Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description details a standard ELISA assay with colorimetric detection and calculation based on provided standards. There is no mention of AI or ML in the device description, intended use, or performance studies.

Therapeutic

No.
This device is an in vitro diagnostic (ELISA) used to detect a biomarker for diagnosis, not to treat a disease.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that Calprest®NG "can be used as an in vitro diagnostic to aid in the diagnosis of Inflammatory Bowel Diseases (IBD)..."

SaMD (Software as a Medical Device)

No

The device description clearly states it is an enzyme-linked immunosorbent assay (ELISA) system, which is a laboratory-based hardware and reagent system, not a software-only device.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

The "Intended Use / Indications for Use" section explicitly states that Calprest®NG is intended to be used as an in vitro diagnostic to aid in the diagnosis of Inflammatory Bowel Diseases (IBD) and differentiate it from Irritable Bowel Syndrome (IBS).

The "Device Description" further describes it as an enzyme-linked immunosorbent assay (ELISA) system, which is a common type of in vitro diagnostic test.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Calprest®NG is a quantitative ELISA for detecting concentration of fecal calprotectin, which can be used as an in vitro diagnostic to aid in the diagnosis of Inflammatory Bowel Diseases (IBD), specifically Crohn's disease and ulcerative colitis, and to differentiate IBD from Irritable Bowel Syndrome (IBS) in conjunction with other clinical and laboratory findings.

Product codes

NXO

Device Description

Calprest®NG is an enzyme-linked immunosorbent assay (ELISA) system with colorimetric detection based on the use of antibodies against calprotectin present in the diluted sample is bound by the antibody adsorbed to the surface of the plastic well. The enzyme conjugated antibody binds to the captured antigen and subsequently the enzyme catalyzes the conversion of the substrate to a colored product. The intensity of the color is proportional to the amount of conjugate bound, and thus to the amount of captured calprotectin. Concentration of calprotectin in the samples is calculated using the provided standards.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies

7.1. Expected Values

Calprotectin Concentration and Interpretation:
27.1 - 120 mg/kg: Abnormal

7.2. Clinical Performance

Study Type: Clinical Performance, using two scenarios for borderline values (considered positive and considered negative).
Sample Size: 273 total subjects.

Borderline values considered Positive

Sensitivity: 94.6% (95% C.I. 89.2% - 97.8%)
Specificity: 90.2% (95% C.I. 84.1% - 94.5%)
PPV: 89.8% (95% C.I. 83.4% - 94.3%)
NPV: 94.9% (95% C.I. 89.7% - 97.9%)

Borderline values considered Negative

Sensitivity: 83.1% (95% C.I. 75.5% - 89.1%)
Specificity: 97.9% (95% C.I. 94.0% - 99.6%)
PPV: 97.3% (95% C.I. 92.3% - 99.4%)
NPV: 86.4% (95% C.I. 80.2% - 91.3%)

7.3. Extraction Reproducibility

Study Type: Extraction Reproducibility
Key Results: %CV ranged from 5.9% to 14.6% for different extracted stool samples (Mean 29.6 to 1973.3 mg/kg).

7.4. Comparison Study

7.4.1. Device Comparison

Comparison against Predicate Device Calprest® (K130945).
Similarities: Intended use, Technology (ELISA), Antigen (Calprotectin), Assay format (Quantitative), Positive and Negative controls (Ready to use), Sample type (Stool), Sample volume (1-5 g stool), Extraction solution (2.5x concentrate), Sample buffer diluent (10x concentrate), Wash buffer (20x concentrate), Platform (96 well microtiter plate), Cut-off (50 mg/kg (µg/g)).
Differences: Calibrator levels, Sample dilution, Incubation time, Conjugate, Substrate, OD Reading, Stop Solution.

7.4.2. Method Comparison

Study Type: Method Comparison between Calprest®NG and Calprest®.
Sample Size: 157 samples.

