Capillary ion electrophoresis of inorganic anions and uric acid in human saliva using a polyvinyl alcohol coated capillary column and hexamethonium chloride as additive of background electrolyte.

A combination of polyvinyl alcohol chemically coated capillary (PVA capillary) and background electrolyte (BGE) with ion-pair reagent (hexamethonium dichloride, HMC) was used on capillary ion electrophoresis-UV detection (CIE-UV) for analysis of Br⁻, I⁻, NO2⁻, NO3⁻, SCN⁻ and uric acid in human saliva. The PVA capillary prepared in our laboratory minimized electro-osmotic flow (EOF) at the BGE in pH 3-10, and did not affect the UV detection at 210 nm by the PVA-layer on capillary wall. Therefore, use of the PVA capillary was suitable for sensitive UV detection for analyte anions, as well as suppression of protein adsorption. In this study, we optimized the BGE of 10 mM phosphate plus 10 mM HMC with applying a voltage of -15 kV. HMC as an additive to BGE could manipulate the electrophoretic mobility of anions, without electrostatic adsorption to the PVA capillary. The CIE-UV could separate and determine analyte anions in human saliva containing proteins by the direct injection without pretreatments such as dilution or deproteinization within 13 min. The relative standard deviations (n=10) were ranged of 0.5-1.6% in migration times, 2.2-6.8% in peak heights and 2.8-8.4% in peak areas. The limits of detection (S/N=3) were ranged of 3.42-6.87 µM. The peak height of anions in this system was gradually decreased through the successive injections of saliva samples, but the problem was successfully solved by periodically conditioning the PVA capillary. The quantifiability of anions in human saliva samples by the CIE-UV was evaluated through the recoveries by standard addition methods and comparison of other representative analytical methods, as well as identification by ion chromatography (IC). From the anion analyses in 12 different saliva samples, the CIE-UV demonstrated that can obtain obvious differences in concentrations of SCN⁻ between of smoker and non-smoker and those of uric acid between male and female with satisfactory results.

Capillary ion electrophoresis-capacitively coupled contactless conductivity detection of inorganic cations in human saliva on a polyvinyl alcohol-coated capillary.

Capillary ion electrophoresis-capacitively coupled contactless conductivity detection (CIE-C4D) with a polyvinyl alcohol chemically coated capillary (PVA capillary) was used to analyze inorganic cations (Na(+), K(+), NH(4)(+), Mg(2+), and Ca(2+)) commonly found in human saliva. The PVA capillary, which was made by our laboratory, minimized electro-osmotic flow in the wide pH range of the background electrolyte (BGE), and the PVA layer adsorbed to capillary wall did not affect the conductimetric background level. In this study, we determined an optimized BGE of 30 mM lactic acid/histidine plus 3 mM 18-crown-6 for the CIE-C4D system using the PVA capillary, which could simultaneously improve the separation of Mg(2+) and Ca(2+) from Na(+) and that of K(+) from NH(4)(+). This system obtained highly reproducible separation of cations in human saliva samples within 8 min at 20 kV without deprotonation. The quantifiability of cations in human saliva samples on the CIE-C4D system was demonstrated through identification by ion chromatography with satisfactory results.