Separation and determination of cysteine enantiomers in plasma after derivatization with 4-fluoro-7-nitrobenzofurazan

Ferré, Sabrina; González‐Ruiz, Víctor; Zangari, Joséphine; Girel, Sergey; Martinou, Jean‐Claude; Sardella, Roccaldo; Rudaz, Serge

doi:10.1016/j.jpba.2021.114539

Cited by 12 publications

(8 citation statements)

References 32 publications

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“…Elution of cysteine from biological samples is critical but it is extremely important as a potential biomarker, scientist has eluted it through UHPLC‐MS [25]. Moreover, recently cysteine has also been separated from plasma by derivatization using 4‐fluoro‐7‐nitrobenzofurazan [26]. Similar to the current study, through a simple HPLC assay, cysteine was eluted with great accuracy.…”

Section: Discussionmentioning

confidence: 64%

High‐performance liquid chromatography‐based assay optimization for the detection of plasma amino acids for applications in metabolic disorders in developing countries

Wasim,

Khan,

Tawab

et al. 2023

Separation Science Plus

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Plasma amino acids are generally analyzed through ion exchange chromatography, a reproducible but time‐consuming method. Here, we report the optimization of a reverse‐phase‐high‐performance liquid chromatography with fluorescence detector (RP‐HPLC‐FLD) assay for the detection and quantification of plasma amino acids for potential applications in metabolic disorders (e.g., aminoacidopathies, a rare group of Inborn Errors of Metabolism). For assay development, initially standard amino acids were derivatized with ortho‐phthalaldehyde‐3‐mercaptopropionic acid (OPA‐3‐MPA) and filtered through a 0.20 μm syringe filter. The excitation and emission wavelengths of 240–450 nm (λex—λem) were used for the detection of amino acids. Chromatographic separation was achieved by gradient RP‐HPLC‐FLD through C18 symmetry column (150 × 4.6 mm, particle size 3.5 μm). HPLC assay was successfully optimized and was able to detect amino acids in the range of 10–400 ng/mL and good linearity (R2 > 0.98) was achieved in the mixture for each standard amino acid. Moreover, the current assay showed great efficiency with two additional advantages: the use of low‐cost mobile phases, and the detection and quantification of amino acids at low level (ng/mL) concentration in biofluids. This assay could be applied for the analysis of human plasma to identify aminoacidopathies in newborn screening programs, and other metabolic disorders.

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Section: Discussionmentioning

confidence: 64%

High‐performance liquid chromatography‐based assay optimization for the detection of plasma amino acids for applications in metabolic disorders in developing countries

Wasim,

Khan,

Tawab

et al. 2023

Separation Science Plus

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“…2 Recent improvements in chromatographic detection methods have improved the separation of these cysteine enantiomers. 16,17 Using a stereospecific bioluminescent luciferase assay, chiral chromatography, and SR À/À mice (serine racemase knockout), we identified endogenous D-Cysteine in mammalian brain and pancreas. 2,7 Our work shows that endogenous D-Cysteine is racemized from Lcysteine by SR. Endogenous D-Cysteine is enriched in the embryonic mouse brain (E9.5) at a concentration of 4.5 mM and decreases significantly with development to approximately 50 μM (Figure 1).…”

Section: D-cysteinementioning

confidence: 99%

“…Analysis and determination of cysteine enantiomers is challenging due to the high reactivity of the SH (thiol) group which is readily oxidized leading to the formation of cystine 2 . Recent improvements in chromatographic detection methods have improved the separation of these cysteine enantiomers 16,17 . Using a stereospecific bioluminescent luciferase assay, chiral chromatography, and SR −/− mice (serine racemase knockout), we identified endogenous D‐Cysteine in mammalian brain and pancreas 2,7 .…”

Section: D‐amino Acids As Novel Neurotransmitters?mentioning

confidence: 99%

Mammalian D‐Cysteine: A new addition to the growing family of biologically relevant D‐amino acids

Roychaudhuri

2023

Chirality

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Mammalian D‐Cysteine is racemized from L‐cysteine by serine racemase, a pyridoxal phosphate (PLP)‐dependent enzyme. Endogenous D‐Cysteine plays a role in neural development by inhibiting proliferation of neural progenitor cells (NPCs) via protein kinase B (AKT) signaling mediated by the FoxO family of transcription factors. D‐Cysteine binds to Myristoylated Alanine Rich C Kinase Substrate (MARCKS) and alters phosphorylation at Ser 159/163 and its translocation from the membrane. By racemizing serine and cysteine, mammalian serine racemase may play important roles in neural development highlighting its importance in psychiatric disorders.

