Determinação simultânea de resíduos de sulfametoxazol e trimetoprima em superfícies de equipamentos de produção

A cleaning validation method was developed and validated, based on swabbing sampling and simultaneous chromatographic determination of sulfamethoxazole (SMX) and trimethoprim (TMP) residues. The method presented limits of detection of 0.06 mg mL-1 for SMX and 0.09 mg mL-1 for TMP. It was considered selective, precise, accurate and robust according to the guidelines from ANVISA, the Brazilian regulatory agency, and International Conference on Harmonization. Mean swab recovery factors of 98.5% for SMX and 97.7% for TMP were obtained for spiked stainless steel plates. The method was successfully applied to the assay of actual swab samples collected from eleven points on an equipment surface.

Produção, purificação e aumento da performance de ciclodextrina glicosiltransferases para produção de ciclodextrinas

Biospecific affinity chromatography was used to purify three cyclodextrin glycosyl transferases (CGTases) obtained from microorganisms isolated of soil. The cyclodextrins (CDs) production by CGTases was evaluated using starches from different sources. CDs were measured through the Complexation Theory and by HPLC. CGTase from Bacillus firmus strain 7B showed the best production (30 mmol/L of β-CD and 4.3 mmol/L of γ-CD), and its cultivation conditions were optimized. The maximum enzymatic activity was achieved using lung peptone, soluble starch and agitation speed of 160 rpm. Studied CGTases were shown quite interesting for the industrial production of CDs.

Validação de métodos cromatográficos de análise: um experimento de fácil aplicação utilizando cromatografia líquida de alta eficiência (CLAE) e os princípios da "Química Verde" na determinação de metilxantinas em bebidas

The validation of analytical methods is an important step in quality control. The main objective of this study is to propose an HPLC experiment to verify the parameters of validation of chromatographic methods, based on green chemistry principles, which can be used in experimental courses of chemistry and related areas.

Efeito da acidez e de modificadores orgânicos na determinação de metilxantinas: um experimento de cromatografia liquida de alta eficiência (CLAE) empregando otimização uni e multivariada

In this work a new experiment using HPLC is proposed in order to explore the role of acidity and the organic modifiers in the determination of methylxanthines in tea and coffee. Multivariate and univariate optimizations of the experimental conditions were used.

Determinação de compostos fenólicos por cromatografia líquida de alta eficiência isocrática durante estacionamento da erva-mate

Different phenolic compound, 5- caffeoylquinic acid (5-CQA), caffeic acid (AC) and rutin (Ru) contents of yerba-mate (Ilex paraguariensis) Brazilian samples of 06 different regions of São Mateus - Paraná, during natural and accelerated industrial storage, were evaluated. For quantification, a reverse phase HPLC isocratic method was developed and validated using methanol:water (35:65 v/v) acidified with 0.5% acetic acid as mobile phase and a photodiode array detector. The six sample global average contents were (34.90 and 36.10 mg g-1) for 5-CQA, (0.18 mg g-1 and 0.23 mg g-1) for AC and (7.12 and 7.18 mg g-1) for Ru, respectively, for the natural and accelerated storage systems. The results showed that the 5-CQA and Ru content are kept constant during the storage while AC content increase only during accelerated storage.

Desenvolvimento e validação de método analítico por cromatografia líquida de alta eficiência (CLAE) para determinação de associação de ampicilinas em pó liofilizado para injeção

An analytical method has been developed and validated for the determination of an association of ampicillins in a lyophilized powder for injection by HPLC. The advantage of chromatographic method other than the microbiological one is that, it is possible to monitor precisely, out-of-specification results in quality control processes and also during stability studies, in which an association of ampicillins is present. The proposed HPLC method was developed by using forced degraded samples, in order to reach a selective analysis of ampicillins when in the presence of their degradation products. It was possible to detect benzatine and through indirect calculation, to determine the ampicillin sodium in the drug sample. The method showed to be selective, accurate, precise, robust and linear (from 45.92 to 36.04 μg mL-1 of total ampicillin and from 14.53 to 43.28 μg mL-1 of benzatine). The accuracy determined from recovery test, gave results in the range of 99.41% of total ampicillin to 100.31% of benzatine. Hence, it can be concluded that the proposed HPLC method is applicable for ampicillins determination.

Determinação simultânea de precursores de serotonina - triptofano e 5-hidroxitriptofano - em café

Epidemiological studies attributed positive effects in the central nervous system (CNS) to coffee. Among possible active constituents, serotonin, a neurotransmitter in the CNS, is present; but dietary sources do not cross the blood-brain barrier. Tryptophan and 5-hidroxytryptophan (5-HTP) are serotonin precursors and can affect brain concentrations. An ion-pair-HPLC, post-column derivatization with o-phthalaldehyde and fluorimetric detection before and after hydrolysis with NaOH and extraction with methanol:water was developed for the simultaneous determination of these compounds. It was selective, sensitive (LOD = 0.3 and 0.2 μg/mL), precise (91.3 and 94.2% recovery for tryptophan and 5-HTP, respectively), and linear from 0.3 to 40 μg/mL for both compounds. It was applied to green and roasted arabica and robusta coffees.

Determination of free urinary cortisol in cushing's syndrome using reversed-phase high performance liquid chromatography

Determination of free urinary cortisol is a test of choice in the diagnosis of Cushing's syndrome. In this study, cortisol was quantified using reversed-phase high-performance liquid chromatography (RP-HPLC) in urine samples previously extracted with ether and using triamcinolone acetonide as internal standard (IS). A BDS-Hypersil-C18® column, water-acetonitrile (72:28; v/v), with a flow rate of 1.0 mL/min and detection at 243 nm were used. This method showed to be both effective and efficient, with sensitivity and linearity ranging from 2.50 to 150 μg/L, and can be used in substitution to the radioimmunoassay technique within this concentration range.

