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Biomedical subjects

M A Raggi

Publications and source records attributed to M A Raggi.

At least 37 records · Page 2Linked to original sources

Quality control of benserazide-levodopa and carbidopa-levodopa tablets by capillary zone electrophoresis.

In modern practice, the treatment of Parkinson's disease and syndrome is carried out using pharmaceutical formulations containing a combination of levodopa and a decarboxylation inhibitor (carbidopa or benserazide). Two pharmaceutical formulations were quantified by capillary zone electrophoresis using two procedures which differed only in the kind of background electrolyte used. One procedure used a 25 mM phosphate buffer, pH 2.5, while the second one used a 25 mM borate buffer, pH 8.5. The electrophoretic analysis was carried out using an uncoated fused- silica capillary, a separation voltage of 20 kV with currents typically less than 60 microA, and spectrophotometric detection at 205 nm. Calibration curves were performed for levodopa (concentration range 1-100 microg/mL), for carbidopa and benserazide (1-50 microg/mL), and the plots of the peak area versus concentration were found to be linear with a correlation coefficient better than 0.9990. Satisfactory results were obtained when commercial tablets were analyzed in terms of accuracy (98-102%), repeatability (0.6-2.0%), and intermediate precision (1.1-2.6%).

Benserazide↗

Quantitation of olanzapine in tablets by HPLC, CZE, derivative spectrometry and linear voltammetry.

Four analytical methods have been developed for the quality control of pharmaceutical formulations containing the novel antipsychotic drug, olanzapine: high performance liquid chromatography (HPLC), capillary zone electrophoresis (CZE), derivative spectrometry and linear voltammetry. All methods require only a simple extraction procedure of olanzapine from the tablets before analysis. HPLC with ultraviolet detection at 260 nm is carried out with a C8 column and a mobile phase constituted of acetonitrile and aqueous tetramethylammonium perchlorate. CZE is performed in an uncoated capillary with phosphate buffer, pH 3.0, as the background electrolyte, with UV detection at 214 nm. Spectrophotometry uses the derivative of the spectrum at 298 nm. In linear voltammetric method (LSV) the current intensity of the oxidation wave at +495 mV is measured. All methods gave similar results in terms of precision and accuracy. For HPLC and CZE, repeatability and intermediate precision, expressed by the RSD was better than 1.8%. The accuracy, resulting from recovery experiments, was between 99.9 and 101.1%. Spectrometry and voltammetry gave slightly higher RSD values (up to 2.9%) and a larger variation of the accuracy (the recovery was between 97.8 and 102.6%). However, the requirements for quantitative analysis are fulfilled for all methods.

Antipsychotic Agents↗

Extractive analysis of aluminum traces in dialysis solutions.

A very sensitive procedure for the fluorimetric determination of aluminum traces in dialysis solutions by means of Mordant Red 19 dyestuff is described with the extraction of the Al complex in isobutylmethylketone. The experimental conditions were studied, in order to obtain the best extraction yield. The emission intensity of the metal chelate, extracted in the organic layer, was measured at 549 nm, exciting at 485 nm. Linearity between emission intensity and Al concentration was found in the 1-30 ng/ml range. The limit of detection was 0.25 ng/ml. The method resulted to be suitable for the determination of Al traces in commercial dialysis solutions for toxicological purposes.

Aluminum↗

Determination of catecholamines in human plasma by high-performance liquid chromatography with electrochemical detection.

In the present study, assays were improved for the determination of catecholamines in human plasma. High-performance liquid chromatography with electrochemical detection was employed for quantitative analysis. The influence of various parameters on chromatographic performance, such as the composition and the pH of the mobile phase, and the detection potential, was investigated. An accurate solid-phase extraction procedure, after catecholamine complexation with diphenylborate, was developed. The efficiency yield for all catecholamines was in the range 92-98%. Relative standard deviation values for repeatability and for intermediate precision were less than 2% and 3%, respectively, for all three analytes.

Calibration↗

Enantiomeric separation of fluoxetine and norfluoxetine in plasma and serum samples with high detection sensitivity capillary electrophoresis.

A capillary electrophoresis method was optimized for the stereoselective analysis of the antidepressant drug fluoxetine and its main demethylated metabolite norfluoxetine using a cyclodextrin-modified sodium phosphate buffer at pH 2.5. The combination of a neutral and a negatively charged cyclodextrin, dimethylated-beta- and phosphated-gamma-respectively, provided the baseline enantiomeric separation of the two compounds. The very low concentrations of chiral selectors employed together with the use of a high sensitivity detection cell of special design (zeta-shaped) in a diode array UV detector allowed us to reach a limit of detection of 0.005 and 0.01 microg/mL for fluoxetine and norfluoxetine, respectively. Analysis of fluoxetine and norfluoxetine standard mixtures showed a reproducibility of migration times and peak area and linearity in the concentration range of 0.1-2.0 microg/mL. The optimized method was applied to the analysis of clinical serum and plasma samples of patients under depression therapy. In all the analyzed samples the enantiomeric forms of fluoxetine and norfluoxetine were easily identified. The fluoxetine and metabolite enantiomeric ratio confirmed the stereoselectivity of the metabolic process of the fluoxetine drug in accordance with the literature data.

