serum: 1.7 mg/dL Plasma: 1.7 mg/dL
Limit of Quantitation (LoQ)	Four independent samples of each matrix spiked at the LoQ concentration were tested on two reagent lots, demonstrating suitable accuracy.	All matrices: 2.9 mg/dL (quantitatively determined with suitable accuracy)
Linearity/Recovery	Serially diluted samples (from high concentration EtOH to drug-free) covering 3 mg/dL to 600 mg/dL, tested in triplicate. Slope and r² values to demonstrate linearity. Recovery percentages.	Urine: Slope: 0.996, Intercept: -0.172, r²: 0.999 (Recovery 97.5% - 102.6%) Serum: Slope: 0.978, Intercept: 2.869, r²: 0.999 (Recovery 96.6% - 104.8%) Plasma: Slope: 0.970, Intercept: 0.307, r²: 0.999 (Recovery 95.1% - 106.1%)
Precision	Performed for 20 days, 2 runs per day in duplicate (N=80 for each matrix) on drug-free samples spiked with EtOH at 25, 75, 125, 150, 175, 200 mg/dL and calibrators/controls at 50, 100, 300 mg/dL. Spiked concentrations confirmed by GC-FID. CV% values for repeatability, between run, and within-laboratory.	Urine: Within-Laboratory CV% range from 1.5% to 1.8% for spiked samples. Serum: Within-Laboratory CV% range from 2.0% to 3.2% for spiked samples. Plasma: Within-Laboratory CV% range from 1.5% to 4.9% for spiked samples.
Specificity and Cross-Reactivity	Structurally and functionally similar compounds spiked into drug-free urine, serum, and plasma at levels yielding the equivalent of 10 mg/dL and 100 mg/dL EtOH. Verified ability of the device to exclusively determine certain drugs.	Low cross-reactivity for most tested compounds (<0.3% to <3.3%). n-Propanol showed higher cross-reactivity (Urine: 16.7% at 10 mg/dL, 9.8% at 100 mg/dL; Serum/Plasma: 11.1% at both 10 mg/dL and 100 mg/dL).
Interference (Structurally Unrelated Compounds)	Structurally unrelated compounds spiked into drug-free urine containing EtOH at 10 mg/dL and 100 mg/dL. Assay performance to be unaffected.	Assay performance was unaffected by all tested compounds at specified concentrations (e.g., Acetaminophen 500,000 mg/dL, Caffeine 500,000 mg/dL, Ibuprofen 100,000 mg/dL, etc.).
Interference (Endogenous Compounds & Urine Preservatives)	Endogenous compounds and urine preservatives spiked into drug-free urine containing EtOH at 10 mg/dL and 100 mg/dL. Assay performance to be unaffected.	Assay performance was unaffected by all tested endogenous compounds (e.g., Ascorbic Acid 1.5 g/dL, Glucose 2.0 g/dL, Hemoglobin 0.3 g/dL) and urine preservatives (e.g., Boric Acid 1% w/v, Sodium Azide 1% w/v).
Interference (Interferents in Serum & Plasma)	Potential interfering substances and anticoagulants spiked into drug-free serum and plasma containing EtOH at 10 mg/dL and 100 mg/dL. Assay performance to be unaffected.	Assay performance was unaffected by all tested compounds in serum/plasma (e.g., Bilirubin total 60-80 mg/dL, Hemoglobin 62.5-1000 mg/dL, Triglycerides 500 mg/dL) and anticoagulants (e.g., Heparin 3 U/mL, EDTA 2 mg/mL).
Interference (pH)	Device performance tested over a range of urine pH values (3.0 to 11.0) with EtOH at 10 mg/dL and 100 mg/dL. No interference observed.	No interference observed at urine pH values ranging from 3.0 to 11.0.
Interference (Specific Gravity)	Device performance tested over a range of physiologically relevant urine specific gravity values (1.000 to 1.030) with EtOH at 10 mg/dL and 100 mg/dL. No interference observed.	No interference observed at urine specific gravity values ranging from 1.000 to 1.030.
Calibration Duration	Drug-free negative urine spiked with EtOH at 10, 100, and 300 mg/dL tested up to 14 days, using an initial two-point calibration. Test results met acceptance criteria.	Calibration is stable for 14 days. Recommended frequency of calibration is 14 days.
Specimen Stability	Urine, serum, and plasma samples spiked with EtOH at 100 mg/dL and 300 mg/dL, stored in industry standard collection containers and tested at various time points and temperatures (up to 30°C and 2°C - 8°C).	Urine: Stable up to 14 days at 30°C, up to 6 months at 2°C - 8°C. Serum: Stable up to 10 days at 30°C, up to 14 days at 2°C - 8°C. Plasma: Stable up to 3 days at 30°C, up to 6 days at 2°C - 8°C.
Method Comparison	Comparison against a predicate device using de-identified, unaltered leftover clinical urine, serum, and plasma samples. Correlation coefficient (r²) and slope/intercept values.	Urine: n=80, Slope: 1.0284, Intercept: -2.7848, r²: 0.9964 (Range 3-600 mg/dL) Serum: n=76, Slope: 1.0067, Intercept: -5.8802, r²: 0.9948 (Range 3-600 mg/dL) Plasma: n=64, Slope: 1.0595, Intercept: -7.3902, r²: 0.9932 (Range 3-600 mg/dL)

