Validation of a stability-indicating RP-LC method for the determination of tigecycline in lyophilized powder

A reversed-phase liquid chromatography (RP-LC) method was validated for the determination of tigecycline in lyophilized powder. The LC method was conducted on a Luna C18 column (250 × 4.6 mm i.d.), maintained at room temperature. The mobile phase consisted of buffer containing sodium phosphate monobasic (0.015M) and oxalic acid (0.015M) (pH 7.0)-acetonitrile (75:25, v/v), run at a flow rate of 1.0 mL/min and using ultraviolet detection at 280 nm. The chromatographic separation was obtained with a retention time of 8.6 min, and was linear in the range of 40-100 μg/mL (r2 = 0.9997). The specificity and stability-indicating capability of the method was proven through forced degradation studies, which also showed no interference of the excipients. The accuracy was 99.01% with a bias lower than 1.81%. The limits of detection and quantitation were 1.67 and 5.05 μg/mL, respectively. Moreover, method validation demonstrated satisfactory results for precision and robustness. The proposed method was applied for the analysis of the lyophilized powder formulation, contributing to improve the quality control and to assure the therapeutic efficacy. © The Author [2012]. Published by Oxford University Press. All rights reserved.

Validation of an alternative analytical method for the quantification of antioxidant activity in plant extracts

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Development and validation of a D-loop mtDNA SNP assay for the screening of specimens in forensic casework

Mitochondrial DNA (mtDNA) analysis is usually a last resort in routine forensic DNA casework. However, it has become a powerful tool for the analysis of highly degraded samples or samples containing too little or no nuclear DNA, such as old bones and hair shafts. The gold standard methodology still constitutes the direct sequencing of polymerase chain reaction (PCR) products or cloned amplicons from the HVS-1 and HVS-2 (hypervariable segment) control region segments. Identifications using mtDNA are time consuming, expensive and can be very complex, depending on the amount and nature of the material being tested. The main goal of this work is to develop a less labour-intensive and less expensive screening method for mtDNA analysis, in order to aid in the exclusion of non-matching samples and as a presumptive test prior to final confirmatory DNA sequencing. We have selected 14 highly discriminatory single nucleotide polymorphisms (SNPs) based on simulations performed by Salas and Amigo (2010) [1] to be typed using SNaPShotTM (Applied Biosystems, Foster City, CA, USA). The assay was validated by typing more than 100 HVS-1/HVS-2 sequenced samples. No differences were observed between the SNP typing and DNA sequencing when results were compared, with the exception of allelic dropouts observed in a few haplotypes. Haplotype diversity simulations were performed using 172 mtDNA sequences representative of the Brazilian population and a score of 0.9794 was obtained when the 14 SNPs were used, showing that the theoretical prediction approach for the selection of highly discriminatory SNPs suggested by Salas and Amigo (2010) [1] was confirmed in the population studied. As the main goal of the work is to develop a screening assay to skip the sequencing of all samples in a particular case, a pair-wise comparison of the sequences was done using the selected SNPs. When both HVS-1/HVS-2 SNPs were used for simulations, at least two differences were observed in 93.2% of the comparisons performed. The assay was validated with casework samples. Results show that the method is straightforward and can be used for exclusionary purposes, saving time and laboratory resources. The assay confirms the theoretic prediction suggested by Salas and Amigo (2010) [1]. All forensic advantages, such as high sensitivity and power of discrimination, as also the disadvantages, such as the occurrence of allele dropouts, are discussed throughout the article. © 2013 Elsevier B.V.

Validation of the English version of the UNESP-Botucatu multidimensional composite pain scale for assessing postoperative pain in cats

Background: A scale validated in one language is not automatically valid in another language or culture. The purpose of this study was to validate the English version of the UNESP-Botucatu multidimensional composite pain scale (MCPS) to assess postoperative pain in cats. The English version was developed using translation, back-translation, and review by individuals with expertise in feline pain management. In sequence, validity and reliability tests were performed.Results: Of the three domains identified by factor analysis, the internal consistency was excellent for 'pain expression' and 'psychomotor change' (0.86 and 0.87) but not for 'physiological variables' (0.28). Relevant changes in pain scores at clinically distinct time points (e.g., post-surgery, post-analgesic therapy), confirmed the construct validity and responsiveness (Wilcoxon test, p < 0.001). Favorable correlation with the IVAS scores (p < 0.001) and moderate to very good agreement between blinded observers and 'gold standard' evaluations, supported criterion validity. The cut-off point for rescue analgesia was > 7 (range 0-30 points) with 96.5% sensitivity and 99.5% specificity.Conclusions: The English version of the UNESP-Botucatu-MCPS is a valid, reliable and responsive instrument for assessing acute pain in cats undergoing ovariohysterectomy, when used by anesthesiologists or anesthesia technicians. The cut-off point for rescue analgesia provides an additional tool for guiding analgesic therapy. © 2013 Brondani et al.; licensee BioMed Central Ltd.

