Amino acid determination in biological fluids by automated ion-exchange chromatography: performance of Hitachi L-8500A.
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 < or = r < or = 0.999). The detection was still linear at 5 mumol/L except for Pro and hydroxyproline (20 mumol/L). The upper limit was at least 2500 mumol/L for 13 AAs and at least 1000 mumol/L for 27 of 29 AAs (anserine = 500, Val = 600 mumol/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.
Le Boucher
J
J
INSERM U402, CHU Saint-Antoine, Paris, France.
Charret
C
C
Coudray-Lucas
C
C
Giboudeau
J
J
Cynober
L
L
D000596:Amino Acids / Q000032:analysis* / Q000097:blood; D001331:Automation; D001826:Body Fluids / Q000737:chemistry*; D002021:Buffers; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods*; D005069:Evaluation Studies as Topic; D006801:Humans; D006909:Hydroxyproline / Q000032:analysis / Q000097:blood; D009555:Ninhydrin; D011392:Proline / Q000032:analysis / Q000097:blood; D012044:Regression Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000596:Amino Acids; D002021:Buffers; D011392:Proline; D009555:Ninhydrin; D006909:Hydroxyproline