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1.
Article in Chinese | WPRIM | ID: wpr-970922

ABSTRACT

OBJECTIVE@#To study the molecular epidemiology of thalassemia in Jiaxing area of Zhejiang province and provide a basis for prenatal diagnosis, genetic counseling and prevention and control of birth defects.@*METHODS@#A total of 24 003 pregnant women who presented at the Jiaxing Maternal and Child Health Care Hospital from April 2017 to September 2021 were enrolled. Capillary hemoglobin electrophoresis in combination with routine blood test were used for primary screening for carriers of thalassemia-associated mutations, and those with positive results were subjected to fluorescence quantitative PCR assay. Prenatal diagnosis was provided for couples with a risk of giving birth to children with intermediate or severe thalassemia.@*RESULTS@#Among the 24 003 pregnant women, 1 211 cases were suspected as carriers of thalassemia-associated mutations, among whom 443 (36.58%) were confirmed by genetic testing. Among these, carriers of α-, β- and α-complex β-globin gene mutations have accounted for 27.31% (121/443), 70.65% (313/443) and 2.04% (9/443), respectively. The result of prenatal diagnosis for an at-risk couple was --SEA/αCSα, and the fetus was predicted to have intermediate or severe thalassemia. Termination of the pregnancy was recommended.@*CONCLUSION@#Hemoglobin electrophoresis combined with routine blood test during pregnancy may be used as a preliminary screening measure for carriers of thalassemia-associated variants. Combined with genetic testing, this will be of great significance for the control of thalassemia in this region.


Subject(s)
Female , Humans , Pregnancy , Electrophoresis, Capillary , Genetic Counseling , Genetic Testing , Mutation , Prenatal Diagnosis , Thalassemia/genetics
2.
Braz. J. Pharm. Sci. (Online) ; 58: e19836, 2022. tab, graf
Article in English | LILACS | ID: biblio-1394041

ABSTRACT

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.


Subject(s)
Chlorpheniramine/antagonists & inhibitors , Electrophoresis, Capillary/methods , Dextromethorphan/antagonists & inhibitors , Ephedrine/antagonists & inhibitors , Pseudoephedrine/antagonists & inhibitors , Aminophenols/antagonists & inhibitors , Acetaminophen/agonists , Buffers , Diagnosis , Methods
3.
São Paulo; s.n; s.n; 2022. 125 p. tab, graf.
Thesis in Portuguese | LILACS | ID: biblio-1416534

ABSTRACT

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


Subject(s)
Mass Spectrometry/methods , Pharmaceutical Preparations/analysis , Electrophoresis, Capillary/methods , Aripiprazole/metabolism , Oxidative Stress , Pharmaceutical Raw Material , Process Optimization
4.
Braz. J. Pharm. Sci. (Online) ; 58: e201045, 2022. tab, graf
Article in English | LILACS | ID: biblio-1420433

ABSTRACT

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.


Subject(s)
Capillaries , Adenosine Triphosphate/agonists , Electrophoresis, Capillary/methods , Dietary Supplements
5.
Braz. J. Pharm. Sci. (Online) ; 58: e20767, 2022. tab, graf
Article in English | LILACS | ID: biblio-1420452

ABSTRACT

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


Subject(s)
Loratadine/antagonists & inhibitors , Electrophoresis, Capillary/methods , Histamine H1 Antagonists/pharmacology , Quality Control , Capillaries/abnormalities , Pharmaceutical Preparations/analysis , Laboratory and Fieldwork Analytical Methods
6.
Rev. Univ. Ind. Santander, Salud ; 53(1): e320, Marzo 12, 2021. tab, graf
Article in Spanish | LILACS | ID: biblio-1365450

ABSTRACT

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.


