A COMPARATIVE STUDY OF PLASMA AND DRIED BLOOD SPOT ACYLCARNITINES IN FATTY ACID OXIDATION DISORDERS AND ORGANIC ACIDURIAS

Azzah Hana Abu Yamin; Muhd Irfan Bukhari  Ahmad Nazri; Nurfarah Nabila Mohd Adhari; Marleena  Mamat; Chew Hui Bein; Moey Lip Hen; Saraswathy Apparow; Salina  Abdul Rahman; Anasufiza  Habib

Issue

Vol. 2 No. 1 (2025): Young Scientists Award (34th MACB Conference 2024)

Issue Published : 16 January 2025

Date Log

Submitted

19 July 2024

Accepted

27 September 2024

Published

16 January 2025

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

A COMPARATIVE STUDY OF PLASMA AND DRIED BLOOD SPOT ACYLCARNITINES IN FATTY ACID OXIDATION DISORDERS AND ORGANIC ACIDURIAS

Azzah Hana AY

Institute for Medical Research

https://orcid.org/0009-0006-1396-1953

Muhd Irfan Bukhari AN

Nurfarah Nabila MA

Marleena M

Chew Hui Bein

https://orcid.org/0000-0002-8223-5852

Moey Lip Hen

https://orcid.org/0000-0002-5516-0592

Saraswathy Apparow

https://orcid.org/0000-0001-8870-7796

Salina Abdul Rahman

https://orcid.org/0000-0002-0851-4124

Anasufiza Habib

https://orcid.org/0000-0001-5529-5115

Corresponding Author(s) : Azzah Hana AY

azzahhana@moh.gov.my

Malaysian Journal of Clinical Biochemistry, Vol. 2 No. 1 (2025): Young Scientists Award (34th MACB Conference 2024)
Article Published : 16 January 2025

Abstract

Analysis of blood acylcarnitines is used in the investigation of fatty acid oxidation disorders (FAOD) and organic acidurias. Dried blood spot (DBS) sample is regarded as the preferred sample matrix for newborn screening programmes due to its certain characteristics. However, there is limited data on the correlation of acylcarnitines in plasma and DBS. This study aims to compare individual acylcarnitines as well as the profile between plasma and DBS in FAOD and organic acidurias. We studied 55 paired plasma and DBS samples of known FAOD and organic acidurias. Individual acylcarnitines from both matrices were analysed as their butyl esters by electrospray ionisation tandem mass spectrometry (ESI-MS/MS) in multiple reaction monitoring (MRM) mode. The similarities and differences in acylcarnitines concentration and profile interpretation between plasma and DBS for individual diseases were described. Free carnitine elevations in patients with CPT1a deficiency were two times higher in DBS than that of plasma. Long-chain acylcarnitines elevations in patients with CACT and CPT2 deficiency were more strikingly demonstrated in plasma than in DBS. Long-chain hydroxylacylcarnitines elevations in MTP/LCHAD deficiency were equally detected on both matrices. Elevations of the main acylcarnitines were seen in both plasma and DBS samples of patients with propionic aciduria, glutaric aciduria type 1, and isovaleric aciduria. In methylmalonic aciduria, 3/19 (15.8%) patients had DBS C3-acylcarnitine within the reference range. DBS is a better sample for detecting CPT1a deficiency, while plasma is better at detecting CACT/CPT2 deficiency. Both plasma and DBS are equally useful in the investigation of suspected LCHAD/MTP deficiency and common organic acidurias.

