1. Investigación

The prevalence of renal stone disease is increasing, although it remains higher in men than in women when matched for age. While still somewhat controversial, several studies have reported an association between renal stone disease and hypertension, but this may be confounded by a shared link with obesity. However, independent of obesity, hyperoxaluria has been shown to be associated with hypertension in stone-formers, and the most common type of renal stone is composed of calcium oxalate. The chloride−oxalate exchanger slc26a6 (also known as CFEX or PAT-1), located in the renal proximal tubule, was originally thought to have an important role in sodium homeostasis and thereby blood pressure control, but it has recently been shown to have a key function in oxalate balance by mediating oxalate secretion in the gut. We have applied two orthogonal analytical platforms (NMR spectroscopy and capillary electrophoresis with UV detection) in parallel to characterize the urinary metabolic signatures related to the loss of the renal chloride− oxalate exchanger in slc26a6 null mice. Clear metabolic differentiation between the urinary profiles of the slc26a6 null and the wild type mice were observed using both methods, with the combination of NMR and CE-UV providing extensive coverage of the urinary metabolome. Key discriminating metabolites included oxalate, m-hydroxyphenylpropionylsulfate (m-HPPS), trimethylamine-N-oxide, glycolate and scyllo-inositol (higher in slc26a6 null mice) and hippurate, taurine, trimethylamine, and citrate (lower in slc26a6 null mice). In addition to the reduced efficiency of anion transport, several of these metabolites (hippurate, m-HPPS, methylamines) reflect alteration in gut microbial cometabolic activities. Gender-related metabotypes were also observed in both wild type and slc26a6 null groups. Urinary metabolites that showed a sex-specific pattern included trimethylamine, trimethylamine-N-oxide, citrate, spermidine, guanidinoacetate, and 2-oxoisocaproate. The gender-dependent metabolic expression of the consequences of slc26a6 deletion might have relevance to the difference in prevalence of renal stone formation in men and women. The different composition of microbial metabolites in the slc26a6 null mice is consistent with the fact that the slc26a6 transporter is found in a range of tissues, including the kidney and intestine, and provides further evidence for the “long reach” of the microbiota in physiological and pathological processes.

In a preliminary study pyroglutamate showed to be over 10 times increased in some lyophilised coffees with respect to brewed or filtered coffees, and probably that increase is related to some stage of the industrial process. Pyroglutamate is known to have a number of remarkable cognitive enhancing effects, which could be also related to the properties of coffee traditionally associated to caffeine. Pyroglutamate improves memory and learning and has anti-anxiety effects in rats. Therefore, a method has been developed and validated for the simultaneous determination of caffeine and pyroglutamate in coffee by capillary electrophoresis. Separation conditions employed MECK conditions with 50 mM borate buffer at pH 9.5 with 130 mM SDS. The applied potential was 10 kV and detection was performed at 200 nm. Afterwards, 10 soluble coffees from the market were measured and caffeine and pyroglutamate levels were compared. Those coffees with higher pyroglutamate with or without caffeine were preliminarily tested for sedative/stimulant properties and cognition enhancing effects in mice. The most relevant finding was a partial reversal of scopolamine-induced amnesia in the passive avoidance paradigm after oral administration of one coffee.

An in-capillary sample preconcentration strategy based on solid phase extraction (SPE) technology coupled with capillary electrophoresis (CE) has been developed taking advantage of both techniques (SPE and CE). An in-line frit-free preconcentration device for capillary electrophoresis containing MCX beads, obtained from the corresponding Waters OASIS® cartridges, was prepared. The retention of the particles was based on the relative diameters of the particles, carefully selected, and the capillaries. An experimental preconcentration factor of 100 was found for the system. Conditions were optimised for 3-nitrotyrosine measurement in rat urine being 4.4 M spiked in the urine the lowest value detectable.

Capillary electrophoresis (CE) poses unique challenges in many different analytical applications, mainly to biological and complex samples and when only small amounts of sample are available, due to its low sample consumption. As a consequence, poor limits of detection are usually observed with this technique, especially with UV photodetectors. Minimal or no sample treatment is desirable in any analytical method to avoid external sources of contamination or errors and to provide a high throughput. On- and in-capillary sample pre-concentration strategies, based on solid-phase extraction (SPE) technology can take advantage of both techniques (SPE and CE), while avoiding sample contamination and tedious manipulations when the sample amount is an issue. Moreover, the combination can provide two-dimensional separations. This review collects the most recent strategies that merge SPE technology built on- and in-capillary pre-concentration for increasing sensitivity and/or selectivity.

A systematic review of the literature on capillary electrophoresis applied to short chain organic acid analysis in body fluids has been conducted with special interest on those acids related to inborn errors of metabolism. The technique is briefly described, as well as the choice of the main analytical parameters: sample pre-treatment, polarity, capillary type, background electrolyte, and detection. The applications described in the literature are listed and the main features of the technique are discussed.

Short chain organic acids play an important role in different areas such as biochemistry, clinical chemistry, or the food industry. The enantiomeric ratio of chiral metabolites is an important parameter for the understanding of metabolic processes and in many cases it can serve diagnostic purposes. On the other hand, the presence of racemates in food products could indicate the use of organic acids as additives; this is not always permitted and needs to be controlled. The short chain of these acids makes difficult the three point interaction generally accepted as necessary for chiral recognition. Relatively recent publications have demonstrated the feasibility of their direct chiral separation in capillary electrophoresis by various techniques utilizing exchange capillary electrophoresis, macrocyclic antibiotics, cyclodextrins, ion-pair method, and transition metal complexes. The present article describes existing methods and strategies proposed to advance these areas.

This review article addresses the different chiral capillary electrophoretic methods that are being used for the study and characterization of foods and food compounds (e.g., amino acids, organic acids, sugars, pesticides). An updated overview, including works published till December 2002, on the principal applications of enantioselective procedures together with their main advantages and drawbacks in food analysis is provided. Some anticipated applications of chiral electromigration methods in food characterization are also discussed.

This review article addresses the different capillary electrophoretic methods that are being used for the study of both short-chain organic acids (including anionic catecholamine metabolites) and fatty acids in biological samples. This work intends to provide an updated overview (including works published until November 2004) on the recent methodological developments and applications of such procedures together with their main advantages and drawbacks. Moreover, the usefulness of CE analysis of organic acids to study and/or monitor different diseases such as diabetes, new-borns diseases or metabolism disorders is examined. The use of microchip devices and CE-MS couplings for organic acid analysis is also discussed.

A capillary electrophoresis method has been developed and validated for acetic, citric, fumaric, lactic, malic, oxalic, succinic, and tartaric acids plus the measurement of nitrate and sulfite ions in white and red wines. The separation was carried out in a neutral coated capillary. Separation was performed at 14 kV of applied potential. Temperature was maintained at 20 C. The background electrolyte used was 200 mM phosphate buffer at pH 7.50. Separation was obtained in less than 13 min. Validation parameters obtained for the method permit it to be considered adequate for routine analysis.