Comparison of methods in oral health education from the perspective of adolescents.

To investigate whether teenagers' knowledge about oral health is influenced by educational methods and to verify the most effective method according to their perception. The study was performed in Araçatuba, São Paulo State, Brazil, with 127 teenagers from a vocational school. It was realised in 3 steps: 1. An evaluation of knowledge about oral health using a self-applied questionnaire. 2. An application of educational methods, where the students were divided into two groups (A and B). Group A participated in three educational activities that involved lectures, individual demonstration, and participatory activity. Group B was divided into three subgroups (B1, B2, B3) and each of them participated in only one of the methods. 3. The acquired knowledge was evaluated. Group A created a focus group to give their opinion about strategies. With regards to knowledge after the application of the different methods in all groups, there was a statistically significant difference concerning periodontitis, gingivitis and herpes. In group A, after the three activities, and in group B2 after the individual demonstration, an association was found between 'healthy teeth' and 'general health' (P = 0.004 and P = 0.022, respectively). After the individual demonstration, an association was shown between variables of acquired knowledge about 'harmful diet' and 'dental caries' (P = 0.002) as well as 'good diet' and 'prevention of oral diseases' (P = 0.032). The favourite method was individual demonstration, due to the contact with educational materials, followed by participatory activity because it encouraged learning in a more dynamic way. Educational methods influenced knowledge about oral health, with individual demonstration proving to be the most effective method for acquiring knowledge. In the adolescents' view, the participatory activity was the preferred method.

Validation of a reference control for an SYBR-Green fluorescence assay-based real-time PCR for detection of bovine herpesvirus 5 in experimentally exposed bovine embryos

The objective of this study was to optimize an internal control to improve SYBR-Green-based qPCR to amplify/detect the BoHV-5 US9 gene in bovine embryos produced invitro and experimentally exposed to the virus. We designed an SYBR-Green-based binding assay that is quick to perform, reliable, easily optimized and compares well with the published assay. Herein we demonstrated its general applicability to detect BoHV-5 US9 gene in bovine embryos produced invitro experimentally exposed to BoHV-5. In order to validate the assay, three different reference genes were tested; and the histone 2a gene was shown to be the most adequate for normalizing the qPCR reaction, by considering melting and standard curves ( p<0.05). On the other hand, no differences were found in the development of bovine embryos invitro whether they were exposed to BoHV-5 reference and field strains comparing to unexposed embryos. The developed qPCR assay may have important field applications as it provides an accurate BoHV-5 US9 gene detection using a proven reference gene and is considerably less expensive than the TaqMan qPCR currently employed in sanitary programs. © 2013 Elsevier Ltd.

Validation of the lactate minimum test as a specific aerobic evaluation protocol for table tennis players

The purpose of this study was to validate the lactate minimum test as a specific aerobic evaluation protocol for table tennis players. Using the frequency of 72 balls·min-1 for 90 sec, an exercise-induced metabolic acidosis was determined in 8 male table tennis players. The evaluation protocol began with a frequency of 40 balls·min-1 followed by an increase of 8 balls·min-1 every 3 min until exhaustion. The mean values that corresponded to the subjects' lactate minimum (Lacmin) were equal to 53.1 ± 1.5 balls·min-1 [adjusted for the time test (Lacmin_time)] and 51.6 ± 1.6 balls·min-1 [adjusted for the frequency of balls (Lacmin_Freq)], which resulted in a high correlation between the two forms of adjustment (r = 0.96 and (P = 0.01). The mean maximum lactate steady state (MLSS) was 52.6 ± 1.6 balls·min-1. Pearson's correlations between Lacmin_time vs. MLSS and Lacmin_freq vs. MLSS were statistically significant (P = 0.03 and r = 0.86, P = 0.03 and r = 0.85, respectively). These findings indicate that the Lacmin test predicts MLSS. Therefore, it is an excellent method to obtain the athletes' anaerobic threshold. Also, there is the advantage that it can be performed in 1 day in the game area. However, the Lacmin value does not depend on the Lacpeak value.

