Determination of bioavailable fluoride in toothpastes marketed in Medellín, Colombia, through capillary electrophoresis / Fluoruro biodisponible en cremas dentales comercializadas en Medellín ­ Colombia mediante electroforesis capilar

Currently, the FDI recommendation (World Dental Federation) for fluoride concen­tration in toothpastes is that the total fluoride content declared by manufacturers be between 1,000 and 1,500 ppm, and of this amount, at least 800 ppm should be bioavai­lable fluoride. The aim of this study was to describe the market of toothpastes marketed in Medellín and to measure the concentration of bioavailable fluoride by capillary elec­trophoresis to verify their compliance with the FDI recommendation for the prevention of dental caries. After sampling the toothpastes available for sale in the 16 communes of the city of Medellin, a single operator measured in triplicate the concentration of bioavailable fluoride in 36 samples of previously blinded toothpastes using the capi­llary electrophoresis (EC) technique. Most of the evaluated toothpastes that declared a content of 1,000­1,500 ppm fluoride met the recommendation of presenting at least 800 ppm bioavailable fluoride. Measurement of bioavailable fluoride in toothpastes with MFP (sodium monofluorophosphate) is recommended, as it is known that binding to the abrasive can decrease its concentration over time.

Actualmente la recomendación de la FDI (Federación Dental Internacional) sobre la concentración de fluoruro en cremas dentales es que el contenido de fluoruro total de­clarado por los fabricantes sea entre 1.000 y 1.500 ppm, y que de esta cantidad al menos 800 ppm sea fluoruro biodisponible. El objetivo de este estudio fue describir el mercado de las cremas dentales comercializadas en Medellín y medir en estas la concentración de fluoruro biodisponible por electroforesis capilar para verificar su cumplimiento de la recomendación de la FDI para la prevención de la caries dental. Luego de realizar un mues­treo en las 16 comunas de la ciudad de Medellín sobre las cremas dentales disponibles a la venta, un solo operador midió por triplicado la concentración de fluoruro biodisponible en 36 muestras de cremas dentales previamente cegadas, mediante la técnica de electroforesis capilar (EC). La mayoría de las cremas dentales evaluadas que declaraban contenido de 1.000 a 1.500 ppm de fluoruro, cumplieron la recomendación de presentar al menos 800 ppm de fluoruro biodisponible. Se recomienda realizar la medición del fluoruro biodisponible en las cremas con MFP, ya que se conoce que la unión al abrasivo puede disminuir su concentración en el tiempo.

Recent applications and chiral separation development based on stationary phases in open tubular capillary electrochromatography(2019-2022) / 药物分析学报

Capillary electrochromatography(CEC)plays a significant role in chiral separation via the double sepa-ration principle,partition coefficient difference between the two phases,and electroosmotic flow-driven separation.Given the distinct properties of the inner wall stationary phase(SP),the separation ability of each SP differs from one another.Particularly,it provides large room for promising applications of open tubular capillary electrochromatography(OT-CEC).We divided the OT-CEC SPs developed over the past four years into six types:ionic liquids,nanoparticle materials,microporous materials,biomaterials,non-nanopolymers,and others,to mainly introduce their characteristics in chiral drug separation.There also added a few classic SPs that occurred within ten years as supplements to enrich the features of each SP.Additionally,we discuss their applications in metabolomics,food,cosmetics,environment,and biology as analytes in addition to chiral drugs.OT-CEC plays an increasingly significant role in chiral separation and may promote the development of capillary electrophoresis(CE)combined with other instruments in recent years,such as CE with mass spectrometry(CE/MS)and CE with ultraviolet light detector(CE/UV).

Application and Prospect of RNA Profiling Analysis in Forensic Body Fluid Identification / 法医学杂志

In forensic physical evidence identification, the accurate identification of the individual origin and their body fluid composition of the biological samples obtained from the crime scene play a critical role in determining the nature of a crime. In recent years, RNA profiling has become one of the fastest developing methods for body fluids identification. Due to the characteristics of tissue or body fluid specific expression, various types of RNA markers have been proven to be promising candidate markers for body fluids identification in previous studies. This review summarizes the research progress of RNA markers in body fluids identification, including the RNA markers that have been effectively verified in current research and their advantages and disadvantages. Meanwhile, this review prospects the application of RNA markers in forensic medicine.

