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Biomedical subjects

P Prognon

Publications and source records attributed to P Prognon.

At least 19 recordsLinked to original sources

Stability and sterility of meglumine gadoterate injection repackaged in plastic syringes.

The analytical and microbiological stability of meglumine gadoterate (Dotarem) repackaged in polypropylene syringe for 3 months at either +4 degrees C or room temperature was studied. For analytical study: six polypropylene syringes (20 ml) were filled with 15 ml of meglumine gadoterate. Three syringes were stored at 4+/-2 degrees C and three at 25+/-2 degrees C, all syringes were kept upright and protected from daylight. Samples were taken on days 0, 6, 14, 30, 45, 60, 75 and 90. Meglumine gadoterate and its degradation product (free Gd3+) concentrations were obtained using a specific HPLC assay. Osmolality and pH determination were made on days 0, 14, 45 and 90. For microbiological study: 28 plastic syringes (5 ml) were filled with 2.5 ml of meglumine gadoterate. Syringes were stored at 25+/-2 degrees C and protected from daylight. At each day of analysis (0, 15, 35, 45, 60, 75 and 90), four syringes were tested as described in European Pharmacopoeia. After 90 days the concentration of gadoterate remained unchanged and no free Gd3+ were detected. The injectable solution of this gadolinium contrast agent was sterile according to European Pharmacopoeia guidelines. The meglumine gadoterate repackaged in polypropylene syringe was stable for 3 months at all the temperatures studied.

Contrast Media↗

Application of the assay of aflatoxins by liquid chromatography with fluorescence detection in food analysis.

HPLC using fluorescence detection has already become the most accepted method for the determination of aflatoxins due to its several advantages over other analytical methods. Both normal- and reversed-phase HPLC can be used. However the reversed-phase HPLC methods are more popular. Liquid chromatographic determination of aflatoxins using fluorescence detection and its application in food analysis is reviewed in this article.

Aflatoxins↗

Postcolumn fluorescence as an alternative to evaporative light scattering detection for ceramide analysis with gradient elution in non-aqueous reversed-phase liquid chromatography.

Ceramide analysis was developed with gradient elution in non-aqueous reversed-phase liquid chromatography with evaporative light scattering detection (ELSD) or postcolumn fluorescence detection. Fluorescence detection (excitation, 360 nm; emission, 425 nm) after postcolumn formation of mixed assemblies between eluted ceramides and 1,6-diphenyl-1,3,5-hexatriene was developed. In comparison with ELSD, fluorescence detection allows a better detection of the minor species ceramide from ceramide type III (commercial mixture of non-hydroxy fatty acid-sphingosine) and appears to be more sensitive for quantitation of ceramides at low concentrations. The fluorescence response is linear over a wide range of injected amount of ceramide III (expressed as stearoyl-phytosphingosine): 10 ng to 1000 ng. The response of ELSD is non linear but can be linearized in double logarithmic coordinates for calculations over a narrow range, e.g. between 10 to 350 ng ceramide III injected. The lower quantitation limits of these two detectors are similar: 5 ng ceramide III was injected.

Ceramides↗

Cerium-doped diosmectite for topical application studies of the cerium-clay interaction.

Cerium is used for its antiseptic and immunomodulatory properties in burn injury. We have developed a cerium-doped clay to replace existing ointments. Adsorption and release of cerium (Ce3+) by diosmectite were studied at 22+/-2 degrees C, in the presence of various other cationic species. Simple spectrofluorimetric determination of cerium was used (lambdaexc=240 nm/lambdaem=360 nm). Cerium binding reached a plateau within 2 min and was a function of the electrolyte content of the solution in contact with the clay. Langmuir isotherm treatment led to a maximal binding capacity of 66 mg of Ce3+ per gram of clay. Partial release occurred within 2 min (19% in the presence of isotonic NaCl solution). The ionic strength of the solution, and the ionic radius and charge of the electrolytes present in the bathing solution significantly influenced cerium release, in contrast to pH and temperature changes. These results strongly point to a cationic exchange mechanism between diosmectite and cerium solution.

Administration, Topical↗

Flourescence-assay on traces of protein on re-usable medical devices: cleaning efficiency.

The cleaning of re-usable medical devices before disinfection or sterilization is recognized as being an essential phase. Detection of residual proteins can be used to validate the process, provided a sufficiently sensitive method is employed. A fluorescent method is presented, using orthophtalaldehyde (OPA) bound to N,N dimethyl-2-mercaptoethylammonium, to demonstrate the presence of amino acids on a medical device following cleaning. The sensitivity of this method (10-5 g/l) was assessed and the applicability of this detection technique is verified, using three types of carriers (steel blades, glass tubes or ceramic penicylinders), three types of contaminants (yeast extract, bovine albumin with native sheep's blood and formaldehyde fixed fibrin). In this context, studies involving formaldehyde-fixed fibrin are more sensitive and are to be recommended.

