LC-MS/MS-Based Separation and Quantification of Marfey’s Reagent Derivatized Proteinogenic Amino Acid <scp>dl</scp>-Stereoisomers

Ayon, Navid J.; Sharma, Amar Deep; Gutheil, William G.

doi:10.1007/s13361-018-2093-9

Cited by 53 publications

(59 citation statements)

References 40 publications

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“…L-Arg and Gly, present in the SCH peptaibols, were included among the derivatized standards for comparison but were absent from the samples, further confirming the differences among these compounds. It should be noted that relatively low absorbance for Aib and putrescine derivatives evident in chromatograms ( Figure S18 ) are likely due to slower derivatization reaction rates required for complete reaction with Marfey’s reagent [ 30 , 31 ].…”

Section: Resultsmentioning

confidence: 99%

Sphaerostilbellins, New Antimicrobial Aminolipopeptide Peptaibiotics from Sphaerostilbella toxica

et al. 2020

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Sphaerostilbella toxica is a mycoparasitic fungus that can be found parasitizing wood-decay basidiomycetes in the southern USA. Organic solvent extracts of fermented strains of S. toxica exhibited potent antimicrobial activity, including potent growth inhibition of human pathogenic yeasts Candida albicans and Cryptococcus neoformans, the respiratory pathogenic fungus Aspergillus fumigatus, and the Gram-positive bacterium Staphylococcus aureus. Bioassay-guided separations led to the purification and structure elucidation of new peptaibiotics designated as sphaerostilbellins A and B. Their structures were established mainly by analysis of NMR and HRMS data, verification of amino acid composition by Marfey’s method, and by comparison with published data of known compounds. They incorporate intriguing structural features, including an N-terminal 2-methyl-3-oxo-tetradecanoyl (MOTDA) residue and a C-terminal putrescine residue. The minimal inhibitory concentrations for sphaerostilbellins A and B were measured as 2 μM each for C. neoformans, 1 μM each for A. fumigatus, and 4 and 2 μM, respectively, for C. albicans. Murine macrophage cells were unaffected at these concentrations.

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Section: Resultsmentioning

confidence: 99%

Sphaerostilbellins, New Antimicrobial Aminolipopeptide Peptaibiotics from Sphaerostilbella toxica

et al. 2020

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“…In 1984, Marfey developed the N α -(5-Fluoro-2,4-dinitrophenyl)- l -alaninamide ( l -FDAA, also named Marfey’s reagent) enabling the indirect chiral resolution of amino acid enantiomers and successfully separated the diastereomeric l - and d -amino acids by HPLC [ 26 ]. This Marfey’s reagent is also compatible with LC/MS analysis [ 27 ]. Fujii et al reported an advanced Marfey’s reagent, N α -(5-Fluoro-2,4-dinitrophenyl)- l -leucinamide ( l -FDLA) [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Measurement of Amino Acid Enantiomers in Aged Mouse Brain Samples by LC/MS/MS Combined with Derivatization Using Nα-(5-Fluoro-2,4-dinitrophenyl)-l-leucinamide (l-FDLA)

2021

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d-amino acids have distinct roles from their l-enantiomer. In particular, some d-amino acids function as agonists or antagonists of neuronal receptors and are involved in higher brain functions. Thus, it is important to precisely measure the levels of these amino acid enantiomers in cells and tissues. Various quantification methods have been developed for measurements of chiral amino acids. However, each method has advantages and disadvantages. Additionally, measuring the amino acid enantiomers in crude biological samples requires a higher selectivity. In this study, we developed a quantification method for amino acid enantiomers using derivatization with Nα-(5-Fluoro-2,4-dinitrophenyl)-l-leucinamide (l-FDLA) followed by liquid chromatography–tandem mass spectrometry (LC/MS/MS) with a conventional reversed-phase column. We simultaneously identified 10 chiral amino acids. Furthermore, we applied this method to investigate murine tissue samples and examined the effect of aging on the amino acid levels in aged brain regions. We found that aging decreased the levels of both d-serine and d-aspartate in the hippocampus. In addition, d-Phenylalanine in the thalamus significantly increased with age. In conclusion, our method is suitable for the quantification of the d-amino acids in crude biological samples and may contribute to elucidating the biological roles of chiral amino acids.

