Capillary electrophoresis with contactless conductivity detection for uric acid determination in biological fluids

The suitability of capillary electrophoresis (CE) with capacitively coupled contactless conductivity detection (C⁴D) for the direct determination of uric acid in human plasma and urine was investigated. It was found that a careful optimization of the buffer composition and pH was necessary to achieve selective determination in the complex sample matrices. An electrolyte solution consisting of 10 mM 2-morpholinoethanesulfonic acid (MES), 10 mM histidine and 0.1 mM hexadecyltrimethylammonium bromide (CTAB), pH 6.0, was finally found suitable for use as running buffer for both sample matrices. The limit of detection (3 S/N) was determined as 3.3 μM. The linearity of the response was tested for the range between 10 and 500 μM and a correlation coefficient of 0.9996 was obtained. Intra- and inter-day variabilities were <10%. Quantitative analysis of urine and plasma samples showed a good correlation with the routine enzymatic method currently used at the University Hospital of Basel.     

Determination of creatine and phosphocreatine in muscle biopsy samples by capillary electrophoresis with contactless conductivity detection

A capillary electrophoresis method with contactless conductivity detection was evaluated as a new approach for quantification of creatine and phosphocreatine in human quadriceps femoris biopsy samples. The running buffers employed consisted of 1 M acetic acid at a pH of 2.3 for the determination of creatine and 50 mM 3-(N-morpholino)propanesulfonic acid/30 mM histidine at a pH of 6.4 for the determination of phosphocreatine in the centrifuged muscle extracts. The limits of detection for creatine and phosphocreatine were found to be 2.5 and 1.0 μM, respectively. Creatine and phosphocreatine were determined in six human muscle biopsy samples and the results were found comparable to those of a standard enzymatic assay. The procedures developed for creatine and phosphocreatine also allow the determination of creatinine as well as adenosine diphosphate and adenosine triphosphate.     

Quantification of plasma lactate concentrations using capillary electrophoresis with contactless conductivity detection

The quantification of plasma lactate and evaluation of the lactate threshold by CE with capacitively coupled contactless conductivity is demonstrated. The only sample preparation needed was deproteinization with a ACN/methanol mixture. A solution of 10 mmol/L 2-morpholinoethanesulfonic acid monohydrate, 10 mmol/L DL-histidine, 70 μmol/L hexadecyltrimethylammonium bromide, pH 6.0 was found suitable as running buffer. Linearity was achieved for the concentration range of 10-1000 μmol/L with a correlation coefficient of 0.9994. The limit of detection (3 S/N) was determined as 3.2 μmol/L. Intra- and inter-day variabilities were less than 7% RSD. The suitability of the method could be demonstrated by analyzing various clinical samples, where the results correlated satisfactorily with those of an established enzymatic method.     

Determination of the enantiomers of α‐hydroxy‐ and α‐amino acids in capillary electrophoresis with contactless conductivity detection

The enantiomers of the anions of five α‐hydroxy acids, namely lactic acid, α‐hydroxybutyric acid, 2‐hydroxycaproic acid, 2‐hydroxyoctanoic acid and 2‐hydroxydecanoic acid, as well as the two α‐amino acids aspartic acid and glutamic acid, were baseline separated and detected by CE with contactless conductivity detection. Vancomycin was employed as chiral selector and could be used with conductivity detection without having to resort to a partial filling protocol as needed when this reagent is used with UV absorbance measurements. The procedure was successfully applied to the determination of the lactic acid enantiomers in samples of milk and yogurt. Linearity was achieved in the concentration range of 10–500 μmol/L with good correlation coefficients (0.9993 and 0.9990 for L ‐ and D ‐lactic acid, respectively). The LODs (3 S/N) for L ‐ and D ‐lactic acid were determined as 2.8 and 2.4 μmol/L, respectively.