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Biomedical subjects

R M Alonso

Publications and source records attributed to R M Alonso.

36 records · Page 2Linked to original sources

Quantitative determination of indapamide in pharmaceuticals and urine by high-performance liquid chromatography with amperometric detection.

A high-performance liquid chromatographic method with amperometric detection for the determination of the diuretic indapamide using a muBondapak C18 column is developed. The mobile phase consists of an acetonitrile-water mixture (45:55, 5 mM) in KH2PO4-K2HPO4 (pH 4.0). The compound is monitored at +1200 mV with an amperometric detector equipped with a glassy carbon working electrode. A liquid-liquid or solid-liquid extraction is performed prior to chromatographic analysis to avoid the interferences found in urine matrix. Percentages of recovery are 88.3 +/- 5.6 and 82.9 +/- 7.8 for liquid-liquid and solid-liquid extraction, respectively. The developed method has a linear concentration range from 25 to 315 ng/mL with a reproducibility in terms of relative standard deviation of 4% for a concentration level of 0.5 microgram/mL and a quantitation limit of 1 ng/mL. The method is applied to the determination of indapamide in tablets and urine obtained from hypertensive patients after the ingestion of Tertensif (indapamide 2.5 mg).

Adult↗

Quantitative determination of the angiotensin-converting enzyme inhibitor cilazapril and its active metabolite cilazaprilat in pharmaceuticals and urine by high-performance liquid chromatography with amperometric detection.

A rapid and simple high-performance liquid chromatographic method with amperometric detection has been developed for the quantitation of cilazapril and its active metabolite and degradation product cilazaprilat in urine and tablets. The chromatographic system consisted of a microBondapak C18 column, using a mixture of methanol-5 mM phosphoric acid (50:50, v/v) as mobile phase, which was pumped at a flow-rate of 1.0 ml/min. The column was kept at a constant temperature of (40+/-0.2) degrees C. Detection was performed using a glassy carbon electrode at a potential of 1350 mV. Sample preparation for urine consisted of a solid-phase extraction using C8 cartridges. This procedure allowed recoveries greater than 85% for both compounds. The method proved to be accurate, precise and sensitive enough to be applied to pharmacokinetic studies and it has been applied to urine samples obtained from four hypertensive patients (detection limit of 50 ng/ml for cilazapril and 40 ng/ml for cilazaprilat in urine). Results were in good agreement with pharmacokinetic data.

Angiotensin-Converting Enzyme Inhibitors↗

Quantitative determination of the beta-blocker labetalol in pharmaceuticals and human urine by high-performance liquid chromatography with amperometric detection.

A rapid and simple high-performance liquid chromatographic (HPLC) method with amperometric detection has been developed for the quantitation of labetalol in urine. The chromatography was performed at 30 degrees C using a reversed-phase column with a base deactivated silica stationary support and an alkylamide bonded phase (Supelcosil ABZ+Plus). A 5 mM acetate buffer (pH 4.5)-acetonitrile (70:30, v/v) mixture was employed as the mobile phase, pumped at a flow-rate of 1 ml/min. Sample preparation was carried out using a simple solid-phase extraction (SPE) procedure, and recoveries higher than 85% were achieved. The method was found to be accurate, precise (R.S.D lower than 8%), and sensitive enough (experimental quantitation limit of 20 ng/ml, detection limit 10 ng/ml) to be applied to doping analysis and pharmacokinetic studies in human urine. The method was applied to the determination of labetalol in pharmaceutical formulations and urine samples obtained from a healthy volunteer after the ingestion of a therapeutic dose of the drug, and the results obtained were in agreement with the pharmacokinetic data.

Adrenergic beta-Antagonists↗

Simultaneous determination of the beta-blocker atenolol and several complementary antihypertensive agents in pharmaceutical formulations and urine by capillary zone electrophoresis.

