Storage stability of five steroids and in dried blood spots for newborn screening and retrospective diagnosis of congenital adrenal hyperplasia
Open accessAbstract
Congenital adrenal hyperplasia (CAH) is a severe inherited disorder of cortisol biosynthesis that is potentially lethal or can seriously affect quality of life. For the first time, we aimed to assess the stability of 21-deoxycortisol (21Deox), 11-deoxycortisol (11Deox), 4-androstenedione (4AD), 17-hydroxyprogesterone (17OHP) and cortisol (Cort), diagnostic for CAH, in dried blood spots (DBSs) during a 1 year storage at different temperatures. Spiked DBS samples were stored at room temperature, 4 °C, -20 °C or -70 °C, respectively and analyzed in triplicates using liquid chromatography-tandem mass spectrometry at Weeks 0, 1, 2, 3 and 4, Month 6 and Year 1. Analyte levels within ±15% vs the baseline were considered stable. Our observations show that 21Deox, 4AD and 17OHP were not significantly changed for 1 year even at room temperature at either analyte levels. In contrast, Cort required storage at 4 °C, -20 °C or -70 °C for long-term stability, being significantly decreased at room temperature from Month 6 (p<0.01) in both the 30(60) nM and the 90(180) nM samples. 11Deox was significantly decreased at room temperature at Year 1 (p<0.01) and only in the 30(60) nM samples. Thus, all biomarkers were stable for up to 1 year at 4 °C, -20 °C or -70 °C and at least for 4 weeks at room temperature. These findings have implications for analyses of stored DBS samples in 2nd-tier assays in newborn screening and for retrospective CAH studies.
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A recent statistical approach using principal component analysis of six steroid levels achieved a positive predictive value of 67%.
“A recent statistical approach using principal component analysis of six steroid levels (17OHP, both first and second-tier, 11-deoxycortisol, androstenedione, 21-deoxycortisol and cortisol) achieved a positive predictive value of 67% (274).”
From Congenital Adrenal Hyperplasia—Current Insights in Pathophysiology, Diagnostics, and Management (2022) · cites this paper as [274]
The Materials and methods > Statistical analysis passage describes ANOVA and Dunnett’s test, not principal component analysis. The Results passage reports steroid storage stability, and searches for the six-steroid approach and a 67% positive predictive value found no passage addressing that claim. Page numbers are unavailable in our copy. (LLM, conf=0.76)
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Cites
Congenital adrenal hyperplasia
El-Maouche, D., Arlt, W., & Merke, D. P. (2017) · The Lancet · doi:10.1016/s0140-6736(17)31431-9 · Cited by 669 · see citing works
Current status of newborn screening worldwide: 2015
Therrell, B. L., Padilla, C. D., Loeber, J. G., Kneisser, I., Saadallah, A., Borrajo, G. J. C., & Adams, J. (2015) · Seminars in Perinatology · doi:10.1053/j.semperi.2015.03.002 · Cited by 602 · see citing works
Plasma steroid-binding proteins: primary gatekeepers of steroid hormone action
Hammond, G. L. (2016) · Journal of Endocrinology · doi:10.1530/joe-16-0070 · Cited by 356 · see citing works
PANG, S., HOTCHKISS, J., DRASH, A. L., LEVINE, L. S., & NEW, M. I. (1977) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jcem-45-5-1003 · Cited by 293 · see citing works
The use of mass spectrometry to analyze dried blood spots
Wagner, M., Tonoli, D., Varesio, E., & Gérard Hopfgartner, D. (2014) · Mass Spectrometry Reviews · doi:10.1002/mas.21441 · Cited by 263 · see citing works
Neonatal screening for congenital adrenal hyperplasia
White, P. C. (2009) · Nature Reviews Endocrinology · doi:10.1038/nrendo.2009.148 · Cited by 240 · see citing works
Congenital Adrenal Hyperplasia
Witchel, S. F. (2017) · Journal of Pediatric and Adolescent Gynecology · doi:10.1016/j.jpag.2017.04.001 · Cited by 187 · see citing works
