[Rapid detection of microalbuminuria with high-performance liquid chromatography]

To develop a rapid detection method for microalbuminuria with size-exclusion high performance liquid chromatography.

With a new mobile phase, samples were injected onto an Agilent Zorbax GF-250 size exclusion chromatographic column. The method was evaluated and urine albumin of 56 diabetic patients was analyzed.

The mobile phase containing 0.1% formic acid and acetonitrile, with 20 μL sample size, 1 mL/min flow rate, 205 nm detection wavelength. The retention time of albumin both in human serum and urine was 1.7 min. The linear range was 5-2000 mg/L. The lower limit of measurement was 2 mg/L. The intra-assay coefficient of variation and the inter-assay coefficient of variation were 3.98% and 4.05% (20 mg/L), 3.55% and 3.60% (200 mg/L), 4.65% and 4.74% (2000 mg/L), respectively. Recovery rates were 95.3%, 98.1%, and 97.2%. Microalbuminuria was detected in 30 samples by high performance liquid chromatography and 15 samples by immunoturbidimetry from 56 patients with diabetic mellitus.

A fast and high sensitivity method, namely size-exclusion high performance liquid chromatography, with mobile phase containing 0.1% formic acid and acetonitrile has been established to analyze microalbuminuria, which can detect more microalbuminuria than other methods and is suitable for clinical routine measurement.