Antidepressive Agents, Second-Generation↗

Spectrophotometric determination of silicate traces in hemodialysis solutions.

Reliable methods for the analysis of silicon are of great importance, because it seems that the silicate anion can reduce aluminum bioavailability in patients undergoing dialysis. Thus, a simple and sensitive spectrophotometric method is described for the determination of silicate traces in dialysis solutions. The method is based on the reaction between silicate ions and excess ammonium molybdate reagent to give a yellow silico-molybdic complex. This complex is then reduced to the heteropoly blue compound by means of ascorbic acid. Absorbance values are measured at 830 nm, and are stable for more than 2 h. A good linearity was obtained up to 300 ng ml(-1) of silicon concentration. The accuracy and the precision of the method were good; relative standard deviation values of 2% intraday and of 3.9% interday for six replicates on 40 ng ml(-1) standard silicate solutions were found. Results of the analysis of some commercial hemodialysis solution samples, obtained by means of the 'standard additions' method, are provided.

Aluminum↗

Experimentally induced aggressiveness in adult children of alcoholics (ACOAs): preliminary behavioral and neuroendocrine findings.

OBJECTIVE: This study was conducted to determine the nature of the reaction of nonalcoholic adult children of alcoholic (ACOA) fathers to the experimental induction of aggression. Of particular interest was the relationship between biochemical factors and personality traits during a stressful event experienced by persons at risk for alcoholism. METHOD: Aggression was induced by a modified free-operant procedure in 14 ACOA and 14 non-ACOA subjects between 18 and 19 years of age with men and women represented in equal numbers. Neurotransmitter-hormonal assays from blood drawn immediately before, and 20 and 30 minutes after, starting the test included norepinephrine (NE), epinephrine (EPI), prolactin (PRL), growth hormone (GH) and cortisol (Cort). Personality traits were assessed by the Minnesota Multiphasic Personality Inventory (MMPI) Tridimensional Personality Questionnaire (TPQ) and the Buss-Durkee Hostility Inventory (BDHI). RESULTS: During the aggression induction session, ACOAs gained (F = 4.6, 1/13 df, p < .05) and subtracted (F = 9.2, 1/13 df, p < .005) significantly less money than non-ACOAs, evidence of lower outward-directed aggressiveness among ACOAs. Higher baseline plasma levels of Cort (F = 9.8, 1/13 df, p < .01) and PRL (F = 4.0, 1/13 df, p < .05) and decreased NE (F = 8.5, 1/13 df, p < .005) and GH (F = 10.9, 1/13 df, p < .001) responses during the experimental session were observed. On personality measures ACOAs scored higher than non-ACOAs on MMPI hysteria (F = 10.8, 1/13 df, p < .005), hypochondria (F = 20.1, 1/13 df, p < .001) and paranoia (F = 4.7, 1/13 df, p < 0.5) subscales, on the TPQ reward dependence (F = 10.9, 1/13 df, p < .005) subscale and on BDHI guilt (F = 15.7, 1/13 df, p < .001) and resentment (F = 6.4, 1/13 df, p < .05) subscales. CONCLUSION: These findings, preliminary in nature, support a hypothesis of inhibition of state and trait aggression in ACOAs in association with monoaminergic and endocrine changes.

Adolescent↗

Controlled insulin release from chitosan microparticles.

This study deals with the production of chitosan microparticles containing insulin by interfacial crosslinkage of chitosan solubilized in the aqueous phase of a water/oil dispersion in the presence of ascorbyl palmitate. The use of ascorbyl palmitate as interfacial crosslinker is based on its amphiphilic properties allowing its disposition at the water/oil interface of the preparative dispersion, thus permitting covalent bond formation with the amino groups of chitosan when its oxidation to dehydroascorbyl palmitate takes place during microparticle preparation. This preparation method produced microparticles characterized by high loading levels of insulin, completely releasing the drug in about 80 h at an almost constant release rate as determined by spectrophotometric and spectrofluorimetric methods. In contrast, the replacement of ascorbyl palmitate by dehydroascorbyl palmitate provided microparticles incompletely releasing the incorporated drug and characterized by a non-constant release rate over time due to the higher lipophilicity of dehydroascorbyl palmitate which hinders its disposition at the water/oil interface and thus decreases the crosslinking efficiency and increases the lipophilicity of the microparticle surface. The efficiency of the spectrofluorimetric and spectrophotometric methods used for determination of the stability and release of the insulin from the chitosan microparticles is also discussed.

