[Update of a selected technique for uric acid determination in plasma and serum. Experimental study and value of derivative spectrophotometry].
A critical study of the candidate reference method for evaluation of uric acid in plasma proposed by the American Association of Clinical Chemistry is followed by testing in six laboratories. The dispersion of results is wide (CV greater than 5%). The importance of turbidity remaining after the deproteinization by trichloracetic acid is clearly demonstrated. This turbidity is really not reproducible from an operation to another one on the same serum. It is very likely responsible of the great dispersion of the results. After that, other deproteinization methods are tried. Ultrafiltration and ultracentrifugation give both defect errors because uric acid is in part bound to proteins. So it is necessary to find another technic which cancels the effects of turbidity on the absorbance readings in the ultra-violet domain. Experimental studies showed that uric acid may be evaluated with a good accuracy by derivative spectrophotometry, in lipid solutions as well as in cloudy ones. Turbidity was created by intralipid suspension additions. Various parameters hitting the method were examined (linearity, smoothing window...), taking as criterion the measure of overloaded serums at different levels. At last, the method is successfully transferred in several sites. In the case of blood serum, the respective influences of derivation and of repetition of final centrifugations are studied in order to estimate the effect of remaining turbidity. By the use of derivative spectrophotometry the improvement of the method of evaluation of uric acid proposed by A.A.C.C. is very noticeable; it reduces the variation coefficient between sites to less than 2%.
Hôpital J. Courmont, CHR Lyon-Sud, Pierre Benite.
P
P
Baltassat
D
D
Brault
M
M
Cambillau
J F
JF
Demelier
D
D
Duchassaing
P
P
Levillain
B
B
Mousson
F
F
Paolaggi
D
D
Porquet
P
P
Laruelle
D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods*; D005069:Evaluation Studies as Topic; D006801:Humans; D013053:Spectrophotometry / Q000379:methods*; D014527:Uric Acid / Q000097:blood*
D014527:Uric Acid
Mise au point d'une technique sélectionnée de dosage de l'acide urique dans le plasma et le sérum. Etude expérimentale et intérêt de la spectrophotométrie dérivée.