Simultaneous determination of paracetamol, dextromethorphan, chlorpheniramine, pseudoephedrine and major impurities of paracetamol and pseudoephedrine by using capillary electrophoresis

Abstract A capillary electrophoresis method was developed for the first time and optimized for the determination of paracetamol, pseudoephedrine, dextromethorphan, chlorpheniramine, 4-aminophenol and ephedrine in tablet formulation. Optimum electrophoretic conditions were achieved by using a background electrolyte of 75 mmol L-1 sodium borate buffer at pH 8.0, a capillary temperature of 30°C, a separation voltage of 30 kV and a pressure injection of the sample at 50 mbar for 10 s. Calibration graphs showed a good linearity with a coefficient of determination (R2) of at least 0.999 for all compounds. Intraday and interday precision (expressed as relative standard deviation (RSD) %) were lower than 1.39% for capillary electrophoresis method. The developed method was demonstrated to be simple and rapid for the determination of paracetamol, pseudoephedrine, dextromethorphan, chlorpheniramine, 4-aminophenol and ephedrine in tablet formulation providing recoveries in the range between 99.62 and 100.57% for all analytes.

Determination of ATP and its metabolites in dietary energy supplements by capillary electrophoresis

Abstract This study aimed to develop a simple and fast capillary electrophoresis (CE) method for the simultaneous determination of adenosine triphosphate (ATP) and its metabolites in dietary energy supplements. Reverse polarity separation mode for faster separation of the three strong negatively charged analytes and capillaries with a 25 µm inner diameter was employed. At -433 V/cm field strength at background electrolyte (BGE) consist with 0.1 M tris-HCl, 0.5 mM tetradecyltrimethylammonium chloride (TTAC) as positively charged surfactant and 0.3 mg/mL hydroxypropylmethylcellulose (HPMC) to reduce the electroosmotic flow (EOF), a complete separation of the three species were achieved in less than 15 minutes. The data acquisition was conducted at a wavelength of 254 nm. Three different commercialised dietary energy supplements were analysed.

Application of capillary zone electrophoresis to determine second-generation H1 antihistaminic drugs, loratadine and rupatadine

Abstract The second generation of H1 antihistamines from the piperidine group are often used for treating allergic diseases due to their action on histaminic receptors, the primary mediator of allergy. Moreover, the antihistamines have anti-inflammatory action, mediated through platelet-activating factor blocking activity. A simple and rapid capillary zone electrophoresis method was developed and validated for the determination of loratadine (LOR) and rupatadine (RUP) in tablets. The analyses were carried out using a fused silica capillary of 50.2 cm (40 cm effective length), 75 µm i.d. The background electrolyte was composed of boric acid 35 mmol/L, pH 2.5. Voltage of 20 kV, hydrodynamic injection of 3447.3 Pa for 3s, temperature at 25 ºC, and UV detection at 205 nm were applied. Electrophoretic separation was achieved at 1.8 and 2.8 min for RUP and LOR, respectively. The method was linear for both drugs in a range of 50.0 to 400.0 µg/mL (r>0.99). The limits of detection and quantification were 46.37 and 140.52 µg/mL, for LOR and 29.60 and 89.69 µg/mL for RUP respectively. The precision was less than 5.0 % for both drugs. The average recovery was approximately 100 %. The proposed novel method can significantly contribute to the rapid detection of counterfeit products and in quality control of drug products containing antihistamines

Aplicação da eletroforese capilar - espectrometria de massas para identificação dos produtos de degradação do aripiprazol em medicamentos / Application of capillary electrophoresis - mass spectrometry to identify aripiprazole degradation products in drugs

Diretrizes internacionais e nacionais como a FDA (Food and Drug Administration), ICH (International Council for Harmonisation) e ANVISA (Agência Nacional de Vigilância Sanitária) estabelecem a exigência de testes de estabilidade para entender melhor a qualidade de um medicamento. O estudo de estabilidade deve ser realizado usando métodos indicativos de estabilidade que possam qualificar e quantificar os insumos farmacêuticos do medicamento, bem como as impurezas e produtos de degradação nele contidos. O aripiprazol é um antipsicótico atípico de segunda geração aprovado para o tratamento de esquizofrenia, transtorno bipolar, depressão e transtornos do espectro do autismo. Os métodos oficiais descritos nas farmacopeias para avaliar o aripiprazol e suas impurezas utilizam a cromatografia líquida de alta eficiência (HPLC) como técnica principal. Nesta pesquisa, objetivou-se desenvolver um método indicativo de estabilidade por eletroforese capilar de zona (CZE) para o aripiprazol na forma farmacêutica de comprimidos, e identificação dos produtos de degradação por espectrometria de massas. O estudo de degradação forçada e a optimização do método desenvolvido por CZE foram realizados utilizando o conceito de delineamento de experimentos (DoE). A separação do aripiprazol de seus produtos de degradação foi conseguida usando uma coluna capilar de sílica fundida (30,2 cm x 75 µm ID), eletrólito de formiato de amônio 6 mmol/L (pH 3) com 5% de metanol sob um potencial de 15 kV e detecção em 214 nm. A capacidade indicativa de estabilidade do método foi investigada pela análise do aripiprazol após ser submetido a condições de estresse ácido, alcalino, térmico, fotolítico e oxidativo, de acordo com as diretrizes ICH. A oxidação foi a principal via de degradação entre as condições de estresse avaliadas. O aripiprazol foi separado dos seus produtos de degradação oxidativa em tempo de corrida abaixo de 5 minutos. O método por CZE mostrou ser linear na faixa de 60 - 140 µg/mL, R2 = 0,9980, precisão calculada como desvio padrão relativo (DPR) menor que 2% e exatidão calculada como recuperação média de 100,93 ± 0,77%. Os resultados obtidos demonstram que o método por HPLC-RP em modo gradiente, separou o aripiprazol e seus produtos de degradação em um tempo de corrida de 30 minutos. Quatro produtos de degradação foram detectados pelo método LC-MS e o principal produto de degradação oxidativo foi identificado. O aripiprazol mostrou-se suscetível à oxidação no grupo piperazina, gerando principalmente o composto aripiprazol-1-N-óxido