Borderline Value considered as Positive:

Positive Agreement: 96.6% (95% C.I. 91.5% - 98.7%)
Negative Agreement: 100.0% (95% C.I. 91.2% - 100.0%)
Overall agreement: 97.5% (95% C.I. 93.9% - 99.0%)

Borderline Value considered as Negative:

Positive Agreement: 96.1% (95% C.I. 89.0% - 98.6%)
Negative Agreement: 96.3% (95% C.I. 89.7% - 98.7%)
Overall agreement: 96.2% (95% C.I. 91.9% - 98.2%)
Deming Regression Analysis (y: Calprest®NG, x: Calprest®): Slope (95% CI) = 0.92 (0.84 to 1.01), Y-intercept (95% CI) = 0.13 (-4.64 to 4.90).

7.5. Precision

7.5.1. Single-Site Precision Evaluation study: Results

Study Type: Single-Site Precision.
Sample Size: 120 for each of 7 samples.
Key Results: Total %CV ranged from 5.5% to 13.5%.

7.5.2. Multisite Precision Evaluation Study: Results

Study Type: Multisite Precision.
Sample Size: 75 for each of 8 samples.
Key Results: Reproducibility %CV ranged from 11.6% to 14.9%.

7.6. Linearity

7.6.1. Matrix Linearity

Test Range: 25.21 - 3066.02 µg/g (mg/kg)
Slope: 1.01 (0.99 - 1.04)
Y-intercept: 9.28 (-26.54 - 45.10)
R2: 0.99
% Recovery Rate (Obtained/Theoretical): 86.91% - 112.84%

7.6.2. Aqueous Linearity

Test Range: 27.81 – 2889.04 µg/g (mg/kg)
Slope: 1.03 (1.01 - 1.06)
Y-intercept: -5.75 (-36.40 - 24.91)
R2: 1.00
% Recovery Rate (Obtained/Theoretical): 93.00 - 118.68

7.7. Calprotectin Recovery

Study Type: Calprotectin Recovery.
Key Results: Recovery (%) ranged from 94.8 to 112.1 across various spiked samples.

7.8. LoB, LoD, LoQ

LoB = 17.5 µg/g (mg/kg)
LoD = 23.4 µg/g (mg/kg)
LoQ = 27.1 µg/g (mg/kg)

7.9. Interfering Substance

Study Type: Interfering Substance Study.
Key Results: No interferences were observed from tested E coli, Salmonella spp, Shigella spp, Yesinia spp, Klebsiella spp, and a number of drugs (Vancomycin, Ciproflaxin hcl, Vitamin E, Prevacid, Azathioprine, Pentasa, Asacol, Prednisone, Multivitamin, Hemoglobin).

Key Metrics

Sensitivity, Specificity, PPV (Positive Predictive Value), NPV (Negative Predictive Value), Precision (%CV), Agreement (Positive, Negative, Overall), Recovery (%).

Predicate Device(s)

K130945

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.5180 Fecal calprotectin immunological test system.

(a)
Identification. A fecal calprotectin immunological test system is anin vitro diagnostic device that consists of reagents used to quantitatively measure, by immunochemical techniques, fecal calprotectin in human stool specimens. The device is intended forin vitro diagnostic use as an aid in the diagnosis of inflammatory bowel diseases (IBD), specifically Crohn's disease and ulcerative colitis, and as an aid in differentiation of IBD from irritable bowel syndrome.(b)
Classification. Class II (special controls). The special control for these devices is FDA's guidance document entitled “Class II Special Controls Guidance Document: Fecal Calprotectin Immunological Test Systems.” For the availability of this guidance document, see § 866.1(e).

Summary

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, arranged in a stacked formation.

