With focus on development and implementation of diagnostic tools to examine hormone systems involved in sex differentiation and maturation, child growth, and reproductive health, the core of the Hormone and Endocrine Disruptor Laboratory Group is the Hormone Laboratory and the Analytical Chemistry Laboratory at Dept. of Growth and Reproduction, Rigshospitalet.
Main research and innovation areas include:
Development and implementation of new clinical biochemical tools
We develop and/or implement hormone assays that are relevant for the patients seen in the out-patient clinic of Dept. of Growth and Reproduction. This includes hormone analyses, which are not commonly available elsewhere (e.g. inhibin B, Insulin-like factor 3 (Insl3), Insulin-like binding proteins) but also methods for more common analytes (e.g. luteinising hormone, oestradiol, testosterone). All our assays are, however, optimised with respect to e.g. sensitivity and measuring range to meet the special requirements of our specific patient groups, which also includes infants and prepubertal children.
Recently we have focused on developing hormone assays based on liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) methodology, which is superior to immunoassays in specificity and provide the possibility of measuring several analytes in the same methods. We have now established LC-MS/MS methods for analysis of several sex steroid hormones1 and Insl32. The latter is currently used to explore Insl3 as a marker of Leydig cell function in both basic and clinical andrology.
Several of our hormone analyses have initially been implemented and used as research tools, which following clinical validation subsequently have become valuable diagnostic tools in the clinic. Our current routine hormone analysis repertoire is listed
here (in Danish). Both the Hormone Laboratory and the Analytical Chemistry Laboratory conducting these clinical analyses are accredited according to the ISO 15189 standard for medical laboratories by the Danish Accreditation fund (DANAK).
Characterisation of hormone patterns during normal and pathological sex development and reproductive health
A hallmark of our method implementation is the establishment of in-house sex- and age-related reference ranges for children, adolescents and adults for the hormones and growth factors analysed in the laboratories3. This work is done in collaboration with PIs of population-based health studies providing samples for analysis from healthy individuals, including studies conducted at Dept. of Growth and Reproduction4 and at Centre from Clinical Research and Prevention5. Sex- and age-related reference material is essential for investigating the natural history and sex differences of a given hormone over different stages of life and it is an indispensable tool for comparison of hormone changes in abnormal development and disease.
Our research also includes investigation of the interaction between hormones and how changes in the profiles of associated hormones is related to different developmental stages and pathological conditions. We strive to translate this information into improved diagnostic tools as e.g. bivariate reference ranges6.
Human biomonitoring and the impact of environmental factors on endocrine systems
Our Analytical Chemistry laboratory was established as an analytical facility serving the human studies on endocrine disruption of reproductive health, which have been carried out at Dept. of Growth and Reproduction since mid-1990’ies. We have accordingly for more than 15 years been involved in human biomonitoring (HBM) of several known and suspected non-persistent endocrine disrupting chemicals (e.g. phthalates, parabens, bisphenols, benzophenones and other simple phenols) measured in several human matrices. Our Analytical Chemistry Laboratory serve(d) as a European reference laboratory in completed (DEMOCOPHES) as well as ongoing (HBM4EU) EU-funded European HBM projects. We have provided some of the first data on exposure to these chemicals in the general Danish population7 - data which also has been used in the regulatory work of the Danish Environment Protection Agency.
About the Laboratories:
The Hormone Laboratory
Group leader Anna-Maria Andersson has since 1992 been the scientific supervisor of the Hormone laboratory and is still part of the laboratory management team. However, since 2019 Trine Holm Johannsen, consultant in clinical biochemistry, is the daily manager of the laboratory, which includes eight biomedical laboratory scientists. The Hormone Laboratory is specialized in hormone analyses by immunoassay methodology and performs annually >60,000 hormone analyses for the clinic and for research projects.
The Analytical Chemistry laboratory
In 2004 the Analytical Chemistry Laboratory was established as a small research unit with focus on human biomonitoring of phthalate metabolites. Since then, the list of analytes including both steroid hormones and several non-persistent environmental chemicals and the laboratory facilities (e.g. three triple-quadropole LC-MS/MS platforms) have expanded under the supervision of chemist Hanne Frederiksen, who joined the team in 2006 as manager of the laboratory. In 2016 biochemist Jakob Albrethsen joined the group bringing experience in LC-MS of peptide hormones to the team. The Analytical Chemistry Laboratory staff also includes two biomedical laboratory scientists, and annually performs >12,000 analyses including from 3 to 36 analytes for the clinic and for research projects.
Current national and international collaborations
Human Biomonitoring for EU (HBM4EU) 2018-2021
The Capital Region of Denmark (Region Hovedstaden) is partner in the European collaboration HBM4EU (hbm4eu.eu ), represented by the Hormone and Endocrine Disruptor Laboratory Group at Dept. of Growth and Reproduction. HBM4EU is a joint effort of 28 countries, the European Environment Agency and the European Commission, co-funded under Horizon 2020. In this project we contribute as analyzing and reference laboratory and with our experience in studies on exposure-health associations. Group leader Anna-Maria Andersson also serves as technical support for the Danish national representative in HBM4EU Governing board.
