PANTEX AM/PM SALIVARY CORTISOL ENZYME IMMUNOASSAY

K102841 · Pantex, Div. Bio-Analysis, Inc. · NHG · May 8, 2012 · Clinical Chemistry

Device Facts

• Record ID: K102841
• Device Name: PANTEX AM/PM SALIVARY CORTISOL ENZYME IMMUNOASSAY
• Applicant: Pantex, Div. Bio-Analysis, Inc.
• Product Code: NHG · Clinical Chemistry
• Decision Date: May 8, 2012
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1205
• Device Class: Class 2

Intended Use

For the in-vitro diagnostic quantitative determination of free and protein bound salivary cortisol in human saliva as an aid in the assessment of Cushing Syndrome and Addison's Disease. Measurements of cortisol in saliva are used in the diagnosis and treatment of disorders of the adrenal gland.

Device Story

The Pantex AM/PM Salivary Cortisol EIA Kit is an enzyme immunoassay for the quantitative measurement of cortisol in human saliva. The device uses a competitive binding principle where cortisol in the saliva sample competes with cortisol-peroxidase conjugate for binding sites on a rabbit anti-cortisol antibody coated onto a 96-well microplate. After incubation and washing, a TMB substrate is added, producing a color change proportional to the amount of enzyme conjugate bound. The optical density is measured at 450 nm using a colorimetric microplate reader; the intensity is inversely proportional to the cortisol concentration in the sample. The kit is intended for use by laboratory professionals to assist clinicians in diagnosing and managing adrenal gland disorders. Results are calculated using a standard curve. The device provides a diagnostic aid for Cushing Syndrome and Addison's Disease.

Clinical Evidence

Performance was established through analytical bench testing. Precision studies (intra-assay, inter-assay, inter-lot) showed CVs ≤10%. Linearity was confirmed across the measuring range (0.1–30 ng/mL). Recovery studies demonstrated accurate cortisol measurement in spiked saliva samples. Interference studies showed no significant impact from caffeine, nicotine, alcohol, food, or gum. Method comparison against the predicate (N=160) yielded a linear regression of Y = 1.0269x + 0.0994 with an r² of 0.9797.

Technological Characteristics

Enzyme immunoassay; goat anti-rabbit gamma globulin coated microplate; TMB substrate; 450 nm detection; measuring range 0.1–30 ng/mL; requires microplate reader with 4-parameter data reduction; storage 2-8°C; shelf-life 9 months.

Indications for Use

Indicated for the quantitative determination of free and protein-bound salivary cortisol in human saliva to aid in the assessment of Cushing Syndrome and Addison's Disease, and for the diagnosis and treatment of adrenal gland disorders.

Regulatory Classification

Identification

A cortisol (hydrocortisone and hydroxycorticosterone) test system is a device intended to measure the cortisol hormones secreted by the adrenal gland in plasma and urine. Measurements of cortisol are used in the diagnosis and treatment of disorders of the adrenal gland.

Predicate Devices

• Salimetrics Salivary Cortisol HS EIA Kit (k011323)

Submission Summary (Full Text)

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

#### K102841 510(k) Summary

| Submitted By: | Pantex, Division of Bio-Analysis, Inc.<br>1701 Berkeley Street<br>Santa Monica, CA 90404<br>USA<br>[phone] |
|----------------------------------|-------------------------------------------------------------------------------------------------------------------|
| Company Contact: | Romulo Garza, Ph.D., President/Senior Scientist |
| Date Summary Prepared: | 4-3-12 |
| Trade Name: | Pantex AM/PM Salivary Cortisol EIA Kit |
| Common Name: | Enzyme immunoassay, cortisol, salivary |
| Regulation Number and Panel: | 862.1205-Clinical Chemistry |
| Classification Product Code: | NHG |
| Classification: | Class II |
| Substantially Equivalent Device: | K011323 Salimetrics HS Salivary Cortisol EIA<br>Kit Item No. 1-3102 (single) 96 well kit |
| Device Description: | |

### A. Test principle.

The basis of the Cortisol Enzyme Immunoassay (EIA) is the quantitative relation between ligand concentration and the proportion of Cortisol (analog) enzyme conjugate bound to the antiserum. For example: Cortisol in the calibrators and unknowns compete with Cortisol coupled to peroxidase for antibody binding sites. After incubation, unbound components are washed away. The reaction between Cortisol peroxidase with the substrate (TMB) produces a blue color. The pre-determined time of incubation the reaction is stopped and a yellow color is formed. The optical density (read at 450 nm) is inversely proportional to the cortisol of calibrators, saliva samples and saliva controls.