International and national guidelines such as the FDA (Food and Drug Administration), ICH (International Council for Harmonization) and ANVISA (National Health Surveillance Agency) establish the requirement for stability tests to better understand quality of a medicine. The stability study must be carried out using stability indicating methods that can qualify and quantify the pharmaceutical ingredients of the drug, as well as the impurities and degradation products contained therein. Aripiprazole is a second-generation atypic antipsychotic drug approved for the treatment of schizophrenia, bipolar disorder, depression, and autism spectrum disorders. The official method described in the pharmacopoeias to evaluate aripiprazole and its impurities is high performance liquid chromatography (HPLC) as the main technique. In this research, the objective was to develop an indicative method of stability by capillary zone electrophoresis (CZE) for aripiprazole in the pharmaceutical form of tablets, and identification of degradation products by mass spectrometry. The forced degradation study and the optimization of the method developed by CZE were carried out using the concept of design of experiments (DoE). The separation of aripiprazole from its degradation products was achieved using a fused silica capillary column (30,2 cm x 75 µm ID), 6 mmol/L ammonium formate electrolyte (pH 3) with 5% methanol under a potential of 15 kV and detection at 214 nm. The indicative stability of the method was investigated by analyzing aripiprazole after being subjected to acid, alkali, thermal, photolytic and oxidative stress conditions, according to the ICH guidelines. Oxidation was the main degradation pathway among the stress conditions evaluated. Aripiprazole was separated from its oxidative degradation products at run times below 5 minutes. The CZE method proved to be linear in the range of 60 - 140 µg/mL, R2 = 0,9980, precision calculated as a relative standard deviation (DPR) of less than 2% and accuracy calculated as a mean recovery of 100,93 ± 0,77%. The results obtained demonstrate that the HPLC-RP method in gradient mode separated aripiprazole and its degradation products in a run time of 30 minutes. Four degradation products were detected by the LC-MS method and the main oxidative degradation product was identified. Aripiprazole was shown to be susceptible to oxidation in the piperazine group, generating mainly the compound aripiprazole-1-N-oxide

Capillary electrophoresis methods for impurity profiling of drugs:A review of the past decade / 药物分析学报

Capillary electrophoresis(CE)is widely used for the impurity profiling of drugs that contain stereo-chemical centers in their structures,analysis of biomolecules,and characterization of bio-pharmaceuticals.Currently,CE is the method of choice for the analysis of foodstuffs and the determination of adulterants.This article discusses the general theory and instrumentation of CE as well as the classification of various CE techniques.It also presents an overview of research on the applications of different CE techniques in the impurity profiling of drugs in the past decade.The review briefly presents a comparison between CE and liquid chromatography methods and highlights the strengths of CE using drug compounds as examples.This review will help scientists,fellow researchers,and students to understand the applications of CE techniques in the impurity profiling of drugs.

The value of multiplex PCR method based on capillary electrophoresis in detecting common serotypes of respiratory adenovirus infection in children / 中华实用儿科临床杂志