A simple capillary zone electrophoresis method is developed for the quantitation of the beta-blocker atenolol and the complementary antihypertensive agents bendroflumethiazide, amiloride, and hydrochlorothiazide in human urine samples. The electrophoretic separation is performed using a 78-cm x 75-micron-i.d. (70-cm effective length) fused-silica capillary. A borate buffer (pH 9) is used as running electrolyte. The sample is hydrostatically introduced for 20 s, and the running voltage is 25 kV at the injector end of the capillary. The analysis of urine samples requires the optimization of solid-phase extraction methods, achieving recoveries > or = 61% for all the drugs and good separation from the urine matrix. The method is successfully applied to the determination of these compounds in pharmaceutical formulations and in urine samples collected after the intake of Neatenol Diu (100 mg atenolol-5 mg bendroflumethiazide) and Kalten (50 mg atenolol-25 mg hydrochlorothiazide-2.5 mg amiloride). The method is validated in terms of reproducibility, linearity, and accuracy.

Adrenergic beta-Antagonists↗

Gas chromatographic-mass spectrometric analysis of the loop diuretic torasemide in human urine.

A fast and reliable gas chromatographic-mass spectrometric (GC-MS) method for the identification and determination of the loop diuretic torasemide in human urine is described. The usefulness of different derivatization procedures and reagents was studied. Flash methylation using trimethylanilinium hydroxide was the most convenient and appropriate procedure. The optimal urine isolation method comprised alkaline liquid-liquid extraction with ethyl acetate. After evaporation of the organic layer to dryness, the solid residue was reconstituted in the derivatizing reagent and was directly injected into the GC-MS system. Samples were analysed in the multiple ion detection mode using electron impact ionization. No interferences from other urinary compounds were found. Torasemide gave rise to a derivative that was identified by GC with Fourier transform infrared detection. There was a 70 +/- 5% recovery of torasemide. The coefficient of variation was 5% at a concentration of 0.05 microgram/ml. The method was used for the determination of torasemide in urine samples obtained from a healthy volunteer that had received a single, 10 mg dose of torasemide.

Diuretics↗

Micellar electrokinetic chromatography as a fast screening method for the determination of the doping agents furosemide and piretanide in urine.

The possibility of using micellar electrokinetic chromatography for the screening of the loop diuretics piretanide and furosemide in urine was studied. A fast and simple method with good repeatability is described. The method was applied to urine samples collected from a healthy volunteer after oral administration of therapeutic doses of each compound. Positive identification in the urine matrix was possible through recording diode array spectra.

Chromatography, High Pressure Liquid↗

Determination of piretanide and furosemide in pharmaceuticals and human urine by high-performance liquid chromatography with amperometric detection.

A high-performance liquid chromatographic method with electrochemical detection (ED) has been developed for the determination of two diuretics: 4-phenoxy-3-(1-pyrrolidinyl)-5-sulfamoylbenzoic acid (piretanide) and 4-chloro-2-furfurylamino-5-sulfamoylbenzoic acid (furosemide). The chromatographic separation was performed on a mu Bondapak C18 column with a mobile phase of acetonitrile-water (40:60) containing 5 mM KH2PO4/K2HPO4 and with a flow-rate of 1 ml/min (69 bar). The temperature was optimized at 30 +/- 0.2 degrees C. The amperometric detector equipped with a glassy carbon electrode was operated at +1200 mV versus Ag/AgCl in the direct current mode. The method was applied to the determination of these compounds in two concentration ranges (ppm and ppb), obtaining a reproducibility in terms of relative standard deviations lower than 1% for within-day and 4% for day-to-day and determination limits of 15 ppb for both compounds. Recoveries greater than 90% were obtained for spiked urine samples, using a liquid-liquid extraction method in the sample clean-up procedure. The LC-ED method was applied to commercially available pharmaceuticals (Seguril, furosemide 40 mg, and Perbilén, piretanide 6 mg) and urine samples obtained from healthy volunteers and hypertensive patients.

Chromatography, High Pressure Liquid↗

High-performance liquid chromatography with amperometric detection applied to the screening of beta-blockers in human urine.