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“…In the case of indirect approaches, with GC-MS, silylation agent such as N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) [22] or perfluorinated anhydrides [13] were used to obtain volatile derivatives prior to their separation in a CSP and (+)-α-methoxy-α-trifluoromethyl-phenylacetyl chloride to obtain diastereomeric amides [23]. In LC, it is common to use (R)-1-boc-2-piperidine carbonyl chloride [24]; ortho-phthalaldehyde (OPA)/ N-isobutyryl-L-cysteine (IBLC) reagent [25]; (R)-1-phenylethyl isothiocyanate ((R)-AMBI) [15], 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide (Marfey´s reagent) [26,27]; 2,3,4,6-tetra-O-acetyl-beta-d-glucopyranosyl isothiocyanate (GITC); (S)-NIFE; 4-(3isothiocyanatopyrrolidin-1-yl)-7-nitrobenzofurazan (NBD-PyNCS) etc. [15,21] or homemade reactant as Biaryl Axially chiral derivatizing agent [28].…”

Section: Introductionmentioning

confidence: 99%

Development and validation of a high performance liquid chromatography-tandem mass spectrometry method for the absolute analysis of 17 α D-amino acids in cooked meals

Barbas-Bernardos

García-Pérez

Lorenzo

et al. 2020

Journal of Chromatography A

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In the nutrition field, there is a lack of understanding about the impact that dietary chiral composition may have on health, especially regarding cooked meals. Chiral amino acids (AAs) are naturally present in food and their proportion may vary quite a lot. Besides, the D-amino acids (D-AAs) are present in very low concentration compared to L-AAs, so very sensitive methods are required for their accurate quantitation. Moreover, some of them have been described as indicators of quality and different food processes. In this research, we propose a robust method for the absolute quantitation and enantiomeric ratio of 17 D-AAs in cooked meals. The AAs were extracted from 1 g of the homogenised meal with methanol, derivatised with (S)-N-(4-nitrophenoxycarbonyl) phenylalanine methoxyethyl ester ((S)-NIFE) and analysed by RP-LC-MS/MS. The separation was carried out with an Acquity BEH C18 (100 mm x 2.1 mm, 1.7 µm) column at 70º C, with 10 mmol/L ammonium bicarbonate in water as eluent A and acetonitrile as eluent B at a 0.3 mL/min flow rate in gradient elution. The MS operated in positive electrospray ionisation method in multiple reaction monitoring (MRM) mode. Isotopically labelled AAs were used as internal standards for the quantitation. The method was validated for 17 D-AAs in the cooked food samples in terms of specificity, linearity, precision, accuracy, matrix effect and stability. LLOQ are 2.0 ng/mL for most of them. Additionally, linearity was also studied for L-AAs. After optimization and validation, the method was applied to real breakfast, lunch and dinner samples of cooked meals (n=18) that were part of a diet with a very high concordance with WHO dietary guidelines. Level of concentration of major and minor D-AAs have been described per total daily intake and within each of the three main meals. This method can be used for quality control purposes as well as to investigate the role of chiral composition in food and clinical outcomes.

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LC-MS/MS-Based Separation and Quantification of Marfey’s Reagent Derivatized Proteinogenic Amino Acid dl-Stereoisomers

Cited by 53 publications

References 40 publications

Sphaerostilbellins, New Antimicrobial Aminolipopeptide Peptaibiotics from Sphaerostilbella toxica

Sphaerostilbellins, New Antimicrobial Aminolipopeptide Peptaibiotics from Sphaerostilbella toxica

Simultaneous Measurement of Amino Acid Enantiomers in Aged Mouse Brain Samples by LC/MS/MS Combined with Derivatization Using Nα-(5-Fluoro-2,4-dinitrophenyl)-l-leucinamide (l-FDLA)

Development and validation of a high performance liquid chromatography-tandem mass spectrometry method for the absolute analysis of 17 α D-amino acids in cooked meals

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