Objective:To establish a common method for detecting serotypes of respiratory adenovirus, and to detect the main serotypes of respiratory human adenovirus (HAdV) infection in children in Wenzhou area.Methods:A multiplex PCR method based on capillary electrophoresis was developed to detect 12 common serotypes of respiratory adenovirus.A total of 1 059 children with acute respiratory infection who were admitted to Yuying Children′s Hospital Affiliated to Wenzhou Medical University from January 2018 to December 2019 with positive infection of HAdV detected by the direct immunofluorescence method were recruited and retrospectively analyzed.Multiplex PCR was performed to determine 12 serotypes of respiratory adenovirus, including HAdV-1, 2, 3, 4, 5, 7, 14, 21, 37, 40, 41 and 55.Meanwhile, some samples were randomly selected to examine the consistency in the detection result by the first-generation sequencing method.Results:A total of 1 059 specimens of respiratory secretions with positive HAdV antigen were collected.Detected by multiplex PCR method, 947 cases (89.4%) were positive for 1 serotype, 13 cases (1.2%) were mixed infection with 2 serotypes, and 24 cases (2.3%) were negative.In addition, 75 cases(7.1%) were positive but could not be serotyped.Among the 947 children with the positive infection of a single serotype, 415 cases (43.8%) were HAdV-3 in subgroup B, 318 cases(33.6%) were HAdV-7, 12 cases (1.2%) were HAdV-55, 2 cases (0.2%) were HAdV-21, 108 cases (11.4%) were HAdV-2 in subgroup C, 70 cases (7.4%) were HAdV-1, 16 cases(1.7%) were HAdV-5, and 6 cases(0.6%) were HAdV-4 in subgroup E. HAdV-14, HAdV-37, HAdV-40 and HAdV-41 were not detected.A total of 51 positive samples of HAdV infection detected by multiplex PCR were randomly selected to compare with the detection result by the first-generation sequencing, which were all consistent.Conclusions:This study successfully established a multiplex PCR based on capillary electrophoresis in diagnosing common serotypes of respiratory adenovirus infection in children.HAdV-3, HAdV-7 of subgroup B and HAdV-2 and HAdV-1 of subgroup C were the main serotypes of respiratory adenovirus infection in children of Wenzhou area.HAdV-14, HAdV-37, HAdV-40 and HAdV- 41 were not detected.

Chiral separation of four acidic drugs by micellar electrokinetic chromatography with ionic liquid-based surfactant / 中国药科大学学报

@#In this paper, micellar electrokinetic chromatography (MEKC) with glucose-β-cyclodextrin (Glu-β-CD) as chiral selector and ionic liquid surfactant N-butyl-N-methyl pyrrolidine lauryl sulfate ([C4MP] [C12SO4]) micelles formed at low pH as a pseudo stationary phase was applied for the chiral separation of four acidic drugs naproxen,warfarin, ketoprofen and ibuprofen.Under the same conditions,significantly improved separation of tested drug enantiomers was achieved with the MEKC system based on [C4MP][C12SO4] compared with the system based on the conventional surfactant sodium dodecyl sulfate (SDS).Several primary parameters affecting enantioseparation such as type and proportion of organic modifier, concentration and pH of the running buffer, concentration of chiral selector,concentration of ionic liquid surfactant and applied voltage were systematically investigated.

Diseño y validación de un ensayo de minisecuenciación múltiple para detectar polimorfismos asociados con Síndrome Metabólico / Design and validation of a multiple minisequencing assay to detect polymorphisms associated with Metabolic Syndrome

Resumen Introducción: Es importante identificar los polimorfismos de interés clínico en patologías complejas como el Síndrome Metabólico. Por esto, las metodologías para su evaluación deben estar diseñadas y validadas correctamente, esto permite optimizar recursos y tiempo en la genotipificación y detección correcta de los alelos presentes en los individuos. Objetivo: Diseñar y validar una PCR múltiple, seguida de detección por minisecuenciación, para la genotipificación de ocho polimorfismos de nucleótido simple ubicados en el gen del Receptor Beta 3-Adrenérgico (rs4994 y rs4998), gen de la Apolipoproteina A5 (rs3135506 y rs2075291), gen de la Adiponectina (rs1501299 y rs2241766) y gen del Receptor Activador de la Proliferación de los Peroxisomas tipo gamma (rs1801282 y rs1800571), asociados con el síndrome metabólico. Materiales y métodos: Se diseñaron 24 cebadores para la amplificación y detección de ocho polimorfismos de nucleótido sencillo ubicados en cuatro genes candidatos a estar asociados con el síndrome metabólico, usando el software Primer3®. Dieciséis fueron diseñados para amplificar los polimorfismos y ocho para detectarlos por minisecuenciación. Las estructuras secundarias entre los cebadores se verificaron con el software Autodimer. Los polimorfismos se amplificaron simultáneamente y los fragmentos amplificados se acoplaron a las sondas diseñadas para detectar por minisecuenciación el alelo presente, por medio de bases marcadas con fluorocromos. Finalmente, los alelos fueron detectados por electroforesis capilar en un analizador genético ABI 310 y se interpretaron con el software GeneMapper®. La validación del multiplex se realizó genotipando 20 muestras de individuos, cada uno de ellos autorizó este procedimiento por medio del consentimiento informado. Resultados: Se obtuvieron los perfiles genéticos de los 20 controles genotipados, a partir de la amplificación múltiple, seguida de minisecuenciación, diseñada y validada para detectar los ocho polimorfismos. Conclusión: Se diseñó y validó un ensayo para la detección simultánea de los polimorfismos, ubicados en cuatro genes asociados con el Síndrome metabólico. Los cuales pueden ser empleados como referencia para futuros estudios poblacionales.