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

Limit of Blank (LoB): 60 blank measurements for urine, 60 for serum, 60 for plasma samples.
Limit of Detection (LoD): 4 samples per matrix, each tested in replicates of 5 (20 measurements per matrix).
Limit of Quantitation (LoQ): 4 independent samples per matrix.
Linearity/Recovery: 12 concentrations for urine, 12 for serum, 12 for plasma, each tested in triplicate.
Precision: 80 samples per matrix (urine, serum, plasma) at each of 10 concentration levels (25, 50, 75, 100, 125, 150, 175, 200, 300 mg/dL, plus 0 mg/dL for urine and 0.4 mg/dL for serum and 0.2 mg/dL for plasma). This implies 800 data points per matrix for precision.
Specificity and Cross-Reactivity: Compounds tested at two concentrations (10 mg/dL and 100 mg/dL equivalent EtOH) in urine, serum, and plasma.
Interference (Structurally Unrelated Compounds): Numerous compounds tested at specific concentrations with EtOH at 10 mg/dL and 100 mg/dL in urine.
Interference (Endogenous Compounds & Urine Preservatives): Various compounds tested with EtOH at 10 mg/dL and 100 mg/dL in urine.
Interference (Interferents in Serum & Plasma) & Anticoagulants: Various compounds and anticoagulants tested with EtOH at 10 mg/dL and 100 mg/dL in serum and plasma.
Interference (pH): Test samples prepared in drug-free urine containing EtOH at 10 mg/dL and 100 mg/dL, tested across a pH range of 3.0 to 11.0.
Interference (Specific Gravity): Test samples prepared in drug-free urine containing EtOH at 10 mg/dL and 100 mg/dL, tested across specific gravity range of 1.000 to 1.030.
Calibration Duration: Drug-free negative urine spiked with EtOH at 10, 100, and 300 mg/dL tested at various time points up to 14 days.
Specimen Stability: Urine, serum, and plasma samples spiked with EtOH at 100 mg/dL and 300 mg/dL, tested at various time points and temperatures.
Method Comparison: 80 de-identified, unaltered leftover clinical urine samples; 76 de-identified, unaltered leftover clinical serum samples; 64 de-identified, unaltered leftover clinical plasma samples.

Data Provenance:
The data provenance specifies "unaltered drug-free negative human urine, serum, and plasma" for many studies and "de-identified, unaltered leftover clinical urine samples," "de-identified, unaltered leftover clinical serum samples," and "de-identified, unaltered leftover clinical plasma samples obtained from clinical testing laboratories" for the method comparison. This indicates the data is retrospective and derived from human biological samples, likely from clinical settings. The country of origin is not explicitly stated, but given the FDA submission, it is typically expected to be from studies conducted under US or internationally recognized standards.

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

This type of device (in vitro diagnostic assay for chemical analysis) does not typically involve human expert consensus for establishing ground truth in the same way an imaging AI algorithm would. Instead, the ground truth for samples (e.g., spiked concentrations, known concentrations in clinical samples) is established through reference methods or highly accurate laboratory techniques.

For the precision study, spiked concentrations were confirmed by Gas Chromatography - Flame Ionization Detector (GC-FID), which is a highly accurate analytical method.

4. Adjudication Method for the Test Set

Not applicable for this type of in vitro diagnostic assay. Ground truth is established by analytical methods, not human adjudication of subjective 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 in vitro diagnostic assay, not an AI-assisted diagnostic imaging or interpretation device that would involve human readers.

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

Yes, the device operates in a standalone (algorithm only) manner. It is an automated clinical chemistry analyzer which quantitatively measures ethyl alcohol. Its performance is evaluated directly through the performance studies described (LoB, LoD, LoQ, Linearity, Precision, Specificity, Interference, etc.). These studies quantify the device's accuracy and reliability in generating results, without human interpretation of the assay's direct output.

7. The Type of Ground Truth Used (expert consensus, pathology, outcomes data, etc.)

The ground truth used for this device is based on:

Known concentrations of spiked ethyl alcohol in various matrices (urine, serum, plasma).
Reference analytical methods, specifically Gas Chromatography - Flame Ionization Detector (GC-FID), to confirm spiked concentrations and potentially for method comparison where samples were cross-validated.
Comparison to a legally marketed predicate device for method comparison, which itself would have established performance characteristics against accepted methods.

8. The Sample Size for the Training Set

Not applicable. This device is an in vitro diagnostic assay based on an enzymatic reaction, not a machine learning or AI model that requires a training set.

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

Not applicable, as there is no training set for this type of device.

Summary

{0}------------------------------------------------

Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA logo on the right. The FDA logo is in blue and includes the letters "FDA" followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in a stacked format.