### B. Kit Description.

The kit consists of a 96 well GARGG (Goat Anti-Rabbit Gamma Globulin) coated microplate (12x8 breakable strip wells), seven ready-to-use calibrators (range 0.1-30 ng/ml) of gravimetrically prepared cortisol from a commercial source (Steraloids) and compared and traced to NIST cortisol, low and high controls, anti-Cortisol (rabbit), 10X concentrated Cortisol (analog)-peroxidase, substrate solution, stop reaction solution and 10X concentrated wash solution.

### C. Intended Use/Indications for Use:

For the in-vitro diagnostic quantitative determination of free and protein bound salivary cortisol in human saliva as an aid in the assessment of Cushing Syndrome and Addison's Disease. Measurements of cortisol in saliva are used in the diagnosis and treatment of disorders of the adrenal gland.

### Predicate Device:

The predicate device for substantial equivalence in this submission is:

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

| Device Name | Salimetrics Salivary Cortisol HS EIA Kit |
|------------------|------------------------------------------|
| Company | Salimetrics |
| 510(k) reference | K011323 |

# Technology Comparison:

| | Predicate Device: | New Device: |
|------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | Salimetrics Salivary Cortisol HS<br>EIA Kit<br>(K011323) | Pantex AM/PM Salivary<br>Cortisol EIA Kit<br>(K102841) pending |
| Indications for use | For the in-vitro diagnostic<br>quantitative determination of free<br>and protein bound salivary<br>cortisol in human saliva as an aid<br>in the assessment of Cushing<br>Syndrome and Addison's Disease.<br>Measurement s of cortisol in<br>saliva are used in the diagnosis<br>and treatment of disorders of the<br>adrenal gland. | For the in-vitro diagnostic<br>quantitative determination of<br>free and protein bound<br>salivary cortisol in human<br>saliva as an aid in the<br>assessment of Cushing<br>Syndrome and Addison's<br>Disease.<br>Measurement s of cortisol in<br>saliva are used in the diagnosis<br>and treatment of disorders of<br>the adrenal gland. |
| Analyte | Free and Protein-bound<br>Cortisol | Free and Protein-bound<br>Cortisol |
| Sample Type | Saliva | Saliva |
| Method | Enzyme immunoassay | Enzyme immunoassay |
| Detection Method | Colormetric microplate reader | Colormetric microplate reader |
| Test Principle | Cortisol in the sample competes<br>with Cortisol-enzyme conjugate<br>for binding sites to antibody<br>bound to a microwell. Unbound<br>components are washed away and<br>enzyme is measured by a colored<br>reaction with the TMB substrate. | Cortisol in the sample<br>competes with cortisol-<br>enzyme conjugate for binding<br>sites to the antibody (rabbit<br>anti cortisol) bound to a<br>GARGG microplate.<br>Unbound components are<br>washed away and enzyme is<br>measured by a colored<br>reaction with the TMB<br>substrate. |
| Calculations | Quantitative determination<br>with standard curve | Quantitative determination<br>with standard curve |
| Quality Control | Use of reference controls is<br>recommended | Use of reference controls is<br>recommended |
| Analytical Measuring<br>Range (AMR) | 0.12 ng/ml - 30.0 ng/ml | 0.1 ng/ml - 30.0 ng/ml |
| Expected Values<br>(Normal range) | N=192<br>Ages: 18-70<br>AM range: 0.8 - 15.5 ng/ml<br>PM range: <0.12 - 3.6 ng/ml | N=152<br>Ages: 23 - 68<br>AM range: 2.58 – 12.69 ng/ml<br>PM range: 0.25 -2.96 ng/ml |
| Limits of Detection | Limit of Blank (LoB)<br>not calculated | Limit of Blank (LoB)<br>0.0392 ng/ml |
| | | |
| | Limit of Detection (LoD)<br>not calculated<br>Limit of Quantitation (LoQ)<br>not calculated | Limit of Detection (LoD)<br>0.0519 ng/ml<br>Limit of Quantitation (LoQ)<br>0.0519 ng/ml |
| Saliva Collection Device | Polypropylene vials and<br>Salimetrics oral swab (SOS), Item<br># 5001.02 | VWR Sample Mailing Tube<br>Cat #16465-260 |
| Interferences | Information not available in<br>Salimetric's package insert | An in-vitro spiking study with<br>high doses of caffeine, food,<br>nicotine, alcohol and chewing<br>gum did not reveal significant<br>interference in the<br>measurement of cortisol in<br>saliva using the Pantex<br>AM/PM Salivary Cortisol EIA<br>Kit Cat #631. |
| Stability and Storage of<br>Kit Reagents (open vial) | Stated as stable at 2°-8°C until the<br>kit's expiration date | The stability of the opened kit<br>reagents were determined to<br>be 31 days when stored at 2°-<br>8°C using Pantex AM/PM<br>Salivary Cortisol EIA Kit Cat<br>#631 |