Keywords

acylcarnitine profile; dried blood spot; fatty acid oxidation disorders; organic acidurias; tandem mass spectrometry

A COMPARATIVE STUDY OF PLASMA AND DRIED BLOOD SPOT ACYLCARNITINES IN FATTY ACID OXIDATION DISORDERS AND ORGANIC ACIDURIAS. (2025). Malaysian Journal of Clinical Biochemistry, 2(1), 1-11. http://journal.macb.org.my/home/article/view/21

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References

Al-Thihli, K., Sinclair, G., Sirrs, S., Mezei, M., Nelson, J., & Vallance, H. (2014). Performance of serum and dried blood spot acylcarnitine profiles for detection of fatty acid β-oxidation disorders in adult patients with rhabdomyolysis. Journal of Inherited Metabolic Disease, 37(2), 207–213. https://doi.org/10.1007/s10545-012-9578-7
Belaramani, K. M., Chan, T. C. H., Hau, E. W. L., Yeung, M. C. W., Kwok, A. M. K., Lo, I. F. M., Law, T. H. F., Wu, H., Wong, S. S. N., Lam, S. W., Ha, G. H. Y., Lau, T. P. Y., Wong, T. K., Or, V. W. C., Wong, R. M. S., Ming, W. L., Chow, J. C. K., Yau, E. K. C., Fu, A., … Fung, C. W. (2024). Expanded Newborn Screening for Inborn Errors of Metabolism in Hong Kong: Results and Outcome of a 7 Year Journey. International Journal of Neonatal Screening, 10(1), 1–14. https://doi.org/10.3390/ijns10010023
Borch, L., Lund, A. M., Wibrand, F., Christensen, E., Søndergaard, C., Gahrn, B., Hougaard, D. M., Andresen, B. S., Gregersen, N., & Olsen, R. K. J. (2012). Normal levels of plasma free carnitine and acylcarnitines in follow-up samples from a presymptomatic case of carnitine palmitoyl transferase 1 (CPT1) deficiency detected through newborn screening in Denmark. In JIMD Reports (Vol. 3, pp. 11–15). Springer. https://doi.org/10.1007/8904_2011_35
Chien, Y. H., & Hwu, W. L. (2023). The modern face of newborn screening. Pediatrics and Neonatology, 64, S22–S29. https://doi.org/10.1016/j.pedneo.2022.11.001
de Sain-van der Velden, M. G. M., Diekman, E. F., Jans, J. J., van der Ham, M., Prinsen, B. H. C. M. T., Visser, G., & Verhoeven-Duif, N. M. (2013). Differences between acylcarnitine profiles in plasma and bloodspots. Molecular Genetics and Metabolism, 110(1–2), 116–121. https://doi.org/10.1016/j.ymgme.2013.04.008
Dowsett, L., Lulis, L., Ficicioglu, C., & Cuddapah, S. (2017). Utility of genetic testing for confirmation of abnormal newborn screening in disorders of long-chain fatty acids: A missed case of carnitine palmitoyltransferase 1A (CPT1A) deficiency. International Journal of Neonatal Screening, 3(2). https://doi.org/10.3390/ijns3020010
Fingerhut, R., Röschinger, W., Muntau, A. C., Dame, T., Kreischer, J., Arnecke, R., Superti-Furga, A., Troxler, H., Liebl, B., Olgemöller, B., & Roscher, A. A. (2001). Hepatic carnitine palmitoyltransferase I deficiency: Acylcarnitine profiles in blood spots are highly specific. Clinical Chemistry, 47(10), 1763–1768. https://doi.org/10.1093/clinchem/47.10.1763
Habib, A., Azize, N. A. A., Rahman, S. A., Yakob, Y., Suberamaniam, V., Nazri, M. I. B. A., Abdullah Sani, H., Ch’ng, G. S., Yin, L. H., Olpin, S., & Lock-Hock, N. (2021). Novel mutations associated with carnitine-acylcarnitine translocase and carnitine palmitoyl transferase 2 deficiencies in Malaysia. Clinical Biochemistry, 98, 48–53. https://doi.org/10.1016/j.clinbiochem.2021.10.002