October 31, 2018

Immunalysis Corporation Wenying (Jessica) Zhu Regulatory Affairs Specialist III 829 Towne Center Drive Pomona, CA 91767

Re: K181553

Trade/Device Name: Immunalysis Ethyl Alcohol Enzyme Assay Regulation Number: 21 CFR 862.3040 Regulation Name: Alcohol test system Regulatory Class: Class II Product Code: DIC Dated: September 19, 2018 Received: September 20, 2018

Dear Wenying (Jessica) Zhu:

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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

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

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part

{1}------------------------------------------------

801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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 comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice

(https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

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

{2}------------------------------------------------

Indications for Use

510(k) Number (if known) K181553

Device Name Immunalysis Ethyl Alcohol Enzyme Assay

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.

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

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

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

{3}------------------------------------------------

K181553 510(k) SUMMARY

A. GENERAL INFORMATION

Applicant Name:	Immunalysis Corporation829 Towne Center DrivePomona, CA 91767Establishment # 2020952
Company Contact:	Wenying (Jessica) ZhuRegulatory Affairs Specialist IIIPhone: (909) 451-6697Email: wzhu@immunalysis.com
Date Prepared:	September 19, 2018

B. DEVICE IDENTIFICATION

Trade or Proprietary Names:	Immunalysis Ethyl Alcohol Enzyme Assay
Common Name:	Ethyl Alcohol Enzyme Assay

C. REGULATORY INFORMATION

Device Classification Name:	Alcohol Test System
Product Codes:	DIC
Regulatory Class:	II
Classification Regulation:	21 CFR 862.3040, Alcohol Test System
Panel:	Toxicology (91)
Predicate Device:	Lin-Zhi Ethyl Alcohol Enzymatic Assay [K032461]

D. DEVICE DESCRIPTION

E. INTENDED USE

{4}------------------------------------------------

Image /page/4/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The background is shaped like a horizontal arrow. The word is written in all capital letters. The image is simple and clear.

Attribute	Predicate DeviceLin-Zhi Ethyl Alcohol EnzymaticAssay [K032461]	Candidate DeviceImmunalysis Ethyl Alcohol EnzymeAssay
Intended Use	For quantitative analyses of ethylalcohol in human urine, serum, orplasma.	Same
Sample Matrix	Urine, serum or plasma	Same
Measured Analytes	Ethyl Alcohol (EtOH)	Same
Test Principle	Enzymatic assay	Same
User Environment	For use in laboratories	Same
Test Methodology	In the presence of nicotinamideadenine dinucleotide (NAD), ADHconverts ethyl alcohol toacetaldehyde and reduces NAD toNADH. The ethyl alcoholconcentration is then directlyproportional to the ADH activity.The enzyme activity is measured at340 nm wavelength.	Same
Instrumentation	Performed on clinical chemistryanalyzers	Same
Reagent Storage	2-8°C until expiration date	Same
Assay Range	3-600 mg/dL	Same

F. COMPARISON WITH PREDICATE

G. PERFORMANCE CHARACTERISTICS

The following laboratory performance studies were performed to determine substantial equivalence of the Immunalysis Ethyl Alcohol Enzyme Assay to the predicate device. Assay performance was established using the Olympus AU400e analyzer.

1. Limit of Blank (LoB)/Limit of Detection (LoD)

Sixty (60) blank measurements were taken from unaltered drug free negative human urine, serum and plasma to calculate Limit of blank (LoB). Four samples of each matrix containing concentrations in the range LoB to 4xLoB were tested in replicates of five on two reagent lots over three days to calculate Limit of Detection (LoD). LoB of the assay is 0.481 mg/dL for urine and 0.668 mg/dL for serum and 0.652 mg/dL for plasma. LoD of the assay is 1.3 mg/dL for urine, 1.7 mg/dL for serum and 1.7 mg/dL for plasma.

2. Limit of Quantitation (LoQ)

Limit of Quantitation (LoQ) is selected at a level equal to or higher than LoD. Four independent samples of each matrix spiked at concentration of LoQ were tested on two reagent lots to verify LoQ. Results demonstrated that drug concentration equal to or above LoQ, 2.9 mg/dL, can be quantitatively determined with suitable accuracy.

{5}------------------------------------------------

3. Linearity/Recovery

A linearity study in the quantitative mode was conducted by spiking a drug free urine, serum and plasma pool with a high concentration of EtOH at upper limit of claimed measuring range. Additional pools were made by serially diluting the high concentration specimen with drug free urine, serum and plasma to achieve concentrations ranging from 3 mg/dL to 600 mg/dL. The high value specimen was evaluated with the predicate device to determine initial high-level value. The 0 mg/dL specimen was made from drug free urine, serum and plasma. Each pool was tested in triplicate to calculate the mean concentration values that were used to calculated drug recovery. Linearity test results are presented in Table 1 to Table 3.