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

### Test Summary:

### Performance Characteristics-

The performance characteristics of Pantex AM/PM Salivary Cortisol Enzyme Immunoassay were based on evaluations by the following analytical performance tests.

- I. Analytical Performance Precision/Repeatability Intra-assay Inter-assay Inter-lot variation Linearity Recovery Traceability, Reagent Stability, Sample Stability and Expected values Detection Limits Analytical Specificity/Cross Reactivity II.Method Comparison
- III. Interferences
- I. Analytical Performance
- a. Precision/Reproducibility
- (i) The intra-assay precision was determined from 20 replicates of low, medium and high samples.

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

| Sample | N | Mean (ng/mL) | Standard<br>Deviation (ng/mL) | %CV |
|--------|----|--------------|-------------------------------|-----|
| Low | 20 | 0.627 | 0.034 | 5.4 |
| Medium | 20 | 3.995 | 0.266 | 6.7 |
| High | 20 | 25.232 | 1.579 | 6.3 |

- (ii) The inter-assay precision was determined from the mean of average duplicates for twelve (12) separate assays.

| Sample | N | Mean (ng/mL) | Standard<br>Deviation (ng/mL) | %CV |
|--------|----|--------------|-------------------------------|-----|
| Low | 12 | 0.587 | 0.037 | 6.3 |
| Medium | 12 | 4.163 | 0.301 | 7.2 |
| High | 12 | 25.126 | 0.712 | 2.8 |

- (iii) The inter-lot or between-lot variation was determined by duplicate measurements of five (5) pools of saliva samples and three (3) saliva controls using three (3) different lots. The results of intra-assay, inter-assay and inter-lot variation concluded a %CV of ≤10% for each sample tested.

| Saliva<br>samples | Lot #012 | Lot #013 | Lot #014 | Inter-lot | Inter-lot | Inter-lot |
|-------------------|-----------------|-----------------|-----------------|-----------------|---------------------|-----------|
| ID | Mean<br>(ng/ml) | Mean<br>(ng/ml) | Mean<br>(ng/ml) | Mean<br>(ng/ml) | Std.Dev.<br>(ng/ml) | CV<br>(%) |
| 20 | 4.65 | 4.45 | 4.79 | 4.64 | 0.164 | 3.5 |
| 21 | 0.67 | 0.61 | 0.71 | 0.67 | 0.049 | 7.4 |
| 22 | 2.02 | 1.95 | 2.09 | 2.02 | 0.069 | 3.4 |
| 23 | 4.75 | 4.69 | 4.76 | 4.73 | 0.041 | 0.9 |
| 24 | 2.01 | 1.99 | 2.04 | 2.01 | 0.026 | 1.3 |
| 25 | 3.64 | 3.67 | 3.71 | 3.68 | 0.036 | 1.0 |
| LC | 0.98 | 0.94 | 1.01 | 0.98 | 0.036 | 3.7 |
| NC | 5.21 | 5.31 | 5.49 | 5.34 | 0.140 | 2.6 |
| HC | 10.79 | 10.13 | 10.52 | 10.48 | 0.329 | 3.1 |

