Evaluating urinary estrogen and progesterone metabolites using dried filter paper samples and gas chromatography with tandem mass spectrometry (GC–MS/MS)

Abstract

Background Measuring concentrations of metabolites of estradiol and progesterone in urine, instead of measuring serum concentrations, is common in research and also is used in patient care. The primary aim of this study was to demonstrate that analysis of urine samples dried on filter paper by gas chromatography with tandem mass spectrometry (GC–MS/MS) provides results similar to serum analyzed by radioimmunoassay (RIA). Secondary aims were to show that collection of four samples during the day (4-spot method) can be substituted for a 24-h collection, and that analysis of urine from dried samples is equivalent to liquid urine samples. Methods This prospective observational study compared results of urine and serum analyses. Urine samples from women throughout the menstrual cycle and single samples from postmenopausal women were evaluated. Urine was collected onto filter paper and dried. Dried urine was extracted, hydrolyzed, and derivatized prior to analysis by GC–MS/MS. Hormone concentrations were normalized to creatinine. Single samples were used to compare results of 24-h urine collection to the 4-spot method from a separate population of women and men. A subset of these samples were used to compare results from dried urine to liquid urine. Results The primary study showed good reliability in the comparisons between the dried urine and serum assays. During the menstrual cycles of a subset of four women, urine metabolite concentrations followed the same pattern as serum concentrations. Comparison of 4-spot to 24-h urine collections and of dried to liquid urine measurements had intraclass correlation coefficients (ICC) greater than 0.95, indicating excellent agreement. Conclusions For estradiol and progesterone, the dried urine assay is a good surrogate for serum testing. The 4-spot method can be used instead of 24-h urine collections and dried urine results are comparable to liquid urine. The dried urine assay is useful for some clinical assessments of hormone disorders and may be useful in large epidemiologic studies due to ease of sample handling. Electronic supplementary material The online version of this article (10.1186/s13065-019-0539-1) contains supplementary material, which is available to authorized users.

Ultradian rhythms in heart rate variability and distal body temperature anticipate onset of the luteinizing hormone surge

“…Despite their namesake, home “ovulation tests” that identify supra-threshold LH concentrations do not measure ovulation, which may occur many days after and occasionally a few days before LH surge onset 71 . Despite this variability, the fertile window is often treated as predictable, with definitions including the 5–6 days prior to the LH surge as a proxy for ovulation 85 , the first day of slippery clear cervical fluid through LH surge onset 86 , the total days of slippery clear cervical fluid 87 , day 10–17 of the cycle 88 , and retrospective measures of salivary ferning 85 , basal body temperature 64 , 89 , and progesterone metabolites (e.g., 90 ). Today, many online and app-based ovulation prediction algorithms are validated using day of cycle or LH data alone, in the absence of hormone measures or USDO 2 , 83 , 84 , 91 .…”

“…In the first week of the cycle, and from 3 days after LH surge completion until the end of the luteal phase, samples were pooled every 2 days (a third day was pooled at the end of cycles in instances where the total number of remaining days after the surge was odd). Urine samples were extracted and analyzed as previously described, with previously established ranges of hormone concentrations expected in urine by phase of cycle and during menopause 90 , 101 . Briefly, creatinine was measured in duplicate using a conventional colorimetric (Jaffe) assay.…”

Ultradian Rhythms in Heart Rate Variability and Distal Body Temperature Anticipate the Luteinizing Hormone Surge Onset

“…week of the cycle, and from 3 days after LH surge completion until the end of the luteal phase, samples were pooled every 2 days (a third day was pooled at the end of cycles in instances where the total number of remaining days after the surge was odd). Urine samples were extracted and analyzed as previously described, with previously established ranges of hormone concentrations expected in urine by phase of cycle and during menopause (Roos et al, 2015;Newman et al, 2019). Briefly, creatinine was measured in duplicate using a conventional colorimetric (Jaffe) assay.…”

Reproductive hormone measurement from minimally invasive sample types: Methodological considerations and anthropological importance

“…Urine samples are consequently best suited for study designs that call for measures of overall reproductive hormone output, such as research questions focused on interpopulation and intrapopulation hormone level variance (Valeggia, 2007). For example, urine samples have been used to monitor the menstrual cycle and timing of ovulation, successfully documenting changes in LH, progesterone, and estradiol (Bouchard, Fehring, & Schneider, 2019; Johnson et al, 2015; Newman, Pratt, Curran, & Stanczyk, 2019; Roos et al, 2015). Likewise, urine samples are commonly collected during sporting events to assess androgen levels and test for doping (Wood & Stanton, 2012).…”