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“…With a teicoplanin core–shell column, fast enantiomer separations in time scales <1 min have been reported for proteinogenic and nonproteinogenic amino acids using LC-UV and injections of single amino acids, yet the simultaneous analysis of all amino acids in a single run has not been shown so far and poses additional challenges even with mass spectrometric (MS) detection . Achiral derivatization has been used to improve the enantioselectivity for CSPs, introduce a strong fluorophore or chromophore for spectroscopic detection, or a moiety with improved ionization efficiency for MS detection and (ideally) characteristic fragment ions for tandem MS (MS/MS) experiments. − The resultant LC separations of complex derivatized amino acid mixtures have typically been on the 5–30 min time scale. To overcome limited chemoselectivity for separation of the challenging suite of Leu and Thr isomers, two-dimensional liquid chromatography (2D-LC) with achiral reversed-phase liquid chromatography (RP-LC) in the first dimension and a chiral column in the second dimension has been suggested but extends the total analysis time (typically >60 min). ,,, Very recently, a three-dimensional liquid chromatography (3D-LC) approach for enantioselective analysis of aliphatic amino acids in urine with RP in the first dimension, a mixed-mode column having remarkable selectivity for the structural isomers of Leu (Leu/Ile/ a Ile) in the second dimension, and an enantioselective column in the third dimension was suggested …”

Section: Introductionmentioning

confidence: 99%

Three-Minute Enantioselective Amino Acid Analysis by Ultra-High-Performance Liquid Chromatography Drift Tube Ion Mobility-Mass Spectrometry Using a Chiral Core–Shell Tandem Column Approach

Jaag,

Valadbeigi,

Causon

et al. 2024

Anal. Chem.

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Fast liquid chromatography (LC) amino acid enantiomer separation of 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) derivatives using a chiral core−shell particle tandem column with weak anion exchange and zwitterionic-type quinine carbamate selectors in less than 3 min was achieved. Enantiomers of all AQC-derivatized proteinogenic amino acids and some isomeric ones (24 in total plus achiral glycine) were baseline separated (R s > 1.5 except for glutamic acid with R s = 1.3), while peaks of distinct amino acids and structural isomers (constitutional isomers and diastereomers of leucine and threonine) of the same configuration overlapped to various degrees. For this reason, drift tube ion mobility-mass spectrometry was added (i.e., LC-IM-MS) as an additional selectivity filter without extending run time. The IM separation dimension in combination with high-resolution demultiplexing enabled confirmation of threonine isomers (threonine, allo-threonine, homoserine), while leucine, isoleucine, and allo-isoleucine have almost identical collisional cross-section ( DT CCS N2 ) values and added no selectivity to the partial LC separation. Density functional theory (DFT) calculations show that IM separation of threonine isomers was possible due to conformational stabilization by hydrogen bond formation between the hydroxyl side chain and the urea group. Generally, the CCS N2 of protonated ions increased uniformly with addition of the AQC label, while outliers could be explained by consideration of intramolecular interactions and additional structural analysis. Preliminary validation of the enantioselective LC-IM-MS method for quantitative analysis showed compliance of accuracy and precision with common limits in bioanalytical methods, and applicability to a natural lipopeptide and a therapeutic synthetic peptide could be demonstrated.

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Separation and determination of cysteine enantiomers in plasma after derivatization with 4-fluoro-7-nitrobenzofurazan

Cited by 12 publications

References 32 publications

High‐performance liquid chromatography‐based assay optimization for the detection of plasma amino acids for applications in metabolic disorders in developing countries

High‐performance liquid chromatography‐based assay optimization for the detection of plasma amino acids for applications in metabolic disorders in developing countries

Mammalian D‐Cysteine: A new addition to the growing family of biologically relevant D‐amino acids

Three-Minute Enantioselective Amino Acid Analysis by Ultra-High-Performance Liquid Chromatography Drift Tube Ion Mobility-Mass Spectrometry Using a Chiral Core–Shell Tandem Column Approach

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