Abstract Introduction: It is important to identify the polymorphisms of clinical interest in complex pathologies such as Metabolic Syndrome. Therefore, the methodologies for its evaluation must be designed and validated correctly, this permits optimization of resources and time in genotyping and correct detection of the alleles present in individuals. Objective: To design and validate a multiplex PCR, followed by detection by minisequencing, for the genotyping of eight single nucleotide polymorphisms located in the Beta 3-Adrenergic Receptor gene (rs4994 and rs4998), Apolipoprotein A5 gene (rs3135506 and rs2075291), Adiponectin gene (rs1501299 and rs2241766) and gamma-type Peroxisome Proliferation Activating Receptor gene (rs1801282 and rs1800571), associated with metabolic syndrome. Materials and methods: Twenty-four primers were designed for the amplification and detection of eight single nucleotide polymorphisms located in four candidate genes to be associated with the metabolic syndrome, using the Primer3® software. Sixteen were designed to amplify the polymorphisms and eight to detect them by minisequencing. The secondary structures between the primers were verified with Autodimer software. The polymorphisms were simultaneously amplified, and the amplified fragments were coupled to probes designed to minisequence the present allele using fluorochrome-labeled bases. Finally, the alleles were detected by capillary electrophoresis using an ABI 310 genetic analyzer and analyzed with the GeneMapper® software. The validation of the multiplex was performed by genotyping 20 individual samples, each of them authorized this procedure through informed consent. Results: The genetic profiles of the 20 genotyped controls were obtained, from multiple amplification, followed by minisequencing, designed and validated to detect the eight polymorphisms. Conclusion: An essay was designed and validated for the simultaneous detection of polymorphisms, located in four genes associated with metabolic syndrome, and can used as a reference for future population studies.

Application of Chlorophyte ChlB Gene and Cyanophyte NIES Gene in the Detection of Drowning-Related Plankton / 法医学杂志

Objective To construct a polymerase chain reaction-capillary electrophoresis （PCR-CE） detection method using ChlB gene and NIES gene, investigate the method's specificity and sensitivity, and to evaluate its application value in drowning diagnosis. Methods The specific primers ChlB and NIES were designed for the conserved sequence of chlorophyte ChlB gene and cyanophyte NIES gene in GenBank to construct PCR-CE detection method; 50 species of standard DNA samples were amplified; the sensitivity was determined by gradient concentration detection of positive standard samples; 25 actual cadaver lung tissue samples （drowned： 20, natural death： 5） were detected, and the simultaneous detection results of microwave digestion-vacuum filtration-automated scanning electron microscopy （MD-VF-Auto SEM） were simultaneously compared. Results The minimum DNA detection concentration of primers ChlB and NIES was 0.161 ng and 0.109 ng, respectively, which could specifically amplify chlorophyte （Chlorella pyrenoidosa） and cyanophyte ［Microcystis aeruginosa （producing and not producing toxin）］ widespread in water. The product fragments were 156 bp and 182 bp, respectively. The results of non-drowning tissues were negative. Conclusion This method has high sensitivity and specificity. It can be applied to the detection of plankton related to drowning and combined with MD-VF-Auto SEM method, can increase the detection range of plankton related to drowning and improve the evidence power of drowning diagnosis.

Residue Analysis of Oxytetracycline in Milk Sample by Two Different Chromatographic Methods and Determination of Ionization Constant Values

In thepresent study, two analytical methods for the residue analysis of oxytetracycline in milk sample have been generated. InHPLC method, the analysis was performed on an X Terra RP-18 column at 25 °C with the mobile phase as methanol: water (20 : 80 )(v/v) modified to pH 5. For the second method capillary electrophoresis system was used. The analysis of oxytetracycline in milk sample could be achieved without using organic modifier in a 58 cm length capillary at a working voltage of 12 kV with 20 mM NaH2PO4-H3PO4(pH 7) by capillary electrophoresis. Tetracycline was used as internal standard in both methods. The results calculatedfrom both methods were compared to each other. The calculated data for drugs was checked with the data predicted by the SPARC on-line pKa estimator

Transferibilidad de marcadores microsatélites de Anas platyrhynchos al pato criollo peruano Cairina moschata domestica / Cross species transferability of microsatellite markers from Anas platyrhynchos to Peruvian Muscovy Duck Cairina moschata domestica

