Dried Urine GCMS Estrogen Hormone Metabolites and LCMS Diurnal Hormones I

Estrone is the predominant oestrogen in postmenopausal women, produced mainly through peripheral aromatisation of androstenedione in adipose tissue. In premenopausal women it serves as a reservoir that can interconvert with oestradiol via 17β-HSD. Dried-urine E1 reflects integrated daily excretion, capturing both parent hormone and conjugated metabolites.

Dried-urine E1 measurement helps assess overall oestrogenic load in postmenopausal women and serves as a baseline for calculating metabolite ratios. Persistently elevated E1 in postmenopause may indicate excessive aromatase activity or exogenous hormone exposure.

Estradiol is the most potent endogenous oestrogen, secreted primarily by ovarian granulosa cells in premenopausal women. After menopause, circulating E2 drops sharply and derives mainly from peripheral conversion of E1. Dried-urine E2 integrates fluctuations over the collection period, smoothing the pulsatile serum pattern.

Urinary E2 tracks production and excretion across the menstrual cycle and is useful for documenting ovarian reserve decline, monitoring hormone therapy, and evaluating menopausal status. Because it is measured in urine, it avoids the transient peaks seen in serum.

Estriol is a weak oestrogen produced in small amounts by peripheral metabolism of E1 and E2 and in large quantities by the fetoplacental unit during pregnancy. In non-pregnant adults E3 excretion is minimal and represents a downstream metabolite.

In non-pregnant contexts E3 contributes little to net oestrogenic activity. Elevated E3 in a dried-urine panel may reflect high overall oestrogen metabolism or exogenous E3 supplementation.

2-Hydroxyestrone is formed by 2-hydroxylation of oestrone via CYP1A1/1A2. It binds weakly to oestrogen receptors and is rapidly conjugated and excreted, producing little net oestrogenic effect. It is often classified as a 'favourable' metabolite in functional endocrinology.

The 2-hydroxylation pathway is considered protective; lower 2-OHE1 relative to other pathways may shift the metabolite profile toward more potent or genotoxic species. The 2:16α-hydroxyestrone ratio is a widely used functional marker.

4-Hydroxyestrone is produced by CYP1B1-mediated 4-hydroxylation. It can form reactive quinones that bind DNA and generate oxidative adducts, giving it a theoretical genotoxic potential.

Elevated 4-OHE1 is hypothesised to increase breast cancer risk through oxidative DNA damage. The 4-OHE1:2-OHE1 ratio is monitored in some functional protocols, though clinical validation of interventions based on this ratio remains limited.

16α-Hydroxyestrone is generated by CYP3A4-mediated 16α-hydroxylation. Unlike 2-OHE1, it retains significant oestrogenic activity and is not rapidly inactivated, acting as a full agonist at the oestrogen receptor.

Elevated 16α-OHE1 contributes to net oestrogenic load and has been associated with proliferative effects in some observational studies. The 2:16α ratio is used to assess the balance between 'protective' and 'proliferative' pathways.

2-Methoxyestrone is the methylated derivative of 2-OHE1, produced by catechol-O-methyltransferase (COMT). Methylation stabilises and inactivates the catechol, facilitating excretion and preventing quinone formation.

The ratio of 2-MeOE1 to 2-OHE1 reflects COMT activity and methylation capacity. Low methylation—resulting in low 2-MeOE1—may allow accumulation of reactive catechols and is a target for nutrient support in functional protocols.

4-Methoxyestrone is the COMT-methylated product of 4-OHE1. Methylation of the 4-catechol inactivates it and reduces its potential to form DNA-damaging quinones.

The 4-MeOE1:4-OHE1 ratio reflects methylation efficiency for the 4-pathway. A low ratio suggests poor methylation, potentially leaving genotoxic 4-OHE1 unmethylated; this is interpreted together with total 4-pathway activity.

