Reaction of Bovine Pancreatic Ribonuclease A with 1,5-Difluoro-2,4-dinitrobenzene

Marfey, Peter S.; Nowak, H; Uziel, Mayo; Yphantis, David A.

doi:10.1016/s0021-9258(18)97213-3

Cited by 48 publications

(2 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To obtain UV spectra of the FDAA derivatives of amino acids, they were measured using photodiode array detection. A typical UV spectrum is shown for the derivative of l -valine in Figure c, together with the UV spectra of the derivative of N -methyl- l -valine (Figure a) and the derivatized reagent, FDAA (Figure b), and is characterized by absorption maxima at 340 and 414 nm, which are derived from the bridge chromophore between the nitro groups of the dinitrobenzene and the amino groups of the amino acid and l -alaminamide …”

Section: Resultsmentioning

confidence: 99%

A Nonempirical Method Using LC/MS for Determination of the Absolute Configuration of Constituent Amino Acids in a Peptide: Elucidation of Limitations of Marfey's Method and of Its Separation Mechanism

et al. 1997

View full text Add to dashboard Cite

As the first step in establishing our proposed method, the advanced Marfey's method, which is planned for the simultaneous determination of the absolute configuration of amino acids in a peptide, we applied Marfey's method to commercially available amino acids, and the separation behavior was examined in detail. Although good resolution of the diastereomeric pair of an individual amino acid was obtained for all amino acids tested and the applicability of the method was confirmed, the (1-fluoro-2,4-dinitrophenyl)-5-l-alaninamide (FDAA) derivative of the l-amino acid was not always eluted prior to its corresponding d-amino acid derivative. Because this proposed method relies on the elution order of a derivatized amino acid with FDAA to determine its absolute configuration, its separation mechanism was carefully investigated using UV and NMR spectral techniques. The results suggested that the resulting conformations of the l- and d-amino acid derivatives are stable and that the resolution between the l- and d-amino acid derivatives is due to the difference in their hydrophobicity, which is derived from the cis- or trans-type arrangement of two more hydrophobic substituents at both α-carbons of an amino acid and l-alanine amide, so that the FDAA derivative of the cis (Z)-type arrangement interacts more strongly with ODS silica gel and has a longer retention time than that of the trans (E)-type arrangement. Therefore, the l-amino acid derivative is usually eluted from the column before its corresponding d-amino acid derivative in Marfey's method. According to this separation mechanism, the elution order of a desired amino acid can be elucidated from the average retention time of the l- and d-amino acid derivatives, and the dl-serine and -asparagine derivatives are critical for Marfey's method.

show abstract

Section: Resultsmentioning

confidence: 99%

A Nonempirical Method Using LC/MS for Determination of the Absolute Configuration of Constituent Amino Acids in a Peptide: Elucidation of Limitations of Marfey's Method and of Its Separation Mechanism

et al. 1997

View full text Add to dashboard Cite

show abstract

“…The rapid loss of enzymatic activity upon reductive methylation is evidence that addition of even a single methyl group to the catalytically vital lysine number 41 (Hirs, 1962) so perturbs the catalytic site as to inactivate the enzyme. Marfey et al (1965) have shown, however, that cross-linking lysine number 41 to lysine number 7 with a bulky dinitrophenyl group results in the loss of only 85 % of the catalytic activity against cytidine 2',3'-cyclic monophosphate. This dif-ference in activity between the cross-linked and methylated enzyme may partly be due to the use of different substrates, a possibility which is presently under study.…”

Section: Discussionmentioning

confidence: 99%

Reductive alkylation of amino groups in proteins

Means¹,

Feeney²

1968

Biochemistry

671

278

View full text Add to dashboard Cite

Marfey's reagent: Past, present, and future uses of 1‐fluoro‐2,4‐dinitrophenyl‐5‐L‐alanine amide

B’Hymer¹,

Montes-Bayón²,

Caruso³

2003

J of Separation Science

View full text Add to dashboard Cite

This article describes some of the uses of Marfey's reagent, 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide (FDAA), a pre-column derivatizing reagent for the separation of enantiomeric isomers of amino acids and amine compounds. An introduction to the basic implementation of this reagent and its historical development are presented. Actual uses in amino acid, short peptide, and pharmaceutical compounds are included, as well as the advantages and disadvantages over other pre-column derivatization techniques and direct chromatographic separations. Applications of current interest, including its use as an orthogonal analysis and in enantiomeric purity analysis of selenoamino acids using element specific detection, are also discussed.

show abstract

Reaction of Bovine Pancreatic Ribonuclease A with 1,5-Difluoro-2,4-dinitrobenzene

Cited by 48 publications

References 8 publications

A Nonempirical Method Using LC/MS for Determination of the Absolute Configuration of Constituent Amino Acids in a Peptide: Elucidation of Limitations of Marfey's Method and of Its Separation Mechanism

A Nonempirical Method Using LC/MS for Determination of the Absolute Configuration of Constituent Amino Acids in a Peptide: Elucidation of Limitations of Marfey's Method and of Its Separation Mechanism

Reductive alkylation of amino groups in proteins

Marfey's reagent: Past, present, and future uses of 1‐fluoro‐2,4‐dinitrophenyl‐5‐L‐alanine amide

Contact Info

Product

Resources

About