Lim, J. S., Tan, E. S., John, C. M., Poh, S., Yeo, S. J., Ang, J. S. M., Adakalaisamy, P., Rozalli, R. A., Hart, C., Tan, E. T. H., Ranieri, E., Rajadurai, V. S., Cleary, M. A., & Goh, D. L. M. (2014). Inborn error of metabolism (IEM) screening in Singapore by electrospray ionization-tandem mass spectrometry (ESI/MS/MS): An 8year journey from pilot to current program. Molecular Genetics and Metabolism, 113(1), 53–61. https://doi.org/10.1016/j.ymgme.2014.07.018
Lim, M. D. (2018a). Review article: Dried blood spots for global health diagnostics and surveillance: Opportunities and challenges. American Journal of Tropical Medicine and Hygiene, 99(2), 256–265. https://doi.org/10.4269/ajtmh.17-0889
Lim, M. D. (2018b). Review article: Dried blood spots for global health diagnostics and surveillance: Opportunities and challenges. American Journal of Tropical Medicine and Hygiene, 99(2), 256–265. https://doi.org/10.4269/ajtmh.17-0889
Lotz-Havla, A. S., Röschinger, W., Schiergens, K., Singer, K., Karall, D., Konstantopoulou, V., Wortmann, S. B., & Maier, E. M. (2018). Fatal pitfalls in newborn screening for mitochondrial trifunctional protein (MTP)/long-chain 3-Hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency. Orphanet Journal of Rare Diseases, 13(1), 1–8. https://doi.org/10.1186/s13023-018-0875-6
Malsagova, K., Kopylov, A., Stepanov, A., Butkova, T., Izotov, A., & Kaysheva, A. (2020). Dried blood spot in laboratory: Directions and prospects. Diagnostics, 10(4). https://doi.org/10.3390/diagnostics10040248
Martín-Rivada, Á., Palomino Pérez, L., Ruiz-Sala, P., Navarrete, R., Cambra Conejero, A., Quijada Fraile, P., Moráis López, A., Belanger-Quintana, A., Martín-Hernández, E., Bellusci, M., Cañedo Villaroya, E., Chumillas Calzada, S., García Silva, M. T., Bergua Martínez, A., Stanescu, S., Martínez-Pardo Casanova, M., Ruano, M. L. F., Ugarte, M., Pérez, B., & Pedrón-Giner, C. (2022). Diagnosis of inborn errors of metabolism within the expanded newborn screening in the Madrid region. JIMD Reports, 63(2), 146–161. https://doi.org/10.1002/jmd2.12265
Merritt, J. L. 2nd, Norris, M., & Kanungo, S. (2018). Fatty Acid Oxidation disorders. Annals of Translational Medicine, 6(24), 473. https://doi.org/10.21037/atm.2018.10.57
Miller, M. J., Cusmano-Ozog, K., Oglesbee, D., & Young, S. (2021). Laboratory analysis of acylcarnitines, 2020 update: a technical standard of the American College of Medical Genetics and Genomics (ACMG). Genetics in Medicine, 23(2), 249–258. https://doi.org/10.1038/s41436-020-00990-1
Millington, D. S., & Stevens, R. D. (2011). Acylcarnitines: Analysis in Plasma and Whole Blood Using Tandem Mass Spectrometry. Methods in Molecular Biology, 708, 55–72. https://doi.org/10.1007/978-1-61737-985-7_3
Reuter, S. E., & Evans, A. M. (2012). Carnitine and Acylcarnitines Pharmacokinetic, Pharmacological and Clinical Aspects. Clin Pharmacokinet, 51, 553–572. https://doi.org/https://doi.org/10.1007/BF03261931
Zhou, D., Cheng, Y., Yin, X., Miao, H., Hu, Z., Yang, J., Zhang, Y., Wu, B., & Huang, X. (2022). Newborn Screening for Mitochondrial Carnitine-Acylcarnitine Cycle Disorders in Zhejiang Province, China. Frontiers in Genetics, 13(March), 1–10. https://doi.org/10.3389/fgene.2022.823687

