Cortisol hormone salivary test
to assess stress levels
Why is it essential to measure cortisol?
Stress and anxiety develop due to social, nutritional factors, lack of sleep hygiene, etc., causing a constant deterioration of our nervous system. The levels of cortisol (stress hormone) and the Beta brain waves (Responsible for wakefulness-stress) of the 70% of the population are at too high levels.
Objectifying the results of stress and anxiety with validated tests, salivary cortisol tests, biochemical or bioelectrical analysis (EEG-ECG) is essential to show the objective changes to the patient regarding their psychosomatic state.
Frequently Asked Questions: Cortisol
Research shows that there are two distinct systems involved in the stress response of humans and other animals. The response and activity of both systems can be measured through various biomarkers in saliva (cortisol and amylase). The classical method to characterize a stress response is to measure cortisol levels, which are increased due to a series of changes in the hypothalamic-pituitary-adrenal (HPA) axis. However, research has shown that this system can be relatively slow to respond to stress (a latency of about 10-15 minutes); while changes in the alpha-amylase response are somewhat more rapid (both markers of sympathetic nervous system (SNS) activity). It is also well known that chronic stress with elevated cortisol levels can lead to immunosuppression.
Types of monitoring and measurements
Daytime measurement of the same subject
In corporate and health and wellness settings, it is more common to look at the diurnal cortisol profile, usually with 5 samples taken throughout the day to establish the profile of our specific patient or study subject.
Typical daytime profile sampling times:
- Wake up
- Wake up +30 minutes
- around 5 p.m.
Measurement in research or evaluation
However, as a research method or to monitor patients, pre and post treatment or weekly measurements can be taken throughout the recovery process. It should always be measured at the same time of day.
What is "Normal"?
"Normal" values are challenging at best. Even with laboratory tests, there are a variety of different kits that can be used and they all give different values for the same sample and/or person. On top of that, the use of the same test kits in different laboratories will still give variability. It is therefore recommended to establish individual analyzes and compare the % of variability in the study samples or in the evaluation of the patients.
Nomura S (2012) Salivary Hormones, Immunes and Other Secretory Substances as Possible Stress Biomarker, Biomarker, Prof. Tapan Khan (Ed.) p 247-27
It should be taken into consideration that cortisol concentrations are highest first thing in the morning, between 6 and 8 am and their lowest point is close to midnight. However, alpha-amylase is inversely proportional, presenting its highest salivary concentration at night, and its lowest in the morning. This is the main reason for the recommendation to perform the measurements at the same time for the same subject, since we can misinterpret the results if we compare measurements taken randomly in the morning and at night.
Sample collection is non-invasive, painless, and comfortable, especially for study subjects or patients. Saliva measurement shows advantages compared to blood sampling, which is more labor intensive, invasive, sometimes painful, carries a higher risk of infection, and in most settings requires medical supervision. Furthermore, for many subjects/patients, the mere aspect of drawing and testing blood is unpleasant.
Collection of saliva samples is possible at any time, day or night, due to the general convenience of collection. Saliva collection can be accomplished in circumstances where blood collection is difficult or inadvisable. It must be taken into account that it must always be collected at the same time on the same subject. This is due to the fact that cortisol and amylase present characteristic concentrations depending on the day of the day. For example, if the first measurement on our subject is performed in the morning at 9:00 am, the rest of the measurements should be performed at the same time.
The measurement we offer is a complement to analyze the evolution of patients with the aim of collecting data and using it for research.
IT IS NOT A DIAGNOSTIC METHOD FOR ENDOCRINE DISEASES
read the manual
Before first use, read the instruction manual carefully. The SOMA Cube is exclusively designed for use in research or to record the evolution of subjects or patients. It must be used on a horizontal surface. During the measurement it should not be moved and should not be exposed to any bright light (for example, sunlight). The device must never be opened (except the battery compartment) otherwise the warranty becomes invalid.
The SOMA Cube works with 3 lithium batteries, which are CR2032 type. Replacement batteries can be purchased at a wide variety of retail stores. The SOMA Cube is supplied with 3 batteries in a separate blister pack and they must be inserted into the SOMA Cube to enable operation.
When inserting the batteries, be careful not to get grease from your fingers in contact with the batteries. Such contamination can lead to further rapid discharge of batteries and reduces their service life. Therefore, the use of gloves or plastic tongs is recommended. In case the device does not start after inserting new batteries, please check the polarity of the batteries and clean them with a dry cloth.
The SOMA Cube device does not have an expiration date. OFC collectors are valid for 24 months. The Buffer is valid for 18 months. And finally, the 12-month LFD foil.
The pack or box includes a SOMA CUBE and several test kits. Likewise, each LFD test kit includes a SOMA OFC MANIFOLD, a SOMA OFC Buffer and a LFD SLIDE. .