Subject(s)
Humans , Electrophoresis, Capillary , Polymorphism, Single Nucleotide , Metabolic Syndrome , Receptors, Adrenergic, beta-3 , PPAR gamma , Adiponectin , Apolipoprotein A-V
7.
Article in Chinese | WPRIM | ID: wpr-880053

ABSTRACT

OBJECTIVE@#To investigate the screening of β-thalassemia among newborns in Wuhan region, so as to explore the influencing factors of Hb A in dried blood spot.@*METHODS@#Concentrations of Hb A,Hb A2,Hb F in the dried blood spots collected from 99 275 neonates in Wuhan region were analyzed by Sebia capillary electrophoresis. The screening result of β-thalassemia was interpretated accroding to the ratio of each group, the suspicious β-thalassemia newborns were recalled and the gene of thalassemia in those newborns was checked.@*RESULTS@#Among 99 275 newborns, 1 408 positive patients were found, and the positive rate of screening was 1.41%. A total of 350 patients with gene mutation were found among 709 β-thalassemia suspicious patients. There were significantly statistical differences of positive predictive value among Hb A levels in different groups and there were also significantly statistical differences of positive predictive values among gestational weeks in different groups. No significantly statistical differences were observed among different genetic defects and phenotypes of heterozygous β-thalassemia in Hb A concentrations. Postnatal day and gestational age were significantly and positively associated with Hb A concentrations.@*CONCLUSION@#The capillary electrophoresis is an effective screening method for β-thalassemia of full-term neonate. Postnatal day and gestational age is associated with the pencentage of Hb A.


Subject(s)
Humans , Infant, Newborn , Electrophoresis, Capillary , Mass Screening , Mutation , Thalassemia , beta-Thalassemia/genetics
8.
Rev. cuba. hematol. inmunol. hemoter ; 36(3): e1164, jul.-set. 2020. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1156440

ABSTRACT

Introducción: En el Instituto de Hematología e Inmunología se realiza el estudio molecular de las leucemias mieloides agudas (LMA). Para las leucemias mieloides agudas no promielocíticas (LPM) se determinan cuatro biomarcadores: los genes de fusión RUNX1-RUNX1T1 y CBF(-MYH11, la duplicación interna en tándem del gen FLT3 (DIT FLT3) y la mutación A del gen NPM1 (NPM1-A). Objetivo: Determinar la frecuencia de estos cuatro biomarcadores, en pacientes cubanos con leucemias mieloides agudas primaria no promielocíticas. Métodos: Se incluyeron 91 pacientes entre niños y adultos, estudiados en el Instituto durante tres años desde el debut. A partir de ARN de sangre medular se obtuvo ADN complementario por transcripción inversa; se amplificaron los fragmentos correspondientes mediante la reacción en cadena de la polimerasa y el producto se analizó por electroforesis capilar. Resultados: El RUNX1-RUNX1T1 apareció en el 24,2 por ciento, fue más frecuente en los pacientes pediátricos y disminuyó significativamente con la edad. El CBFβ-MYH11 solo se encontró en adultos (4,8 por ciento). La NPM1-A con 41 por ciento fue mayoritaria entre los adultos. La DIT FLT3 se observó en el 21,6 por ciento y no mostró relación con la edad. NPM1-A y DIT FLT3 fueron las aberraciones con mayor presencia simultánea. Conclusiones: Por primera vez se describe la frecuencia de los cuatro biomarcadores moleculares en los pacientes cubanos con leucemias mieloides agudas primaria no promielocíticas; su comportamiento fue similar a lo descrito por otros autores, aunque se encontraron algunas particularidades(AU)


Introduction: At the Institute of Hematology and Immunology, the molecular study of acute myeloid leukemias (AML) is carried out. For nonpromyelocytic acute myeloid leukemias, four biomarkers are determined: the RUNX1-RUNX1T1 and CBF(-MYH11 fusion genes, the internal tandem duplication of the FLT3 gene (DIT FLT3), and the A mutation of the NPM1 gene (NPM1-A). Objective: To determine the frequency of these four biomarkers in Cuban patients with nonpromyelocytic primary acute myeloid leukemias. Methods: 91 patients were included, children and adults, who were studied at the Institute for three years from their disease debut. Complementary DNA was obtained from medullary blood RNA by reverse transcription. The corresponding fragments were amplified by polymerase chain reaction and the product was analyzed by capillary electrophoresis. Results: RUNX1-RUNX1T1 appeared in 24.2 percent; it was more frequent in pediatric patients and decreased significantly with age. CBFβ-MYH11 was found only in adults (4.8 percent). NPM1-A, accounting for 41 percent, represented the majority among adults. FLT3 DIT was observed in 21.6 por ciento and was not related to age. NPM1-A and DIT FLT3 were the disorders with the greatest concurrence. Conclusions: For the first time, the frequency of the four molecular biomarkers is described in Cuban patients with primary non-promyelocytic acute myeloid leukemias. Its characterization was similar to that described by other authors, although some peculiarities were found(AU)


