Janaína Marques Rodrigues

The validation of a new high throughput method for determination of chloramphenicol in milk using liquid–liquid extraction with low temperature partitioning (LLE-LTP) and isotope-dilution liquid chromatography tandem mass spectrometry (ID-LC-MS/MS)

Chloramphenicol (CAP) is an antibiotic banned for treatment of food-producing animals. The minimum required performance limit (MRPL) of 0.3 μg kg−1 for analytical methods was set by the European Community for the detection of its residues in different matrices, including milk. A highly sensitive analytical method using isotope-dilution liquid chromatography tandem mass spectrometry (ID-LC-MS/MS) and liquid–liquid extraction with low temperature partitioning (LLE-LTP) was developed and validated for CAP residues in milk. Selected validation parameters such as selectivity, working range and linearity, trueness (recoveries), precision (repeatability and intermediate precision), limit of detection (LOD) and limit of quantification (LOQ) were evaluated. The validation procedures were based on the new Eurachem Guide and the European Union Commission Decision 2002/657/EC. The matrix effect was evaluated by the ion suppression test and by comparison of the matrix-matched analytical curve and the solvent standard analytical curve. A linear working range between 0.1 and 5.0 μg kg−1 was observed. The homoscedasticity was demonstrated by the Cochran test, the coefficient of determination (R2) was higher than 0.99 and the residual plot was free of trends. The limits of detection and quantification were 0.015 μg kg−1 and 0.05 μg kg−1, respectively. Mean recoveries evaluated at three levels (0.3, 1.0 and 3.0 μg kg−1) ranged from 94% to 114% with RSDs lower than 6.7% (repeatability). For intermediate precision, different analytes were compared and the RSDs were lower than 7.3%. The method was accurate and reproducible, and was successfully applied to the evaluation of CAP residues in milk samples.

Final report on key comparison CCQM-K55.b (aldrin): An international comparison of mass fraction purity assignment of aldrin

Under the auspices of the Organic Analysis Working Group (OAWG) of the Comit? Consultatif pour la Quantit? de Mati?re (CCQM) a key comparison, CCQM K55.b, was coordinated by the Bureau International des Poids et Mesures (BIPM) in 2010/2011. Nineteen national measurement institutes and the BIPM participated. Participants were required to assign the mass fraction of aldrin present as the main component in the comparison sample for CCQM-K55.b which consisted of technical grade aldrin obtained from the National Measurement Institute Australia that had been subject to serial recrystallization and drying prior to sub-division into the units supplied for the comparison. Aldrin was selected to be representative of the performance of a laboratorys measurement capability for the purity assignment of organic compounds of medium structural complexity [molar mass range 300 Da to 500 Da] and low polarity (pKOW < ?2) for which related structure impurities can be quantified by capillary gas phase chromatography (GC). The key comparison reference value (KCRV) for the aldrin content of the material was 950.8 mg/g with a combined standard uncertainty of 0.85 mg/g. The KCRV was assigned by combination of KCRVs assigned by consensus from participant results for each orthogonal impurity class. The relative expanded uncertainties reported by laboratories having results consistent with the KCRV ranged from 0.3% to 0.6% using a mass balance approach and 0.5% to 1% using a qNMR method. The major analytical challenge posed by the material proved to be the detection and quantification of a significant amount of oligomeric organic material within the sample and most participants relying on a mass balance approach displayed a positive bias relative to the KCRV (overestimation of aldrin content) in excess of 10 mg/g due to not having adequate procedures in place to detect and quantify the non-volatile content?specifically the non-volatile organics content?of the comparison sample. There was in general excellent agreement between participants in the identification and the quantification of the total and individual related structure impurities, water content and the residual solvent content of the sample. The comparison demonstrated the utility of 1H NMR as an independent method for quantitative analysis of high purity compounds. In discussion of the participant results it was noted that while several had access to qNMR estimates for the aldrin content that were inconsistent with their mass balance determination they decided to accept the mass balance result and assumed a hidden bias in their NMR data. By contrast, laboratories that placed greater confidence in their qNMR result were able to resolve the discrepancy through additional studies that provided evidence of the presence of non-volatile organic impurity at the requisite level to bring their mass balance and qNMR estimates into agreement. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Validation of a liquid chromatographic method for determination of related substances in a candidate certified reference material of captopril

This paper describes the validation of a reversed-phase high performance liquid chromatography method (RP-HPLC) with diode array detection (DAD) for determination of related substances (impurities from organic synthesis and degradation products) of captopril according to the Brazilian Pharmacopeia IV. The aim of this study was to guarantee the method accuracy for quantification of related substances, an essential requisite to determine, using the mass balance approach, the captopril content in the first Brazilian certified reference material (CRM) of an active pharmaceutical ingredient (API), developed by Inmetro. The captopril instability in solution is discussed and the captopril content determined by mass balance is compared to the results from titration and differential scanning calorimetry (DSC).

