Plasma amino-acid determinations by reversed-phase HPLC: Improvement of the orthophthalaldehyde method and comparison with ion exchange chromatography
1992

Improving Amino Acid Testing with HPLC

Sample size: 27 publication Evidence: moderate

Author Information

Author(s): Frédéric Ziegler, Jacques Le Boucher, Colette Coudray-Lucas, Luc Cynober

Primary Institution: Laboratoire de Biochimie, UER des Sciences, Pharmaceutiques et Biologiques, Université Paris XI

Hypothesis

Is reversed-phase HPLC a better method for determining plasma amino acid concentrations compared to ion exchange chromatography?

Conclusion

Reversed-phase HPLC is a rapid and sensitive method for determining amino acids in plasma, outperforming traditional ion exchange chromatography in speed and sensitivity.

Supporting Evidence

  • RP-HPLC analysis was about three times faster than HP-IEC.
  • Sensitivity of RP-HPLC was 100-fold better than HP-IEC.
  • An excellent correlation (p < 0.003) was found between RP-HPLC and HP-IEC results.
  • Most amino acid concentrations were similar between the two methods.
  • RP-HPLC separated 24 amino acids, including the internal standard norvaline.

Takeaway

This study shows that a new way to test amino acids in blood is much faster and better at finding small amounts than the old method.

Methodology

The study compared reversed-phase HPLC with ion exchange chromatography for amino acid determination in plasma samples from healthy subjects.

Limitations

Some amino acids like aspartic acid, proline, and cysteine could not be identified by the RP-HPLC method.

Participant Demographics

27 healthy subjects (15 males, 12 females) aged 28 ± 5 years.

Statistical Information

P-Value

p<0.003

Statistical Significance

p<0.05

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