Disinfection↗

Fluorescence and mass spectrometry studies of meta-tetra(hydroxyphenyl)chlorin photoproducts.

The meta-tetra(hydroxyphenyl)chlorin (m-THPC), a second-generation sensitizer used in photodynamic therapy (PDT), is currently under clinical trial. In vivo fluorometry provides direct evidence that photobleaching processes are induced at the tumor site during PDT. Photoproduct formation has thus to be taken into account to fully understand PDT treatment. A preliminary step is to determine the fluorescence characteristics of photoproducts formed in solution. Solutions of m-THPC irradiated at 514 nm have been separated by HPLC using absorption and fluorescence detection. Six main photoproducts have been isolated. According to matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOFMS) results, five fluorescent photoproducts emitting at 652 nm have been attributed to three mono-, one di- and one tri-hydroxy derivatives (m/z 697, 713 and 729, respectively). Fluorescence characteristics of mono-hydroxy forms were found to be similar to those of m-THPC, whereas fluorescence yields in di- and tri-hydroxy derivatives were very low. Another product, corresponding to a MALDI-TOF MS main signal at m/z 542, showed an absorption spectrum maximum at 522 nm while a weak fluorescence was detected at 480 nm. The loss of the Soret band suggests that this photoproduct results from the opening of the reduced pyrrole ring. The part played by each of these products in the photobleaching phenomenon of m-THPC is discussed.

Chromatography, High Pressure Liquid↗

Interaction between cyclodextrins and aflatoxins Q1, M1 and P1. Fluorescence and chromatographic studies.

The fluorescence properties of the aflatoxins M1, Q1, P1 in solution and the effect of various cyclodextrins (alpha-, beta-, gamma-, hydroxypropyl-beta- and alpha-beta-heptakis-di-O-methyl-beta-) on their fluorescence emission were studied. Among the aflatoxins, a substantial enhancement of the fluorescence emission of aflatoxin Q1 in the presence of aqueous solutions of alpha-, beta-, hydroxypropyl-beta, and alpha-beta-heptakis-di-O-methyl-beta-cyclodextrin, was observed. On the contrary, gamma-cyclodextrin proved to be inefficient to enhance the fluorescence properties of this compound. No important fluorescence enhancement was found for aflatoxins P1 or M1 for any of the cyclodextrin derivatives tested. The complex formation constant (Kf) of these compounds with beta-cyclodextrin was chromatographically determined, and from the results obtained, we can conclude that Kf cannot be used alone to explain the fluorescence increase. Thermodynamic studies showed that delta-H and delta-S parameters, associated with the partition of aflatoxins in RP-HPLC, increased when beta-cyclodextrin was added to the eluent.

Aflatoxin M1↗

Determination of total cysteamine in human serum by a high-performance liquid chromatography with fluorescence detection.

A convenient, reliable and rapid method for determination of total cysteamine in human plasma by high-performance liquid chromatography with fluorescence detection is reported. This assay involves reduction of samples with dithiothreitol, derivatization of total cysteamine by addition of monobromobimane and protein precipitation by perchloric acid. The calibration curve was linear in the range 2-150 nmol ml-1 and the detection limit was 0.5 nmol ml-1. This method was successfully applied for a pharmacokinetic study of three cysteamine derivatives in healthy volunteers without any interference from coexisting substances.

Biological Availability↗

Photochemically induced fluorimetric detection of tianeptine and some of its metabolites. Application to pharmaceutical preparation.

The photochemically induced fluorescence (PIF) properties of tianeptine and some of its metabolites were investigated in acidic (pH 2.3) water-alcohol mixtures at room temperature. Two PIF methods were developed, including bulk solution and flow injection analysis (FIA). Linear calibration plots were established over a concentration range of more than one order of magnitude. Limits of detection ranged from 15 ng ml-1 for FIA-PIF to 25 ng ml-1 in bulk solution. The RSDs were between 3 and 5%. The PIF methods were applied to the determination of tianeptine in a pharmaceutical preparation with recoveries varying from 96 to 106% in bulk solutions and from 98 to 106% for FIA-PIF.

Antidepressive Agents, Tricyclic↗

Liquid chromatographic determination using lanthanides as time-resolved luminescence probes for drugs and xenobiotics: advantages and limitations.