- (iv) Repeatability
This study was conducted during 4 days of familiarization period and 20 days of testing. Two assays were performed daily with a minimum of 2 hours between assays. Three (3) different reagents lots and three (3) saliva pools were used for the study (Low, medium and high concentration). The pools were aliquoted and frozen until day of assay.

| Repeatability Low Concentration | |
|----------------------------------------|--------|
| Concentration (ng/ml) | 0.600 |
| Standard Deviation | 0.0141 |
| (I) (User Variance/Claim Variance) x R | 62.695 |
| (II) Critical Chi-square | 65.171 |
| Claim Rejected (I>II) | No |
| Claim Accepted (I Yes | |

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

| Precision Low Concentration Pool | | |
|----------------------------------|-------------------------|---------------------------------|
| | Standard Deviation, SD) | (% Coefficient of Variation CV) |
| Within Run | 0.0224 | 3.79 |
| Between Run | 0.0462 | 7.80 |
| Repeatability | 0.0162 | 2.73 |
| Total Device Precision | 0.0538 | 9.09 |

.

and the comments of the comments of the comments of

-

| Repeatability Medium Concentration | |
|----------------------------------------|-----------|
| Concentration (ng/ml) | 4.0 ng/ml |
| Standard Deviation | 0.0894 |
| (I) (User Variance/Claim Variance) x R | 63.073 |
| (II) Critical Chi-square | 65.171 |
| Claim Rejected (I>II) | No |
| Claim Accepted (I Yes | |

| Precision Medium Concentration Pool | | |
|-------------------------------------|-------------------------|---------------------------------|
| | Standard Deviation, SD) | (% Coefficient of Variation CV) |
| Within Run | 0.1475 | 3.60 |
| Between Run | 0.0514 | 1.26 |
| Repeatability | 0.1025 | 2.50 |
| Total Device Precision | 0.1869 | 4.56 |

| Repeatability High Concentration | |
|----------------------------------------|----------|
| Concentration (ng/ml) | 25 ng/ml |
| Standard Deviation | 0.5477 |
| (I) (User Variance/Claim Variance) x R | 62.035 |
| (II) Critical Chi-square | 65.171 |
| Claim Rejected (I>II) | No |
| Claim Accepted (I Yes | |

| Precision High Concentration Pool | | |
|-----------------------------------|------------------------|---------------------------------|
| | Standard Deviation, SD | (% Coefficient of Variation CV) |
| Within Run | 0.4442 | 1.176 |
| Between Run | 0.2915 | 1.15 |
| Repeatability | 0.62276 | 2.46 |
| Total Device Precision | 0.8185 | 3.24 |

.

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

### b. Linearity

Ten (10) sample concentrations that span the assay measuring range were performed Per EP6-A, Evaluation of the Linearity of Quantitative Measurement Procedures.

S=10 samples (dilutions) Concentration = (C1*V1+C10*V10)/(V1+V10)

| | C1<br>ng/ml | V1<br>ng/ml | C10<br>ng/ml | V10<br>ng/ml | Calculated<br>Concentration<br>ng/ml | Obtained<br>Concentration<br>ng/ml | Recovery<br>% |
|----|-------------|-------------|--------------|--------------|--------------------------------------|------------------------------------|---------------|
| 1 | 0.093 | * | | | 0.100 | 0.093 | 93.0 |
| 2 | 0.093 | 0.889 | 33.788 | 0.111 | 3.833 | 3.729 | 97.3 |
| 3 | 0.093 | 0.778 | 33.788 | 0.222 | 7.573 | 7.620 | 100.6 |
| 4 | 0.093 | 0.667 | 33.788 | 0.333 | 11.313 | 10.842 | 95.8 |
| 5 | 0.093 | 0.556 | 33.788 | 0.444 | 15.054 | 14.350 | 95.3 |
| 6 | 0.093 | 0.444 | 33.788 | 0.556 | 18.827 | 18.313 | 97.3 |
| 7 | 0.093 | 0.333 | 33.788 | 0.667 | 22.568 | 21.547 | 95.5 |
| 8 | 0.093 | 0.222 | 33.788 | 0.778 | 26.308 | 24.694 | 93.9 |
| 9 | 0.093 | 0.111 | 33.788 | 0.889 | 30.048 | 30.459 | 101.4 |
| 10 | | | | * | 35.000 | 33.788 | 96.5 |