Expected Concentration(mg/dL)	Mean ObservedConcentration(mg/dL)	Recovery (%)
0.0	0.0	N/A
3.0	3.1	102.2
12.0	12.0	100.3
72.0	73.9	102.6
132.0	132.2	100.2
192.0	195.4	101.8
252.0	250.3	99.3
312.0	308.0	98.7
372.0	363.0	97.6
432.0	421.3	97.5
492.0	484.4	98.5
552.0	565.8	102.5
Slope		0.996
Intercept		-0.172
r2		0.999

Table 1. Linearity/Recovery Test Results - Urine

Table 2. Linearity/Recovery Test Results - Serum

Expected Concentration(mg/dL)	Mean ObservedConcentration(mg/dL)	Recovery (%)
0.0	0.0	N/A
3.0	3.0	101.1
23.2	23.1	99.4
83.2	87.2	104.8
143.2	146.1	102.0
203.2	199.8	98.3
263.2	262.2	99.6
323.2	317.8	98.3
383.2	385.9	100.7

{6}------------------------------------------------

Image /page/6/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The background is shaped like a horizontal arrow. The word is written in all capital letters and is centered on the arrow. The arrow is pointing to the right.

Expected Concentration(mg/dL)	Mean ObservedConcentration(mg/dL)	Recovery (%)
443.2	441.4	99.6
503.2	493.6	98.1
563.2	544.3	96.6
Slope	0.978
Intercept	2.869
r²	0.999

Table 3. Linearity/Recovery Test Results - Plasma

Expected Concentration(mg/dL)	Mean ObservedConcentration(mg/dL)	Recovery (%)
0.0	0.0	N/A
3.0	3.0	98.9
9.9	10.5	106.1
69.9	69.7	99.8
129.9	127.5	98.2
189.9	188.4	99.2
249.9	242.3	96.9
309.9	299.9	96.8
369.9	351.9	95.1
429.9	410.0	95.4
489.9	470.4	96.0
549.9	547.0	99.5
Slope	0.970
Intercept	0.307
r2	0.999

4. Precision

Precision study was performed for 20 days, 2 runs per day in duplicate on drug free urine, serum and plasma (N=80 for each matrix) spiked with EtOH to concentrations of 25 mg/dL, 75 mg/dL, 125 mg/dL, 150 mg/dL, 175 mg/dL and 200 mg/dL and calibrators and controls at concentrations of 50 mg/dL, 100 mg/dL and 300 mg/dL. The spiked concentrations were confirmed by Gas Chromatography - Flame Ionization Detector (GC-FID). The study established assay reproducibility and verified the precision performance in the claimed measuring range. Precision test results are presented in Table 4 to Table 6.

{7}------------------------------------------------

Image /page/7/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The word is written in all capital letters and is centered in the image. The background is a solid red color, and the letters are a bright white color. The image is simple and easy to read.

Concentration(mg/dL)	# ofSample	MeanConc.(mg/dL)	Repeatability		Between Run		Within-Laboratory
	Sample	(mg/dL)	SD	CV%	SD	CV%	SD	CV%
0	80	0.0	0.1	N/A	0.0	N/A	0.1	N/A
25	80	24.5	0.3	1.2	0.3	1.1	0.5	1.8
50	80	49.0	0.4	0.9	0.6	1.2	0.8	1.5
75	80	74.0	0.7	0.9	1.0	1.3	1.1	1.5
100	80	99.6	0.8	0.8	1.6	1.6	1.6	1.6
125	80	121.3	1.2	1.0	1.5	1.2	1.8	1.5
150	80	146.1	1.3	0.9	1.9	1.3	2.4	1.7
175	80	169.1	1.4	0.9	2.0	1.1	2.7	1.6
200	80	193.9	1.8	0.9	2.6	1.3	3.4	1.7
300	80	292.0	2.2	0.8	3.8	1.3	4.6	1.6

Table 4. Precision - Urine

Table 5. Precision – Serum

Concentration(mg/dL)	# ofSample	MeanConc.(mg/dL)	Repeatability		Between Run		Within-Laboratory
			SD	CV%	SD	CV%	SD	CV%
0	80	0.4	0.2	N/A	0.1	N/A	0.2	N/A
25	80	25.8	0.2	0.9	0.5	2.0	0.8	3.2
50	80	49.3	0.3	0.7	1.0	2.0	1.0	2.0
75	80	75.7	0.4	0.5	1.4	1.8	2.2	3.0
100	80	99.6	0.5	0.5	2.0	2.1	2.1	2.1
125	80	125.9	0.8	0.6	2.4	1.9	3.9	3.1
150	80	148.4	1.0	0.6	2.6	1.7	3.8	2.6
175	80	172.3	1.0	0.6	3.1	1.8	5.1	3.0
200	80	195.2	1.3	0.7	3.4	1.8	4.9	2.5
300	80	290.4	1.4	0.5	5.5	1.9	6.0	2.1

Table 6. Precision - Plasma

Concentration(mg/dL)	# ofSample	MeanConc.(mg/dL)	Repeatability		Between Run		Within-Laboratory
			SD	CV%	SD	CV%	SD	CV%
0	80	0.2	0.1	N/A	0.1	N/A	0.1	N/A
25	80	24.3	0.2	0.7	0.2	1.0	1.2	4.9
50	80	49.9	0.3	0.6	0.5	0.9	0.7	1.5
75	80	69.8	0.4	0.5	0.7	1.0	1.8	2.6

{8}------------------------------------------------

Image /page/8/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The background is shaped like a horizontal hexagon. The text is centered in the hexagon. The font is sans-serif and bold.