A high-performance liquid chromatographic method with electrochemical detection has been developed for the determination of six beta-blockers: atenolol, nadolol, timolol, metoprolol, oxprenolol, and alprenolol. The chromatographic separation was performed using a mu Bondapack C18 column, a mobile phase of acetonitrile-water (40:60), containing 5 mM KH2PO4/K2HPO4 proved to be optimal at a 1.3 ml/min flow-rate, and a pH of 6.5. The temperature was optimized at 30 +/- 0.2 degrees C. The amperometric detector, equipped with a glassy carbon electrode, was operated at 1300 mV versus Ag/AgCl in the direct current mode. The method was applied to the determination of these compounds at two concentration levels: ppm and ppb (ng/ml), obtaining relative standard deviations lower than 5% at ppm levels and lower than 10% at ppb levels, and quantitation limits ranging from 15 ppb to 500 ppb. The method was applied to the screening of beta-blockers in spiked urine samples, with a total elution time lower than 12 min, obtaining the best recoveries for timolol and metoprolol (never greater than 93%). These recoveries together with the low limits of quantitation achieved, allows its application to doping analysis in human urine.

Acetonitriles↗

Comparison between the effects of VIP and the novel peptide PACAP on the exocrine pancreatic secretion of the rat.

The effect of intravenous infusion of pituitary adenylate cyclase-activating peptide (PACAP) 27, a novel regulatory peptide that shows a close structural and chemical similarity to vasoactive intestinal peptide (VIP), on the rat exocrine pancreatic secretion was studied. PACAP and VIP stimulated the flow rate of exocrine pancreatic secretion (p < 0.05). However, protein output and amylase secretion were mainly stimulated by PACAP. Intravenous infusion of VIP increased the plasma levels of secretin (p < 0.05). On the other hand, PACAP released neither secretin nor VIP. Our results show: (a) both PACAP and VIP stimulate exocrine pancreatic secretion, (b) PACAP stimulation of pancreatic amylase and protein secretion is greater than that induced by VIP, and (c) PACAP probably exerts a direct effect on exocrine pancreas whereas some of the actions of VIP might be mediated by secretin.

Amylases↗

Effects of acute intravenous ethanol on basal exocrine pancreatic secretion in rat: cholinergic involvement.

The effect of intravenous infusion of ethanol on the basal exocrine pancreatic secretion of anesthetized rats was studied. The cholinergic involvement on the actions of ethanol was also studied using previously atropinized animals. During the stimulation period, pancreatic flow rate was significantly increased by intravenous ethanol in both un-atropinized (199% compared with basal) and atropinized rats (195% compared with basal). Pancreatic protein output was also increased during ethanol administration in both groups of animals (171% and 165% compared with basal in, respectively, un-atropinized and atropinized rats). After the administration of ethanol, in the poststimulation period, pancreatic flow rate was further increased only in the atropinized group of rats (290% compared with basal), whose values were significantly higher than those of ethanol-treated un-atropinized animals (195% compared with basal). A similar profile of response was observed in pancreatic protein output. Since intravenous ethanol did not stimulate either secretin or VIP release to portal plasma, the present results point to a direct effect of this substance on the exocrine pancreas. Furthermore, atropine revealed the existence of an inhibitory cholinergic effect of ethanol on the exocrine pancreas. In summary, results show that the effect of intravenous ethanol on the basal exocrine pancreatic secretion is dual and antagonistic.

Acetylcholine↗

Mechanisms involved in the control of exocrine pancreatic secretion in the interdigestive state in the rabbit.

