Rapid quantification of underivatized amino acids in plasma by hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass‐spectrometry

Prinsen, Hubertus C.M.T.; Schiebergen-Bronkhorst, B. G. M.; Roeleveld, Martin W.; Jans, Judith; Velden, Monique G.M. de Sain–van der; Visser, Gepke; Hasselt, Peter M. van; Verhoeven‐Duif, Nanda M.

doi:10.1007/s10545-016-9935-z

Cited by 139 publications

(87 citation statements)

References 22 publications

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“…The derivatives could be detected by HPLC-ICP-MS, but the LODs (12-128 fmol/injection) and precision for the Orbitrap MS method are higher than those of the HPLC-ICP-MS method (440-1100 fmol/injection) [17]. Thirty-six amino acids, including Hcy, Met, Hcy-Cys disulfide, Cys, Met-sulfone, Met-sulfoxide, HcyHcy, and CysCys, were determined after post-column derivatization with ninhydrin [76]. The derivatized amino acids were separated on a hydrophilic interaction liquid chromatography column with an analysis time of 18 min and LODs of 0.1 μmol/L.…”

Section: Derivatization Methodsmentioning

confidence: 99%

HPLC Analysis of Homocysteine and Related Compounds

Wada¹,

Nakamura²,

Nakashima³

2018

Non-Proteinogenic Amino Acids

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Homocysteine (Hcy), a sulfur-containing amino acid, is a representative intermediate metabolite of methionine (Met) to cysteine (Cys) via several intermediates. An elevated level of Hcy in plasma plays an important role in diseases such as neural tube defects and Down syndrome. Homocystinuria is the most common inborn error of sulfur metabolism and is caused by mutations in the metabolic enzymes of Hcy. These errors can be caused by abnormal levels of Met metabolites and classified on the basis of plasma Met levels. Additionally, Hcy and related compounds such as glutathione play an important role in maintaining homeostasis. Therefore, the simultaneous determination of Hcy and/or related compounds is required for appropriate clinical management of several diseases. The sulfur-containing amino acids and their derivatives in biological samples are quantified sensitively using high-performance liquid chromatography methods coupled with various detection methods such as UV/Vis, fluorescence, chemiluminescence, electrochemical, mass spectrometry, and tandem mass spectrometry. In this chapter, we review recent advances in these analytical methods and their applications.

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Section: Derivatization Methodsmentioning

confidence: 99%

HPLC Analysis of Homocysteine and Related Compounds

Wada¹,

Nakamura²,

Nakashima³

2018

Non-Proteinogenic Amino Acids

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“…Cells were harvested at T = 0.5, 4 and 12 h. Uniformly labelled 13 C 6 -glucose (99%) was purchased from Cambridge Isotope Laboratories, Inc. (MA, USA). To quantify the intracellular 13 C 3 -serine and 13 C 2 -glycine, we adapted the LC-MS/MS method described by Prinsen et al (2016).…”

Section: Metabolic Flux Analysis: Serine De Novo Biosynthesismentioning

confidence: 99%

“…Amino acid concentrations were determined using the UPLC-MS/MS method described by Prinsen et al (2016). Apart from adapting the range of the calibrators and quality control (QC) samples to resemble the concentrations in the samples, no further adaptations were needed for sample preparation or analysis of the amino acids.…”

Section: Amino Acidsmentioning

confidence: 99%

Vitamin B6 is essential for serine de novo biosynthesis

Ramos

Pras‐Raves

Gerrits

et al. 2017

J of Inher Metab Disea

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Pyridoxal 5′-phosphate (PLP), the metabolically active form of vitamin B6, plays an essential role in brain metabolism as a cofactor in numerous enzyme reactions. PLP deficiency in brain, either genetic or acquired, results in severe drug-resistant seizures that respond to vitamin B6 supplementation. The pathogenesis of vitamin B6 deficiency is largely unknown. To shed more light on the metabolic consequences of vitamin B6 deficiency in brain, we performed untargeted metabolomics in vitamin B6-deprived Neuro-2a cells. Significant alterations were observed in a range of metabolites. The most surprising observation was a decrease of serine and glycine, two amino acids that are known to be elevated in the plasma of vitamin B6 deficient patients. To investigate the cause of the low concentrations of serine and glycine, a metabolic flux analysis on serine biosynthesis was performed. The metabolic flux results showed that the de novo synthesis of serine was significantly reduced in vitamin B6-deprived cells. In addition, formation of glycine and 5-methyltetrahydrofolate was decreased. Thus, vitamin B6 is essential for serine de novo biosynthesis in neuronal cells, and serine de novo synthesis is critical to maintain intracellular serine and glycine. These findings suggest that serine and glycine concentrations in brain may be deficient in patients with vitamin B6 responsive epilepsy. The low intracellular 5-mTHF concentrations observed in vitro may explain the favourable but so far unexplained response of some patients with pyridoxine-dependent epilepsy to folinic acid supplementation.

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“…Directly after collection, blood samples were centrifuged during 10 minutes at 1200 g and a temperature of 4°C, and plasma was transferred to 1 mL microvial tubes and stored at -80°C until laboratory analyses. Plasma tyrosine concentrations were quantified by hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass-spectrometry as described by Prinsen and colleagues [33].…”

Section: Plasma Tyrosine Concentrationsmentioning

confidence: 99%

Dose-Dependent Effects of Oral Tyrosine Administration on Plasma Tyrosine Levels and Cognition in Aging

Rest¹,

Bloemendaal²,

Heus³

et al. 2017

Preprint

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Abstract:The effects of tyrosine on plasma response and cognition in aging are unknown. We assessed the dose-dependent response to tyrosine administration in older adults in both plasma tyrosine concentrations and working memory performance. In this double blind randomized cross-over trial 17 older adults (aged 60-75 years) received a single administration of 100, 150 or 200 mg/kg body weight of tyrosine. For comparison, 17 young adults (aged 18-35 years) received a dose of 150 mg/kg body weight of tyrosine. Tyrosine plasma concentrations were determined before and 90, 120, 150, 180, 210 and 240 minutes after tyrosine intake. Working memory was assessed using the N-back task at 90 minutes after tyrosine administration. Older adults showed a dose-dependent increase in plasma tyrosine concentrations (p<.001), and the plasma response was higher than for young adults with the same dose (p<.001). Load-dependent working memory performance decreased with higher doses of tyrosine (p=.048), especially in older adults with greater dose-dependent plasma tyrosine responses (p=.035). Our results show an age-related increase in plasma tyrosine response, which was associated with a dose-dependent decline in cognitive functioning in older adults.

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Rapid quantification of underivatized amino acids in plasma by hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass‐spectrometry

Cited by 139 publications

References 22 publications

HPLC Analysis of Homocysteine and Related Compounds

HPLC Analysis of Homocysteine and Related Compounds

Vitamin B6 is essential for serine de novo biosynthesis

Dose-Dependent Effects of Oral Tyrosine Administration on Plasma Tyrosine Levels and Cognition in Aging

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