SOMA OFC SPONGE OR COLLECTOR
The oral fluid collector consists of a specially formulated synthetic sponge-like material. The sponge is composed of a polymer attached to a plastic tube containing a volume adequacy indicator (which changes color when the volume is complete). The collector is designed to collect 0.5 ml oral liquid.
BOTTLE SOMA OFC OR Buffer
The SOMA OFC Buffer or bottle contains a buffer substance, made up of sodium phosphate, salts, detergents and preservatives. It has a number of key properties to make it an easy-to-use oral fluid collection tool for research and patient assessment. Once the sponge is placed in the Buffer, it is stable at 37 °C for three weeks. It is recommended that continued storage (weeks) be in a refrigerator or (months) in a freezer.
Important: do not ingest the buffer.
LFD SOMA SHEET
The SOMA LDF sheet is the component that will be used for the incubation of the sample and its subsequent measurement with the SOMA LFD Reader cube. As the LFD sheet can be affected by moisture, it is important to check that there has been no color change in the silica gel or silica gel sachet that is also packed inside each foil bag with the LFD. If the silica gel has changed color (orange to green), then the LFD is unlikely to be suitable for use and should be ruled out of use. All SOMA LFD sheets have colored test lines corresponding to the LFD type; this is to more easily differentiate between LFDs when they are out of packaging.
SOMA LFD READER CUBE OR SOMA CUBE
The pack includes the SOMA LFD READER CUBE OR SOMA CUBE. The cube-shaped device is the cortisol and amylase meter, which will perform the analysis on the LFD slide. A plastic support is included where the LFD sheet is placed for its measurement. The SOMA LFD CUBE is an electronic device that allows measurement on a timer, that is, a specific configured incubation of the sample is required, for 10 minutes. At the end of the incubation period, the CUBE starts the measurement and displays the result.
- The Oral Fluid Collector is designed for single use only.
- Do not chew or suck on the oral fluid collector.
- Do not place the collector in the mouth after it has been in the Collection Solution or Buffer.
Samples can be collected quickly and easily using the collector and analyzed within minutes of collection using the SOMA LFD Cube Reader. Both the collector, the buffer and the LFD sheet are used as a separate collection device per person.
1. Take the SOMA OFC COLLECTOR out of the bag.
2. Place the COLLECTOR in the mouth, on top of the tongue and close the mouth, drawing the collected saliva into the COLLECTOR (do not suck on it!). This method ensures reduced variability due to IgA secretion at different rates from various salivary glands distributed throughout the mouth.
3. Continue collecting until the volume sufficiency indicator has come on. in BLUE color (it is perceived in a plastic tube that holds the collector). This will normally take 20-50 seconds for most people, but can take several minutes if you are dehydrated, or the flow rate is very low. In these rare cases, please be patient and wait for the color change.
4. Once the oral fluid has been collected, it should be placed in the Buffer bottle (unscrew the largest cap first), holding the plastic tube and inserting the collector into the bottle, in the direction shown below. Be careful not to spill any drops from the bottle.
5. Replace the top of the Buffer bottle firmly and secure its closure.
6. The bottle should then be mixed for a period of at least two minutes. This should be done in a rhythmic up and down, or back and forth motion. Mixing is important to allow complete extraction of the assay. Do not shake too vigorously.
7. Then remove the LFD sheet from its bag by tearing the notches on both sides. The control line will appear on the slide itself as an orange/red line which is predefined before any analysis is performed.
8. Hold the Buffer bottle perpendicular to the surface where the LFD is at rest (for good repeatable performance, it should be a flat level). Place the Buffer in the area marked with a circle located on the LFD sheet and deposit 2 to 3 drops. Within a few seconds RED colored liquid will begin to appear in the rectangular test window on the slide and it will run. You will notice that the original color tints will be washed out by the swatches. In the unlikely event that the red color does not appear within 90 seconds, add one more drop. You should start timing the test from when the reddish color begins in the rectangular test window (exactly 15 timed minutes).
You will notice that within the test window there is the formation of three red lines, one control (C) and two test lines (T). Scanning for Cortisol LFD before or after 15 minutes will add variability to your readings, so it is important to be consistent and adept at reading at exactly 15 minutes.
9. Basic measurement: (recommended) Once the 15 minutes have passed, we proceed to read the results with the LFD Reader cube or SOMA cube. The sheet must be placed in the corresponding space in the included plastic support, in which the direction that the sheet should take can be seen. Make sure the LFD and the bottom of the SOMA Cube are flush and on a flat surface. And we proceed to the basic operation of the SOMA cube is the following:
- When the device is turned off, the screen is blank.
- To turn on the device, press the button briefly for less than 1 second.
- After activation, an audible alarm appears, the display shows "ON".