The European reference Network on rare endocrine conditions (Endo-ERN)
Endo-ERN (endo-ern.eu ) is a virtual network involving reference centers across Europe with the aim to improve access to high-quality healthcare for patients with hormonal disorders. Dept. of Growth and Reproduction is one of these reference centers and head of department, Prof. Anders Juul, is furthermore pediatric chair of Work Package 5 (Diagnostics and Laboratory Analysis) within Endo-ERN. In this function he is assisted by group member Trine Holm Johannsen. The Endo-ERN network is a channel for dissemination of innovation carried out within the Hormone and Endocrine Disruptor Laboratory group. Endo-ERN is based on the EU 3rd Health Program
Danish National Centre on Endocrine Disruptors (CEHoS)
Group leader Anna-Maria Andersson has been leader of the Danish National Centre on Endocrine Disrupters (cehos.dk ) since it started in 2008. She and group member Hanne Frederiksen have, together with other colleagues at Dept. of Growth and Reproduction, carried out several project under CEHoS including HBM projects. The Centre, which is a partnership between Rigshospitalet, the Technical University of Denmark and University of Southern Denmark, reports directly to the Danish Environment Protection Agency.
Odense Child Cohort
We have a longstanding collaboration with the PI of Odense Child Cohort8, Prof. Tina Kold Jensen. We have on several occasions analysed hormones and endocrine disrupting chemicals in samples collected in the study.
Current members of the group:
- Anna-Maria Andersson, MSc, Ph.D, Research director and Group leader
- Hanne Frederiksen, MSc, Ph.D, senior scientist and chemist, Leader of the Analytical Chemistry Laboratory
- Trine Holm Johansen, M.D, Ph.D. Consultant in Clinical Biochemistry, Leader of the Hormone Laboratory
- Jakob Christian Albrethsen, MSc, Ph.D, senior scientist and biochemist
Affiliated biomedical laboratory scientists:
- Stine Ehlern Andersen
- Ole Nielsen
- Irina Safonenko
- Marianne Seilund
- Maiken Probst
- Mariann Bech Henriksen
- Farshid Ghazi
- Helle Bekker Sørensen
- Majken Marie Rasmussen
- Karina Skou
1Søeborg T, Frederiksen H, Johannsen TH, Andersson AM, Juul A. Isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of ten steroid metabolites in serum. Clin Chim Acta. 2017 May;468:180-186.
Søeborg T, Frederiksen H, Fruekilde P, Johannsen TH, Juul A, Andersson AM. Serum concentrations of DHEA, DHEAS, 17α-hydroxyprogesterone, Δ4-androstenedione and testosterone in children determined by TurboFlow-LC-MS/MS. Clin Chim Acta. 2013 Apr 18;419:95-101.
2Albrethsen J, Frederiksen H, Andersson AM, Anand-Ivell R, Nordkap L, Bang AK, Jørgensen N, Juul A. Development and validation of a mass spectrometry-based assay for quantification of insulin-like factor 3 in human serum. Clin Chem Lab Med. 2018 Oct 25;56(11):1913-1920
3See e.g. Johannsen TH, Main KM, Ljubicic ML, Jensen TK, Andersen HR, Andersen MS, Petersen JH, Andersson AM, Juul A. Sex Differences in Reproductive Hormones During Mini-Puberty in Infants with Normal and Disordered Sex Development. J Clin Endocrinol Metab. 2018;103(8):3028-3037.
Mouritsen A, Søeborg T, Hagen CP, Mieritz MG, Johannsen TH, Frederiksen H, Andersson AM, Juul A. Longitudinal changes in serum concentrations of adrenal androgen metabolites and their ratios by LC-MS/MS in healthy boys and girls. Clin Chim Acta. 2015 Oct 23;450:370-5..
6Andersson AM, Petersen JH, Jørgensen N, Jensen TK, Skakkebæk NE. Serum inhibin B and follicle-stimulating hormone levels as tools in the evaluation of infertile men: significance of adequate reference values from proven fertile men. J Clin Endocrinol Metab. 2004 Jun;89(6):2873-9. Aksglaede L, Andersson AM, Jørgensen N, Jensen TK, Carlsen E, McLachlan RI, Skakkebaek NE, Petersen JH, Juul A. Primary testicular failure in Klinefelter's syndrome: the use of bivariate luteinizing hormone-testosterone reference charts. Clin Endocrinol (Oxf). 2007 Feb;66(2):276-81.
7See e.g. Frederiksen H, Jensen TK, Jørgensen N, Kyhl HB, Husby S, Skakkebæk NE, Main KM, Juul A, Andersson AM. Human urinary excretion of non-persistent environmental chemicals: an overview of Danish data collected between 2006 and 2012.Reproduction. 2014 Mar 4;147(4):555-65
9Aksglaede, L. et al. Changes in Anti-Müllerian Hormone (AMH) throughout the Life Span: A Population-Based Study of 1027 Healthy Males from Birth (Cord Blood) to the Age of 69 Years. J. Clin. Endocrinol. Metab.
95, 5357–5364 (2010).
Hagen, C. P. et al. Serum levels of anti-Müllerian hormone as a marker of ovarian function in 926 healthy females from birth to adulthood and in 172 Turner syndrome patients. J. Clin. Endocrinol. Metab.
95, 5003–10 (2010).