Mass spectrometry signal enhancement by reductive amination

Liu, Meng-Chieh; Lin, Yi-Reng; Huang, Mei-Fang; Tsai, De-Cheng; Liang, Shih‐Shin

doi:10.1016/j.ijms.2015.06.010

Cited by 6 publications

(5 citation statements)

References 23 publications

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“…Dimethyl-labeling (DM) is frequently used as a labeling strategy, − while the most prominent application is the multiplex quantification of proteins . In a different approach the reductive methylation of amines was performed prior to modification of carboxyl groups to analyze proteins’ carboxy termini …”

Section: Introductionmentioning

confidence: 99%

ArgC-Like Digestion: Complementary or Alternative to Tryptic Digestion?

et al. 2017

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Enzymatic digestion of complex protein samples is often performed by use of multiple proteases to improve protein identification and characterization. Combining trypsin with ArgC is one option to enhance sequence coverage in bottom-up proteomics. However, the low selectivity of this endoprotease derogates from the benefit of the combination. Our approach here is to mimic ArgC digestion by chemically modifying all lysine residues in proteins so that trypsin can only cleave C-terminal to arginine. Four different amine modifications, dimethylation, acetylation, propionylation, and carbethoxylation, were tested, and the protocols were optimized. A nearly complete conversion of the primary amines was achieved for all modifications. Tryptic digestion of Escherichia coli lysate proteins after acylation of lysine residues shows the most significant improvement compared with data received from ArgC digest. After propionylation, 9216 unique peptides identified 1439 proteins, which, compared with a conventional tryptic digestion, represents the identification of 150 additional proteins due to a reasonable reduction of the sample complexity and higher fragmentation efficiencies of the peptides. It is therefore concluded that the Arg-C like digestion should no longer be regarded as a complementary approach but forms a viable and superior alternative to the conventional trypsin digestion.

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Section: Introductionmentioning

confidence: 99%

ArgC-Like Digestion: Complementary or Alternative to Tryptic Digestion?

et al. 2017

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“…The integrated and statistical results, including the average integrated area, relative standard deviation, and signal enhancement, are shown in Table 2 . We have previously discussed the factor that enhances signals via reductive amination [ 40 ]. We estimated that the factor is the amine group showing a higher electronic density after modification with two methyl groups, causing the lone pair in the amine group to show a more basic quality.…”

Section: Resultsmentioning

confidence: 99%

“…Herein, we used isotopic formaldehyde and sodium cyanoborohydride to generate two methyl groups on the amine functional group of TRP, 5-HTP, and serotonin. This modification, with cheap and easily obtained materials, showed signal enhancement and helped target molecules to be monitored [ 40 ]. Eventually, we establish the derivatized method by estimating calibration curves, the limit of detection (LOD), the limit of quantification (LOQ), intra-day precision, and inter-day precision.…”

Section: Introductionmentioning

confidence: 99%

Assisted Reductive Amination for Quantitation of Tryptophan, 5-Hydroxytryptophan, and Serotonin by Ultraperformance Liquid Chromatography Coupled with Tandem Mass Spectrometry

et al. 2023

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Herein, we used isotopic formaldehyde and sodium cyanoborohydride via reductive amination to label two methyl groups on primary amine to arrange the standards (h2-formaldehyde-modified) and internal standards (ISs, d2-formaldehyde-modified) of tryptophan and its metabolites, such as serotonin (5-hydroxytryptamine) and 5-hydroxytryptophan. These derivatized reactions with a high yield are very satisfactory for manufacturing standards and ISs. This strategy will generate one or two methyl groups on amine to create different mass unit shifts with 14 vs. 16 or 28 vs. 32 in individual compounds for biomolecules with amine groups. In other words, multiples of two mass units shift are created using this derivatized method with isotopic formaldehyde. Serotonin, 5-hydroxytryptophan, and tryptophan were used as examples to demonstrate isotopic formaldehyde-generating standards and ISs. h2-formaldehyde-modified serotonin, 5-hydroxytryptophan, and tryptophan are standards to construct calibration curves, and d2-formaldehyde-modified analogs such as ISs spike into samples to normalize the signal of each detection. We utilized multiple reaction monitoring modes and triple quadrupole mass spectrometry to demonstrate the derivatized method suitable for these three nervous biomolecules. The derivatized method demonstrated a linearity range of the coefficient of determinations between 0.9938 to 0.9969. The limits of detection and quantification ranged from 1.39 to 15.36 ng/mL.

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“…The internal standards were used to normalize PHE and TYR signals, [18][19][20][21][22][23] and formaldehydemodified PHE and TYR were detected by mass spectrometry signal enhancement. 24 We compared the concentrations of plasma on filter paper and crude plasma samples and used HPLC-MS/MS to monitor derivatized PHE and TYR in the plasma samples. The results showed no obvious differences.…”

Section: Introductionmentioning

confidence: 99%

Quantification of derivatized phenylalanine and tyrosine in dried blood spots using liquid chromatography with tandem spectrometry for newborn screening of phenylketonuria

Duh,

Liang,

Shen

et al. 2024

Eur J Mass Spectrom (Chichester)

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Phenylketonuria (PKU) is an autosomal genetic disorder caused by a deficiency of the phenylalanine hydroxylase (PAH) enzyme. The lack of PAH results in the inability of phenylalanine (PHE) to transform into tyrosine (TYR). Consequently, this leads to the accumulation of PHE in the blood samples of newborns causing metabolic diseases such as irreversible neurological problems. An analysis was required for determining the values of PHE and TYR in blood samples from newborn babies. In this study, therefore, we developed a derivatized method to monitor PHE and TYR in plasma samples using liquid phase chromatography linked with quadrupole mass spectrometry. Accessible formaldehyde isotopes and cyanoborohydride were used to react with PHE and TYR amino groups to generate h2-formaldehyde-modified PHE and TYR (as standards) and d2-formaldehyde-modified PHE and TYR (as internal standards). We used tandem mass spectrometry for multiple reaction monitoring. We demonstrated a derivatized method suitable for the PKU screening of newborns. The recoveries for PHE and TYR were 85% and 90%, respectively. Furthermore, we compared the values of PHE and TYR in different human plasma sample storage methods, including direct plasma and dried blood spots, and the results showed no significant difference.

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Mass spectrometry signal enhancement by reductive amination

Cited by 6 publications

References 23 publications

ArgC-Like Digestion: Complementary or Alternative to Tryptic Digestion?

ArgC-Like Digestion: Complementary or Alternative to Tryptic Digestion?

Assisted Reductive Amination for Quantitation of Tryptophan, 5-Hydroxytryptophan, and Serotonin by Ultraperformance Liquid Chromatography Coupled with Tandem Mass Spectrometry

Quantification of derivatized phenylalanine and tyrosine in dried blood spots using liquid chromatography with tandem spectrometry for newborn screening of phenylketonuria

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