Subject(s)
Humans , Male , Female , Biomarkers , Leukemia, Myeloid, Acute/genetics , Polymerase Chain Reaction , DNA, Complementary , Reverse Transcription , Electrophoresis, Capillary , Cuba
9.
Rev. chil. pediatr ; 91(4): 568-572, ago. 2020. graf
Article in Spanish | LILACS | ID: biblio-1138672

ABSTRACT

OBJETIVO: Alertar al personal de la salud sobre la importancia de la detección temprana de las he- moglobinopatías, dado que es el trastorno monogénico recesivo más frecuente. Pacientes y MÉTODO: Estudio retrospectivo del resultado de eletroforesis capilar (CE) de 152 pacientes entre 0 y 18 años que durante el año 2017 fueron evaluados por sospecha de hemoglobinopatías en un Hospital Universitario de Colombia. La información se tomó de los registros médicos y del Laboratorio de Hematología y Hemostasia, asegurando la privacidad de los datos y aprobado por el Comité de Ética local. RESULTADOS: De 152 pacientes, 48,6% tenía entre 7 y 18 años. La frecuencia de hemoglobinopatías fue de 42,7%. La variante más frecuente fue el rasgo de células falciformes (Hb S) con 14,5%. El hematólogo fue el profesional que más frecuentemente solicitó EC. DISCUSIÓN: Se detectó que las hemoglobinopatías se diagnostican usualmente en niños mayores de siete años. Esto puede favorecer las complicaciones y progresión de la enfermedad, y aumento en los costos de la salud. Se requiere más información y educación a los médicos generales y pediatras para un diagnóstico más temprano.


OBJECTIVE: The objective of this study is to spread awareness among health personnel about the importance of early detection of hemoglobinopathies since it is the most frequent monogenic recessive disorder worldwide. PATIENTS AND METHOD: Retrospective study of the results of capillary electropho resis (CE) of 152 patients aged between 0 and 18 years who were evaluated in 2017 due to suspected hemoglobinopathies in a University Hospital in Colombia. The information was collected from me dical records and the Hematology and Hemostasis Laboratory, ensuring data privacy and approved by the local Ethics Committee. RESULTS: Of 152 patients, 48.6% were aged between 7 and 18. The frequency of hemoglobinopathies was 42.7%. The most frequent hemoglobin variant was the sickle cell trait (Hb S) with 14.5%. The hematologist was the professional who most frequently requested CE. DISCUSSION: We found that hemoglobinopathies are usually diagnosed late in pediatric patients. This may favor complications and progression of the disease and increase healthcare costs. More information and education are required for general physicians and pediatricians in order to achieve early diagnosis.


Subject(s)
Humans , Male , Female , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Practice Patterns, Physicians'/statistics & numerical data , Delayed Diagnosis/statistics & numerical data , Hemoglobinopathies/diagnosis , Prognosis , Quality of Life , Retrospective Studies , Colombia/epidemiology , Electrophoresis, Capillary , Early Diagnosis , Developing Countries , Hemoglobinopathies/epidemiology , Hospitals, University
10.
Braz. j. med. biol. res ; 53(1): e9001, Jan. 2020. tab, graf
Article in English | LILACS | ID: biblio-1055477

