The salivary cortisol test requires patients to collect saliva samples at home four times during one day, in the morning, noon, afternoon, and at night. Things that may compromise the sample are discouraged, such as specific behaviors. For example, smoking, posture, and eating can all influence salivation and may thus introduce artifact into the sample. As a result, patients are given very specific directions for collecting their saliva sample (described in detail in Methods section of this paper). A chart review was performed to examine the patterns of cortisol secretion and levels of DHEA in patients suspected of dysregulated glucose metabolism.
Data was reviewed and collected from 29 patient charts from a naturopathic primary care clinic in Portland, OR. Informed consent for records review was obtained upon admission to the clinic. All data was coded to remove any identifiable information. In naturopathic clinics, individuals who are suspected of cortisol/DHEA dysregulation for reasons related to prediabetes or type 2 diabetes often undergo a clinical laboratory test called the Adrenal Stress Index™ (ASI™, Diagnos-Techs, Kent WA). ASI™ measures cortisol, DHEA, sIgA, and anti-gluten antibodies from saliva collected during the day. According to Diagnos-Techs, the analytic sensitivity observational of this test is 0.8 nM to 1.0 nM and the specificity of the immunoassay to cortisol is at 99% or greater.(25) In diabetics and those at risk for diabetes, naturopathic physicians often do this test to help establish etiology of the disease. The chart review was done on patients who were predominantly untreated or uncontrolled diabetics, and patients presenting with symptoms that indicated insulin resistance and/or adrenal dysfunction whom had an ASI recorded in their chart. Additional patient care, number of visits, and information on medication intake was unobtainable for review.

Patients were instructed to follow the prementioned salivary collection protocol, but compliance was not assessed in the charts. Salivary samples were self-collected by the patient at four intervals in one day (between 7-8 AM, 11 AM-12 PM, 4-5 PM, and 11 PM-midnight). Patients were instructed not to eat or drink, use antacids, bismuth or mouthwash, or brush their teeth or smoke for 30-60 minutes before collecting the sample. They were also instructed not to eat more than one tablespoon of chocolate, onions, garlic, cabbage, cauliflower, or broccoli, or to drink coffee, tea, or caffeine on the day of collection. Patients were instructed to maintain a typical exercise regimen and activity level to obtain representative daily results. A sample consisted of saliva collected on a cotton roll held in the mouth until saturated and then placed in a 5 mL tube. Samples are refrigerated and mailed within 3 days. Samples are considered stable for a week at room temperature. The ASI™ tests were evaluated at Diagnos-Techs’ lab in Kent, WA.

The saliva samples were analyzed for cortisol by ELISA. DHEA and DHEA(S) were analyzed by ELISA using pooled samples from the noon and afternoon time points. Data were entered and analyzed in Microsoft Excel. A total of 29 ASI™ tests in 29 patients were found. For 28 of these patients, DHEA levels were also available. Serum fasting blood glucose levels were available for 20 of these patients.