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Licensed Unlicensed Requires Authentication Published by De Gruyter February 6, 2020

Investigating the suitability of high-resolution mass spectrometry for newborn screening: identification of hemoglobinopathies and β-thalassemias in dried blood spots

Thomas Wiesinger, Thomas Mechtler, Markus Schwarz, Xiaolei Xie, Regine Grosse, Paulina Nieves Cobos, David Kasper and Zoltan Lukacs


A fast and reliable method for the determination of hemoglobinopathies and thalassemias by high-resolution accurate mass spectrometry (HRAM/MS) is presented. The established method was verified in a prospective clinical study (HRAM/MS vs. high-pressure liquid chromatography [HPLC]) of 5335 de-identified newborn samples from the Hamburg area. The analytical method is based on a dual strategy using intact protein ratios for thalassemias and tryptic digest fragments for the diagnosis of hemoglobinopathies. Due to the minimal sample preparation and the use of flow injection, the assay can be considered as a high-throughput screening approach for newborn screening programs (2 min/sample). Using a simple dried blood spot (DBS) extraction (tryptic digest buffer), the following results were obtained: (1) a carrier incidence of 1:100 newborns (35 FAS, nine FAC, eight FAD and two FAE), and (2) no homozygous affected patient was detected. Using the HRAM/MS protocol, an unknown Hb mutation was identified and confirmed by genetic testing. In addition to greater specificity toward rare mutations and β-thalassemia, the low price/sample (1–2€) as well as an automated data processing represent the major benefits of the described HRAM/MS method.


ARCHIMEDlife Science GmbH was supported by Gina Tan, Claudio DeNardi and Bradley Hart from Thermo Fisher Scientific.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


1. Kohne E, Kleihauer E. Hemoglobinopathies: a longitudinal study over four decades. Dtsch Arztebl Int 2010;107:65–71.Search in Google Scholar

2. Aguilar Martinez P, Angastiniotis M, Eleftheriou A, Gulbis B, Del Mar Mañú Pereira M, Petrova-Benedict R, et al. Haemoglobinopathies in Europe: health & migration policy perspectives. Orphanet J Rare Dis 2014;9:97.10.1186/1750-1172-9-97Search in Google Scholar

3. Taher AT, Weatherall DJ, Cappellini MD. Thalassaemia. Lancet 2018;391:155–67.10.1016/S0140-6736(17)31822-6Search in Google Scholar

4. Higgs DR. The molecular basis of α thalassemia. Cold Spring Harb Perspect Med 2013;3:a011718.10.1101/cshperspect.a011718Search in Google Scholar PubMed PubMed Central

5. Hoppe CC. Prenatal and newborn screening for hemoglobinopathies. Int J Lab Hematol 2013;35:297–305.10.1111/ijlh.12076Search in Google Scholar PubMed

6. Hoppe CC. Newborn screening for hemoglobin disorders.Hemoglobin 2011;35:556–64.10.3109/03630269.2011.607905Search in Google Scholar PubMed

7. Grosse R, Lukacs Z, Cobos PN, Oyen F, Ehmen C, Muntau B, et al. The prevalence of sickle cell disease and its implication for newborn screening in Germany (Hamburg Metropolitan area). Pediatr Blood Cancer 2016;63:168–70.10.1002/pbc.25706Search in Google Scholar PubMed

8. Lobitz S, Klein J, Brose A, Blankenstein O, Frömmel C. Newborn screening by tandem mass spectrometry confirms the high prevalence of sickle cell disease among German newborns. Ann Hematol 2019;98:47–53.10.1007/s00277-018-3477-4Search in Google Scholar PubMed

9. Pattloch D. Sichelzellkrankheit unter Neugeborenen in Deutschland: Eine Studie an Routinedaten der AOK. Das Gesundheitswesen 2018.10.1055/a-0719-5165Search in Google Scholar PubMed

10. Greene DN, Vaughn CP, Crews BO, Agarwal AM. Advances in detection of hemoglobinopathies. Clin Chim Acta 2015;439:50–7.10.1016/j.cca.2014.10.006Search in Google Scholar PubMed

11. Daniel Y, Turner C, Farrar L, Dalton RN. A comparison of IEF and MSMS for clinical hemoglobinopathy screening in 40,000 newborns. Blood 2008;112:2387.10.1182/blood.V112.11.2387.2387Search in Google Scholar

12. Moat SJ, Rees D, King L, Ifederu A, Harvey K, Hall K, et al. Newborn blood spot screening for sickle cell disease by using tandem mass spectrometry: implementation of a protocol to identify only the disease states of sickle cell disease. Clin Chem 2014;60:373–80.10.1373/clinchem.2013.210948Search in Google Scholar PubMed