*Targets of low and high sample concentrations

#### c. Recovery:

Ten (10) saliva samples containing different levels of endogenous cortisol were spiked with known quantities of cortisol and assayed.

| Sample | Endogenous<br>(ng/ml) | Added<br>(ng/ml) | Expected<br>(ng/ml) | Observed<br>(ng/ml) | Recovery<br>(%) |
|--------|-----------------------|------------------|---------------------|---------------------|-----------------|
| 1 | 0.493 | 0.250 | 0.743 | 0.739 | 99.5 |
| 2 | 0.878 | 0.500 | 1.378 | 1.291 | 93.7 |
| 3 | 1.551 | 1.000 | 2.551 | 2.641 | 103.5 |
| 4 | 1.850 | 2.000 | 3.850 | 3.958 | 102.8 |
| 5 | 0.936 | 4.000 | 4.936 | 4.951 | 100.3 |
| 6 | 1.042 | 8.000 | 9.042 | 9.394 | 103.9 |
| 7 | 0.691 | 16.000 | 16.691 | 17.165 | 102.8 |
| 8 | 0.622 | 20.000 | 20.622 | 19.997 | 97.0 |
| 9 | 2.057 | 24.000 | 26.057 | 24.938 | 95.7 |
| 10 | 0.348 | 28.000 | 28.348 | 28.943 | 102.1 |

d. Traceability/Reagent Stability/Sample Stability/Expected Values

The calibrators and controls are prepared from stock cortisol (Steraloids) and are gravimetrically weighed and prepared. Concentration of stock cortisol (Steraloids) concentrations were confirmed by comparison to NIST cortisol (y=1.029 x-0.2195. R20.9924)

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

### Summary of the stability results

Real time stability studies are conducted to determine the reagent and kit shelf life (expiration date). Expiration date of the Pantex AM/PM Salivary Cortisol EIA Kit. Cat 631, is determined by results of shelf life studies and is based on the reagent that has the shorter assigned expiration day.

- 1. The reagents stored at 2-8°C are stable for 9 months; therefore, the expiration date of the kit components is established at 9 months from the manufacturing date. We are basing the 9 month stability claim based on the results obtained with the real time stability study when stored at 2-8°C, supported by the reagents stored at room temperature (20 -28°C) and in reference to the stress chart that predicts reagents year stability at +5℃, by Kennon, L. "Use of models in determining chemical pharmaceutical stability".
### Sample stability

| 1 ปี 1 ปี 2017 คนนิสเทคร กิน ที่ 11 ของแล้ว ค.ศ. 2017 ค.ศ. 2010 พ.ศ. 25 เมตร พ.ศ. 2010 พ.ศ. 25 | | | | | | |
|------------------------------------------------------------------------------------------------|--------------|---------|---------|------------------|----------------|--|
| Storage | Room | 37 °C | 2-8 °C | <-15 °C | <- 15 ° C | |
| | Temperature | | | (7 freeze / thaw | (Long term) | |
| | 20 - 30 °C | | | cycles) | | |
| Stability | Up to 7 days | Up to 7 | Up to 7 | Up to 7 days | Up to 180 days | |
| | | days | days | | | |

Four (4) stress conditions on freshly collected saliva revealed the following:

Open vial and working Cortisol-HRP Conjugate solution stability determination.

| Condition | Stability | Storage Temperature |
|-----------------------------------------|-----------|---------------------|
| Reagents. Open vial stability | 31 days | 2-8 °C |
| Working Cortisol-HRP conjugate solution | 31 days | 2-8 °C |

# Expected Reference Values:

The reference range was re-established by testing 152 male saliva samples and 152 female samples to have an equal number of male and female samples. The reference range and median were recalculated using CLSI C28-A3 as a guide. The following tables indicate the summary of the results