Ascorbic Acid↗

Determination of fluoxetine and norfluoxetine in human plasma by high-pressure liquid chromatography with fluorescence detection.

Fluoxetine is an atypical antidepressant drug, which selectively inhibits the neuronal reuptake of serotonin, and is widely used in the treatment of depressive disorders. The aim of this research is the development of an HPLC method with fluorescence detection for the monitoring of fluoxetine plasma levels. The determination requires no more than 250 microl of plasma, which undergo solid phase extraction (SPE), then are injected in the HPLC. For the analytical separation a reversed phase C8 column (150 x 4.6 mm I.D.) was used, while the mobile phase was a mixture of acetonitrile and water containing perchloric acid and tetramethylammonium perchlorate (flow rate: 1 ml min(-1)). The very low levels of analytes in plasma required the employment of a fluorescence detector (lambda(exc) = 230 nm, lambda(em)=290 nm), which also granted a good selectivity. Fluoxetine is revealed as a single peak at a retention time of 9.7 min, while norfluoxetine, the main metabolite of fluoxetine, is revealed at a retention time of 8.1 min. Linearity was obtained over the concentration range 8-200 ng ml(-1) for both substances. The method seems suitable, in accuracy and precision, for the determination of fluoxetine plasma levels of patients; furthermore, it is rapid and sensitive.

Calibration↗

Analytical methods for the quality control of Prozac capsules.

Some analytical methods (two spectrophotometric and two chromatographic procedures) for the determination of fluoxetine in Prozac capsules are described. All of them are applied to the samples after extracting the drug with a methanol water mixture. The direct and derivative spectrophotometric methods are simple and reliable; the derivative method gives better recovery and lessens interference. Both methods show linearity in the 5-30 microg ml(-1) range of the fluoxetine concentration range. Both HPLC methods (spectrophotometric and spectrofluorimetric detection) use a tetramethylammonium perchlorate buffer-acetonitrile mixture as the mobile phase and a C8 reversed phase column. The UV detection is performed at 226 nm, while the fluorimetric detection is performed by exciting at 230 nm and revealing the emission at 290 nm. The HPLC method with UV detection is more precise, but the procedure with fluorimetric detection is more sensitive.

Antidepressive Agents, Second-Generation↗

Toxicological assessment of liquorice: biliary excretion in rats.

Glycyrrhizin (G) and its aglycone, glycyrrhetic acid (GA) have been prescribed for several therapeutic purposes. However, side effects have pointed out the problem of the toxicity of G. On the contrary, it was recently shown that the pure aqueous liquorice extract (LE), which also contains G, produces reduced adverse effects in rat and human, as compared to pure G, this is likely be related to differences in G bioavailability and the resulting pharmacokinetics of G and GA. Using a sensitive HPLC procedure for the determination of G and GA in rat bile, pharmacokinetics of G and GA in bile have been determined. The results of the analysis showed significantly lower concentrations of G in bile samples from rats treated with LE compared to pure G. Furthermore, LE presented a significant choleretic effect after both oral and i.v. administration, which increases the excretion rate of G. In case of GA, all the concentrations were very low, often below the detection limit. The results prompted us to assess the risk associated with liquorice intake and to determine the daily amount of pure liquorice root extract that can be safely consumed.

Administration, Oral↗

[The choleretic effects of licorice: identification and determination of the pharmacologically active components of Glycyrrhiza glabra].

Recent studies indicate that licorice extract, when administered per os or i.v., causes an evident choleretic effect in rats. Aim of this research is to identify and quantify those licorice constituents which are responsable for the observed choleresis. The quali-quantitative analysis of umbelliferon (7-idroxycoumarin), was at first performed by a fluorimetric method, subsequently by a more selective HPLC method. Moreover, this HPLC method allows the determination of glycyrrhizin, an important licorice constituent. Unlike the glycyrrhizin, which is present in a fairly large amount, umbelliferon resulted to be present at a very low concentration (at trace level), both in licorice and in bile. Research is in progress, aiming to determine the substances, beyond glycyrrhizin, which are responsable for the choleretic effect of licorice.

Animals↗

Effects of prolonged ingestion of graded doses of licorice by healthy volunteers.