References

Al-Thihli, K., Sinclair, G., Sirrs, S., Mezei, M., Nelson, J., & Vallance, H. (2014). Performance of serum and dried blood spot acylcarnitine profiles for detection of fatty acid β-oxidation disorders in adult patients with rhabdomyolysis. Journal of Inherited Metabolic Disease, 37(2), 207–213. https://doi.org/10.1007/s10545-012-9578-7

Belaramani, K. M., Chan, T. C. H., Hau, E. W. L., Yeung, M. C. W., Kwok, A. M. K., Lo, I. F. M., Law, T. H. F., Wu, H., Wong, S. S. N., Lam, S. W., Ha, G. H. Y., Lau, T. P. Y., Wong, T. K., Or, V. W. C., Wong, R. M. S., Ming, W. L., Chow, J. C. K., Yau, E. K. C., Fu, A., … Fung, C. W. (2024). Expanded Newborn Screening for Inborn Errors of Metabolism in Hong Kong: Results and Outcome of a 7 Year Journey. International Journal of Neonatal Screening, 10(1), 1–14. https://doi.org/10.3390/ijns10010023

Borch, L., Lund, A. M., Wibrand, F., Christensen, E., Søndergaard, C., Gahrn, B., Hougaard, D. M., Andresen, B. S., Gregersen, N., & Olsen, R. K. J. (2012). Normal levels of plasma free carnitine and acylcarnitines in follow-up samples from a presymptomatic case of carnitine palmitoyl transferase 1 (CPT1) deficiency detected through newborn screening in Denmark. In JIMD Reports (Vol. 3, pp. 11–15). Springer. https://doi.org/10.1007/8904_2011_35

Chien, Y. H., & Hwu, W. L. (2023). The modern face of newborn screening. Pediatrics and Neonatology, 64, S22–S29. https://doi.org/10.1016/j.pedneo.2022.11.001

de Sain-van der Velden, M. G. M., Diekman, E. F., Jans, J. J., van der Ham, M., Prinsen, B. H. C. M. T., Visser, G., & Verhoeven-Duif, N. M. (2013). Differences between acylcarnitine profiles in plasma and bloodspots. Molecular Genetics and Metabolism, 110(1–2), 116–121. https://doi.org/10.1016/j.ymgme.2013.04.008

Dowsett, L., Lulis, L., Ficicioglu, C., & Cuddapah, S. (2017). Utility of genetic testing for confirmation of abnormal newborn screening in disorders of long-chain fatty acids: A missed case of carnitine palmitoyltransferase 1A (CPT1A) deficiency. International Journal of Neonatal Screening, 3(2). https://doi.org/10.3390/ijns3020010

Fingerhut, R., Röschinger, W., Muntau, A. C., Dame, T., Kreischer, J., Arnecke, R., Superti-Furga, A., Troxler, H., Liebl, B., Olgemöller, B., & Roscher, A. A. (2001). Hepatic carnitine palmitoyltransferase I deficiency: Acylcarnitine profiles in blood spots are highly specific. Clinical Chemistry, 47(10), 1763–1768. https://doi.org/10.1093/clinchem/47.10.1763

Habib, A., Azize, N. A. A., Rahman, S. A., Yakob, Y., Suberamaniam, V., Nazri, M. I. B. A., Abdullah Sani, H., Ch’ng, G. S., Yin, L. H., Olpin, S., & Lock-Hock, N. (2021). Novel mutations associated with carnitine-acylcarnitine translocase and carnitine palmitoyl transferase 2 deficiencies in Malaysia. Clinical Biochemistry, 98, 48–53. https://doi.org/10.1016/j.clinbiochem.2021.10.002