Concentration(mg/dL)	# ofSample	MeanConc.(mg/dL)	Repeatability		Between Run		Within-Laboratory
100	80	100.5	0.5	0.5	1.0	1.0	1.6	1.6
125	80	118.3	0.6	0.5	1.0	0.9	2.7	2.2
150	80	155.8	0.9	0.6	1.2	0.8	4.1	2.6
175	80	170.6	0.9	0.5	1.5	0.9	3.2	1.9
200	80	197.6	1.1	0.6	1.4	0.7	3.9	2.0
300	80	292.7	2.0	0.7	2.7	0.9	5.2	1.8

5. Specificity and Cross-Reactivity

Structurally and functionally similar compounds were spiked into drug free urine, serum and plasma at levels that will yield a result that is equivalent to the concentration of 10 mg/dL and 100 mg/dL. The study verified assay performance relative to the ability of the device to exclusively determine certain drugs, in quantitative mode. Cross-reactivity test results are presented in Table 7 to Table 9.

Table 7. Cross-Reactivity - Urine

	10 mg/dL		100 mg/dL
Compound	Compound Conc.(mg/dL)	Cross-Reactivity(%)	Compound Conc.(mg/dL)	Cross-Reactivity(%)
Lot #1
Acetaldehyde	3,000	<0.3	3,000	<3.3
Acetone	3,000	<0.3	3,000	<3.3
n-Butanol	500	2.0	3,000	<3.3
Ethylene Glycol	3,000	<0.3	3,000	<3.3
Isopropanol	3,000	<0.3	3,000	<3.3
Methanol	3,000	<0.3	3,000	<3.3
n-Propanol	60	16.7	1,025	9.8
Propylene Glycol	3,000	<0.3	3,000	<3.3
Lot #2
Acetaldehyde	3,000	<0.3	3,000	<3.3
Acetone	3,000	<0.3	3,000	<3.3
n-Butanol	500	2.0	3,000	<3.3
Ethylene Glycol	3,000	<0.3	3,000	<3.3
Isopropanol	3,000	<0.3	3,000	<3.3
Methanol	3,000	<0.3	3,000	<3.3
n-Propanol	60	16.7	1,025	9.8
Propylene Glycol	3,000	<0.3	3,000	<3.3
Lot #3

{9}------------------------------------------------

Image /page/9/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The word is centered and in all capital letters. The background is a red hexagon shape with the word inside.

Compound	10 mg/dL		100 mg/dL
Acetaldehyde	3,000	<0.3	3,000	<3.3
Acetone	3,000	<0.3	3,000	<3.3
n-Butanol	500	2.0	3,000	<3.3
Ethylene Glycol	3,000	<0.3	3,000	<3.3
Isopropanol	3,000	<0.3	3,000	<3.3
Methanol	3,000	<0.3	3,000	<3.3
n-Propanol	60	16.7	1,025	9.8
Propylene Glycol	3,000	<0.3	3,000	<3.3

Table 8. Cross-Reactivity - Serum

Compound	10 mg/dL		100 mg/dL
	Compound Conc.(mg/dL)	Cross-Reactivity(%)	Compound Conc.(mg/dL)	Cross-Reactivity(%)
Acetaldehyde	3,000	<0.3	3,000	<3.3
Acetone	3,000	<0.3	3,000	<3.3
n-Butanol	500	2.0	3,000	<3.3
Ethylene Glycol	3,000	<0.3	3,000	<3.3
Isopropanol	3,000	<0.3	3,000	<3.3
Methanol	3,000	<0.3	3,000	<3.3
n-Propanol	90	11.1	900	11.1
Propylene Glycol	3,000	<0.3	3,000	<3.3

Table 9. Cross-Reactivity - Plasma

Compound	10 mg/dL		100 mg/dL
	Compound Conc.(mg/dL)	Cross-Reactivity(%)	Compound Conc.(mg/dL)	Cross-Reactivity(%)
Acetaldehyde	3,000	<0.3	3,000	<3.3
Acetone	3,000	<0.3	3,000	<3.3
n-Butanol	500	2.0	3,000	<3.3
Ethylene Glycol	3,000	<0.3	3,000	<3.3
Isopropanol	3,000	<0.3	3,000	<3.3
Methanol	3,000	<0.3	3,000	<3.3
n-Propanol	90	11.1	900	11.1
Propylene Glycol	3,000	<0.3	3,000	<3.3

6. Interference – Structurally Unrelated Compounds in Urine

Structurally unrelated compounds were evaluated in quantitative mode by spiking the potential interferent into drug free urine containing EtOH at 10 mg/dL and 100 mg/dL. All potential interferents analyzed verified that assay performance is unaffected by externally ingested

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

Image /page/10/Picture/0 description: The image contains the word "IMMUNALYSIS" in white text on a red background. The text is bold and appears to be in all capital letters. The red background is a solid color, and the text is centered on the background.

compounds. The levels of structurally unrelated compounds that did not interfere in the assay are presented in Table 10.

