Open Access
JIMD Reports, volume 63, issue 4, pages 341-350
Method development and validation for analysis of phenylalanine, 4‐hydroxyphenyllactic acid and 4‐hydroxyphenylpyruvic acid in serum and urine
Publication type: Journal Article
Publication date: 2022-04-03
Journal:
JIMD Reports
Quartile SCImago
Q3
Quartile WOS
—
Impact factor: —
ISSN: 21928304, 21928312, 21928312
Endocrinology, Diabetes and Metabolism
Internal Medicine
Biochemistry, Genetics and Molecular Biology (miscellaneous)
Abstract
Alkaptonuria (AKU) is a rare debilitating autosomal recessive disorder of tyrosine (TYR) metabolism which results in a deficiency of the enzyme homogentisate 1,2-dioxygenase activity. Several studies have reported the metabolic changes in homogentisic acid (HGA) concentrations and subsequent deposition of an ochronotic pigment in connective tissues, especially cartilage. Treatment with nitisinone (NTBC) reduces urinary and circulating HGA, but its mode of action results in hypertyrosinaemia. The effect of NTBC on other metabolites in the TYR pathway has not been reported. Modification of the current reverse phase liquid chromatography tandem mass spectrometry methods for serum and urine to include phenylalanine (PHE), hydroxyphenyllactate (HPLA) and hydroxyphenylpyruvate (HPPA) has been validated. HPPA and HPLA (negative ionisation) eluted at 2.8 and 2.9 min respectively on an Atlantis C18 column with PHE (positive ionisation) eluting earlier at 2.4 min. Intra- and inter-assay accuracy was between 96.3% and 100.3% for PHE; 96.6% and 110.5% for HPLA and 95.0% and 107.8% for HPPA in both urine and serum. Precision, both inter- and intra-assay, was <10% for all analytes in both serum and urine. No significant issues with carry-over, stability or matrix interferences were seen in either the urine or serum assays. Measurement of serum and urine from AKU patients has demonstrated a robust, fully validated assay, appropriate for monitoring of patients with AKU and for demonstrating metabolite changes, following NTBC therapy.
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Hughes A. T. et al. Method development and validation for analysis of phenylalanine, 4‐hydroxyphenyllactic acid and 4‐hydroxyphenylpyruvic acid in serum and urine // JIMD Reports. 2022. Vol. 63. No. 4. pp. 341-350.
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Hughes A. T., Milan A. M., Shweihdi E., Gallagher J., RANGANATH L. Method development and validation for analysis of phenylalanine, 4‐hydroxyphenyllactic acid and 4‐hydroxyphenylpyruvic acid in serum and urine // JIMD Reports. 2022. Vol. 63. No. 4. pp. 341-350.
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TY - JOUR
DO - 10.1002/jmd2.12287
UR - https://doi.org/10.1002/jmd2.12287
TI - Method development and validation for analysis of phenylalanine, 4‐hydroxyphenyllactic acid and 4‐hydroxyphenylpyruvic acid in serum and urine
T2 - JIMD Reports
AU - Hughes, Andrew T
AU - Shweihdi, Ella
AU - Gallagher, James
AU - Milan, Anna M.
AU - RANGANATH, L.
PY - 2022
DA - 2022/04/03 00:00:00
PB - Springer Nature
SP - 341-350
IS - 4
VL - 63
SN - 2192-8304
SN - 2192-8312
SN - 2192-8312
ER -
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@article{2022_Hughes,
author = {Andrew T Hughes and Ella Shweihdi and James Gallagher and Anna M. Milan and L. RANGANATH},
title = {Method development and validation for analysis of phenylalanine, 4‐hydroxyphenyllactic acid and 4‐hydroxyphenylpyruvic acid in serum and urine},
journal = {JIMD Reports},
year = {2022},
volume = {63},
publisher = {Springer Nature},
month = {apr},
url = {https://doi.org/10.1002/jmd2.12287},
number = {4},
pages = {341--350},
doi = {10.1002/jmd2.12287}
}
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Hughes, Andrew T., et al. “Method development and validation for analysis of phenylalanine, 4‐hydroxyphenyllactic acid and 4‐hydroxyphenylpyruvic acid in serum and urine.” JIMD Reports, vol. 63, no. 4, Apr. 2022, pp. 341-350. https://doi.org/10.1002/jmd2.12287.