Novel liquid chromatographic determination of cystatin C in human urine.
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 urine biological matrix. Separation and detection were accomplished by ion pair liquid chromatography and UV detection. Gradient elution mode was utilized to elute CC with a UV detection of 224nm. The analysis time was 14min per sample using Ace C8 (150x4.6mm i.d., 5mum) as a chromatographic column with a flow rate of 1.0mL/min. Calibration curve with good linearity (r(2)=0.99) within the range from 0.390 to 0.001mg/mL was obtained. Limits of detection and quantification were 0.001 and 0.002mg/mL, respectively. Inter-assay and intra-assay variabilities were <15% for all levels and <20% at the limit of quantification level. Major advantages of the method: specific where no false positive results might be obtained and fast where sample pretreatment needs only 1h.
Al-Musaimi
Othman Ibrahem Yousef
OI
Analytical Research Department, Hikma Pharmaceuticals-Amman, Department of Chemistry, University of Jordan, Amman, Jordan.
Fayyad
Manar Khalid
MK
Mishal
Adel Khalil
AK
D000328:Adult; D002853:Chromatography, Liquid / Q000379:methods*; D055316:Cystatin C / Q000652:urine*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged
D055316:Cystatin C