Lanthanide sensitized luminescence is a very attractive alternative to UV detection and other luminescence techniques, i.e., fluorescence and phosphorescence, in separation science for the detection of drugs and xenobiotics because of the large Stokes shift, narrow emission bands and long lifetime. Some published applications of HPLC determination with lanthanide (Ln3+) sensitized luminescence detection are reviewed. Advantages and limitations of this technique are discussed. Normal-phase (NP) HPLC is not influenced by the quenching effect of water whereas reversed-phase (RP) HPLC is applicable to more compounds than NP-HPLC. However, pH adjustment and the quenching effect of water on Ln3+ luminescence are the main drawbacks of RP-HPLC. Elution properties and the need for pH adjustment are two arguments for selecting the mode of addition of Ln3+, i.e., pre- or post-column in the HPLC system. Sensitized Ln3+ luminescence detection is a much more specific method of detection than UV or fluorescence detection after HPLC separation but nevertheless, in some cases, does not always exhibit a significant increase in analytical performance when the donor itself is a strong fluorophore. The development of more powerful excitation sources could improve the limit of detection of the Ln3+ sensitized detection technique. This review suggests that it would be useful to obtain predicting factors about the drug to establish whether the latter is suitable to be measured using an HPLC-Ln3+ approach.

Chromatography, High Pressure Liquid↗

Study protocol: stability of morphine injected without preservative, delivered with a disposable infusion device.

BACKGROUND AND OBJECTIVE: Morphine hydrochloride, a major analgesic drug, is being increasingly administered using portable disposable infusion devices. The objective of this study was to investigate the stability of morphine in such a system at two concentrations (2.50 and 5.00 mg/ ml) over a 30-day period. METHOD: High-performance liquid chromatography of stored morphine solutions. RESULTS: The best stability was observed with disposable infusion devices filled with a morphine solution containing sodium metabisulphite as a preservative. No breakdown products were detected after 1 month of storage at room temperature, in light or darkness. On the other hand, 2.50 and 5.00 mg/ml morphine solutions without sodium metabisulphite, stored in the infusion device led to the formation of 0.205% and 0.235% of pseudomorphine, respectively, after 6 days of storage in the light, and 1.50% and 0.94% after 30 days storage. CONCLUSION: Morphine hydrochloride solutions stored in disposable infusion devices degraded very slowly, particularly when preserved with sodium metabisulphite. The solutions are stable over 5 days, the maximum period of storage normally required when using disposable infusers.

Analgesics, Opioid↗

Rapid semi-quantitative fluorimetric determination of citrinin in fungal cultures isolated from cheese and cheese factories.

A new rapid semi-quantitative fluorimetric assay for citrinin production testing in mould cultures has been developed. The chemical structure of the citrinin makes it a weak native fluorophore. This fluorescence can be strongly enhanced in an acidic environment. A standard curve where the concentration of HCl needed to show the yellow fluorescence signal of different concentrations of citrinin was established, thus providing a semi-quantitative method to prove the capacity of toxin production of fungal cultures. Two Penicillium strains from the Spanish National Collection of Type Cultures, were studied for the toxin production on YES broth at 25 degrees C for 21 d. The culture was assayed daily for the presence/absence and quantification of citrinin by adding the HCl concentration set, and also quantified by RP-HPLC as a confirmation procedure. Experiments demonstrate that 5 d are necessary to show the presence of citrinin. As an illustration, a total of 48 strains of Penicillium isolated from cheese and cheese factories were analysed with the proposed method.

Cheese↗

Simple and sensitive high-performance liquid chromatography--fluorescence method for the determination of citrinin application to the analysis of fungal cultures and cheese extracts.

A new and highly sensitive method for the detection of the important mycotoxin, citrinin, has been developed. Spectroscopic studies demonstrate that the fluorescence of this metabolite is influenced by the pH of the environment. This fact was exploited in the chromatographic determination of citrinin with fluorescence detection. The proposed method, based on the addition of 1 M hydrochloric acid as an acidic post-column reagent, has a limit of detection of 0.9 center dot 10(-7) M. Analytical validation shows that linearity can be assumed from 2 center dot 10(-7) to 10(-4) M citrinin. The repeatability and reproducibility are satisfactory, with R.S.D. = 5.1% (n = 9, c = 10(-5) M) and R.S.D. = 7.2% (n = 9, c = 10(-5) M). The method was also applied to the determination of this mycotoxin produced by mould cultures isolated from soft cheese and also from soft cheese and also from cheese extracts spiked with citrinin. The specificity of the method is demonstrated and the necessity for post-column acidification is illustrated on real samples.

Cheese↗

Postcolumn excitation of aflatoxins using cyclodextrins in liquid chromatography for food analysis.

