Novel liquid chromatographic determination of cystatin C in human urine

摘要

A rapid and sensitive method for quantitative determination of cystatin C (CC) protein in human urine via HPLC was developed and validated. Acetone has been used as a precipitating agent of CC protein from the h urine biological matrix. Separation and detection were accomplished by ion pair liquid chromatoragphy and UV detection. Gradient elution mode was utilized to elute CC with a UV detection of 224 nm. The analysis time was 14 min per sample using Ace C8 (150 x 4.6 mm i.d., 5 mu m) as a chromatographic column with a flow rate of 1.0 mL/min. Calibration curve with good linearity (r(2) = 0.99) within the range from 0.390 to 0.001 mg/mL was obtained. Limits of detection and quantification were 0.001 and 0.002 mg/mL., respectively. Inter-assay and intra-assay variabilities were < 15% for all levels and < 20% at the limit of quantification level. Major advantages of I-lie method: specific where no false positive results might be obtained and fast where sample pretreatment needs only 1 h.