Licorice can induce a hypermineralocorticoid syndrome. Current literature usually refers to the effects of sweets containing glycyrrhizin, but little is known about the consequences of a prolonged intake of "pure licorice". We administered graded daily doses of dried, aqueous extract of licorice root, containing 108, 217, 380 and 814 mg of glycyrrhizin, to 4 groups of 6 healthy volunteers of both sexes for 4 weeks. No significant effects occurred in groups 1 and 2. After 2 weeks, side effects leading to withdrawal from the protocol occurred in a female in group 3 (headache), a male with a family history of hypertension in group 4 (arterial hypertension), and a female also taking oral contraceptives in group 4 (hypertension, hypokalaemia and peripheral edema). In group 4, transient reduction in kalaemia and increase in body weight were found after 1 and 2 weeks, respectively. A depression of plasma renin activity occurred in groups 3 and 4. In healthy subjects, only the highest doses of licorice led to untoward effects. These were favoured by subclinical disease or oral contraceptives, and were less common and pronounced than what has been reported after the intake of glycyrrhizin taken as such or as a flavouring agent in confectionery products.

Adult↗

Interaction of licorice on glycyrrhizin pharmacokinetics.

The effects of components of aqueous licorice root extract (LE) on the pharmacokinetics of glycyrrhizin (G) and glycyrrhetic acid (GA) were investigated in rats and humans. The aim of this work was to define the role of pharmacokinetics in G toxicity. In the procedure, G and GA were detected in biological fluids by means of recently improved HPLC methods. Significantly lower G and GA plasma levels were found in rats and humans treated with LE compared to the levels obtained with those in which G alone was administered. The pharmacokinetic curves showed significant differences in the areas under the plasma-time curve (AUC), Cmax, and Tmax parameters. The data obtained from urine samples are in agreement with the above results and confirm a reduced bioavailability of G present in LE compared to pure G. This should be attributed to the interaction during intestinal absorption between the G constituent and the several components in LE. The modified bioavailability could explain the various clinical adverse effects resulting from the chronic oral administration of G alone as opposed to LE.

Administration, Oral↗

HPLC determination of glycyrrhizin and glycyrrhetic acid in biological fluids, after licorice extract administration to humans and rats.

Simple and sensitive HPLC methods were developed for the determination of glycyrrhyzin (G) and its main metabolite glycyrrhetic acid (GA) in biological samples, in order to investigate the pharmacokinetic behaviour of G after oral administration of licorice extract (LE) or G to humans and rats. The analysis have been carried out by HPLC with UV detector (251 nm), after a careful pretreatment of the samples. These methods are suitable in terms of precision and accuracy for the G and GA determination in plasma and urine of human volunteers and in bile, plasma ad urine of rats.

Animals↗

Bioavailability of glycyrrhizin and licorice extract in rat and human plasma as detected by a HPLC method.

The pharmacokinetic behaviour of glycyrrhizin (1) was investigated in order to evaluate the difference in bioavailability after oral administration of licorice extract (LE) or glycyrrhizin (1) to rats and humans. For this study, two reliable HPLC methods were developed for the dosage of the levels of 1 and of its metabolite, the glycyrrhetic acid (2) in plasma samples. The determinations were carried out by HPLC on a reversed phase column with UV detector (251 nm), after a careful extraction step of 1 and 2 from the biological matrix. These methods afford good accuracy and satisfactory precision and they allow the determination of both compounds at levels as low as 200 ng/ml. The analytical results improved the knowledge of 1 pharmacokinetics, showing a significantly reduced bioavailability, when administered as LE compared to 1 when administered as such.

Animals↗

Inhibition of poly(ADP-ribose) polymerization preserves the glutathione pool and reverses cytotoxicity in hydrogen peroxide-treated lymphocytes.

DNA damage caused by oxygen radicals activates poly(ADP-ribosyl) polymerase (pADPRP), a nuclear enzyme that utilizes NAD+ as substrate. It has been demonstrated that pharmacological inactivation of pADPRP rescues human lymphocytes damaged by oxygen radicals, but not those damaged by equitoxic doses of ionizing radiation. In the present paper we demonstrate that the NAD+ pool decreases after both damaging treatments and is preserved in a similar fashion by pADPRP inhibition. On the contrary, the ATP pool, cell energy charge and reduced thiols are decreased only by the administration of oxygen radicals, and are preserved if poly(ADP)ribosylation is inhibited. In fact, treatment with oxidant agents depletes the cell energy pools owing to the simultaneous demands of the glutathione (GSH)/NADPH cycle and pADPRP-driven NAD+ consumption, while in irradiated cells only the latter mechanism operates. We suggest that, when pADPRP is inhibited, enough energy is available for the preservation of cell thiols, thereby allowing oxidant-treated cells to survive and undergo mitosis. Thus, GSH and energy shortage appear to be the main cause of cell death in oxidant-injured cells.

Adenosine Triphosphate↗