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

November 10, 2016

Eurospital S. p. A. C/O Ms. Catherine L. Wong U.S. Agent 2832 Via Pacheco Palos Verdes Estates, CA 90274

Re: K160447

Trade/Device Name: Calprest NG Regulation Number: 21 CFR 866.5180 Regulation Name: Fecal Calprotectin Immunological Test System Regulatory Class: II Product Code: NXO Dated: September 29, 2016 Received: October 3, 2016

Dear Ms. Wong:

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

1

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,

Kelly Oil

FOR

Leonthena R. Carrington, MS, MBA, MT(ASCP) Director Division of Immunology and Hematology 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) K160447

Device Name Calprest®NG

Indications for Use (Describe)

Calprest®NG is a quantitative ELISA for detecting concentration of fecal calprotectin, which can be used as an in vitro diagnostic to aid in the diagnosis of Inflammatory Bowel Diseases (IBD), specifically Crohn's disease and ulcerative colitis, and to differentiate IBD from Irritable Bowel Syndrome (IBS) in conjunction with other clinical and laboratory findings.

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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Image /page/3/Picture/0 description: The image shows the logo for Eurospital. The word "Eurospital" is written in a sans-serif font in a light blue color. To the right of the word is a graphic of curved lines in light blue and green, resembling a stylized plant or wave pattern. The logo is clean and modern, with a focus on the company name and a simple, abstract design.

510(k) Summary

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

The assigned 510(k) number is: K160447

1. Submitter's name, address, telephone, contact person, and the date the summary was prepared:

Submitter's Name:	Eurospital S.p.A.
Submitter's Address:	Via Flavia, 122 – I 34147 Trieste, Italy
Submitter's Telephone Number:	Tel. +39 0408997285
Submitter's Contact Name:	Claudia Brandolin, Quality Assurance Manager

1. Name of the device, including the trade or proprietary name, the common or usual name and the classification name:

Proprietary Name:	Calprest®NG
Common Name:	Fecal calprotectin immunological test system
Classification Name:	Calprotectin, Fecal; 21 CFR 866.5180
Class II
Product Code: NXO

1. ldentification of the legally marketed device to which the submitter claims substantial equivalence:

Predicate Device Name:	Calprest®
510(k) Number:	K130945
Regulation Number:	21CFR866.5180
Regulation Class:	Class II

4. Description of the device

Calprest®NG is an enzyme-linked immunosorbent assay (ELISA) system with colorimetric detection based on the use of antibodies against calprotectin present in the diluted sample is bound by the antibody adsorbed to the surface of the plastic well. The enzyme conjugated antibody binds to the captured antigen and subsequently the enzyme catalyzes the conversion of the substrate to a colored product. The intensity of the color is proportional to the amount of conjugate bound, and thus to the amount of captured calprotectin. Concentration of calprotectin in the samples is calculated using the provided standards.

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5. Statement of the intended use of the device

Calprest®NG is a quantitative ELISA for detecting concentration of fecal Device Intended Use: calprotectin, which can be used as an in vitro diagnostic to aid in the diagnosis of Inflammatory Bowel Diseases (IBD), specifically Crohn's disease and ulcerative colitis, and to differentiate IBD from Irritable Bowel Syndrome (IBS) in conjunction with other clinical and laboratory findings.

6. Summary of the technological characteristics of the device

Calprest NG and Calprest Test are manufactured by Eurospital S.p.A. Trieste, Italy. Comparing to Calprest , the differences of Calprest NG include the name of the test on the labels, detection antibody, the use of a Horse-radish peroxidase / TMB conjugate/substrate system, the provided Stop solution, the concentration of calibrators and controls in the kit and the dynamic range of the assay.

7. Summary of performance characteristics

7.1. Expected Values

Calprotectin Concentration	Interpretation	Follow-Up
27.1 - 120 mg/kg	Abnormal	Repeat as clinically indicated