Lim, J. S., Tan, E. S., John, C. M., Poh, S., Yeo, S. J., Ang, J. S. M., Adakalaisamy, P., Rozalli, R. A., Hart, C., Tan, E. T. H., Ranieri, E., Rajadurai, V. S., Cleary, M. A., & Goh, D. L. M. (2014). Inborn error of metabolism (IEM) screening in Singapore by electrospray ionization-tandem mass spectrometry (ESI/MS/MS): An 8year journey from pilot to current program. Molecular Genetics and Metabolism, 113(1), 53–61. https://doi.org/10.1016/j.ymgme.2014.07.018

Lim, M. D. (2018a). Review article: Dried blood spots for global health diagnostics and surveillance: Opportunities and challenges. American Journal of Tropical Medicine and Hygiene, 99(2), 256–265. https://doi.org/10.4269/ajtmh.17-0889

Lim, M. D. (2018b). Review article: Dried blood spots for global health diagnostics and surveillance: Opportunities and challenges. American Journal of Tropical Medicine and Hygiene, 99(2), 256–265. https://doi.org/10.4269/ajtmh.17-0889

Lotz-Havla, A. S., Röschinger, W., Schiergens, K., Singer, K., Karall, D., Konstantopoulou, V., Wortmann, S. B., & Maier, E. M. (2018). Fatal pitfalls in newborn screening for mitochondrial trifunctional protein (MTP)/long-chain 3-Hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency. Orphanet Journal of Rare Diseases, 13(1), 1–8. https://doi.org/10.1186/s13023-018-0875-6

Malsagova, K., Kopylov, A., Stepanov, A., Butkova, T., Izotov, A., & Kaysheva, A. (2020). Dried blood spot in laboratory: Directions and prospects. Diagnostics, 10(4). https://doi.org/10.3390/diagnostics10040248

Martín-Rivada, Á., Palomino Pérez, L., Ruiz-Sala, P., Navarrete, R., Cambra Conejero, A., Quijada Fraile, P., Moráis López, A., Belanger-Quintana, A., Martín-Hernández, E., Bellusci, M., Cañedo Villaroya, E., Chumillas Calzada, S., García Silva, M. T., Bergua Martínez, A., Stanescu, S., Martínez-Pardo Casanova, M., Ruano, M. L. F., Ugarte, M., Pérez, B., & Pedrón-Giner, C. (2022). Diagnosis of inborn errors of metabolism within the expanded newborn screening in the Madrid region. JIMD Reports, 63(2), 146–161. https://doi.org/10.1002/jmd2.12265

Merritt, J. L. 2nd, Norris, M., & Kanungo, S. (2018). Fatty Acid Oxidation disorders. Annals of Translational Medicine, 6(24), 473. https://doi.org/10.21037/atm.2018.10.57

Miller, M. J., Cusmano-Ozog, K., Oglesbee, D., & Young, S. (2021). Laboratory analysis of acylcarnitines, 2020 update: a technical standard of the American College of Medical Genetics and Genomics (ACMG). Genetics in Medicine, 23(2), 249–258. https://doi.org/10.1038/s41436-020-00990-1

Millington, D. S., & Stevens, R. D. (2011). Acylcarnitines: Analysis in Plasma and Whole Blood Using Tandem Mass Spectrometry. Methods in Molecular Biology, 708, 55–72. https://doi.org/10.1007/978-1-61737-985-7_3

Reuter, S. E., & Evans, A. M. (2012). Carnitine and Acylcarnitines Pharmacokinetic, Pharmacological and Clinical Aspects. Clin Pharmacokinet, 51, 553–572. https://doi.org/https://doi.org/10.1007/BF03261931

Zhou, D., Cheng, Y., Yin, X., Miao, H., Hu, Z., Yang, J., Zhang, Y., Wu, B., & Huang, X. (2022). Newborn Screening for Mitochondrial Carnitine-Acylcarnitine Cycle Disorders in Zhejiang Province, China. Frontiers in Genetics, 13(March), 1–10. https://doi.org/10.3389/fgene.2022.823687