Compound	Compound Conc. (mg/dL)
Acetaminophen	500,000
6-Acetylcodeine	100,000
6-Acetylmorphine	100,000
Acetylsalicylic Acid	500,000
Alphenal	100,000
Alprazolam	100,000
7-Aminoclonazepam	40,000
7-Aminoflunitrazepam	100,000
7-Aminonitrazepam	100,000
Amitriptyline	100,000
Amobarbital	100,000
S-(+)-Amphetamine	100,000
Aprobarbital	100,000
Barbital	100,000
Benzoylecgonine	100,000
Benzylpiperazine	100,000
Bromazepam	100,000
4-bromo 2-5, dimethoxyphenethylamine	100,000
Buprenorphine	50,000
Bupropion	100,000
Butabarbital	100,000
Butalbital	100,000
Caffeine	500,000
Cannabidiol	100,000
Cannabinol	100,000
Carbamazepine	100,000
Carisoprodol	100,000
Chlordiazepoxide	100,000
Chlorpromazine	100,000
Clobazam	100,000
Clomipramine	100,000
Clonazepam	100,000
Clozapine	100,000
Cocaine	100,000
Codeine	100,000
Compound	Compound Conc. (mg/dL)
Cotinine	100,000
Cyclobenzaprine	100,000
Cyclopentobarbital	100,000
Demoxepam	100,000
Desalkylflurazepam	100,000
Desipramine	100,000
Dextromethorphan	100,000
Diazepam	100,000
Digoxin	100,000
Dihydrocodeine	100,000
Diphenhydramine	500,000
Dehydronorketamine	40,000
delta9 THC	100,000
Doxepin	100,000
Doxylamine	100,000
ecgonine	100,000
ecgonine methyl ester	100,000
EDDP	100,000
EMDP	100,000
1R,2S Ephedrine	100,000
1S,2R Ephedrine	100,000
Ethyl-β-D-Glucuronide	50,000
Ethylmorphine	100,000
Fenfluramine	100,000
Fentanyl	100,000
Flunitrazepam	100,000
Fluoxetine	100,000
Flurazepam	100,000
Haloperidol	100,000
Heroin	100,000
Hexobarbital	100,000
hydrocodone	100,000
hydromorphone	100,000
11-hydroxy-delta9 THC	100,000
Ibuprofen	100,000
Imipramine	100,000
Ketamine	100,000
Lamotrigine	100,000
Levorphanol Tartrate	100,000
Compound	Compound Conc. (mg/dL)
Lidocaine	100,000
Lorazepam	100,000
Lorazepam Glucuronide	50,000
Lormetazepam	100,000
LSD	100,000
Maprotiline	100,000
MDA	100,000
MDEA	100,000
MDMA	100,000
Meperidine	100,000
Meprobamate	100,000
Methamphetamine	100,000
Methadone	500,000
Methaqualone	100,000
Methoxetamine	100,000
Methylone	100,000
Methylphenidate	100,000
Midazolam	100,000
Morphine	100,000
Morphine-3-glucuronide	100,000
Morphine-6-glucuronide	100,000
N-Desmethyltapentadol	100,000
Nalorphine	100,000
Naloxone	100,000
Naltrexone	100,000
Naproxen	100,000
Nitrazepam	100,000
11-nor-9-carboxy-delta9-THC	100,000
Norbuprenorphine	50,000
Norcodeine	100,000
Nordiazepam	100,000
Norketamine	100,000
Normorphine	100,000
Norproxyphene	100,000
Norpseudoephedrine	50,000
Nortriptyline	100,000
N-desmethyltramadol	100,000
N-desmethylvenlafaxine	100,000
O-desmethyltramadol	100,000
Compound	Compound Conc. (mg/dL)
O-desmethylvenlafaxine	100,000
Olanzapine	100,000
Oxycodone	100,000
Oxymorphone	100,000
Oxazepam	100,000
PCP	100,000
Pentazocine	100,000
Pentobarbital	100,000
Phenazepam	100,000
Phenobarbital	100,000
Phentermine	100,000
Phenylephrine	100,000
Phenylpropanolamine	100,000
Phenytoin	100,000
PMA	100,000
Prazepam	100,000
Propranolol	100,000
Propoxyphene	100,000
Protriptyline	100,000
R,R Pseudoephedrine	100,000
S,S Pseudoephedrine	100,000
Ritalinic Acid	100,000
Salicylic Acid	100,000
Sertraline	100,000
Sufentanil Citrate	50,000
Talbutal	50,000
Tapentadol	100,000
Temazepam	100,000
Theophylline	100,000
Thiopental	100,000
Thioridazine	100,000
Tramadol	100,000
Trazadone	100,000
Triazolam	100,000
Trifluoromethylphenyl-piperazine	100,000
Trimipramine	100,000
Venlafaxine	100,000
Verapamil	100,000
Zolpidem Tartrate	100,000

Table 10. Non-Interfering Structurally Unrelated Compounds in Urine

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

Image /page/11/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The word is in all capital letters and is centered in the image. The background is a solid red color. The image is simple and easy to read.

