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Home / Applications / High Speed Analysis of Dabsyl Amino Acids in Stout Beer using UHPLC

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  • Technique

High Speed Analysis of Dabsyl Amino Acids in Stout Beer using UHPLC

By Jiawei Xu

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January 5, 2024

Introduction

There are a wide variety of HPLC methods that can be used for amino acid analysis, two examples are a separation by reversed-phase column with precolumn derivatization and UV-Visible detection, and a post-column derivatization with separation by ion-exchange column and fluorescence detection. The precolumn derivatization method using Dabsyl chloride as reagent is commonly used as Dabsyl chloride reacts easily with both  primary and secondary amino acids, and the derivatized amino acids are stable, for easy handling, good accuracy and high sensitivity. In order to enable an easier derivatization method, a DAB-Label kit is available from JASCO.

Here, the amino acids in Stout beer were measured using DAB-Label derivatization kit by Ultra High-performance Liquid Chromatography (UHPLC) with UV-Visible detection.

Jasco XLC-3000

Experimental

Figure 1 shows the dabsylation procedure and in figure 2, the reaction mechanism is illustrated.

  1. Dilute the sample by dabsylation buffer*.
  2. Take 20 μL.
  3. Add 40 μL of dabsylation reagent and agitate.
  4. Warm at 70ºC for 12 min.(During warming agitate several times.)
  5. After cooling, add 440 μL of dilution buffer and agitate.
    * Included in DAB-Label kit
Figure 2. Reaction mechanism of dabsylation

Keywords

430023X

Results

Figure 3 shows the chromatogram of 21 components of a dabsylation amino acid standard mixture. 21 components including gamma-aminobutyric acid(GABA) – suppressive neurotransmitter of central nerve, taurine, ornithine, etc. were separated within 7.5 minutes.

Figure 3. Chromatogram of 21 components of a dabsylated amino acid standard mixture. 1: Aspartic acid, 2: Glutamic acid, 3: Serine, 4: Threonine, 5: Arginine, 6: Glycine, 7: Alanine, 8: Proline, 9: Taurine, 10: Valine, 11: GABA (c-aminobutyric acid), 12: Methionine, 13: Isoleucine, 14: Leucine, 15: Phenylalanine, 16: Cystine, 17: Hydroxylysine, 18: Ornithine, 19: Lysine, 20: Histidine, 21: Tyrosine

Figure 4 shows the chromatogram of dabsylated amino acids in Stout beer.

Figure 4. Chromatogram of dabsylated amino acids in Stout beer.
The peak numbers are the same as in figure 3. Preparation: Stout beer was diluted by 50-fold using dabsylation buffer and dabsyated according to procedure shown in figure 1.
This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.

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About the Author

Jiawei Xu

JASCO Application Note

High Speed Analysis of Dabsyl Amino Acids in Stout Beer using UHPLC

Introduction

There are a wide variety of HPLC methods that can be used for amino acid analysis, two examples are a separation by reversed-phase column with precolumn derivatization and UV-Visible detection, and a post-column derivatization with separation by ion-exchange column and fluorescence detection. The precolumn derivatization method using Dabsyl chloride as reagent is commonly used as Dabsyl chloride reacts easily with both  primary and secondary amino acids, and the derivatized amino acids are stable, for easy handling, good accuracy and high sensitivity. In order to enable an easier derivatization method, a DAB-Label kit is available from JASCO.

Here, the amino acids in Stout beer were measured using DAB-Label derivatization kit by Ultra High-performance Liquid Chromatography (UHPLC) with UV-Visible detection.

Jasco XLC-3000

Experimental

Figure 1 shows the dabsylation procedure and in figure 2, the reaction mechanism is illustrated.

  1. Dilute the sample by dabsylation buffer*.
  2. Take 20 μL.
  3. Add 40 μL of dabsylation reagent and agitate.
  4. Warm at 70ºC for 12 min.(During warming agitate several times.)
  5. After cooling, add 440 μL of dilution buffer and agitate.
    * Included in DAB-Label kit
Figure 2. Reaction mechanism of dabsylation

Results

Figure 3 shows the chromatogram of 21 components of a dabsylation amino acid standard mixture. 21 components including gamma-aminobutyric acid(GABA) – suppressive neurotransmitter of central nerve, taurine, ornithine, etc. were separated within 7.5 minutes.

Figure 3. Chromatogram of 21 components of a dabsylated amino acid standard mixture. 1: Aspartic acid, 2: Glutamic acid, 3: Serine, 4: Threonine, 5: Arginine, 6: Glycine, 7: Alanine, 8: Proline, 9: Taurine, 10: Valine, 11: GABA (c-aminobutyric acid), 12: Methionine, 13: Isoleucine, 14: Leucine, 15: Phenylalanine, 16: Cystine, 17: Hydroxylysine, 18: Ornithine, 19: Lysine, 20: Histidine, 21: Tyrosine

Figure 4 shows the chromatogram of dabsylated amino acids in Stout beer.

Figure 4. Chromatogram of dabsylated amino acids in Stout beer.
The peak numbers are the same as in figure 3. Preparation: Stout beer was diluted by 50-fold using dabsylation buffer and dabsyated according to procedure shown in figure 1.

Keywords

430023X

This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.
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