7.2. Clinical Performance

| Borderline values
considered Positive | Calprest®NG | IBD$ | | Total | Borderline values
considered Negative | IBD | | Total |
|------------------------------------------|-------------|----------|-------------------------|-----------------|------------------------------------------|----------|--------------------------|-------|
| | | Positive | Negative | | Calprest®NG | Positive | Negative | |
| Positive | | 123 | 14 | 137 | Positive | 108 | 3 | 111 |
| Negative | | 7 | 129 | 136 | Negative | 22 | 140 | 162 |
| Total | | 130 | 143 | 273 | Total | 130 | 143 | 273 |
| Sensitivity | | 94.6% | 95%C.I.§ | (89.2% - 97.8%) | Sensitivity | 83.1% | (95% C.I. 75.5% - 89.1%) | |
| Specificity | | 90.2% | 95%C.I. (84.1% - 94.5%) | | Specificity | 97.9% | (95% C.I. 94.0% - 99.6%) | |
| PPV* | | 89.8% | 95%C.I. (83.4% - 94.3%) | | PPV* | 97.3% | (95% C.I. 92.3% - 99.4%) | |
| NPV** | | 94.9% | 95%C.I. (89.7% - 97.9%) | | NPV** | 86.4% | (95% C.I. 80.2% - 91.3%) | |

$ IBD: Inflammatory Bowel Disease

PPV: Positive Predictive Value

** NPV: Negative Predictive Value
Ş CI: Confidence Interval

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7.3. Extraction Reproducibility

Values are reported in mg/kg

Extracted stool sample No.	1	2	3	4
Mean (mg/kg)	29.6	52.7	227.0	1973.3
SD	4.3	5.8	15.5	117.3
% CV	14.6	11.1	6.8	5.9

7.4. Comparison Study

7.4.1.Device Comparison
Reagents: Calprest"NG, code 9069, (manufactured by Eurospital S.p.A.)
Predicate device: Calprest® code 9031 (K130945, manufactured by Eurospital S.p

Similarities
Item	New device
Calprest®NG, code 9069	Predicate (K130945)
Calprest®, code 9031
Intended use	Calprest®NG is a quantitative ELISA
for detecting concentration of fecal
calprotectin, which can be used as an
in vitro diagnostic to aid in the
diagnosis of Inflammatory Bowel
Diseases (IBD), specifically Crohn's
disease and ulcerative colitis, and to
differentiate IBD from Irritable Bowel
Syndrome (IBS) in conjunction with
other clinical and laboratory findings.	Calprest® is a quantitative ELISA kit for
detecting concentration of fecal
calprotectin which can be used as in
vitro diagnostic to aid in the diagnosis
of Inflammatory Bowel Diseases (IBD,
Crohn's disease and ulcerative colitis)
and to differentiate IBD from Irritable
Bowel Syndrome (IBS) in conjunction
with other clinical and laboratory
findings.
Technology	ELISA	ELISA
Antigen	Calprotectin	Calprotectin
Assay format	Quantitative	Quantitative
Positive and Negative
controls	Ready to use	Ready to use
Sample type	Stool	Stool
Sample volume	1-5 g stool	1-5 g stool
Extraction solution	2.5x concentrate	2.5x concentrate
Sample buffer diluent	10x concentrate	10x concentrate
Wash buffer	20x concentrate	20x concentrate
Platform	96 well microtiter plate	96 well microtiter plate
Cut-off	50 mg/kg (µg/g)	50 mg/kg (µg/g)

Differences
Item	New device	Predicate (K130945)
	Calprest®NG, code 9069	Calprest®, code 9031
Calibrators	6 levels:
0, 2.5, 12.5, 25, 50, 150 ng/ml	6 levels:
6.25, 12.5, 25, 50, 100, 200 ng/ml
Sample dilution	1:20000	1: 2500

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Differences
Item	New device	Predicate (K130945)
	Calprest®NG, code 9069	Calprest®, code 9031
Incubation time	60-30-15 minutes at Room
Temperature (RT)	45-45-30 minutes at RT
Conjugate	HRP Horse Radish Peroxidase (IgG
mouse)	Alkaline phosphatase (IgG, rabbit)
Substrate	TMB	pNPP
OD Reading	450 nm	405 nm
Stop Solution	H2SO4 (0.5M)	None

