Establishment of Reference Value of 20 Amino Acids for Toddlers by High Performance Liquid Chromatography Tandem Mass Spectrometry
Abstract
BACKGROUND: Amino acids are one of the essential metabolites, especially the 20 amino acids that are preserved as the building blocks of protein. Alterations in amino acid concentrations are related to disease such as inborn error of metabolism, cancer, as well as nutritional status. Hence, it is necessary to define reference values of 20 plasma-free amino acids for Indonesian toddlers and to establish a robust measurement technique using chromatography with tandem mass spectrometry (MS).
METHODS: This study was a cross-sectional preliminary study to establish reference values. The sample was prepared by mixing plasma with 20% sulfosalicylic acid. Plasma-free amino acids were measured with high-performance liquid chromatography (HPLC) non-derivatization technique using column XTerra for chromatographic separation coupled with tandem MS. Amino acids reference values were taken from 101 healthy Indonesian toddlers aged 1-3 years old. Since amino acids data were not Gaussian distributed, the lower and upper of the reference value was established from the 5th percentile and the 95th percentile, respectively.
RESULTS: Analysis for 20 amino acids was validated. The accuracy ranged from 90.53-105.39% and the precision ranged from 0.06-3.80%. The limit of detection range was 1-2 nmol/mL, and the limit of quantification range was 2-4 nmol/mL. The result was linear, with R2 higher than 0.998. There was no significant difference between boys and girls for all amino acids except for glycine.
CONCLUSION: HPLC with tandem MS method can be used to evaluate amino acids in clinical practice. The reference values obtained are specific for aged 1-3 years old from urban areas in Indonesia. The study suggests that for each population, the reference values for amino acids should be established.
KEYWORDS: amino acids, high-performance liquid chromatography, tandem mass spectrometry, reference values, Indonesia
References
Aliu E, Kanungo S, Arnold GL. Amino acid disorders. Ann Transl Med. 2018; 6(24): 471, CrossRef.
Schuck PF, Malgarin F, Cararo JH, Cardoso F, Streck EL, Ferreira GC. Phenylketonuria pathophysiology: On the role of metabolic alterations. Aging Dis. 2015; 6(5): 390-9, CrossRef.
Blackburn PR, Gass JM, Vairo FPE, Farnham KM, Atwal HK, Macklin SK, et al. Maple syrup urine disease: Mechanisms and management. Appl Clin Genet. 2017; 10: 57-66, CrossRef.
Semba RD, Shardell M, Sakr Ashour FA, Moaddel R, Trehan I, Maleta KM, et al. Child stunting is associated with low circulating essential amino acids. EBioMedicine. 2016; 6: 246-52, CrossRef.
Panigoro SS, Kurniawan A, Ramadhan, Sukartini N, Herqutanto, Paramita RI, et al. Amino acid profile of luminal A and B subtype breast cancer. Indones Biomed J. 2023; 15(3): 269-76, CrossRef.
Kusumastuti AC, Ardiaria M, Hendrianingtyas M. Effect of zinc and iron supplementation on appetite, nutritional status and intelligence quotient in young children. Indones Biomed J. 2018; 10(2): 133-9, CrossRef.
Mak CM, Lee HC, Chan AY, Lam CW. Inborn errors of metabolism and expanded newborn screening: Review and update. 2013; 50(6): 142-62, CrossRef.
Uaariyapanichkul J, Chomtho S, Suphapeetiporn K, Shotelersuk V, Punnahitananda S, Chinjarernpan P, et al. Age-related reference intervals for blood amino acids in Thai pediatric population measured by liquid chromatography tandem mass spectrometry. J Nutr Metab. 2018; 2018: 5124035, CrossRef.
Rutherfurd S, Gilani G. Amino acid analysis. Curr Protoc Protein Sci. 2009; 11: 11.9.1-37, CrossRef.
Kaspar H, Dettmer K, Gronwald W, Oefner PJ. Advances in amino acid analysis. Anal Bioanal Chem. 2009; 393(2): 445-52, CrossRef.
Schwarz EL, Roberts WL, Pasquali M. Analysis of plasma amino acids by HPLC with photodiode array and fluorescence detection. Clin Chim Acta. 2005; 354(1-2): 83-90, CrossRef.