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

Image /page/12/Picture/0 description: The image is a red arrow pointing to the right with the word "IMMUNALYSIS" written in white letters inside the arrow. The word is written in all capital letters and is centered in the arrow. The arrow is a bright red color, and the letters are a bright white color. The image is simple and easy to read.

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

Image /page/13/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The background is shaped like a rounded rectangle with pointed ends, similar to a road sign. The text is bold and centered within the shape.

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

7. Interference - Endogenous Compounds and Urine Preservatives

Endogenous compounds and urine preservatives were evaluated in quantitative mode by spiking the potential interferent into drug free urine containing EtOH at concentration of 10 mg/dL and 100 mg/dL. Assay performance is unaffected by all the internally existing physiological conditions tested. The levels of endogenous compounds and urine preservatives that did not interfere in the assay are presented in Table 11 to Table 12.

Compound	10 mg/dL and 100 mg/dLCompound Conc.
Ascorbic Acid	1.5 g/dL
Bilirubin	0.02 g/dL
Creatinine	0.5 g/dL
Galactose	0.01 g/dL
γ-Globulin	0.5 g/dL
Glucose	2.0 g/dL
Hemoglobin	0.3 g/dL
Human Serum Albumin	0.5 g/dL
Oxalic Acid	0.1 g/dL
Riboflavin	0.0075 g/dL
Sodium Chloride	6.0 g/dL
Urea	6.0 g/dL

Table 11. Non-interfering Endogenous Compounds - Urine

Table 12. Non-interfering Urine Preservatives - Urine

Compound	10 mg/dL and 100 mg/dL
	Compound Conc.
Boric Acid	1% w/v
Sodium Azide	1% w/v
Sodium Fluoride	1% w/v
Compound	10 mg/dL	100 mg/dL
	Compound Conc.	Compound Conc.
Acetaminophen	20 mg/dL	20 mg/dL
Amikacin	8 mg/dL	8 mg/dL
Ampicillin	5.3 mg/dL	5.3 mg/dL
Ascorbic Acid	6 mg/dL	6 mg/dL
Bilirubin total	60 mg/dL	80 mg/dL
Bilirubin direct	60 mg/dL	80 mg/dL
Caffeine	6 mg/dL	6 mg/dL
Carbamazepine	3 mg/dL	3 mg/dL
Chloramphenicol	5 mg/dL	5 mg/dL
Chlordiazepoxide	1 mg/dL	1 mg/dL
Chlorpromazine	0.2 mg/dL	0.2 mg/dL
Cholesterol	30 mg/dL	503 mg/dL
Cimetidine	2 mg/dL	2 mg/dL
Creatinine	30 mg/dL	30 mg/dL
Dextran	6000 mg/dL	6000 mg/dL
Diazepam	0.51 mg/dL	0.51 mg/dL
Digoxin	6.1 mg/dL	6.1 mg/dL
Erythromycin	6 mg/dL	6 mg/dL
Ethosuximide	25 mg/dL	25 mg/dL
Furosemide	6 mg/dL	6 mg/dL
Gentamicin	1 mg/dL	1 mg/dL
Hemoglobin	62.5 mg/dL	1,000 mg/dL
Heparin	3 U/mL	3 U/mL
Ibuprofen	50 mg/dL	50 mg/dL
IgG	5,000 mg/dL	5,000 mg/dL
Intralipid	187.5 mg/dL	750 mg/dL
Lactate Dehydrogenase	1,000 U/L	237,500 U/L
Lactate	150 mg/dL	901 mg/dL
Lidocaine	1.2 mg/dL	1.2 mg/dL
Lithium	2.2 mg/dL	2.2 mg/dL
Mannitol	500 mg/dL	500 mg/dL
Penicillin	25 U/mL	25 U/mL
Pentobarbital	8 mg/dL	8 mg/dL
Phenobarbital	10 mg/dL	10 mg/dL
Phenytoin	5 mg/dL	5 mg/dL
Primidone	4 mg/dL	4 mg/dL
Propoxyphene	0.16 mg/dL	0.16 mg/dL
HSA (albumin)	37.5 mg/dL	1,250 mg/dL
Compound	10 mg/dL	100 mg/dL
	Compound Conc.	Compound Conc.
Protein (total)	50 mg/dL	1,500 mg/dL
Salicylic Acid	60 mg/dL	60 mg/dL
Theophylline	4 mg/dL	4 mg/dL
Triglycerides	500 mg/dL	500 mg/dL
Urea	500 mg/dL	500 mg/dL
Uric Acid	20 mg/dL	20 mg/dL
Valproic Acid	50 mg/dL	50 mg/dL

