Simultaneous determination of uric acid and creatinine in urine by an eco-friendly solvent-free high performance liquid chromatographic method

Simultaneous Determination of Creatinine and Uric Acid in Human Urine by High-Performance Liquid Chromatography

“…The chosen wavelength of 205 nm provides a higher sensitivity with a clean chromatogram than the wavelength of 220 or 235 nm employed in previous studies. [19][20][21] The peak purity was confirmed by constant signal ratios (relative absorbances at different wavelength) across the peak profile of creatinine, uric acid and the internal standard.…”

“…11,[13][14][15] To avoid interferences in the sample matrices, many HPLC methods for the isolation and quantitation of creatinine, uric acid, and other metabolites in biological fluids have been described, including ion-exchange, normal-phase, reversed-phase, and reversed-phase ion-pair chromatography. [16][17][18][19][20][21] Reversed-phase HPLC methods with a highly aqueous mobile phase have been increasingly attractive in light of the recent movement of green analytical chemistry, which encourages the development of analytical methods without or with reduced consumption of organic solvent, energy and time. [17][18][19][22][23][24] However, many studies have reported that the use of a highly aqueous mobile phase (>90% water) with C18 and C8 columns causes an anomalous chromatographic behavior, and a retention loss of analytes.…”

“…[16][17][18][19][20][21] Reversed-phase HPLC methods with a highly aqueous mobile phase have been increasingly attractive in light of the recent movement of green analytical chemistry, which encourages the development of analytical methods without or with reduced consumption of organic solvent, energy and time. [17][18][19][22][23][24] However, many studies have reported that the use of a highly aqueous mobile phase (>90% water) with C18 and C8 columns causes an anomalous chromatographic behavior, and a retention loss of analytes. 19,25 These anomalies have been attributed to (i) aggregation of the bonded hydrocarbon chains in the presence of a highly aqueous mobile phase, rendering them inaccessible to analytes, and (ii) extrusion of the highly aqueous mobile phase from the pores of the particles of the stationary phase.…”

“…[17][18][19][22][23][24] However, many studies have reported that the use of a highly aqueous mobile phase (>90% water) with C18 and C8 columns causes an anomalous chromatographic behavior, and a retention loss of analytes. 19,25 These anomalies have been attributed to (i) aggregation of the bonded hydrocarbon chains in the presence of a highly aqueous mobile phase, rendering them inaccessible to analytes, and (ii) extrusion of the highly aqueous mobile phase from the pores of the particles of the stationary phase. [25][26][27] Our preliminary experiments in this study showed a rapid loss in the retention time of creatinine and uric acid on the C18 column with highly aqueous mobile phases reported in some recent publications.…”

“…[25][26][27] Our preliminary experiments in this study showed a rapid loss in the retention time of creatinine and uric acid on the C18 column with highly aqueous mobile phases reported in some recent publications. 19,21 In this paper, a simple, fast, accurate and environmentally friendly reversed-phase HPLC method is described for the simultaneous determination of creatinine and uric acid in human urine. The method was optimized through gradient elution to remove hydrophilic and hydrophobic residues, and to maintain the C18 column resolution following separation of the analytes using an aqueous mobile phase of phosphate buffer.…”