Resumen El pato criollo peruano (Cairina moschata domestica) es una de las especies de mayor importancia económica en la alimentación humana. Las especies de patos forman grupos genéticos complejos y difíciles de reconocer, por lo que el uso marcadores microsatélites (SSR) identificados en una especie relacionada como Anas platyrhynchos, representa una opción atractiva, de menor costo y útil para resolver temas relacionados con la conservación de la diversidad genómica, flujo génico e hibridación entre poblaciones. El objetivo de la investigación fue evaluar la transferibilidad de 24 SSR identificados para A. platyrhynchos a las poblaciones peruanas de C. moschata doméstica y determinar el grado de polimorfismo (PIC) de los marcadores transferibles. Para ello, se obtuvo ADN a partir de plumas alares usando el método cloroformo-alcohol isoamílico. Los SSR se construyeron con una secuencia adicional de 19 pb (cola M13) y se utilizaron fluoróforos 6-FAM, VIC, NED y PET para su etiquetado. Los fragmentos amplificados fueron visualizados en geles de agarosa 2% y separados por electroforesis capilar en un secuenciador automático ABI 3130XL. Los resultados mostraron 7 SSRs con un valor PIC alto (PIC>0.5) y que el marcador CMO211 se expresaba con un tamaño molecular menor del de la referencia. En conclusión, el presente trabajo demostró que el 75% de los SSR diseñados para A. platyrhynchos son transferibles a C. moschata domestica; y que sólo 7 fueron altamente informativos. Demostrando así que los SSRs son útiles en la detección de polimorfismos en especies relacionadas y pueden ser usados para mejorar las poblaciones peruanas de patos criollos.

Abstract Peruvian Muskovy duck (Cairina moschata domestica) is one of the most economically important species in human nutrition. Duck species form complex genetic groups which are difficult to recognize, thus the use microsatellite markers (SSRs) identified already in Anas platyrhynchos (related species), represents a very attractive option for its cheapness and usefulness for solving issues related to conservation of genomic diversity, gene flow and hybridization between population. The main goal of this work was to evaluate the degree of polymorphism (PIC) and the transferability of 24 SSRs identified for A. platyrhynchos to C. moschata domestica. In this study, DNA collected from wing feathers was extracted using the chloroform-isoamyl alcohol method. SSRs were constructed with an additional 19 bp sequence (M13 tail) and 6-FAM, VIC, NED and PET fluorophores were used for their labeling. The amplified fragments were visualized on 2% agarose gels and separated by capillary electrophoresis in an automatic ABI 3130XL sequencer. Results showed 7 SSR with high PIC value (PIC> 0.5) and the CMO211 marker expressed in a smaller molecular size that the one used as reference. In conclusion, we showed that 75% of the SSR designed for A. platyrhynchos were transferable to C. moschata domestica as well as we found only 7 SSR highly informative, thus we proved that SSR are highly useful for detecting polymorphisms in related species and improved the Peruvian populations of Muskovy ducks.

Chiral discrimination of amlodipine from pharmaceutical products using capillary electrophoresis

The present work describes the development of a capillary electrophoresis (CE) method for the chiral discrimination of amlodipine (AML) enantiomers using cyclodextrine (CD) derivatives as chiral selectors. A large number of native and derivatized, neutral and ionized CD derivatives were screened to find the optimal chiral selector; and carboximethyl-β-CD (CM-β-CD) was selected for the enantiomeric discrimination. A factorial analysis study was performed by orthogonal experimental design in which several factors were varied at the same time to optimize the separation method. The optimized method (25 mM phosphate buffer, pH = 9.0, 15 mM CM-β-CD, 15 ºC, + 25 kV, 30 mbar/1 second, detection wavelength 230 nm) was successfully applied for the baseline separation of AML enantiomers within 5 minutes. Successful validation and application of the proposed CE method suggest its routine use in enantioselective control of AML in pharmaceutical preparations.

