RESUMEN
Biodiesel is an important alternative fuel produced from animal fats or vegetable oils. According to several world regulatory agencies, the acceptable threshold content for free glycerol in biodiesel is 200 mg kg-1. Exceeding concentrations may yield high amounts of acrolein after combustion. The analytical methods for glycerol determination are usually preceded by conventional liquid-liquid extraction that may affect precision and accuracy and impair analytical frequency. In this work, a multi-pumping flow system was proposed for the online dispersive liquid-liquid extraction of free glycerol from biodiesel followed by spectrophotometric determination. The analyte was transferred to the aqueous phase by mixing the sample and water under a pulsed flow regime. The emulsion was directed towards a retention column to remove the organic phase before chemical derivatization. Glycerol was oxidized by NaIO4 to generate formaldehyde, which reacted with acetylacetone in an ammonium acetate medium to yield 3,5-diacetyl-1,4-dihydrolutidine (λMAX = 412 nm). The optimization of the main parameters of the system was carried out by multivariate methods. The screening of variables was performed by fractional factorial design 24-1. The models for free glycerol determination and extraction were refined by central composite and full factorial 23, respectively. In both cases, the validation was carried out by analysis of variance, which yielded a satisfactory F-test value. After optimization, a linear range from 3.0 to 50.0 mg L-1 glycerol was observed. The detection limit, coefficient of variation and determination frequency were estimated to be 2.0 mg L-1 (n = 20; 99.7% confidence level), 4.2-6.0% (n = 20) and 16 h-1, respectively. The efficiency of the process was estimated to be 66%. After each extraction, the retention column (filled with 185 mg of glass microfiber) was washed with 50% ethanol solution to avoid carry-over effects. The comparative analyses of samples by the proposed and reference methods demonstrated the accuracy of the developed procedure at a 95% confidence level. Recoveries between 86 and 101% also indicated that the proposed procedure is accurate, suitable and reliable for online extraction and determination of free glycerol in biodiesel.
Asunto(s)
Glicerol , Microextracción en Fase Líquida , Animales , Glicerol/análisis , Microextracción en Fase Líquida/métodos , Biocombustibles/análisis , Agua/química , Extracción Líquido-LíquidoRESUMEN
Drug quality assessment and stress testing are important to ensure both treatment efficacy and patient safety. High performance liquid chromatography may be considered a standard technique for pharmaceutical analysis, showing good precision and accuracy, but it also involves relatively high cost and low analytical frequency. Flow injection analysis presents high sample throughput, lower cost and might be used for selective drug analysis with an appropriate assay and/or detector. In this paper, for the first time, photoreactions promoted by UV radiation were employed for reagentless spectrophotometric determination of hydrochlorothiazide. Optimized parameters led to a linear range of 50 to 500 mg L-1, estimated limit of detection of 3.0 mg L-1 and 24 determinations per hour. The use of diluted NaOH solution as a carrier allowed solubilization of hydrochlorothiazide and analysis without organic solvents. The presence of the most common excipients was evaluated and no significant interferences were observed. The results from the analysis of samples by the proposed and by the reference procedures demonstrated accuracy and matching results. The proposed in-line photolysis of the pharmaceutical, performed in 5 min, is a promising alternative to the conventional hydrolytic forced degradation, which requires elevated temperature and prolonged time period. To evaluate the degree of photoconversion, a capillary zone electrophoresis method was developed, which performed well for separations manifesting good analytical frequency and reduced amount of waste. The combination of in-line photodegradation followed by separation by capillary electrophoresis is a promising approach for the stress test of hydrochlorothiazide in pharmaceutical formulations.
Asunto(s)
Hidroclorotiazida , Preparaciones Farmacéuticas , Cromatografía Líquida de Alta Presión , Electroforesis Capilar , HumanosRESUMEN
Sodium cysteamine phosphate is a prodrug derivative of cysteamine that can be used in cystinosis treatment. Although titrimetric assays are very well established and precise, iodimetric determination of sodium cysteamine phosphate requires considerably more carefulness and time from the analyst than usual. The possibility to assess sodium cysteamine phosphate by CE was evaluated by means of the quantification of its oxidation product, cystamine, which is a more suitable substance to be used as primary standard than sodium cysteamine phosphate. Apparently, this approach should be straightforward, but systematic differences between the results obtained with CE and titrimetric assays were noticed. MS and CE-MS were employed to aid in the investigation of the possible causes of imprecision of the sodium cysteamine phosphate titration and CE determination. For this purpose, a simple and inexpensive ESI source was constructed. It was observed that cystamine is not the final product of the cysteamine and/or sodium cysteamine phosphate iodine-oxidation and other species besides cystamine may be formed depending on the reaction conditions, which explains the difficulties observed in the sodium cysteamine phosphate quantification.