The effect of rapid wash-out of the duodenum with phosphate buffered saline on exocrine pancreatic secretion and plasma levels of secretin, VIP, gastrin and CCK was studied. Furthermore, the possible nervous role in this effect was checked after atropine and hexamethonium treatment. Rapid wash-out significantly increased protein output (35.0 micrograms/min, in the control group without duodenal perfusion and 72.15 micrograms/min, in the perfused group) and the plasma levels of CCK (from 5.2 to 13.17 fmol/ml). Intravenous infusion of atropine significantly reduced the protein output (from 78.19 to 32.45 micrograms/min) and the plasma levels of CCK (from 10.1 to 5.55 fmol/ml), with no change in the remaining parameters in the intraduodenally perfused group. Intravenous administration of hexamethonium significantly stimulated hydroelectrolyte secretion (from 6.99 to 15.15 microliters/min) and the plasma levels of VIP (from 4.8 to 7.3 fmol/ml) and reduced the protein output (from 61.47 to 30.75 micrograms/min) and the plasma levels of CCK (from 14.56 to 6.25 fmol/ml) in the intraduodenally perfused group. Our results suggest that, in the interdigestive state, the exocrine pancreatic secretion of the rabbit is tonically inhibited. This inhibition can be divided into two different mechanisms: on the one hand there is a decrease in enzyme secretion produced by a duodenal factor and mediated by CCK and muscarinic mechanisms and on the other, there is an inhibition of hydroelectrolyte secretion with no duodenal participation which is probably controlled by nervous non-muscarinic mechanisms and VIP involvement.

Animals↗

Separation and determination of aminohalogenbenzophenones by high-performance liquid chromatography with electrochemical detection.

A high-performance liquid chromatographic method with electrochemical detection has been developed for the determination of three aminohalogenbenzophenones: 2-amino-2',5-dichlorobenzophenone, 2-amino-5-chlorobenzophenone and 2-amino-5-bromo-2'-fluorobenzophenone, metabolites of benzodiazepinooxazoles and other psychotropic drugs. A mobile phase of methanol-water (65:35), containing 5 mM KH2PO4 appeared to be the optimal when a 4-microns, 60-A Nova-Pak C18 column and a flow-rate of 0.75 ml/min (130 bar) were used. The temperature was optimized at 30 degrees C. The amperometric detector, equipped with glassy carbon electrode, was operated at 1.3 V versus Ag/AgCl in the DC mode. The method was applied to the determination of these compounds at two concentration levels: ppm and ppb (ng/cm3) using 2-amino-5-chlorobenzophenone as internal standard. The limit of determination was 750 pg/ml of biological fluid for each compound, and recoveries greater than 97% were obtained for spiked samples of urine and serum, using C18 Sep-Pak cartridges in the sample clean-up procedure.

Anti-Anxiety Agents↗

[Social self-positioning as indicator of socioeconomic status].

OBJECTIVE: Self-perceived class results from directly questioning subjects about his or her social class. The aim of this investigation was to analyse self-perceived class in relation to other indicator variables of socioeconomic level. METHODS: Data from the 1994 Catalan Health Interview Survey, a cross-sectional survey of a representative sample of the non-institutionalised population of Catalonia was used. We conducted a discriminant analysis to compute the degree of right classification when different socioeconomic variables potentially related to self-perceived class were considered. All subjects who directly answered the questionnaire were included (N = 12,245). With the aim of obtaining the discriminant functions in a group of subjects and to validate it in another one, the subjects were divided into two random samples, containing approximately 75% and 25% of subjects (analysis sample, n = 9,248; and validation sample, n = 2,997). RESULTS: The final function for men and women included level of education, social class (based in occupation) and equivalent income. This function correctly classified 40.9% of the subjects in the analysis sample and 39.2% in the validation sample. Two other functions were selected for men and women separately. In men, the function included level of education, professional category, and family income (39.2% of classification in analysis sample and 37.2% in validation sample). In women, the function (level of education, working status, and equivalent income) correctly classified 40.3% of women in analysis sample whereas the percentage was 38.9% in validation sample. The percentages of right classification were higher for the highest and lowest classes. CONCLUSION: These results show the utility of a simple variable to self-position within the social scale. Self-perceived class is related to education, income, and working determinants.

Adolescent↗