- Successively press the button until you reach the “RUN” option (shown on the screen).
- After a few seconds, an audible alarm starts and the result is displayed showing “C” followed by the cortisol value and “T” for alpha-amylase.
It should be taken into account that if the measurement is carried out again through the same process on the same sheet that has already been analyzed, it may show variability in the results. Therefore it is recommended to record the data.
It is recommended to use this option for its simplicity and speed, except when carrying out a study or investigation with a large sample that requires saving large amounts of data. However, the types of samples with recording are detailed below.
RFID measurement: There is another form of measurement where you can record the data on the RFID card included in the pack. After the incubation time on the slide (15 minutes) place the slide in its space in the plastic support and press the button on the SOMA CUBE for more than 1 second. The screen will show RFID. Each timer measurement can be stopped by pressing the button during the measurement.
- Place the lot-specific RFID card on top of the SOMA Cube reader taking care that the lot number displayed on the RFID card is exactly the same as the lot ID for the LFDs you are measuring.
- After a successful wireless transmission of the data, an audible signal is initiated. The display shows "TEST". The test must be placed in the cavity of the plastic holder, as explained in basic option 9a. Make sure the LFD and the bottom of the SOMA Cube are flush and on a flat surface.
- Start the measurement by briefly pressing the button. The device measures and the display shows "RUN". After a few seconds, an audible alarm starts and the result is displayed.
10. Storage of results of the tests in the SOMA CUBE itself (If storage is not required, proceed with point 11). If the button is pressed for more than 1 sec. the measurement results will be stored. An audible alarm appears, the display shows “SAVE”. The reader has an internal memory to store more than 100 readings. If the internal memory is full and a new result must be saved, the reader overwrites the first saved result. Each new saved result will overwrite the saved results in chronological order.
Once the results are stored or recorded, press the button again to start the next measurement. The display will show “ON” again, as when the SOMA cube was turned on. Thereafter, proceed with the instructions above from point 9.
11. If storage of results is not required, short press the button for less than 1 second. To not store the result. “ON” appears on the screen; continue with point 9 if another measurement of another sample is required.
12. If the device is turned on and does not wake up for about 50 seconds, the SOMA Cube automatically turns off to save battery. Please note that there is no active function to turn off the SOMA CUBE.
ATTENTION!! For RFID measurement. It is important that the SOMA CUBE ID displayed is the same as the batch ID. Number shown on the LFD foil bag label; this is because the calibration characteristics coincide and are programmed in the reader for each batch of sheets manufactured. If the lot is changed or the ID does not match it will not be able to be recorded with the RFID card supplied with your LFD test kits. If you are using LFDs from two lots in the same test session, be sure to use the correct RFID card for your LFDs.
The device could not read the information on the RFID card.
Press the button briefly (<1 sec.), the display will show 'ON' and continue with point 9a or 9b in the instructions above. If the error occurs multiple times, please contact the dealer directly.
The device could not find a control line or the control line signal is very weak. The most common cause of this is insufficient sample applied to the LFD slide from the OFC Buffer or the test has not been run correctly. This can happen if there has been a large air bubble within one of the two drops, or only one drop was added to the LFD.
Check that the LFD, if it is positioned correctly under the hub (section: measurement procedure, point 8). Then press the briefly (<1 sec), the device displays “ON” and repeat with 9a or 9b of the above instructions. If the error does not disappear, select a new LFD sheet and repeat the measurement procedure.
IT DOES NOT TURN ON
If, despite pressing the button, there is no information visible on the screen, the cause may be that the batteries are discharged.
Open the battery compartment and replace all 3 batteries with new ones as described in the “Battery Power in SOMA Cube” section near the beginning of this manual. If the device still does not react after changing the batteries, please contact the dealer.
These are the most common errors that can appear in SOMA CUBE.
Put the SOMA cube in the “ON” position according to step 9. Press the button twice short (<1 sec.). The year, time and date will appear on the screen. Press the button for about 1 second, a flashing screen appears with the first specification of time and then year. By repeatedly short (<1 sec) pressing the button, the displayed value can be changed. When the desired value (for example, year) is reached, press the button for a longer time (> 1 second), the required year will be stored and information will be displayed next time. Repeat these steps to successively go to year, month, day, hour, and minute. Press the button once more, the reader will show “ON” and now it is ready to use. Repeat this process with each battery change.
The SOMA Cube has the ability to transfer data to a PC or laptop. The dedicated USB cable and Cube Data Reader software are used for this.
In case of any defect, it may be necessary to return the device to the distributor or manufacturer. As the device may have been contaminated by debris or substances, the surface must be disinfected before shipping. Therefore use a suitable cleaning disinfectant. The disinfectant must be of good quality and not attack the material of the device (for example, Mikrozid® AF Liquid products or similar). Disinfection must be documented.