7.4.2.Method Comparison

| Borderline Value

considered as Positive	Calprest®
	Pos	Neg
Calprest®NG	Pos	113	0	113
	Neg	4	40	44
		117	40	157
Positive Agreement		96.6%	(95% C.I. 91.5% - 98.7%)
Negative Agreement		100.0%	(95% C.I. 91.2% - 100.0%)
Overall agreement		97.5%	(95% C.I. 93.9% - 99.0%)

| Borderline Value

considered as Negative	Calprest®
		Pos	Neg
Calprest®NG	Pos	73	3	76
	Neg	3	78	81
		76	81	157
Positive Agreement	96.1%	(95% C.I. 89.0% - 98.6%)
Negative Agreement	96.3%	(95% C.I. 89.7% - 98.7%)
Overall agreement	96.2%	(95% C.I. 91.9% - 98.2%)

Deming Regression Analysis (y: Calprest®NG, x: Calprest®)

	Slope (95% CI)	Y-intercept (95% CI)
Y = 0.92x + 0.13	0.92 (0.84 to 1.01)	0.13 (-4.64 to 4.90)

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7.5. Precision

| Sample
ID°# | N | mean
(mg/kg) | Within Run
SD | Within Run
%CV | Between Run
SD | Between Run
%CV | Between Day
SD | Between Day
%CV | Between Operator
SD | Between Operator
%CV | Total
SD | Total
%CV |
|----------------|-----|-----------------|------------------|-------------------|-------------------|--------------------|-------------------|--------------------|------------------------|-------------------------|-------------|--------------|
| 1 | 120 | 170.88 | 3.40 | 2.0% | 6.77 | 4.0% | 5.51 | 3.2% | 0.80 | 0.5% | 9.40 | 5.5% |
| 2 | 120 | 741.60 | 17.38 | 2.3% | 43.12 | 5.8% | 55.37 | 7.5% | 0.00 | 0.0% | 72.30 | 9.7% |
| 3 | 120 | 248.74 | 6.44 | 2.6% | 11.98 | 4.8% | 12.21 | 4.9% | 0.00 | 0.0% | 18.28 | 7.3% |
| 4 | 120 | 42.60 | 1.28 | 3.0% | 4.30 | 10.1% | 0.00 | 0.0% | 0.55 | 1.3% | 4.52 | 10.6% |
| 5 | 120 | 1193.10 | 52.91 | 4.4% | 73.33 | 6.1% | 118.78 | 10.0% | 60.32 | 5.1% | 161.01 | 13.5% |
| 6 | 120 | 1006.17 | 41.69 | 4.1% | 39.67 | 3.9% | 55.80 | 5.5% | 28.24 | 2.8% | 84.98 | 8.4% |
| 7 | 120 | 2267.29 | 223.08 | 9.8% | 83.22 | 3.7% | 155.52 | 6.9% | 58.33 | 2.6% | 290.31 | 12.8% |

7.5.1.Single-Site Precision Evaluation study: Results

7.5.2.Multisite Precision Evaluation Study: Results

| Sample
ID# | N | mean
(mg/kg) | Repeatability | | Within-Site Precision | | Reproducibility | |
|---------------|----|-----------------|---------------|-------|-----------------------|-------|-----------------|-------|
| | | | SD | %CV | SD | %CV | SD | %CV |
| A | 75 | 433.80 | 13.43 | 3.1% | 63.99 | 14.8% | 64.55 | 14.9% |
| B | 75 | 36.54 | 3.69 | 10.1% | 5.11 | 14.0% | 5.11 | 14.0% |
| C | 75 | 1180.64 | 71.08 | 6.0% | 156.16 | 13.2% | 156.16 | 13.2% |
| D | 75 | 682.76 | 35.88 | 5.3% | 49.57 | 7.3% | 78.95 | 11.6% |
| E | 75 | 1629.44 | 96.29 | 5.9% | 214.76 | 13.2% | 226.84 | 13.9% |
| F | 75 | 62.08 | 4.19 | 6.8% | 5.64 | 9.1% | 8.68 | 14.0% |
| G | 75 | 356.84 | 12.00 | 3.4% | 30.07 | 8.4% | 51.93 | 14.6% |
| H | 75 | 2152.20 | 173.58 | 8.1% | 308.13 | 14.3% | 312.19 | 14.5% |