Clinicopathological features and types of microsatellite instability in 1394 patients with colorectal cancer / 南方医科大学学报

OBJECTIVE@#To explore the clinicopathological features and types of genic mutations in DNA mismatch repair (MMR) in colorectal cancer (CRC).@*METHODS@#Immunohistochemistry was used to determine the expression of MMR proteins in 1394 patients with CRC, and PCR-capillary electrophoresis (PCR-CE) was used to detect microsatellite instability (MSI) in 106 cases of defective MMR (dMMR), 46 cases of proficient MMR (pMMR) with heterogeneous expression and 147 randomly selected cases of pMMR. The relationship between the expressions of MMR proteins and the clinicopathological features of the patients was evaluated. The consistency between the results of immunohistochemistry and PCR-CE was assessed.@*RESULTS@#Immunohistochemical staining showed an incidence of dMMR of 7.6% in the patients. The main type of dMMR was co-deletion of MLH1 and PMS2, accounting for 55.7% of the total dMMR cases. The deletion of MMR proteins was significantly correlated with the patients' age, tumor location, tumor size, gross type, histological type, degree of differentiation, lymph node status and TNM stage (@*CONCLUSIONS@#The main type of dMMR is co-deletion of MLH1 and PMS2 in patients with colorectal cancer. dMMR colorectal cancer has typical clinicopathological features and a lower incidence in China than in Western countries. The results of immunohistochemistry and PCR-CE are highly consistent for detecting dMMR in colorectal cancer patients.

Development of analytical method for quantification of antineoplastic in drug delivery systems

Nos últimos anos têm crescido cada vez mais o número de pesquisas envolvendo nanotecnologia para obtenção de medicamentos com liberação controlada, pois esses sistemas podem: proteger o fármaco de incompatibilidades tanto biológicas quanto físico-químicas assim como controlar a biodisponibilidade do fármaco. Embora com todas essas vantagens não existem métodos in vitro realmente capazes de prever com precisão a liberação dos fármacos por esses sistemas, por esse motivo, é muito importante o desenvolvimento de métodos de liberação in vitro para determinar a cinética de liberação desses sistemas.O presente trabalho teve como objetivo desenvolver e validar os métodos de eletroforese capilar (CE) e cromatografia líquida de alta eficiência (HPLC) para determinar a eficiência de encapsulação do fármaco imatinibe em nanopartículaspreviamente elaboradas e caracterizadas, assim como estudar sua liberação in vitro por CE. As nanopartículas foramdesenvolvidas pelo método de nanoprecipitaçãoe caracterizadas quanto ao tamanho, potencial zeta, morfologia e eficiência de encapsulação. A eletroforese capilar é uma técnica alternativa muito promissora em relação ao HPLC devido ao seu baixo custo, menor tempo de corrida e menos poluente ao meio ambiente. Os métodos de quantificação por CE e HPLCforam desenvolvidose validadossegundo as diretrizes do ICH, Farmacopeia Americana e ANVISA, permitindo desenvolver um estudo de liberação.As nanoesferas desenvolvidas apresentaram diâmetro médio próximo a 150nm, com índice de polidispersão menor que 0,1 e aproximadamente 90% de eficiência de encapsulação. Ambos métodos se mostraram lineares com coeficientes de determinação superiores a 0,99, os métodos se mostraram precisos (%DPR< 2), exatos(101,0±4,2% e 98,0±2,5% para HPLC e CE, respectivamente)e seletivos.O método de CE permitiu desenvolver um método de estudo de liberação independente das membranas de diálise

In recent years, there has been a growing number of researches involving nanotechnology to obtain controlled release drugs, these systems can: protect the drug against biological and physico-chemical incompatibilities; controlling the bioavailability of the drug. Although with all these advantages there are no in vitro methods really capable of accurately predicting drugs release by such systems, therefore, the development of in vitro release methods to determine the release kinetics of such systems is very important. The objective of the present work was to develop and validate capillary electrophoresis (CE) and HPLC methods to determine the encapsulation efficiency of the imatinib drug in previously elaborated and characterized nanoparticles, as well as to study its release in vitro by CE method. The nanoparticles were synthesized using the nanoprecipitation method and characterized by size, zeta potential, morphology and encapsulation efficiency. Capillary electrophoresis is a very promising alternative to HPLC because of its low cost, less runtime and less polluting environment. The CE and HPLC methodswere developed and validated according ICH, American Pharmacopoeia and ANVISA guidelines.Developed nanospheres had an average diameter close to 150nm, with polydispersity index less than 0.1 and approximately 90% encapsulation efficiency. Both methods were linear with determination coefficients higher than 0.99, the methods were precise (%RSD < 2), accurate (101.0±4,2% and 98.0±2,5% for HPLC and CE, respectively) and selective. Capillary electrophoresis method allowed to develop a drug release study independent of dialysis membranes

Development of a generic method for the determination of proton-pump inhibitors by capillary zoneelectrophoresis

A generic capillary zone electrophoresis method was developed for the analysis of four proton pump inhibitors: omeprazole, pantoprazole, lansoprazole and rabeprazole. During preliminary analysis screening of phosphate buffers at different pH levels was performed, in order to determine the optimum pH domain suitable for the simultaneous determination of all studied compounds. A face centered central composite design was employed for the optimization of separation conditions. The effect of buffer concentration, pH and applied voltage was studied; resolution between peaks and migration time of the last compound were considered as responses. Other factors as system temperature, injection parameters, capillary length, were held constant during the optimization process. The optimized conditions consisted of 40mM phosphate background electrolyte at pH 5.0, +25 kV applied voltage and 20 °C temperature. The migration order of the analytes was as follows: rabeprazole, omeprazole, lansoprazole and pantoprazole. Full resolution of all analytes was achieved within 9 minutes. The method was validated and proved to be suitable in terms of repeatability, sensitivity, linearity, accuracy and robustness. Determinations from commercially available pharmaceutical formulation were performed for omeprazole; good reproducibility and recovery were obtained.

