A simple, rapid and reliable high performance liquid chromatography method for the simultaneous determination of creatinine and uric acid in plasma and urine

Abstract

A simple, rapid and reliable method for the simultaneous determination of creatinine (Cr) and uric acid (UA) in plasma and urine has been developed using high performance liquid chromatography with ultraviolet detection. Separation was achieved by an Agilent ZORBAX SB-C18 column and an optimized mobile phase consisting of 20 mmol L À1 sodium acetate, 30 mmol L À1 acetic acid and 1% methanol.The total time of analysis was 4 min. Good linear responses were found with correlation coefficient (r) >0.999 for both Cr and UA within the concentration range of relevant levels. The recoveries of Cr and UA were 102 AE 1% and 99.3 AE 2.1% for plasma; and 100 AE 2% and 102 AE 4% for urine. CVs of repeatability and intermediate precision of all analytes were less than 5%. The limit of detection of the developed method was: Cr, 0.09 mmol L À1 and UA, 0.04 mmol L À1 . The proposed HPLC method could be used for not only reference purpose but also routinely in clinical laboratory testing.

Graphitic carbon nitrides modified hollow fiber solid phase microextraction for extraction and determination of uric acid in urine and serum coupled with gas chromatography-mass spectrometry

“…Therefore, establishing a simple, direct and accurate technique to monitor UA in biofluid is of great importance for diagnosing diseases and clinical study. In recent years, many analytical methods have been proposed to measure UA in biofluid, such as gas chromatography (GC) [6][7][8], enzyme-linked immunosorbent assay (ELISA) [9], fluorescence (FL) [10], liquid chromatography (LC) [11], electrochemistry [2,[12][13][14]. In a typical biofluid analysis, a long processing stage of sample pretreatment is invariably essential before instrumental analysis [15,16].…”

Simultaneous determination of plasma creatinine, uric acid, kynurenine and tryptophan by high‐performance liquid chromatography: method validation and in application to the assessment of renal function

“…Our previous work demonstrated that UA was stable enough to satisfy the need for automated analysis of a large number of samples in HClO 4 ‐treated plasma during the analytical process (Zhao, ). In this work, we tested the stability of Cr, UA, KYN and TRP in plasma after going through freeze–thaw cycles.…”

“…Several high‐performance liquid chromatography (HPLC) methods have been developed for the simultaneous determination of Cr and UA (Seki et al ., ; Yang, ; Zhao, ; Zhiri et al ., ) or KYN and TRP (Iizuka et al ., ; Laich et al ., ; Vignau et al ., ; Zhang et al ., ; Zhao et al ., ) in plasma or serum using a single mobile phase with constant elution. In reverse‐phase HPLC, the volume of organic solvent contained in the mobile phase for the separation of Cr and UA (Zhao, ) was much lower than that for the separation of KYN and TRP (Zhang et al ., ; Zhao et al ., ). In our previous work, we have successfully developed an HPLC method for the simultaneous determination of urinary Cr, TRP and TRP‐related metabolites by HPLC with ultraviolet (UV) and fluorescence detection through a gradient elution program so as to utilize Cr as an internal standard for normalizing the excretion of TRP and TRP‐related metabolites in urine (Zhao et al ., ).…”

Quantitative analysis of biomarkers of liver and kidney injury in serum and urine using ultra-fast liquid chromatography with tandem mass spectrometry coupled with a hydrophilic interaction chromatography column: Application to monitor injury induced by E

“…To date, although some approaches, such as CE , GC , HPLC–UV , LC–MS , have been reported to determine one or several of the biomarkers, separately, none of comprehensive methods were reported to determine all of them for simultaneous monitoring of liver and kidney injury, nor applications to injury induced by EPR. Besides, CE may lack robustness for routine tests of biosamples.…”