| Sample

ID#	Repeatability 95% CIs		Within-Site Precision 95% CIs		Reproducibility 95% CIs
	SD	%CV	SD	%CV	SD	%CV
A	11.395 to 16.346	2.6% to 3.8%	46.388 to 103.087	10.7% to 23.8%	47.256 to 101.796	10.9% to 23.5%
B	3.132 to 4.493	8.6% to 12.3%	4.166 to 6.611	11.4% to 18.1%	4.166 to 6.611	11.4% to 18.1%
C	60.329 to 86.539	5.1% to 7.3%	117.995 to 230.930	10.0% to 19.6%	119.470 to 225.503	10.1% to 19.1%
D	30.452 to 43.682	4.5% to 6.4%	40.416 to 64.137	5.9% to 9.4%	49.284 to 193.644	7.2% to 28.4%
E	81.724 to 117.230	5.0% to 7.2%	162.277 to 317.595	10.0% to 19.5%	168.946 to 345.240	10.4% to 21.2%
F	3.559 to 5.105	5.7% to 8.2%	4.634 to 7.222	7.5% to 11.6%	5.419 to 21.291	8.7% to 34.3%
G	10.186 to 14.611	2.9% to 4.1%	22.561 to 45.073	6.3% to 12.6%	31.114 to 149.229	8.7% to 41.8%
H	147.319 to 211.322	6.8% to 9.8%	240.599 to 428.660	11.2% to 19.9%	242.642 to 437.935	11.3% to 20.3%

7.6. Linearity

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7.6.1.Matrix Linearity

| Test Range
µg/g (mg/kg) | Slope (95% CI) | Y-intercept (95% CI) | R2 | % Recovery Rate
(Obtained/Theoretical) |
|----------------------------|--------------------|-----------------------|------|-------------------------------------------|
| 25.21 - 3066.02 | 1.01 (0.99 - 1.04) | 9.28 (-26.54 - 45.10) | 0.99 | 86.91% - 112.84% |

7.6.2. Aqueous Linearity

| Test Range µg/g
(mg/kg) | Slope (95% CI) | Y-intercept (95% CI) | R2 | % Recovery Rate
(Obtained/Theoretical) |
|----------------------------|--------------------|------------------------|------|-------------------------------------------|
| 27.81 – 2889.04 | 1.03 (1.01 - 1.06) | -5.75 (-36.40 - 24.91) | 1.00 | 93.00 - 118.68 |

Image /page/8/Figure/4 description: The image contains two scatter plots, each displaying a linear relationship between 'Theoretical conc' on the x-axis and 'Observed value' on the y-axis. The plot on the left shows a linear equation of y = 1.014x + 9.2789. The plot on the right shows a linear equation of y = 0.9687x + 6.0221 with an R-squared value of 1.

Matrix linearity graph

Aqueous linearity graph

7.7. Calprotectin Recovery

	Calprotectin Recovery Data
Sample Description	#1	#2	#3	#4	#5	#6	#7
Baseline (mg/kg)	9.3	51.7	162.7	307.7	458.3	1204.0	1975.2
Spike Value (mg/kg)	32.3	32.3	32.3	115.0	115.0	115.0	115.0
Theoretical (Baseline+Spike) (mg/kg)	41.7	84.0	195.0	422.7	573.3	1319.0	2090.2
Observed (Baseline+Spike) (mg/kg)	42.3	79.7	200.7	425.3	592.3	1397.7	2343.8
Recovery (%)	101.6	94.8	102.9	100.6	103.3	106.0	112.1

7.8. LoB, LoD, LoQ

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LoB = 17.5 µg/g (mg/kg) LoD = 23.4 µg/g (mg/kg) LoQ = 27.1 µg/g (mg/kg)

7.9. Interfering Substance

A number of potential interfering foods, pharmaceutical substances were tested. No interferences were observed.

