DUTCH Plus Hormone Test: A Comprehensive Guide

Many patients with ongoing issues like fatigue, poor sleep, anxiety, or mood changes may have underlying hormonal or stress-related factors contributing to their symptoms. Standard blood tests may not fully capture diurnal hormone variation or metabolite patterns.

The DUTCH Plus test (Dried Urine Test for Comprehensive Hormones) is a non-invasive laboratory assessment that collects dried urine and saliva samples to examine hormone and adrenal function over the course of a day. 

It provides information on hormone levels, their breakdown products (metabolites), and daily cortisol patterns, including the cortisol awakening response (CAR), to support clinical evaluation of endocrine health.

This guide provides integrative and functional medicine practitioners with clear, practical information on how the DUTCH Plus test works, how to interpret the results, and how to use the findings to inform personalized treatment plans.

What the DUTCH Plus measures

The DUTCH Plus test is designed to provide a multi-faceted overview using a combination of salivary and dried urine samples collected across one day. It includes the following components:

• Sex hormone metabolites: Measures forms of estrogen (estrone, estradiol, estriol), progesterone (via alpha- and beta-pregnanediol), testosterone metabolites (such as 5α-DHT), and adrenal androgens like DHEA-S.
• Adrenal hormones: Assesses cortisol and cortisone at multiple time points throughout the day to evaluate daily patterns and the cortisol awakening response (CAR).
• Organic acids:Includes markers associated with biochemical pathways related to oxidative stress (8-OHdG), neurotransmitter metabolism (HVA, VMA), melatonin (6-OHMS), gut microbial activity (indican), and neuroinflammatory activity (quinolinate).
• Creatinine correction: Adjusts hormone values to account for variations in urine concentration, supporting more accurate interpretation.

Topically applied hormones, particularly vaginal formulations, may influence urinary hormone levels and affect interpretation. For accurate assessment, it is advisable to consider the timing of application relative to sample collection and to evaluate results alongside the patient's clinical presentation.

How it works

The DUTCH Plus test involves non-invasive sample collection completed at home over a 24-hour period. Patients provide multiple dried urine samples on filter paper and salivary samples at designated time points, including immediately upon waking. Collected materials are returned to the laboratory using pre-packaged shipping supplies.

The test utilizes liquid chromatography–tandem mass spectrometry (LC-MS/MS), a highly specific and sensitive analytical method for quantifying hormone metabolites.

Compared to single-point serum or saliva tests, DUTCH Plus may offer a broader assessment. It assesses hormone production, metabolite pathways, and diurnal hormone rhythms in a single panel.

A distinguishing feature is its measurement of the CAR, assessed through salivary samples collected immediately upon waking, and again approximately 30 and 60 minutes afterward. 

This allows for a more detailed assessment of hypothalamic-pituitary-adrenal (HPA) axis activity during the critical early-morning window.

Clinical applications of DUTCH Plus

The DUTCH Plus test provides a comprehensive view of hormone activity, adrenal patterns, and biochemical pathways involved in metabolism and cellular processes. 

Accurate outcomes rely on proper sample timing, preparation, and handling, in line with pre-test protocols. 

To ensure meaningful clinical application, results should be interpreted within the full context of the patient’s health history, presenting symptoms, and lifestyle factors.

Hormonal imbalances and symptom mapping

The DUTCH Plus test can assist in evaluating patterns of hormone levels and metabolism. Clinically observed patterns may include:

• Estrogen excess or increased estrogen exposure: has been associated with conditions such as uterine fibroids and breast tenderness, while other symptoms, such as premenstrual syndrome (PMS) and migraines, are more closely linked to hormonal fluctuations rather than absolute hormone levels.
• Low progesterone: May correlate with symptoms such as sleep disturbances, anxiety, or spotting during the luteal phase.
• Androgen excess: Often observed in conditions like polycystic ovary syndrome (PCOS), and may present with acne or increased hair growth in androgen-sensitive areas.

By assessing both hormone levels and their metabolic pathways, clinicians can better understand not only hormone presence but also how efficiently the body is processing and eliminating them.

Adrenal dysfunction and stress adaptation

The DUTCH Plus test provides insight into cortisol and cortisone production across the day, offering a window into HPA-axis activity and stress regulation:

• Cortisol awakening response (CAR): A blunted CAR may reflect altered stress reactivity and HPA-axis function. An exaggerated CAR may indicate elevated anticipatory stress.
• Flattened diurnal cortisol patterns: May suggest HPA-axis dysregulation, potentially observed in cases of prolonged stress or prolonged physiological strain.
• Elevated evening cortisol: Can be associated with poor sleep quality, nighttime awakenings, or glucose regulation challenges.
• Optional midnight cortisol: Late-night sampling may provide additional data in cases of disrupted sleep.