AM Expected Values:

| Subjects<br>(Number) | Subjects<br>(Gender) | Age<br>(Years) | AM Median<br>(ng/mL) | AM Range<br>(ng/mL) |
|----------------------|------------------------|----------------|----------------------|---------------------|
| 152 | 76 Males<br>76 Females | 23-68 | 6.70 | 2.58 - 12.69 |

# PM Expected Values:

| Subjects<br>(Number) | Subjects<br>(Gender) | Age<br>(Years) | PM Median<br>(ng/mL) | PM Range<br>(ng/mL) |
|----------------------|------------------------|----------------|----------------------|---------------------|
| 152 | 76 Males<br>76 Females | 23-68 | 0.58 | 0.25 - 2.96 |

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

### e. Detection Limits

The LoB (limit of the blank), the LoD (limit of detection) and the LoO (limit of quantitation) were determined by generating one hundred and twenty (120) measurements each of "cortisol free free salvia and low level (<0.1 ng/ml) cortisol samples (Reference, CLSI EP17-A, protocols for Determination of Limits of Detection and Limits of Quantitation).

| Limits of Blank<br>(LoB)<br>ng/mL | Limits of Detection<br>(LoD)<br>ng/mL | Limits of Quantitation<br>(LoQ)<br>ng/mL |
|-----------------------------------|---------------------------------------|------------------------------------------|
| 0.0392 | 0.0519 | 0.0519 |

f. Analytical Specificity/Cross Reactivity

Cross-reactivity was determined by testing those compounds most like to interfere with the Pantex AM/PM Salivary Cortisol E1A Kit. The specificity of the antiserum was evaluated by evaluating the cross-reactivity expressed as ratios of concentration of unlabeled cortisol over the compound that displaces 50% of cortisol-enzyme conjugate from the antiserum.

| Compound | Spiked Concentration | % Cross-reactivity |
|-------------------------------------|----------------------|--------------------|
| C-21 Steroids | | |
| Cortisol | 10,000 ng/ml | 100.0000 |
| 17-OH-Progesterone | 10,000 ng/ml | 0.0284 |
| Pregnenolone | 10,000 ng/ml | 0.0038 |
| 17-OH-Pregnenolone | 10,000 ng/ml | 0.0066 |
| Progesterone | 10,000 ng/ml | 0.0079 |
| Desoxycorticosterone | 10,000 ng/ml | 0.0517 |
| 11-Desoxycortisol | 10,000 ng/ml | 1.8133 |
| Dexamethasone | 10,000 ng/ml | 0.0164 |
| Cortisone | 10,000 ng/ml | 0.7600 |
| Corticosterone | 10,000 ng/ml | 1.0847 |
| Aldosterone | 10,000 ng/ml | 0.0070 |
| C-19 Steroids | | |
| Androstenedione | 10,000 ng/ml | 0.0038 |
| Testosterone | 10,000 ng/ml | 0.0042 |
| 5αDHT | 10,000 ng/ml | 0.0019 |
| DHEA-SO4 | 10,000 ng/ml | 0.0031 |
| Androstanedione | 10,000 ng/ml | 0.0028 |
| C-18 Steroids | | |
| Estradiol 17β | 10,000 ng/ml | 0.0024 |
| Estradiol 17α | 10,000 ng/ml | 0.0003 |
| Estrone | 10,000 ng/ml | 0.0010 |
| Estriol | 10,000 ng/ml | 0.0015 |
| Other structurally related steroids | | |
| Dehydroisoandrosterone | 1000 ng/ml | 0.0076 |
| 6αmethyl-17-Hydroxyprogesterone | 1000 ng/ml | 0.1427 |
| 6β Hydroxycortisol | 1000 ng/ml | 1.7177 |
| Prednisone | 1000 ng/ml | 1.0874 |
| Prednisolone | 1000 ng/ml | 25.9001 |

At >10% cross reaction prednisolone is a potential interfering substance.