Asunto(s)
Cistamina/química , Cisteamina/química , Espectrometría de Masa por Ionización de Electrospray/métodos , Oxidación-ReducciónRESUMEN
Ritonavir is a synthetic peptidomimetic human immunodeficiency virus (HIV) protease inhibitor employed in the treatment of AIDS since 1996. Synthetic precursors are potential impurities in the final product. In the present work a micellar electrokinetic chromatography (MEKC) method for the separation of Ritonavir from three available synthetic precursors was developed. The optimized separation is performed in a background electrolyte composed of sodium tetraborate (pH 9.6; 15mM) containing sodium dodecylsulfate (30mM) and acetonitrile (18%, v/v). Mass spectrometry was used to confirm the identity of the tested substances. Good repeatability was observed for migration time (RSD about 0.4%) and peak area (RSD about 0.8%). The limits of detection (LOD) obtained allow the determination of two of the impurities at levels as low as 0.005% m/m, and one at a level of 0.3% m/m.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar/métodos , Contaminación de Medicamentos , Micelas , Ritonavir/análisis , Cromatografía Capilar Electrocinética Micelar/normas , Inhibidores de la Proteasa del VIH/análisis , Inhibidores de la Proteasa del VIH/química , Ritonavir/químicaRESUMEN
A capillary zone electrophoresis (CZE) method was developed aiming the analysis of Hippeastrum hybrid agglutinin (HHA) samples. HHA is presently being tested as a vaginal microbicide to prevent HIV transmission. It acts by direct binding to mannose residues that are abundantly present on the HIV gp120 envelope and so interrupts the virus entry process. The final CZE method employs 50mM sodium tetraborate (pH 9.9) as background electrolyte. In this condition, a cluster of about 30 isoform peaks is obtained, with very repeatable patterns. RSDs in the order of 0.2% for the migration time and detection sensitivity in the order of 70microgml(-1) were achieved.
Asunto(s)
Aglutininas/análisis , Electroforesis Capilar/métodos , Liliaceae/química , Concentración de Iones de Hidrógeno , Peso Molecular , Reproducibilidad de los ResultadosRESUMEN
A capillary electrophoresis method for determination of impurities in sodium cysteamine phosphate-an alternative drug to use in place of cysteamine (Cystagon), Mylan Laboratories Inc.) in the treatment of cystinosis-was developed. The administration of cysteamine, divided in four doses due to the short half-life of this drug, is a helpful treatment, but several patients show intolerance, due to the very unpleasant odor and taste of cysteamine. Sodium cysteamine phosphate is less organoleptic aversive and also has a larger active time, allowing the compression of the doses to 2 per day, increasing the acceptance of the drug. In the developed method the two main decomposition products of sodium cysteamine phosphate, cystamine and cysteamine, can be determined with LOQs of 30 microg/ml (0.2%) and 16 microg/ml (0.1%), respectively. The background electrolyte is 15 mM ammonium acetate (pH 8.85) with 10% methanol and the separation takes less than 4 min. UV detection is performed at 195 nm. This volatile method was developed with the purpose of further hyphenation to a mass spectrometer.
Asunto(s)
Cisteamina/análisis , Contaminación de Medicamentos , Electroforesis Capilar/métodos , Concentración de Iones de HidrógenoRESUMEN
The separation and detection of common mono- and disaccharides by capillary electrophoresis (CE) with contactless conductivity detection (CCD) is presented. At high values of pH, the sugars are converted to anionic species that can be separated by CE and indirectly detected by CCD. The main anionic species present in the running electrolytes are hydroxide and phosphate, which have greater mobility than the ionized sugars, and, thus, the indirect detection is possible. The method was applied to analysis of glucose, fructose, and sucrose in soft drinks, isotonic beverages, fruit juice, and sugarcane spirits. Galactose was used as internal standard in all cases. Plate numbers range from ca. 70,700 to 168,200 and the limits of detection from 13 to 31 microM.