Determination of "true" serum creatinine by high-performance liquid chromatography combined with a continuous-flow...

We coupled a high-performance liquid chromatograph with a continuous-flow microanalyzer to produce a system for specifically determinig "true" creatinine in urine and serum specimens. A selective microbore pellicular cation-exchange column and a single eluting sodium citrate buffer are used to separate the noncreatinine but Jaffé-positive constituents from the creatinine in normal and experimental specimens. The effluent is analyzed continuously, on-line, the alkaline picrate complex being developed and measured in the microanalyzer. Physiological samples, reference standards, and internal control specimens are assayed in 6-min intervals subsequent to the initial injection. The relationship between concentration and peak are is linear for creatinine standards between 5 and 10 mg/liter. Specimen volumes ranging from 1 to 25 microliters, and containing creatinine in amounts exceeding 5 ng per injected sample, can be assayed with this system.