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

- II. Method Comparison Studies
Tests were conducted for comparison between the Pantex AM/PM Salivary Cortisol EIA Kit, Cat and the predicate assay , Salimetrics HS Salivary Cortisol results of 160 samples were compared . Comparison of the Pantex AM/PM Salivary Cortisol EIA Kit (new device) and the Salimetrics HS Salivary Cortisol EIA (predicate) demonstrated acceptable regression and correlation statistics and appears to be substantially equivalent to the FDA cleared predicate device.

| Pantex AM/PM Salivary Cortisol EIA versus Salimetrics HS Salivary Cortisol EIA | |
|--------------------------------------------------------------------------------|------------------------|
| Linear Regression equation | $Y = 1.0269x + 0.0994$ |
| Correlation (r²) | 0.9797 |

### III. Interferences Studies.

An in-vitro experiment was performed by spiking three (3) levels of Cortisol (low, medium and high) with high concentrations of five (5) potentially interfering substances: alcohol, coffee (as caffeine), cigarette (as nicotine) and food and gum extracts. The results obtained appear to demonstrate no significant interference of the substances tested in this study with the measurement of Cortisol in saliva using the Pantex AM/PM Salivary Cortisol EIA Kit, Cat #631.

In-vitro experiment results:

| Pools | Potential Interferent Caffeine Added (ug/mL) | Obtained Value (ng/mL) | Recovery from Control (%) |
|-------------|----------------------------------------------|------------------------|---------------------------|
| Low Pool | 0 | 1.420 | 100 |
| Low Pool | 800 | 1.444 | 101.7 |
| Low Pool | 400 | 1.438 | 101.3 |
| Low Pool | 200 | 1.477 | 104.0 |
| Middle Pool | 0 | 4.932 | 100 |
| Middle Pool | 800 | 5.143 | 104.3 |
| Middle Pool | 400 | 5.063 | 102.7 |
| Middle Pool | 200 | 4.539 | 92.0 |
| High Pool | 0 | 28.302 | 100 |
| High Pool | 800 | 26.010 | 91.9 |
| High Pool | 400 | 26.811 | 94.7 |
| High Pool | 200 | 27.563 | 97.4 |

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

| Pools | Potential Interferent<br>Food Added<br>(mg/mL) | Obtained Value<br>(ng/mL) | Recovery from<br>Control<br>(%) |
|-------------|------------------------------------------------|---------------------------|---------------------------------|
| Low Pool | 0.000 | 1.345 | 100 |
| | 426 | 1.340 | 99.6 |
| | 213 | 1.332 | 99.0 |
| | 106.5 | 1.291 | 96.0 |
| Middle Pool | 0.000 | 4.739 | 100 |
| | 426 | 4.730 | 99.8 |
| | 213 | 4.834 | 102.0 |
| | 106.5 | 4.835 | 102.0 |
| High Pool | 0.000 | 26.447 | 100 |
| | 426 | 27.495 | 104.0 |
| | 213 | 26.283 | 99.4 |
| | 106.5 | 28.055 | 106.1 |

| Pools | Potential Interferent NicotineAdded (ug/mL) | Obtained Value (ng/mL) | Recovery from Control (%) |
|-------------|---------------------------------------------|------------------------|---------------------------|
| Low Pool | 0 | 1.362 | 100 |
| | 800 | 1.436 | 101.7 |
| | 600 | 1.437 | 101.3 |
| | 400 | 1.4345 | 104.0 |
| | 200 | 1.317 | 96.7 |
| Middle Pool | 0 | 4.871 | 100 |
| | 800 | 5.243 | 107.6 |
| | 600 | 5.258 | 107.9 |
| | 400 | 5.087 | 104.4 |
| | 200 | 5.155 | 105.8 |
| High Pool | 0 | 25.503 | 100 |
| | 800 | 27.033 | 106.0 |
| | 600 | 26.397 | 103.5 |
| | 400 | 25.642 | 100.6 |
| | 200 | 24.928 | 97.8 |

.

.

