GC/MS has served as a reference approach in urinary steroid profiling since the 1960s—excellent chromatographic resolution and utility for inborn errors of metabolism. LC/MS/MS offers high specificity, sensitivity, and throughput, driving rapid adoption over the past decade.^4–5 Many labs leverage both. Compared to immunoassays, mass-spec methods minimize cross-reactivity and inter-assay variability for low-level analytes.

• Same analytical platform (mass-spec), different collection dynamics → different reference ranges; values cannot be directly equated point-for-point.
• Dry urine filter paper shows recovery comparable to liquid urine (no meaningful matrix effect); discussion below references RRM (reference range median) to orient trends.
• Expect 10–15% variance (sometimes more) due to timing/half-life effects. Trends should align even when absolute %RRM differ.

EQ = E3 / (E1 + E2). E3 (estriol) signals via ERβ and is generally protective.^6–8 Optimal EQ > 1.5 (many aim > 2.0). Mary’s EQ is < 1.5 on both 24-hour and dry—same directional signal. Clinical note: Iodine is sometimes used to support EQ; defer to clinician judgment.

• 2OHE1 often considered protective.^10–11 Mary: 91% (24-hr) and 92.4% (dry) of RRM → healthy.
• 16αOHE1 can raise cancer risk when high but supports bone density; too low elevates osteopenia risk.^12–13 Mary: outside optimal in both; dry uses a broader range/left-shifted “optimal,” yet still suboptimal.
• 4OHE1 more reactive; elevated values merit attention. Mary: 116% (24-hr) and 108% (dry) of RRM → somewhat elevated and consistent; consider Mg and methylation support.¹⁹

2/16α ratio target ≈ 2–4.^14–18 Mary: 2.1 and 2.5 → within range. Given family history + absolute 16α elevation, consider DIM/I3C to reduce 16αOHE1 while maintaining bone-protective context.

2MeO-E1 derives from 2OHE1; a 2MeO-E1 : 2OHE1 ratio ≥ ~0.5 is often desirable. Mary: below 0.5 on both tests → methylation support may be warranted. 2MeO-E2 is highly protective but low abundance; Mary’s values are below median on both. Compounded 2MeO-E2 exists but is a clinician-directed decision.

Progesterone exhibits different kinetics than estrogens (longer stabilization/clearance). PdG is a reliable urinary marker of progesterone production;²⁵ progesterone itself is not measured in urine due to excretion chemistry. Mary’s PdG: 69% (24-hr) and 68.8% (dry) of RRM—remarkably consistent, indicating robust luteal output for age.

Reminder: PdG is a marker of progesterone activity over time, not an instantaneous serum progesterone value.

Cortisol↔cortisone interconvert throughout the day; a 24-hour collection “averages,” while dry captures time-points. More telling are the metabolites that account for ~half of daily output:^26–28 THE, 5-THF, THF. Their sum (“Adrenal Reserve”) ideally ≥ 100% of RRM.

Mary’s Adrenal Reserve: 53% (24-hr), 58% (dry) → below optimal; tracks with AM/PM fatigue.

5-THB well below RRM; THB above median → pattern consistent with low 5-reductase activity across both methods. Other mineralocorticoids appear within normal range.

Saliva reflects blood ≈1 hour prior; urine reflects ≈5+ hours prior. Thus, a “healthy” urinary curve peaks at the second morning time-point. Mary’s first-morning free cortisol/cortisone are the day’s highest → suggests elevated overnight production and daytime decline, aligning with fatigue/sleep disruption.

Dry urine cannot directly measure melatonin (light-sensitive on paper), so it reports 6-sulfatoxymelatonin (robust metabolite marker). 24-hour urine—stored in amber—allows direct melatonin measurement plus the metabolite. Mary: 6-sulfatoxy-melatonin 85% (24-hr) vs. 167% (dry); direct melatonin 147% (24-hr). Different numerically, but together indicate adequate overnight production.

Mary’s aldosterone is healthy. Literature notes low aldosterone has been linked with age-related hearing decline in some individuals; select reports describe improvement with supplementation in specific contexts.^30–32

Mary’s FT4 = 786 (near optimal), FT3 = 402 (below optimal). A widened FT4→FT3 gap can suggest reduced 5′-deiodinase activity and increased rT3. Potential contributors: chronic stress, elevated cortisol, heavy metals. Selenium and improved psychosocial environment may support conversion.

GH = 4514, high within a healthy aging reference derived from adults 18–35 → consistent with Mary’s youthful appearance and potentially supportive of healthy aging trajectories.

Low-normal range; no current symptoms (e.g., social withdrawal, trauma features). Given the hormone’s role in bonding, stress buffering, and potential bone effects,³⁴ re-evaluate later if osteopenia emerges.

• Eliassen AH, Ziegler RG, Rosner B, et al. Reproducibility of urinary estrogens/metabolites. Cancer Epidemiol Biomarkers Prev. 2009.
• Falk RT, Xu X, Keefer L, et al. LC-MS variability/reproducibility for urinary estrogens. CEBP. 2008.
• Faupel-Badger JM, Fuhrman BJ, Xu X, et al. MS vs. RIA/ELISA for urinary estrogen. CEBP. 2010.
• Krone N, Hughes BA, Lavery GG, et al. GC/MS in clinical steroid studies. J Steroid Biochem Mol Biol. 2010.
• Grebe SKG, Singh RJ. LC/MS/MS in the clinical lab—trends. Clin Biochem Rev. 2011.
• Additional citations as referenced (ERβ/E3, 2/16α, methylation, adrenal metabolites, aldosterone/hearing, oxytocin/bone, etc.).

For the full reference list (25+), contact Client Services or request the PDF.

Patients: Talk with your licensed provider about whether a 24-hour or dry profile best fits your goal. Meridian Valley Lab provides laboratory services only and cannot advise patients directly.

Practitioners: Contact Client Services to review cases, interpretation, and panel selection.