Yi P, Liu L, Mei H, Zeng F, Huang Z, Niu H. Establishment of reference range of plasma amino acids for younger Chinese children by reverse phase HPLC. J Pediatr Endocrinol Metab. 2011; 24(9-10): 733-8, CrossRef.
Meiliana A, Wijaya A. Nutrigenetics, nutrigenomic and precision nutrition. Indones Biomed J. 2020; 12(3): 189-200, CrossRef.
U.S. Department of Health and Human Services Food and Drug Administration. M10 Bioanalytical Method Validation and Study Sample Analysis: Guidance for industry. Maryland: ICH; 2022, article.
Mengerink Y, Kutlán D, Tóth F, Csámpai A, Molnár-Perl I. Advances in the evaluation of the stability and characteristics of the amino acid and amine derivatives obtained with the o-phthaldialdehyde/3- mercaptopropionic acid and o-phthaldialdehyde/Nacetyl-l-cysteine reagents: High-performance liquid chromatography-mass spectrometry study. J Chromatogr A. 2002; 949: 99-124, CrossRef.
Bartolomeo MP, Maisano F. Validation of a reversed-phase HPLC method for quantitative amino acid analysis. J Biomol Tech. 2006; 17(2): 131-7, PMID.
Krumpochova P, Bruyneel B, Molenaar D, Koukou A, Wuhrer M, Niessen WM, et al. Amino acid analysis using chromatography-mass spectrometry: An inter platform comparison study. J Pharm Biomed Anal. 2015; 114: 398-407, CrossRef.
Prinsen HCMT, Schiebergen-Bronkhorst BGM, Roeleveld MW, Jans JJM, de Sain-van der Velden MGM, Visser G, et al. Rapid quantification of underivatized amino acids in plasma by hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass-spectrometry. J Inherit Metab Dis. 2016; 39(5): 651-60, CrossRef.
Arlinda D, Oktoberia I, Karyana M. Validation of a liquid chromatography/tandem mass spectrometry assay for the quantification of plasma dihydroartemisinin. Mol Cell Biomed Sci. 2021; 5(3): 127-36, CrossRef.
Giordano G, Gucciardi A, Pirillo P, Naturale M. Quantification of underivatized amino acids on dry blood spot, plasma, and urine by HPLC-ESI-MS/MS. Methods Mol Biol. 2019; 2030: 153-72, CrossRef.
van Vliet K, van Ginkel WG, van Dam E, de Blaauw P, Koehorst M, Kingma HA, et al. Dried blood spot versus venous blood sampling for phenylalanine and tyrosine. Orphanet J Rare Dis. 2020; 15(1): 82, CrossRef.
Markofski MM, Volpi E. Protein metabolism in women and men: Similarities and disparities. Curr Opin Clin Nutr Metab Care. 2011; 14(1): 93-7, CrossRef.
Lepage N, McDonald N, Dallaire L, Lambert M. Age-specific distribution of plasma amino acid concentrations in a healthy pediatric population. Clin Chem. 1997; 43(12): 2397-402, CrossRef.
Cruz AF, Barbosa TMCC, Adelino TER, Lima WP, Mendes MO, Valadares ER. Amino acid reference intervals by high performance liquid chromatography in plasma sample of Brazilian children. J Bras Pathol Med Lab. 2016; 52: 70-7, CrossRef.
Mayo Clinic Laboratories [Internet]. Amino acids, quantitative, plasma [cited 2017 May 15]. Available from: https://www.mayocliniclabs.com/.
Widodo Y, Sandjaja S, Sumedi E, Khouw I, Deurenberg P. The effect of socio-demographic variables and dairy use on the intake of essential macro- and micronutrients in 0.5-12-year-old Indonesian children. Asia Pac J Clin Nutr. 2016; 25(2): 356-67, CrossRef.
Menteri Kesehatan Republik Indonesia. Peraturan Menteri Kesehatan Republik Indonesia Nomor 28 Tahun 2019. Jakarta; Menteri Kesehatan Republik Indonesia: 2019, article.
DOI: https://doi.org/10.18585/inabj.v16i2.2902
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