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Related Experiment Videos

Amino acid analysis utilizing phenylisothiocyanate derivatives.

S A Cohen1, D J Strydom

  • 1Waters Chromatography Division of Millipore Corporation, Milford, Massachusetts 01757.

Analytical Biochemistry
|October 1, 1988
PubMed
Summary

New liquid chromatography techniques enable highly sensitive amino acid analysis using phenylisothiocyanate, achieving detection limits under 1 pmol for protein and complex samples. This offers a fast, accurate alternative to traditional methods.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry

Background:

  • Liquid chromatography (LC) advancements enable novel amino acid analysis techniques.
  • Precolumn derivatization is crucial for enhancing sensitivity and detection limits.

Purpose of the Study:

  • To introduce and evaluate new LC techniques for amino acid analysis.
  • To achieve low detection limits and fast analysis times for various sample types.

Main Methods:

  • Utilized precolumn derivatization with phenylisothiocyanate (PITC).
  • Employed advanced liquid chromatography for separation and detection.
  • Analyzed complex biological and food matrices.

Main Results:

  • Achieved routine detection limits below 1 picomole (pmol).

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  • Enabled analysis of submicrogram protein samples.
  • Demonstrated rapid analysis times: 10 min (post-hydrolysis) and 1 hour (physiologic samples).
  • Provided accurate and reproducible quantitation in plasma, urine, feed, and food.

    • Conclusions:

    • The developed method offers a sensitive and efficient alternative to traditional ion-exchange chromatography for amino acid analysis.
    • This technique provides flexibility without compromising accuracy or desirable features of established methods.