Flow injection green method for the quantitative analysis of ketoconazole in pharmaceutical preparations

A flow injection method for the quantitative analysis of ketoconazole in tablets, based on the reaction with iron (III) ions, is presented. Ketoconazole forms a red complex with iron ions in an acid medium, with maximum absorbance at 495 nm. The detection limit was estimated to be 1×10--4 mol L-1; the quantitation limit is about 3×10--4 mol L-1 and approximately 30 determinations can be performed in an hour. The results were compared with those obtained with a reference HPLC method. Statistical comparisons were done using the Student's t procedure and the F test. Complete agreement was found at the 0.95 significance level between the proposed flow injection and the HPLC procedures. The two methods present similar precision, i.e., for HPLC the mean relative standard deviation was ca. 1.2% and for FIA ca. 1.6%.

Determination of 5-aminosalicylic acid in pharmaceutical formulations by square wave voltammetry at pencil graphite electrodes

An analytical method for the determination of the anti-inflammatory drug 5-aminosalicylic acid (5-ASA) in pharmaceutical formulations using square wave voltammetry at pencil graphite electrodes was developed. After the optimization of the experimental conditions, calibration curves were obtained in the linear concentration range from 9.78 × 10-7 to 7.25 × 10-5 mol L-1 resulting in a limit of detection of 2.12 ± 0.05 x 10-8 mol L-1. Statistical tests showed that the concentrations of 5-ASA in commercial tablets and enemas obtained with the proposed voltammetric method agreed with HPLC values at a 95% confidence level.

Uso de CLAE no controle de qualidade em produtos comerciais de Nim: reprodutibilidade da ação inseticida

The Neem tree, Azadirachta indica, provides many useful compounds that are used as pesticides. However, the efficiency in field of products like neem oil can be committed because they have not been observed reproductive content of secondary metabolic like azadirachtin. Based on reverse-phase high-performance liquid chromatography (HPLC) a new method was developed to permit the rapid quantitative analysis of azadirachtin from seeds, extracts and oil of Neem. In the present study it was evaluated the azadirachtin quantitative variation among various Neem's extracts and seeds showing the importance of quality control for reproduction of the insecticide efficiency, using S. frugiperda as target insect.

Degradação eletroquímica do cloranfenicol em reator de fluxo

This paper reports a study of electrochemical degradation of the chloramphenicol antibiotic in aqueous medium using a flow-by reactor with DSA® anode. The process efficiency was monitored by chloramphenicol concentration analysis with liquid chromatography (HPLC) during the experiments. Analysis of Total Organic Carbon (TOC) was performed to estimate the degradation degree and Ion Chromatography (IC) was performed to determinate inorganic ions formed during the eletrochemical degradation process. In electrochemical flow-by reactor, 52% of chloramphenicol was degraded, with 12% TOC reduction. IC analysis showed the production of chloride ions (25 mg L-1), nitrate ions (6 mg L-1) and nitrite ions (4.5 mg L-1).

Development and validation of a LC-method for the determination of phenols in a pharmaceutical formulation containing extracts from Stryphnodendron adstringens

A sensitive RP-HPLC method with UV detection successfully measured phenol(s) in an ointment containing 3% Stryphnodendron adstringens extract. Chromatography used acetonitrile (0.05% trifluoroacetic acid):water (0.05% trifluoroacetic acid) (v/v), flow rate 0.8 mL min-1. Quantitation was accomplished by the external-standard method. Linearity for 2.00 to 16.00 μg mL-1 (gallic acid) and 1.14 to 18.24 μg mL-1 (gallocatechin) was established. Intra- and inter-day precision levels were under 5%. LOD and LOQ were 0.231 and 0.770 μg mL-1 (gallic acid) and 0.151 and 0.504 μg mL-1 (gallocatechin), respectively. Determination of phenols was unaffected by product excipients.

Otimização e validação de método para determinação de ácidos orgânicos em vinhos por cromatografia líquida de alta eficiência

The organic acids (tartaric, malic, citric, lactic and succinic) are de main components responsible for the acidity in the wine. This method for simultaneous determination of organic acids and interfering peaks in wines can be achieved in 16 min. The sample preceded by a dilution and filtration step. The chromatographic separation required one reversed phase column, isocratic mobile phase (acetonitrila, formic acid in water) and detection wavelength was set at 212 nm. The validation confirmed good repeatability, recovery and application in red and white wines.

Synchronous scanning phosphorimetry for the selective determination of chrysene: a metrological study

Room-temperature phosphorimetry was used to quantify trace levels of chrysene in sugar-cane spirits and in fish bile. A selective phosphorescence enhancer (AgNO3) and synchronous scanning allowed the detection of ng amounts of chrysene. Accuracy (113 ± 17%) and selectivity was evaluated using the CRM-NIST-1647d - Priority Pollutant Polycyclic Aromatic Hydrocarbons in acetonitrile. Analysis of sugar-cane spirit samples enabled recovery of 108 ± 18% which agreed with the one achieved using HPLC. Method's uncertainty was equivalent to 3.4 ng of the analyte, however, the analyte pre-concentration (SPE) improved sensibility and minimized the relative uncertainty. Characterization and homogeneity studies in fish bile were also performed.