13. Daniel YA, Henthorn J. Newborn screening for sickling and other haemoglobin disorders using tandem mass spectrometry: a pilot study of methodology in laboratories in England. J Med Screen 2016;23:175–8.10.1177/0969141316631008Search in Google Scholar PubMed

14. Yu C, Huang S, Wang M, Zhang J, Liu H, Yuan Z, et al. A novel tandem mass spectrometry method for first-line screening of mainly beta-thalassemia from dried blood spots. J Proteom 2017;154:78–84.10.1016/j.jprot.2016.12.008Search in Google Scholar PubMed

15. Rai DK, Landin B, Alvelius G, Griffiths WJ. Electrospray tandem mass spectrometry of intact-chain hemoglobin variants. Anal Chem 2002;74:2097–102.10.1021/ac011068fSearch in Google Scholar PubMed

16. Edwards RL, Creese AJ, Baumert M, Griffiths P, Bunch J, Cooper HJ. Hemoglobin variant analysis via direct surface sampling of dried blood spots coupled with high-resolution mass spectrometry. Anal Chem 2011;83:2265–70.10.1021/ac1030804Search in Google Scholar PubMed PubMed Central

17. Daniel YA, Turner C, Haynes RM, Hunt BJ, Neil Dalton R. Rapid and specific detection of clinically significant haemoglobinopathies using electrospray mass spectrometry-mass spectrometry. Br J Haematol 2005;130:635–43.10.1111/j.1365-2141.2005.05646.xSearch in Google Scholar PubMed

18. Théberge R, Dikler S, Heckendorf C, Chui DH, Costello CE, McComb ME. MALDI-ISD mass spectrometry analysis of hemoglobin variants: a top-down approach to the characterization of hemoglobinopathies. J Am Soc Mass Spectrom Chem 2015;26:1299–310.10.1007/s13361-015-1164-4Search in Google Scholar PubMed PubMed Central

19. Helmich F, Van Dongen JL, Kuijper PH, Scharnhorst V, Brunsveld L, Broeren MA. Rapid phenotype hemoglobin screening by high-resolution mass spectrometry on intact proteins. Clin Chim Acta 2016;460:220–6.10.1016/j.cca.2016.07.006Search in Google Scholar PubMed

20. Municie HL, James SC. Alpha and beta thalassemia. Am Fam Physician 2009;80:339–44.Search in Google Scholar

21. Aliyeva G, Asadov C, Mammadova T, Gafarova S, Abdulalimov E. Thalassemia in the laboratory: pearls, pitfalls, and promises. Clin Chem Lab Med 2018;57:165–74.10.1515/cclm-2018-0647Search in Google Scholar PubMed

22. Moat SJ, Rees D, George RS, King L, Dodd A, Ifederu A, et al. Newborn screening for sickle cell disorders using tandem mass spectrometry: three years’ experience of using a protocol to detect only the disease states. Ann Clin Biochem. 2017;54:601–11.10.1177/0004563217713788Search in Google Scholar PubMed

23. de Souza Carrocini GC, Zamaro PJ, Bonini-Domingos CR. What influences Hb fetal production in adulthood? Rev Bras Hematol Hemoter 2011;33:231–6.10.5581/1516-8484.20110059Search in Google Scholar PubMed PubMed Central

24. Edoh D, Antwi-Bosaiko C, Amuzu D. Fetal hemoglobin during infancy and in sickle cell adults. Afr Health Sci 2006;6:51–4.Search in Google Scholar

25. Lehotay DC, Hall P, Lepage J, Eichhorst JC, Etter ML, Greenberg CR. LC–MS/MS progress in newborn screening. Clin Biochem 2011;2011:21–31.10.1016/j.clinbiochem.2010.08.007Search in Google Scholar PubMed

26. Sankaran VG, Orkin SH. The switch from fetal to adult hemoglobin. Cold Spring Harb Perspect Med 2013;3: a011643.10.1101/cshperspect.a011643Search in Google Scholar PubMed PubMed Central

27. Modell B, Darlison M, Birgens H, Cario H, Faustino P, Giordano PC, et al. Epidemiology of haemoglobin disorders in Europe: an overview. Scand J Clin Lab Invest 2007;67:39–70.10.1080/00365510601046557Search in Google Scholar PubMed

Supplementary Material

The online version of this article offers supplementary material (

Received: 2019-08-08
Accepted: 2019-12-23
Published Online: 2020-02-06
Published in Print: 2020-04-28

©2020 Walter de Gruyter GmbH, Berlin/Boston

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