·

·

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

| Pools | Potential Interferent<br>Gum Added<br>(mg/mL) | Obtained Value<br>(ng/mL) | Recovery from<br>Control<br>(%) |
|-------------|-----------------------------------------------|---------------------------|---------------------------------|
| Low Pool | 0 | 1.289 | 100 |
| | 270 | 1.346 | 104.4 |
| | 135 | 1.286 | 99.8 |
| | 67.5 | 1.248 | 96.8 |
| | 33.75 | 1.280 | 99.3 |
| Middle Pool | 0 | 4.843 | 100 |
| | 270 | 4.957 | 102.4 |
| | 135 | 4.845 | 100 |
| | 67.5 | 4.712 | 97.3 |
| | 33.75 | 4.764 | 98.4 |
| High Pool | 0 | 28.367 | 100 |
| | 270 | 29.419 | 103.7 |
| | 135 | 29.247 | 103.1 |
| | 67.5 | 27.870 | 98.3 |
| | 33.75 | 28.216 | 99.5 |

| Pools | Potential Interferent<br>Ethanol Added<br>(%) | Obtained Value<br>(ng/mL) | Recovery from<br>Control<br>(%) |
|-------------|-----------------------------------------------|---------------------------|---------------------------------|
| Low Pool | 0 | 1.268 | 100 |
| | 0.025 | 1.267 | 104.4 |
| | 0.050 | 1.230 | 99.8 |
| | 0.100 | 1.213 | 96.8 |
| Middle Pool | 0 | 4.539 | 100 |
| | 0.025 | 4.403 | 102.4 |
| | 0.050 | 4.320 | 100 |
| | 0.100 | 4.490 | 97.3 |
| High Pool | 0 | 26.272 | 100 |
| | 0.025 | 28.362 | 108.0 |
| | 0.050 | 28.744 | 109.4 |
| | 0.100 | 28.750 | 109.4 |

# Concluding Statement:

Taken together, the performance characteristics, comparison studies with a predicate device and acceptable statistical performance studies in this 510(k) submission demonstrates that the Pantex AM/PM Salivary Cortisol EIA Kit, Cat #631, is safe and effective for its intended use and is substantially equivalent to the predicate device.

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

Image /page/11/Picture/0 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 an image of a stylized eagle with three heads, representing the department's mission to protect the health of all Americans and provide essential human services.

# DEPARTMENT OF HEALTH & HUMAN SERVICES

Food and Drug Administration

10903 New Hampshire Avenue Silver Spring, MD 20993

Pantex, Division of Bio-Analysis, Inc c/o Romulo Garza 1701 Berkeley Street Santa Monica, CA 90404

MAY - 8 2012

Re: k102841 ·

Trade Name: Pantex AM/PM Salivary Cortisol Enzyme Immunoassay Regulation Number: 21 CFR §862.1205 Regulation Name: Cortisol (hydrocortisone and hydroxycorticosterone) test system Regulatory Class: Class II Product Codes: NHG Dated: April 3, 2012 Received: May 2, 2012

Dear Dr. Garza:

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.

If your device is classified (see above) into either class II (Special Controls) or class III -(PMA), it may be subject to such additional controls. Existing major regulations affecting (1 wr 1), it can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

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

### Page 2

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please 11 Jou ability of In Vitro Diagnostic Device Evaluation and Safety at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding postmarket surveillance, please contact CDRH's Office of Surveillance and Biometric's (OSB's) Division of Postmarket Surveillance at [phone]. For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/Medical

Devices/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 Small Manufacturers, International and Consumer Assistance at its toll-free number [phone] or [phone] or at its Internet address http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm

Sincerely yours,

N.

Steven H. Liao, Ph.D.

Courney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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

# Indications for Use

K102841 510(k) Number (if known):

Device Name: Pantex AM/PM Salivary Cortisol Enzyme Immunoassay

#### Indications For Use:

For the in-vitto diagnostic quantitative determination of free and protein bound salivary cortisol in human saliva as an aid in the assessment of Cushing Syndrome and Addison's Disease. Measurements of cortisol in saliva are used in the diagnosis and treatment of disorders of the adrenal gland.

× Prescription Use (21 CFR Part 801 Subpart D) .

And/Or

Over the Counter Use (21 CFR Part 801 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Device Evaluation and Safety (OIVD)

Ruto Chem

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) 102841

Page 1 of 1