State of the Science in Dried Blood Spots
Freeman, J. D., Rosman, L. M., Ratcliff, J. D., Strickland, P. T., Graham, D. R. M., & Silbergeld, E. K. (2017) · Clinical Chemistry · doi:10.1373/clinchem.2017.275966 · Cited by 179 · see citing works
Stability of salivary steroids: the influences of storage, food and dental care
Gröschl, M. (2001) · Steroids · doi:10.1016/s0039-128x(01)00111-8 · Cited by 176 · see citing works
Abu‐Rabie, P., & Spooner, N. (2009) · Analytical Chemistry · doi:10.1021/ac901985e · Cited by 118 · see citing works
Adam, B. W., Hall, E. M., Sternberg, M., et al. (2011) · Clinical Biochemistry · doi:10.1016/j.clinbiochem.2011.09.010 · Cited by 96 · see citing works
Expanded newborn screening: social and ethical issues
Dhondt, J. (2010) · Journal of Inherited Metabolic Disease · doi:10.1007/s10545-010-9138-y · Cited by 62 · see citing works
Saedi, S. A., Dean, H., Dent, W., Stockl, E., & Cronin, C. (1996) · PEDIATRICS · doi:10.1542/peds.97.1.100 · Cited by 55 · cited 1× · see citing works
“Of note, the analytical techniques in these previous studies (radioimmunoassay, DELFIA, HPLC-diode array detection) are reportedly prone to cross-reactions and/or have relatively low selectivities [2,27] when compared with LC-MS/MS, which is the technique of choice for the determination of steroids in CAH [1,2,4–6].”
MethodCongenital adrenal hyperplasia.
Witchel, S., & Azziz, R. (2011) · Journal of Pediatric and Adolescent Gynecology · doi:10.1016/j.jpag.2010.10.001 · Cited by 50 · see citing works
Determination of a steroid profile in heel prick blood using LC-MS/MS
Boelen, A., Ruiter, A. F., Claahsen-van der Grinten, H. L., Endert, E., & Ackermans, M. T. (2016) · Bioanalysis · doi:10.4155/bio.16.6 · Cited by 49 · see citing works
de Kock, N., Acharya, S. R., Ubhayasekera, S. J. K. A., & Bergquist, J. (2018) · Scientific Reports · doi:10.1038/s41598-018-35007-0 · Cited by 46 · see citing works
Chen, Z., Zhang, Q., Chen, S., Wang, W., Liu, G., & Deng, H. (2019) · Steroids · doi:10.1016/j.steroids.2019.05.010 · Cited by 43 · see citing works
The Long-Term Stability in Whole Blood of 14 Commonly-Requested Hormone Analytes
Diver, M. J., Hughes, J. G., Hutton, J. L., West, C. R., & Hipkin, L. J. (1994) · Annals of Clinical Biochemistry International Journal of Laboratory Medicine · doi:10.1177/000456329403100606 · Cited by 39 · see citing works
Hofman, L. F., Klaniecki, J. E., & Smith, E. K. (1985) · Clinical Chemistry · doi:10.1093/clinchem/31.7.1127 · Cited by 37 · see citing works
Mass spectrometry theory and application to adrenal diseases
Wooding, K. M., & Auchus, R. J. (2013) · Molecular and Cellular Endocrinology · doi:10.1016/j.mce.2012.12.026 · Cited by 37 · see citing works
Gaudl, A., Kratzsch, J., & Ceglarek, U. (2019) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2019.105389 · Cited by 37 · see citing works
Multi-matrix assay of cortisol, cortisone and corticosterone using a combined MEPS-HPLC procedure
Saracino, M. A., Iacono, C., Somaini, L., Gerra, G., Ghedini, N., & Raggi, M. A. (2013) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2013.10.008 · Cited by 30 · see citing works
Biagini, D., Antoni, S., Lomonaco, T., Ghimenti, S., Salvo, P., Bellagambi, F. G., Scaramuzzo, R. T., Ciantelli, M., Cuttano, A., Fuoco, R., et al. (2019) · Talanta · doi:10.1016/j.talanta.2019.120236 · Cited by 30 · see citing works
Choi, R., Park, H., Oh, H. J., Lee, K., Song, J., & Lee, S. (2019) · Annals of Laboratory Medicine · doi:10.3343/alm.2019.39.3.263 · Cited by 29 · see citing works
Stability of 17α-Hydroxyprogesterone in Dried Blood Spots after Autoclaving and Prolonged Storage
Török, D., Mühl, A., Votava, F., Heinze, G., Sólyom, J., Crone, J., Stöckler‐Ipsiroglu, S., & Waldhauser, F. (2002) · Clinical Chemistry · doi:10.1093/clinchem/48.2.370 · Cited by 26 · see citing works