Attention: Please note that a returned device will not be accepted without any proof of disinfection and maintenance and cleaning of the plastic holder.
The SOMA Cube is maintenance free. Before each measurement, it must be checked for impurities, liquids or residues. For cleaning, we recommend a cloth together with a cleaning fluid, e.g. for glasses products.
Independent Validation Papers
- Coad S, et al., (2015) Validity and reliability of a novel immunoassay for Individual Profiling in Applied Sports Science. Research in Sports Medicine 23(2): 1-11.
- Fisher R, et al., (2015) The validity and reliability of a salivary cortisol Point of Care Test. Journal of Athletic Enhancement 4:4-10
- MacDonald L, et al., (2017) Reliability of salivary cortisol and Immunoglobulin A measurements from the IPRO before and after sprint cycling exercise. Journal of Sports Medicine and Physical Fitness (in Press)
Peer-reviewed Validation papers:
- Dunbar J, et al., (2015) Evaluation of a new point of care quantitative reader for salivary analysis in the English Premier League soccer environment. Brit J Sports Med Oct 2015
- Dunbar J, et al., (2015) Investigating the use of a Point of Care salivary Amylase Test in the English Premier League soccer environment. 8th World Congress of Science and Football, Denmark.
- Dunbar J, et al., (2015) Investigating a dual sIgA and alpha amylase Point of Care test in the sporting environment. Proceedings 12th Symposium Intl Soc Ex Imunol.
- Dunbar J, et al., (2013) Investigating the use of a Point of Care salivary IgG test in the sporting environment. Proceedings 11th Symposium Intl Soc Ex Imunol.
- Dunbar J, et al., (2013) Investigating the use of a Point of Care salivary cortisol test in the professional football environment. UKSCA Conference, East Midlands, UK.
- Dunbar J, & Jehanli A, (2011) Investigating the use of a point of care sIgA test in English Premier League Soccer players. . Proceedings 10th Symposium Intl Soc Ex Imunol.
- Jehanli A, et al., (2011) Development and validation of an oral fluid collection device and its use in the immunoassay of salivary steroids and immunoglobulins in sports persons. Proceedings 10th Symposium Intl Soc Ex Imunol
Papers using Soma technology
- MacDonald L, & Minihan C, (2017) Mindfulness training attenuates the increase in salivary cortisol concentration associated with competition in highly trained wheelchair basketball players. Journal of Sports Sciences.
- Anton-Solanos A, et al., (2016) Physiological and cognitive responses to and Antarctic Expedition: A case report. International Journal of Sports Physiology and Performance. Vol 11(8): 1053-1059.
- Coad S, et al., (2016) Seasonal Analysis of Mucosal Immunological Function and Physical Demands in Professional Australian Rules Footballers. Int. J. Sports Physiol. Perf. 11(5) 574-580.
- Edmonds R, et al., (2016) Cardiac Autonomic and Salivary Responses to a Repeated Training Bout in Elite Swimmers. Sports, 4, 13, 2-14
- Owen A, et al., (2016) High Intensity Training and Salivary Immunoglobuin A responses in Professional Top-Level Soccer Players. J Str. Cond. Res. 30(9): 2460-9
- Dunbar J, et al., (2016) Salivary Cortisol is Highly Correlated With Training Intensity in English Premier League Players; in International Research in Science and Soccer II Eds., Favero T, et al. Routledge pp 104-109
- Coad S, et al., (2015) Physical Demands and Salivary Immunoglobulin A Responses of Elite Australian Rules Football Athletes to Match Play. Int. J. Sports Physiol. Perf. 10(5) 613-617
- Dimitriou L, et al., (2015) Influence of Montmorency cherry juice blend on indices of exercise-induced stress and upper respiratory tract symptoms following marathon running. J Int Soc. Sp. Nutr. 12(22)
- Edmonds R, et al., (2015) Effect of chronic training on heart rate variability, salivary IgA and salivary alpha-amylase in elite swimmers with a disability. PLoS One, 10, e0127749
- Morgans R, et al., (2015) Pre-Match Salivary IgA in Soccer Players From the 2014 World Cup Qualifying Campaign. International Journal of Sports Physiology and Performance 10(3): 401-403
- Dunbar J, et al., (2014) Secretary IgA Responses to a Premier League Soccer Season. Proceedings of the 4th World Conference of Science and Soccer, Portland, USA.
- Morgans R, et al., (2014) Intensive Winter Fixture Schedule Induces a Transient Fall in Salivary IgA in English Premier League Soccer Players. Research in Sports Medicine 22: 346-354
- Dunbar J, et al., (2013) Mucosal Immunity and Self-reported Upper Respiratory Symptoms in a cohort of Premier League Academy Soccer Players: Proceedings 11th Symposium Intl Soc Ex Imunol.