8. Interference - Interferents in Serum and Plasma

Potential interfering substances in serum and plasma matrices were evaluated in quantitative mode by spiking the compound into drug free serum and plasma containing EtOH at concentration of 10 mg/dL and 100 mg/dL, respectively. Additional anticoagulants were evaluated by spiking into drug free plasma containing EtOH at concentration of 10 mg/dL and 100 mg/dL. Assay performance is unaffected by all the internally existing physiological conditions tested. The levels of substances that did not interfere in the assay are presented in Table 13 and Table 14.

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

Image /page/15/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The word is written in all capital letters and is centered in the image. The background is a solid red color, and the letters are a bright white color. The image is simple and easy to read.

Table 13. Non-interfering Substances – Serum and Plasma

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

Image /page/16/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The word is centered in the image and is written in all capital letters. The background is a bright red color, and the letters are a slightly lighter shade of white. The image is simple and easy to read.

Table 14. Non-interfering Anticoagulants -Plasma

Compound	10 mg/dLCompound Conc.	100 mg/dLCompound Conc.
Heparin	3 U/mL	3 U/mL
EDTA	2 mg/mL	2 mg/mL
Oxalic Acid	2 mg/mL	2 mg/mL
Sodium Citrate	3.8 g/dL	3.8 g/dL
Sodium Fluoride	2 mg/mL	2 mg/mL

9. Interference - pH

To evaluate potential interference from the effect of urine pH, device performance in the quantitative mode was tested using a range of urine pH values. All test samples were prepared in drug-free urine containing EtOH at concentration of 10 mg/dL and 100 mg/dL. No interference was observed at urine pH values ranging from 3.0 to 11.0.

10. Interference - Specific Gravity

To evaluate potential interference from the specific gravity of urine, device performance in the quantitative mode was tested using a range of physiologically relevant urine specific gravity values. All test samples were prepared in drug free urine containing EtOH at concentration of 10 mg/dL and 100 mg/dL. No interference was observed at urine specific gravity values ranging from 1.000 to 1.030.

11. Calibration Duration

Drug free negative urine spiked with EtOH at concentration of 10 mg/dL, 100 mg/dL and 300 mg/dL were tested in quantitative mode at time points up to 14 days. At the initial time point, a two-point calibration curve was established. This calibration was used through the duration of this study. The test results met acceptance criteria at each time point. The recommended frequency of calibration is 14 days.

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

Image /page/17/Picture/0 description: The image shows the word "IMMUNALYSIS" in white letters on a red background. The word is written in all capital letters and is centered in the image. The background is a solid red color. The image is simple and easy to read.

12. Specimen Stability

Drug free negative urine, serum and plasma spiked with EtOH at concentration of 100 mg/dL and 300 mg/dL were stored in industry standard collection containers and tested by assay kit at each time point at 30°C and at 2°C - 8°C. Test results indicated that urine samples containing EtOH are stable for up to 14 days stored at ambient temperature up to 30℃, up to 6 months stored at 2°C - 8°C; serum samples containing EtOH are stable for up to 10 days stored at ambient temperature up to 30°C, up to 14 days stored at 2°C - 8°C; plasma samples containing EtOH are stable for up to 3 day stored at ambient temperature up to 30°C, up to 6 days stored at 2°C - 8°C.

13. Method Comparison

Eighty (80) de-identified, unaltered leftover clinical urine samples, seventy-six (76) de-identified, unaltered leftover clinical serum samples and sixty-four (64) de-identified, unaltered leftover clinical plasma samples obtained from clinical testing laboratories were analyzed for EtOH with the Immunalysis Ethyl Alcohol Enzyme Assay in quantitative mode and compared to results of predicate device. The instruments used were an Olympus AU 400e. Method comparison test results are presented in Table 15.

Table 15. Method Comparison Test Results

Matrix	n	Slope	Intercept	r²	CandidateDevice Range(mg/dL)	PredicateDeviceRange(mg/dL)
Urine	80	1.0284	-2.7848	0.9964	3-600	3-600
Serum	76	1.0067	-5.8802	0.9948	3-600	3-600
Plasma	64	1.0595	-7.3902	0.9932	3-600	3-600

CONCLUSION

The information provided in this pre-market notification demonstrates that the Immunalysis Ethyl Alcohol Enzyme Assay is substantially equivalent to the legally marketed predicate device for its intended use.

Regulation Number and Section

§ 862.3040 Alcohol test system.

(a)
Identification. An alcohol test system is a device intented to measure alcohol (e.g., ethanol, methanol, isopropanol, etc.) in human body fluids (e.g., serum, whole blood, and urine). Measurements obtained by this device are used in the diagnosis and treatment of alcohol intoxication and poisoning.(b)
Classification. Class II.