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Clinical Chemistry 43: 1421-1428, 1997;
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(Clinical Chemistry. 1997;43:1421-1428.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

Amino acid determination in biological fluids by automated ion-exchange chromatography: performance of Hitachi L-8500A

Jacques Le Boucher1,2,a, Christelle Charret2, Colette Coudray-Lucas1,2,3, Jacqueline Giboudeau3 and Luc Cynober1,2,3

1 INSERM U402,
2 GRENEMH,
3 Laboratoire de Biochimie A, CHU Saint-Antoine, 75571 Paris Cedex 12, France.
a Address correspondence to this author at: INSERM U402, CHU Saint-Antoine 27, rue de Chaligny, 75571 Paris Cedex 12, France. Fax +33 1.44.75.99.80.

The Hitachi L-8500A is a newly available apparatus for amino acid (AA) analysis that allows automatic on-line mixing of the ninhydrin reagent. The within-run precision (human plasma pools at three different concentrations) showed CVs <3.8% except for the lowest concentration of citrulline (4.4%), Tyr (4.5%), and {alpha}-aminobutyric acid (7.6%), and for the intermediate concentration of Asp (8.7%). Between-run precision (CV) was <3.1% for 17 AAs and <8.0% for 24 of 25 AAs (CV Asp = 12.0%). For retention times, within-run precision was <0.4% and between-run precision <1.8%. Excellent relations were found between the results from the Hitachi L-8500A and the widely used Beckman 6300 analyzer (0.929<= r <=0.999). The detection was still linear at 5 µmol/L except for Pro and hydroxyproline (20 µmol/L). The upper limit was at least 2500 µmol/L for 13 AAs and at least 1000 µmol/L for 27 of 29 AAs (anserine = 500, Val = 600 µmol/L). Values from 100 human plasma samples agreed with previously published data. We conclude that the results obtained with the Hitachi L-8500A are satisfactory when compared with those of other AA analyzers utilizing the same method. Furthermore, the Hitachi L-8500A displays several advantages including programming flexibility, microsample capacity, low noise plotting, ammonia filtering, and manual repacking of the analytical column.


Key Words: indexing terms: ninhydrin reaction • cation-exchange chromatography • analytical performance evaluation




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