Development and Validation of A Stability-Indicating Mekc Method for Determination of Flucloxacillin Sodium in Capsules

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Content Validation of the Operational Definitions of the Nursing Diagnoses of Activity Intolerance, Excess Fluid Volume, and Decreased Cardiac Output in Patients With Heart Failure

Objectives To consensually validate the operational definitions of the nursing diagnoses activity intolerance, excessive fluid volume, and decreased cardiac output in patients with decompensated heart failure. Method Consensual validation was performed in two stages: analogy by similarity of defining characteristics, and development of operational definitions and validation with experts. Results A total of 38 defining characteristics were found. Operational definitions were developed and content-validated. One hundred percent of agreement was achieved among the seven experts after five rounds. Ascites was added in the nursing diagnosis excessive fluid volume. Conclusion The consensual validation improves interpretation of human response, grounding the selection of nursing interventions and contributing to improved nursing outcomes. Implications for Practice Support the assessment of patients with decompensated heart failure. Objetivos Realizar a validacAo consensual das definicoes operacionais dos diagnosticos de enfermagem Intolerancia a atividade, Volume de liquidos excessivo e Debito cardiaco diminuido em pacientes com insuficiencia cardiaca descompensada. Metodo ValidacAo consensual em duas etapas: Analogia de semelhanca das caracteristicas definidoras e desenvolvimento de definicoes operacionais e validacAo com expertst. Resultados Foram encontradas 38 caracteristicas definidoras para os diagnosticos de enfermagem. Suas definicoes operacionais foram desenvolvidas e seu conteudo validado. Os resultados mostram que houve 100% de concordancia entre os sete experts apos cinco rodada. As definicoes operacionais foram classificadas com base no nivel de concordanica. Ascite foi acrescentada ao diagnostico Volume de liquidos excessivo. ConclusAo A validacAo consensual melhora a interpretacAo das respostas humanas, embasando a selecAo de intervencoes de enfermagem e contribuindo para melhorar os resultados. Implicacoes Para A Pratica Apoio a avaliacAo dos pacientes com insuficiencia cardiaca descompensada.

Validation of an HPLC Method for the Determination of Dibucaine Encapsulated in Solid Lipid Nanoparticles and Nanostructured Lipid Carriers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Validation of analytical methodology for quantification of cefazolin sodium pharmaceutical dosage form by high performance liquid chromatography to be applied for quality control in pharmaceutical industry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Multivariate Development and Validation of a Stability-Indicating HPLC Method for the Determination of Glimepiride in Tablets

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Theoretical Analysis and Experimental Validation of the Scholte Wave Propagation in Immersed Plates for the Characterization of Viscous Fluids

This work reports the study of an attractive interfacial wave for application in ultrasonic NDE techniques for inspection and fluid characterization. This wave, called quasi-Scholte mode, is a kind of flexural wave in a plate in contact with a fluid which presents a good sensitivity to the fluid properties. In order to explore this feature, the phase velocity curve of quasi-Scholte mode is experimentally measured in a plate in contact with a viscous fluid, showing a good agreement with theory.

Translation, cross-cultural adaptation and validation of the Portuguese version of the DYMUS questionnaire for the assessment of dysphagia in multiple sclerosis

Translation, cross-cultural adaptation and validation of the DYMUS questionnaire for the assessment of dysphagia in multiple sclerosis. The original English version of the DYMUS was translated using the forward-backward technique, cross-culturally adaptated, pilot-tested in 40 patients, and then applied to 100 multiple sclerosis patients to assess the reliability and construct validity. Construct validity was assessed by Mann–Whitney test and Spearman’s correlation coefficient (rs). The internal consistency of the questionnaire was evaluated using Cronbach’s alpha coefficient and inter-item correlation. DYMUS-BR internal consistency was high (Cronbach’s alpha= 0.72); Cronbach’s alpha was 0.65 for the ‘dysphagia for solids’ subscale and 0.67 for the ‘dysphagia for liquids’ subscale and positive inter-item correlations was found between all items, except for weight loss question. Significant association (p<0.001) and correlation (rs = 0,357; p = 0.01) was found between DYMUS-BR and dysphagia self-assessment. The DYMUS-BR questionnaire maintained the characteristics of that originally described, demonstrating to be a reliable, valid, easy and consistent tool to be used by health professionals for preliminary selection of Brazilian MS patients who need more specific instrumental analyses of swallowing.