Development studies of captopril certified reference material

Este artigo descreve os estudos realizados com o candidato a Material de Referencia Certificado (MRC) de captopril, primeiro MRC de farmacos no Brasil, incluindo a determinacao de impurezas (orgânicas, inorgânicas e volateis), testes de homogeneidade, testes de estabilidade de curta e longa duracao, calculo do teor de captopril por balanco de massa e estimativa da incerteza de medicao associada ao valor certificado.

Development studies of a new metronidazole certified reference material

This paper presents the results of the studies carried out with the candidate certified reference material (CRM) of metronidazole, first CRM of this active pharmaceutical ingredient (API) available on the market and second Brazilian CRM of an API. The investigation includes the determination of organic impurities, inorganic impurities and volatiles, validation of the HPLC-DAD method, stability studies under transport and storage conditions, homogeneity testing, calculation of metronidazole content by mass balance, confirmation of the certified value by differential scanning calorimetry (DSC), and estimation of measurement uncertainties.

Certified reference material to water content determination in bioethanol fuel

Bioethanol is a strategic biofuel in Brazil. Thus, a strong metrological basis for its measurements is required to ensure the quality and promote its exportation. Recently, Inmetro certified a reference material for water content in bioethanol. This paper presents the results of these studies. The characterization, homogeneity, short-term stability and long-term stability uncertainty contributions values were 0.00500, 0.0166, 0.0355 and 0.0391 mg g-1, respectively. The certificated value for water content of bioethanol fuel was (3.65 ± 0.11) mg g-1. This CRM is the first and up to now the unique in the world.

The measurement uncertainty estimation in chromatographic analisys: approach on the quantification of ethyl carbamate in " cachaça"

The measurement uncertainty is useful to estimate the confidence of analytical results. Nowadays, a result without the uncertainty statement cannot be considered reliable, but the scientific literature still lacks examples of the estimate of the measurement uncertainty. This paper presents a practical and reliable description of the measurement uncertainty estimation of the analytical determination of ethyl carbamate in cachaca by GC-IDMS. The isotope dilution technique (ID) associated with GC-MS was used to improve the accuracy. The uncertainty estimated corresponds to 10% of the mass fraction of ethyl carbamate (115 ± 11) ng/g, which is in agreement with ppb level.

Determination of PAHs: A Practical Example of Validation and Uncertainty Assessment

The objective of this study was to present a reliable and practical example of method validation and uncertainty assessment with an analytical method for the determination of polycyclic aromatic hydrocarbons (PAHs) in urban dust. The method was gas chromatography-mass spectrometry in combination with isotope dilution principle to achieve better accuracy for the results. The method performance parameters for five PAHs were determined (phenanthrene, fluoranthene, benzo[a]anthracene, benzo[a]pyrene and benzo[ghi]perylene); this method was used in the key comparison of CCQM-K50b for PAHs in particulate matter. The limits of detection and quantification were lower than 0.075 and 0.250 µg/g, respectively. The linear correlation coefficients were greater than 0.99. The major uncertainty contributions resulted from the accuracy of each analyzed PAH and the repeatability of the process. Certified reference material (National Institute of Standards and Technology SRM 1649a, urban dust) was used to determine the accuracy and precision of the method. The obtained results were satisfactory and agreed with all evaluated performance parameters.

Quantitative nuclear magnetic resonance for purity assessment of polycyclic aromatic hydrocarbons

The application of 1H quantitative nuclear magnetic resonance (1H qNMR) for purity determination of organic compounds is well documented in the literature. The aim of this work is to determine if the 1H qNMR method produces consistent values for the purity of polycyclic aromatic hydrocarbons (PAH) and is sufficiently accurate for the certification of reference materials. For this purpose, 15 different commercial PAH standards had their purity evaluated by 1H qNMR. The purity values and the associated expanded uncertainty of the 15 analyzed PAH ranged from (97.21 ± 0.28)% to (99.52 ± 1.10)%. The expanded measurement uncertainties were acceptable for the certification of reference materials and the purity values were in the expected range, confirming, therefore, that qNMR is appropriate for this type of analysis.

Development and validation of a method for detection and quantification of ochratoxin A in green coffee using liquid chromatography coupled to mass spectrometry.

Um metodo utilizando Cromatografia Liquida de Alta Eficiencia-Espectrometria de Massas Sequencial (CLAE-EM/EM) com curva de calibracao em matriz foi desenvolvido e validado para a determinacao de ocratoxina A (OTA) em cafe verde. A linearidade foi demonstrada entre 3,0 e 23,0 ng.g-1. As recuperacoes medias variaram entre 90,45% e 108,81%; o desvio padrao relativo sob condicoes de repetitividade e precisao intermediaria foram de 5,39% e 9,94% e de 2,20% e 14,34%, respectivamente. Os limites de deteccao e quantificacao foram 1,2 ng.g-1 e 3,0 ng.g-, respectivamente. O metodo desenvolvido foi adequado, contribuiu para o campo de analises em micotoxinas e sera usado para a futura producao de Material de Referencia Certificado (MRC) para OTA em cafe.