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. 2006 Nov;39(11):1071-9.
doi: 10.1016/j.clinbiochem.2006.08.009. Epub 2006 Sep 1.

Urinary beta-luteinizing hormone and beta-follicle stimulating hormone immunoenzymometric assays for population research

Eleanor Brindle  1 , Rebecca C Miller, Jane B Shofer, Nancy A Klein, Michael R Soules, Kathleen A O'Connor
Affiliations

Urinary beta-luteinizing hormone and beta-follicle stimulating hormone immunoenzymometric assays for population research

Eleanor Brindle et al. Clin Biochem. 2006 Nov.

Abstract

Objective: We developed assays for measurement of urinary betaLH and betaFSH under collection and storage conditions typical of non-clinical research settings.

Design and methods: IEMAs for free betaLH and total betaFSH were validated by standard methods. Stability of urinary betaLH and betaFSH was tested across freeze-thaws and stored long term at 4 degrees C or -20 degrees C, or short term at room temperature, and with heating to dissociate the subunits.

Results: The IEMAs exhibited acceptable parallelism, specificity, recovery (averaging 100% for betaLH, 97% for betaFSH), imprecision (maximum within-run and between run CVs, respectively, 4.8% and 25.7% for betaLH, 5.6% and 17.0% for betaFSH), and minimum detectable dose (2.5 pmol/L for betaLH, 6.8 pmol/L for betaFSH). Urine and serum measures were highly correlated (r=0.95 for LH, 0.86 for FSH). There was no consistent decline with any storage type. Dissociation of subunits by heating was needed for betaLH, but not betaFSH.

Conclusion: These IEMAs measure free betaLH and total betaFSH, overcoming inter-individual variability in, and collection and storage effects on, subunit dissociation, without the need for urine preservatives.

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Figures

Figure 1
Figure 1
βLH and βFSH assay parallelism. Log concentration of urinary βLH (top panel) and urinary βFSH (bottom panel) plotted against log urine volume. Error bars are ± 2 SE.
Figure 2
Figure 2
Mean (± 2 SE) gonadotropin profiles of 30 cycles from daily paired urine and serum specimens. Top panel: serum intact LH and urinary βLH; Bottom panel: serum intact FSH and urinary βFSH. Cycles are aligned by day of serum LH peak (day 0). Number of observations varies by cycle day, with a minimum of 4 observations on cycle days -17 and 16 and a maximum of 28 to 30 observations for cycle days -10 through 11. pmol/L SG Adj. = urinary hormone concentration adjusted for specific gravity.
Figure 3
Figure 3
Long-term stability of βLH and βFSH in urine specimens. Log urinary βLH (Panel A) and urinary βFSH (Panel B) measured after 0, 1, 2, 4, 8, 16 (βFSH only), 32, and 64 weeks of storage refrigerated (R) or frozen at −20°C (F). Panel A, βLH, includes an additional treatment where specimens were heated to dissociate subunits before storage in the refrigerator (HR). Values at 16 weeks of storage are missing for βLH because laboratory errors led to assay failure. Error bars are ± 2 SE.
Figure 4
Figure 4
Effects of short-term room temperature storage and repeated freeze-thaw cycles. Log urinary βLH (Panel A) and urinary βFSH (Panel B) measured after 0, 1, 2, 4, or 8 days at room temperature (RT), and 0, 1, 2, 4 or 8 freeze thaw (FT) cycles. Error bars are ± 2 SE.
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