Measurement of fluorescence increase was used for the comparative quantification of the effect that several cyclodextrins (alpha-, beta-, heptakis-2,6-beta-omicron-dimethyl- and gamma-) produce on the fluorescent response of aflatoxins B1 and G1. This constitutes a new chromatographic method with stability of the mobile phase, and shows general improvements in the chromatographic conditions with respect to other methods (especially those using an iodine reservoir as a postcolumn reactor). A C18-type column was used, with methanol-water (60:40, v/v) as the mobile phase. The excitation phase of the natural fluorescence of aflatoxins, a 10(-2) M solution of each cyclodextrin, was introduced postcolumn. The determination of the elution order aflatoxin G2 > G1 > B2 > B1 was performed for each phase in less than 15 min. As expected using an aqueous-alcoholic medium, an increase in the fluorescence response of aflatoxins with an unsaturated furanic ring was found to occur with all the cyclodextrins studied, except gamma-cyclodextrin. The observed increase was larger for heptakis-2,6-beta-omicron-dimethyl- than for beta-cyclodextrin (to our knowledge, the only cyclodextrin previously described in the literature to serve for the determination of aflatoxins). The difference is of the order of 70.1-fold in the case of aflatoxin G1 and 45.2-fold in the case of aflatoxin B1. The detection limit in the mobile phase used was determined (for aflatoxin B1) for beta-cyclodextrin and 2,6-beta-omicron-dimethylcyclodextrin (signal-to-noise ratio 1:3) to be 4 and 9 mg 1(-1), respectively.

Aflatoxin B1↗

Effects of pH and solvent on the fluorescence properties of biomedically important benzamides. Application to determination in drugs and in human urine.

The fluorescence properties of five substituted benzamides, including alizapride, metoclopramide, sulpiride, sultopride and tiapride, were investigated at several pH values and in various solvents (dimethyl sulfoxide, ethanol, ethylene glycol, methanol, propan-2-ol, tetrahydrofuran and water). Except for alizapride, the fluorescence intensities were found to be higher at acidic (1-6) than at alkaline (8-12) pH values. Using the optimum solvent (aqueous solutions) and appropriate pH conditions, linear spectrofluorimetric calibration curves were established over a concentration range of about two orders of magnitude, with correlation coefficients larger than 0.996. Limits of detection were between 1 and 13 ng ml-1, depending on the compound. The method was applied to the determination of benzamides in pharmaceutical preparations and in human urine, with recoveries ranging from 94 to 108% and from 93 to 104%, respectively.

Benzamides↗

[Contribution to the study of phenothiazine detection by inhibition of phosphorescence in liquid chromatography].

Diacetyl, in deaerated solutions, is a major room temperature phosphorescent compound. Its phosphorescence quenching is proposed in order to quantify results on phenothiazine derivatives. It is established from the luminescent data on phenothiazines that diacetyl phosphorescence can be quenched quantitatively by these compounds. This detection mode is applied to liquid chromatography limitations as well as the mechanism of the quenching are discussed.

Chromatography, High Pressure Liquid↗

Roles of divalent cations and pH in mechanism of action of nitroxoline against Escherichia coli strains.

The antibacterial activity of nitroxoline (NIT), an antibiotic used in the treatment of acute or recurrent urinary tract infections caused by Escherichia coli, is decreased in the presence of Mg2+ and Mn2+ but not Ca2+. In order to elucidate the interaction between this drug and the divalent cations, spectrophotometric studies based on the natural absorption of the nitroxoline moiety were conducted. In the presence of the divalent metal ions, a shift in the NIT A448 suggested the formation of drug-ion complexes, for which the stability followed the order Mn2+ > Mg2+ > Ca2+. A clear correlation was found between the chelating property and antibacterial activity of NIT; both were pH dependent. A convenient colorimetric method for the determination of NIT uptake by bacterial cells was also developed. Uptake was energy independent and showed biphasic kinetics: a rapid association with cells and then a slower increase in cell-associated NIT which reached a plateau. NIT uptake was reduced in the presence of magnesium. The implications of metal ion complexation and pH on the clinical efficacy of NIT are discussed.

Anti-Infective Agents, Urinary↗

[Microbiological consequences of chelation of bivalent metal cations by nitroxoline].

The present work deals with the physico-chemical and microbiological investigations made in order to explain the role of divalent cations in the mechanism of the action of nitroxoline (NIT), an antibiotic used in the treatment of uncomplicated urinary tract infections. Preliminary studies reported that bacteriostatic and bactericidal activities of NIT on Escherichia coli strains are decreased in the presence of Mg2+ and Mn2+ but not with Ca2+. A spectrophotometric study, conducted in order to elucidate the interaction between metal ions and NIT, showed the formation of drug-metal ion complexes. In addition, we examined the relationships between the metal ions-chelating property of NIT and its effects on the decrease of the mannose sensitive hemagglutination titer and the production of siderophores from E. coli. The results suggested that these antimicrobial activities of NIT would also be due to the complexation of metal ions by this molecule.

Anti-Infective Agents, Urinary↗