Monostori, P. B., Szabó, P. T., Mărginean, O., Bereczki, C., & Karg, E. (2015) · Hormone Research in Paediatrics · doi:10.1159/000439380 · Cited by 25 · see citing works
Self-citationMommers, J. H. M., Mengerink, Y., Ritzen, E., et al. (2013) · Analytica Chimica Acta · doi:10.1016/j.aca.2013.03.001 · Cited by 24 · see citing works
Török, D., Mühl, A., Votava, F., Heinze, G., Sólyom, J., Crone, J., Stöckler‐Ipsiroglu, S., & Waldhauser, F. (2002) · PubMed · Cited by 23 · see citing works
Use of filter paper for sample collection and transport in steroid pharmacology
Howe, C. J., & Handelsman, D. J. (1997) · Clinical Chemistry · doi:10.1093/clinchem/43.8.1408 · Cited by 23 · see citing works
Proposed definitive methods for measurement of plasma testosterone and 17α-hydroxyprogesterone
Massé, R., & Wright, L. A. (1996) · Clinical Biochemistry · doi:10.1016/0009-9120(96)00037-9 · Cited by 19 · see citing works
Lando, V. S., Batista, M. C., Nakamura, I. T., Mazi, C. R., Mendonca, B. B., & Brito, V. N. (2008) · Journal of Medical Screening · doi:10.1258/jms.2008.007086 · Cited by 18 · see citing works
Elvers, L. H., Loeber, J. G., Dhondt, J., et al. (2007) · Journal of Inherited Metabolic Disease · doi:10.1007/s10545-007-0622-y · Cited by 12 · see citing works
Metabolic profiles derived from residual blood spot samples: A longitudinal analysis
Murphy, M. S. Q., Hawken, S., Cheng, W., Wilson, L. A., Lamoureux, M., Henderson, M., Potter, B. K., Little, J., Chakraborty, P. K., & Wilson, K. R. (2018) · Gates Open Research · doi:10.12688/gatesopenres.12822.1 · Cited by 9 · see citing works
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Research works printed but not linked: 1
- [20]Guideline on bioanalytical method validation. European Medicines Agency Reference number: EMEA/CHMP/EWP/192217/2009 Rev. 1 Corr. 2. Effective from: February 1st 2012. https://www.ema.europa.eu/en/bioanalytical-method-validation (Accessed: January 24th 2020).
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Congenital Adrenal Hyperplasia—Current Insights in Pathophysiology, Diagnostics, and Management
Claahsen - van der Grinten, H. L., Speiser, P. W., Ahmed, S. F., Arlt, W., Auchus, R. J., Falhammar, H., Flück, C. E., Guasti, L., Huebner, A., Kortmann, B. B. M., et al. (2022) · Endocrine Reviews · doi:10.1210/endrev/bnab016 · Cited by 556 · see citing works
- Authors
- Grecsó, N., Zádori, A., Szécsi, I., Baráth, Á., Galla, Z., Bereczki, C., & Monostori, P.
- Affiliations
- University of Szeged (HU)
- Corresponding author
- Grecsó, Nóra
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2020-05-29
- Volume
- 15
- Issue
- 5
- Pages
- e0233724
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC7259505
- OpenAlex
- W3032371537
- Cited by
- 26
- References
- 33
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 5 · 2023: 5 · 2022: 1 · 2021: 8 · 2020: 1
- FWCI
- 1.22
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- PLoS ONE (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- SZTE Publicatio Repozitórium (University of Szeged) (acceptedVersion, other-oa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- University of Szeged Open Access Fund (4581)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Sexual Differentiation and Disorders
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
- Keywords
- congenital adrenal hyperplasia, dried blood spots, sample storage stability, 21-deoxycortisol, 11-deoxycortisol, 4-androstenedione, 17-hydroxyprogesterone, cortisol, steroid biomarkers, newborn screening, retrospective diagnosis, storage temperature
- MeSH terms
- Adrenal Hyperplasia, Congenital, Female, Humans, Infant, Newborn, Male, Mass Screening, Pregnenediones, Preservation, Biological, Androstenediol, Dried Blood Spot Testing
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