2-Hydroxyestradiol is the 2-hydroxylated metabolite of oestradiol, formed by the same CYP enzymes that hydroxylate oestrone. It contributes to the overall 2-hydroxylation pathway.

2-OHE2 is grouped with 2-OHE1 in the 'favourable' pathway. Its measurement adds granularity to the metabolite profile, particularly in premenopausal women with higher E2 levels.

Cortisol is the principal glucocorticoid, secreted by the adrenal cortex in a diurnal rhythm that peaks shortly after waking. Dried-urine morning cortisol reflects the integrated overnight and early-morning secretion captured in the first void.

Morning cortisol is a surrogate for the cortisol awakening response and overall HPA-axis activation. Low values may suggest adrenal insufficiency or HPA dysregulation; high values point to stress, Cushing syndrome, or erratic circadian rhythm.

Cortisol secretion falls sharply in the evening, reaching its nadir around midnight. Dried-urine evening cortisol captures the end-of-day integrated excretion and reflects circadian amplitude.

Elevated evening cortisol suggests loss of the normal diurnal rhythm and is seen in chronic stress, sleep disorders, and Cushing syndrome. The morning:evening ratio is used to assess circadian integrity.

Cortisone is the inactive 11-keto metabolite of cortisol, produced by 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2). It circulates at higher concentrations than cortisol and is converted back to active cortisol by 11β-HSD1 in target tissues.

Dried-urine cortisone parallels cortisol but reflects both direct secretion and peripheral metabolism. The cortisol:cortisone ratio indicates 11β-HSD activity and tissue glucocorticoid exposure.

Evening cortisone reflects the integrated inactive glucocorticoid excretion at the end of the circadian cycle.

Evening cortisone is interpreted with evening cortisol to assess circadian rhythm and the balance of active to inactive glucocorticoid. Elevated evening cortisone with normal evening cortisol may indicate increased peripheral inactivation.

Testosterone in women is secreted by the ovaries and adrenals and through peripheral conversion of androstenedione. Dried-urine testosterone integrates both free and conjugated metabolites, providing a measure of total androgen exposure.

Elevated testosterone points to polycystic ovary syndrome, congenital adrenal hyperplasia, or androgen-secreting tumours. Low testosterone is associated with reduced libido, fatigue, and loss of muscle mass, and may occur with ovarian insufficiency or adrenal dysfunction.

Progesterone is secreted by the corpus luteum after ovulation and by the adrenal glands. Dried-urine progesterone includes both the parent hormone and its metabolite pregnanediol glucuronide, giving an integrated measure of luteal-phase production.

Low luteal progesterone indicates anovulation or luteal insufficiency, a common cause of short cycles, premenstrual symptoms, and infertility. Elevated values are seen with exogenous supplementation or, rarely, progesterone-secreting tumours.

Dehydroepiandrosterone is the most abundant adrenal androgen, serving as a precursor for peripheral testosterone and oestrogen synthesis. Dried-urine DHEA reflects total daily secretion.

DHEA declines sharply with age and is a marker of adrenal reserve. Low DHEA may accompany adrenal insufficiency, chronic stress, or ageing; elevated DHEA is seen in adrenal hyperplasia and some virilising tumours.

Melatonin is secreted by the pineal gland at night and rapidly metabolised to 6-sulfatoxymelatonin, which is excreted in urine. First-morning urine aMT6s reflects integrated overnight melatonin production.

Low melatonin is associated with poor sleep onset, circadian misalignment, and reduced antioxidant capacity. High values are uncommon outside of exogenous supplementation. Melatonin output declines with age and is suppressed by light exposure at night.

Creatinine is a stable metabolite of muscle creatine phosphate, excreted at a relatively constant rate. It is used as a denominator to normalise urinary analytes for hydration variation.

Creatinine-adjusted results (per mg creatinine) allow comparison across samples with different dilutions. Very low creatinine (<30 mg/dL) suggests overhydration or collection error; very high (>300 mg/dL) suggests dehydration or concentrated urine.