| Pool | No
addition
µg/g
(mg/kg) | E coli | | Salmonella spp | | Shigella spp | | Yesinia spp | | Klebsiella spp | |
|------|-----------------------------------|-----------------|-------|-----------------|-------|-----------------|-------|-----------------|-------|-----------------|-------|
| | | µg/g
(mg/kg) | Δ% | µg/g
(mg/kg) | Δ% | µg/g
(mg/kg) | Δ% | µg/g
(mg/kg) | Δ% | µg/g
(mg/kg) | Δ% |
| A | 2068.6 | 2009.7 | 97.2 | 2100.1 | 101.5 | 2026.6 | 98.0 | 2021.1 | 97.7 | 2062.5 | 99.7 |
| B | 674.7 | 665.4 | 98.6 | 687.9 | 102.0 | 642.9 | 95.3 | 649.7 | 96.3 | 699.0 | 103.6 |
| C | 256.0 | 268.5 | 104.9 | 269.9 | 105.4 | 255.0 | 99.6 | 265.8 | 103.8 | 265.6 | 103.8 |
| D | 79.3 | 82.1 | 103.6 | 87.5 | 110.4 | 72.0 | 90.8 | 78.4 | 98.9 | 73.2 | 92.4 |
| E | 37.2 | 37.3 | 100.1 | 35.2 | 94.5 | 37.5 | 100.8 | 40.6 | 109.2 | 38.8 | 104.3 |

| Pool | No
addition | Addition of Drugs, Nutrients and Hemoglobin - Table 2a | | | | | | | | | | |
|------|-----------------|--------------------------------------------------------|------|-----------------|------|-----------------|------|--------------|------|--------------|------|--|
| | | Vancomycin | | Ciproflaxin hcl | | Vitamin E | | Prevacid | | Azathioprine | | |
| | µg/g
(mg/kg) | µg/g
(mg/kg) | Δ% | µg/g
(mg/kg) | Δ% | µg/g
(mg/kg) | Δ% | µg/g (mg/kg) | Δ% | µg/g (mg/kg) | Δ% | |
| A | 2089.5 | 2197.4 | 105% | 2261.1 | 108% | 2097.6 | 100% | 2085.5 | 100% | 1986.0 | 95% | |
| B | 603.8 | 626.5 | 104% | 623.2 | 103% | 600.3 | 99% | 589.1 | 98% | 590.5 | 98% | |
| C | 279.2 | 269.6 | 97% | 290.3 | 104% | 290.4 | 104% | 265.1 | 95% | 277.3 | 99% | |
| D | 82.4 | 79.7 | 97% | 88.1 | 107% | 86.0 | 104% | 83.8 | 102% | 83.9 | 102% | |
| E | 35.9 | 33.3 | 93% | 33.2 | 92% | 34.2 | 95% | 34.7 | 97% | 37.4 | 104% | |

| | No
addition
µg/g
(mg/kg) | Addition of Drugs, Nutrients and Hemoglobin - Table 2b | | | | | | | | | | |
|------|-----------------------------------|--------------------------------------------------------|-----------------|---------|-----------------|---------|--------------|---------|--------------|---------|------|--|
| Pool | Pentasa | | Asacol | | Prednisone | | Multivitamin | | Hemoglobin | | | |
| | µg/g
(mg/kg) | Δ% | µg/g
(mg/kg) | Δ% | µg/g
(mg/kg) | Δ% | µg/g (mg/kg) | Δ% | µg/g (mg/kg) | Δ% | | |
| A | 2089.5 | 2260.9 | 108% | 1937.50 | 93% | 2151.30 | 103% | 2284.80 | 109% | 2119.30 | 101% | |
| B | 603.8 | 586.7 | 97% | 599.80 | 99% | 592.90 | 98% | 589.90 | 98% | 583.70 | 97% | |
| C | 279.2 | 279.4 | 100% | 271.70 | 97% | 280.10 | 100% | 272.60 | 98% | 275.40 | 99% | |
| D | 82.4 | 80.0 | 97% | 75.90 | 92% | 84.60 | 103% | 83.90 | 102% | 80.60 | 98% | |
| E | 35.9 | 34.1 | 95% | 33.00 | 92% | 35.70 | 100% | 33.90 | 95% | 36.80 | 103% | |