Organic acid insights

Organic acid markers included in DUTCH Plus expand its functional assessment capacity by providing biochemical context for neurotransmitter activity, nutrient metabolism, and detoxification status:

• Neurotransmitter metabolites: Elevated vanilmandelate (also known as Vanillylmandelic acid or (VMA)) and homovanillate (HVA) may be associated with catecholamine metabolism, though interpretation is context-dependent.
• Nutrient markers: Methylmalonic acid (MMA) may suggest B12 insufficiency, while xanthurenate can reflect suboptimal vitamin B6 status.

Integrating DUTCH Plus into clinical practice

The DUTCH Plus test is a non-invasive tool used to explore hormone rhythms and adrenal activity. 

When thoughtfully applied, it can offer valuable insights into individual wellness patterns. 

Integrating this tool effectively involves choosing appropriate candidates, ensuring accurate sample collection, and interpreting findings within a holistic care framework.

Who might benefit

The DUTCH Plus test may offer helpful context for individuals who:

• Experience persistent fatigue, mood changes, or disrupted sleep
• Are transitioning through hormonal phases, such as perimenopause or menopause
• Are pursuing fertility support or addressing hormone-related health concerns
• Operate in high-demand environments or are seeking recovery from chronic stress
• Are participating in integrative care strategies for mental, emotional, or physical wellness

Best practices for sample collection

Reliable results depend on following clear preparation guidelines:

• Collection timing may be adapted based on cycle variability or symptom tracking
• Patients may be advised to pause substances like caffeine, alcohol, nicotine, and certain supplements beforehand
• Saliva samples should be chilled or frozen promptly; dried urine strips must be thoroughly air-dried before mailing

Interpreting results in context

DUTCH Plus data should always be reviewed alongside a full clinical picture:

• Look for patterns in hormone output, cortisol rhythm, and metabolite balance
• Tailor recommendations to support nutrition, lifestyle, stress resilience, and overall wellbeing
• Consider retesting several weeks after changes to monitor progress and refine care

Hormone support considerations

For individuals using non-oral hormone modalities, DUTCH Plus may support tracking of:

• Hormone exposure patterns and metabolite pathways
• Adjustments to timing, dosage, or delivery based on patient feedback and overall trends

Frequently asked questions (FAQs)

This section addresses commonly asked questions to support clinical decision-making when considering or interpreting the DUTCH Plus test.

How does DUTCH Plus differ from serum hormone testing?

Serum testing measures hormone levels at a single point in time, typically reflecting total or bound hormones. In contrast, DUTCH Plus provides additional information by assessing hormone production, metabolism, and daily rhythm patterns.

What might a blunted cortisol awakening response (CAR) suggest?

A reduced CAR may reflect alterations in stress reactivity. This pattern is sometimes observed in individuals with low energy, prolonged stress exposure, or disrupted sleep patterns.

Should patients stop taking supplements before testing?

Certain supplements—such as hormone therapies, adaptogens, melatonin, or those targeting neurotransmitters—can affect results. It's important for patients to follow all preparation guidelines provided by the testing lab or their practitioner.

Can DUTCH Plus inform decisions around hormone or adaptogen use?

DUTCH Plus may help clarify how the body is responding to or metabolizing hormones, which can support personalized care planning. Any changes should always be made within a broader clinical framework.

Is DUTCH Plus appropriate for use in adolescents or older adults?

The test can be used in these populations with careful clinical oversight. Age-specific hormonal changes should be considered during interpretation.

Key takeaways

• DUTCH Plus is a non-invasive test that uses dried urine and saliva to assess daily hormone patterns, adrenal function, and related metabolites.
• It provides a comprehensive view of sex hormones, cortisol rhythms, and biochemical markers linked to stress, sleep, and nutrient status.
• A standout feature is the Cortisol Awakening Response (CAR), which captures HPA axis activity shortly after waking.
• The test may provide additional clinical context for symptoms like fatigue, mood shifts, sleep disruption, and hormonal transitions.
• Results should always be interpreted within the full clinical context, including history, symptoms, and lifestyle factors.
• Accurate results depend on proper sample timing, preparation, and handling, guided by clear pre-test instructions.