Determination of uric acid in plasma and allantoic fluid of chicken embryos by capillary electrophoresis

Abstract

Capillary electrophoresis with diode array detection (DAD) was used to determine uric acid (UA) in chicken plasma and the allantoic fluid of chicken embryos. Complete separation of uric and ascorbic acids was attained in less than 10 min in the optimized BGE containing 60 mM MES + 30 mM Tris + 0.001% (w/v) polybrene (pH 6.1). The limit of UA detection (0.2 mg/L) was found to be low enough for sensitive analysis of native plasma and allantoic fluid samples. Range of linearity (1-200 mg/L), repeatability for peak area (CV <4.1%) and migration time (CV < 2.5%), as well as recovery of UA from biological samples (97-100%), were found to be satisfactory. The method was applied to detect the elevated UA concentrations (hyperuricemia) in chicken embryos with induced unilateral renal agenesis. CE/DAD analysis of the chicken plasma can be carried out with a relatively small volume of samples (1 microL).

Theoretical Study of Uric Acid and its Ions in Aqueous Solution

“…There have been many measurements of the UV absorption spectra of uric acid and its anions in aqueous solution, some with specific pH controlled by buffers and some without specified pH values [37,[42][43][44][45][46][47][48][49][50][51][52]. In Supplementary table, we summarize the measurements in order to assess the reliability of calculated vertical excitation energies and present the midrange value for each transition, after rejecting some likely outliers, indicated in italics.…”

Determination of uric acid in human urine by capillary zone electrophoresis with indirect laser‐induced fluorescence detection

“…Concerning the quantitative analysis of UA, several efforts have been taken by means of HPLC and CE. But the detection methods of these investigations are mainly in the domain of conventional techniques including UV 4–8, photodiode array 9–12, electrochemical detection (ED) 13–19 and chemiluminescence (CL) 20, 21. So far, LIF detection has never been used as the detection method for the determination of UA in CE separation.…”

Tear analysis of ascorbic acid, uric acid and malondialdehyde with capillary electrophoresis

“…Although MDA is detected but not quantifi ed in tears of healthy individuals, the method might be useful for monitoring high MDA concentrations in pathological situations. The time necessary for complete separation of ascorbic and uric acid (18 min) is longer than the analysis time that has been reported (4-10 min) for CE plasma analysis of ascorbic and uric acid (Cheng et al, 1999;Zinellu et al, 2004Zinellu et al, , 2005Matejckov et al, 2007;Ruiz-Jiménez et al, 2007;Yao et al, 2007). This represents the compromise of migration times in favor of sensitivity; indeed our LLOQ values (0.386 and 3.522 mg/L for uric and ascorbic acid, respectively) are lower than those reported for CE with UV detection [10 and 2.5 mg/L for uric and ascorbic acid in Zinellu et al (2004) and 5 and 2.5 mg/L, respectively, in Zinellu et al (2005)].…”

“…Capillary electrophoresis is an analytical technique of high separation capacity, requiring only a few nanoliters of sample for each analysis. Capillary electrophoresis has been used for the determination of ascorbic and uric acid in plasma and urine samples with ultraviolet or electrochemical detection (Cheng et al, 1999;Zinellu et al, 2004;Zinellu et al, 2005;Matejckov et al, 2007;Ruiz-Jiménez et al, 2007;Yao et al, 2007). Although it is an ideal technique for analysis of low volume bio-logic samples such as tears, it has not been applied to the determination of these low molecular weight antioxidants and MDA in tears.…”