Standardization and application on ribotyping library of Clostridioides difficile in China / 中华流行病学杂志

Objective@#To establish a standard operation procedure (SOP) for ribosome genotyping (ribotyping) on Clostridioides (C.) difficile, supplement and verify ribotyping typing library, so as to improve the comparability of data between different laboratories and to develop surveillance network of C. difficil in China.@*Methods@#Molecular typing of 54 reference strains from the United States and Europe of C. difficile were performed by using the SOP referencing correspondence from abroad and from our laboratory with a BioNumerics 7.6 software to estimate the reference library of types of C. difficile. Identification of 374 clinical and animal isolates of C. difficile from 13 cities in China between 2010 and 2018, to supplement the library information. Kappa test was used to evaluate the consistency.@*Results@#Results of capillary electrophoresis of reference strains appeared clear and stable, which guaranteed the clustering results being fast and accurate. Results from the supplementary typing showed that there were 84 types of isolates, of which 25 RT types were consistent with reference strains from abroad, while 58 RT types were different from referenced types. In the 40 referenced types, 15 RT types were not found in this study. In the consistency evaluation, the Kappa value was 0.891 and (P<0.01), showing the two Molecular typing as consistent and with close resemblance.@*Conclusions@#The result of capillary electrophoresis by applying SOP for ribotyping on C. difficile base on QIAxcel capillary electrophoresis system, appeared clear and stable. The standardized library seemed more easily used for comparability and data sharing between the laboratories.

Analysis of respiratory pathogens for children respiratory tract infection by capillary electrophoresis-based multiplex PCR / 中国小儿急救医学

Objective@#To investigate the effect of capillary electrophoresis-based multiplex PCR (CEMP) in detecting pathogens for children respiratory tract infection, and to provide scientific basis for clinical diagnosis and treatment rapidly and accurately.@*Methods@#The cases were defined according to the national monitoring program of febrile respiratory syndrome during the 12th Five-Year Plan, and the samples were collected from nasopharyngeal swabs, bronchoalveolar lavage fluid and sputum of children with respiratory tract infection hospitalized in Changchun Children′s Hospital from January 2017 to February 2018.Multiplex PCR amplification was performed by one-step method, then PCR products were separated by DNA length size with capillary electrophoresis and pathogens were analyzed by "Genemapper software" software.Detecting pathogens included Influenza A virus (InfA), Human Adenovirus (HADV), Boca virus (Boca), Human Rhinovirus (HRV), Novel InfA-09H1 (InfA-09H1) and Seasonal Influenza virus H3N2 (InfA-H3N2), Parainfluenza virus (HPIV), Human metapneumonia virus (HMPV), Influenza B virus (InfB), Mycoplasma pneumoniae (Mp), Chlamydia pneumoniae (CP), Human Coronavirus (HCOV), Human Respiratory Syncytial virus (HRSV).@*Results@#The effective detection rate of the CEMP assay was 95.71%.The positive detection rate of respiratory tract pathogens was 62.84% and the mixed infection rate was 9.61%.The mixed infection was mainly InfA and HRSV.The highest three positive rates were named InfA, HRSV and Mp.The positive rate of HRSV was significantly higher in the 0-3 age group than that in older group.Different pathogens were detected in different age groups, and the high-occurrence season of respiratory tract infection with virus was from December to March of the next year.InfA-09H1 was the main prevalent influenza virus in January, February and March 2017, InfA-H3N2 was the main prevalent influenza virus in November and December 2017, and the outbreak of InfB was happened in Changchun in late 2017 and early 2018.HRSV was detected only in the coldest season in Changchun from November to March of the next year.Different pathogens were detected in different respiratory infection.HRSV was the main pathogen detected in pneumonia; InfA-03H2 and HPIV were the main pathogens detected in acute bronchitis; HRV and InfA were the main pathogens detected in upper respiratory tract infection.@*Conclusion@#CEMP is an efficient, rapid and accurate method for the detection of pathogens in patients with respiratory tract infections, and it will have a broad application prospect to develop reagents suitable for clinical diagnosis.