ABSTRACT

Due to its various structures in bio-compounds, snake venom is the indisputable result of evolutionary stages of molecules with an increasingly complex structure, high specificity, and of great importance for medicine because of their potential. The present study proposed an underpinning examination of venom composition from nine species of venomous snakes using a useful and replicable methodology. The objective was the extension of the evaluation of protein fractions in the field up to 230 kDa to permit possible identification of some fractions that are insufficiently studied. The gel capillary electrophoresis method on the chip was performed using an Agilent 2100 bioassay with the 80 and 230-LabChip Protein kits. Interpretation of electrophoresis was performed using the Protein 2100 expert (Agilent) test software as follows: a) Protein 80 (peak size scale): 1.60, 3.5, 6.50, 15.00, 28.00, 46.00, 63.00, 95.00 kDa; b) Protein 230 (peak size scale): 4.50, 7.00, 15.00, 28.00, 46.00, 63.00, 95.00, 150.00, 240.00 kDa. The screening revealed the presence of compounds with a molecular weight greater than 80 kDa, in the case of Vipera aspis and Vipera xantina palestinae. For V. aspis, a 125 kDa molecular weight pro-coagulant protein was identified, known as being involved in the reduction of plasma clotting time without any direct activity in the fibrinogen coagulation process. The samples examined on the Protein 230-LabChip electrophoresis chip can be considered as a novelty with possible uses in medicine, requiring further approaches by advanced proteomics techniques to confirm the intimate structural features and biological properties of snake venoms.


Subject(s)
Animals , Viper Venoms/chemistry , Proteins/chemistry , Viperidae/classification , Viper Venoms/analysis , Proteins/isolation & purification , Proteins/analysis , Electrophoresis, Capillary , Proteome/classification , Proteome/chemistry , Proteomics/methods
11.
Journal of Experimental Hematology ; (6): 1316-1320, 2020.
Article in Chinese | WPRIM | ID: wpr-827119

ABSTRACT

OBJECTIVE@#To explore the abnormal hemoglobinopathy in couples of child-bearing age in Chongqing.@*METHODS@#A total of 34 800 subjects of child-bearing age were screened for thalassemia by using capillary electrophoresis from January 2015 to September 2018. PCR-flow cytometry fluorescence hybridization assay was used to detect the common thalassemia gene deletions and mutations.@*RESULTS@#8 kinds of abnormal hemoglobinopathy were detected in 200 cases from 34 800 subjects of child-bearing age, the detection rate was 0.57% in couples of child-bearing age in Chongqing: Among 200 cases of abnormal hemoglobin pathy, Hb E was found in 90 cases (accounting for 45.0%), and Hb D in 25 cases (accounting for 12.5%). Hb NewYork was found in 25 cases (accounting for 12.5%). HbJ-bangkok was found in 25 cases (accounting for 12.5%), and Hb Q-Thailand in 16 cases (accounting for 8.0%). Hb Hope was detected in 15 cases (accounting for 7.5%). Hb S was detected in 3 cases (accounting for 1.5%). Hb Hasharon was detected in 1 case (accounting for 0.5%). The mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) of Hb E and Hb Q-Thailand were lower than normal reference intervals.@*CONCLUSION@#The detection rate of abnormal hemoglobinopathy in Chongqing is higher than the average level in China. Capillary electrophoresis can effectively screen abnormal hemoglobinopathy, which is great significant for aristogenesis and improvement of population quality.


Subject(s)
Child , Humans , China , Electrophoresis, Capillary , Hemoglobinopathies , Hemoglobins, Abnormal , Thailand , Thalassemia
12.
Journal of Experimental Hematology ; (6): 1012-1018, 2020.
Article in Chinese | WPRIM | ID: wpr-827169

ABSTRACT

OBJECTIVE@#To develop an automated chimeric analysis and reporting platform based on short tandem repeat (STR) and capillary electrophoresis methods for allogeneic hematopoietic stem cell transplantation (allo-HSCT) so as to improve work efficiency.@*METHODS@#Apache, MySQL, PHP and HTML5 were used to build the database and interface. The STR locus geno typing and chimeric analysis logic and flow were set up on the basis of STR rules and capillary electrophoresis. STR genotyping and 194 times of chimeric testing data of 100 patients after allo-HSCT were used to test the platform for automatic STR locus genotyping, chimeric calculation and report generation.@*RESULTS@#The established platform could realize the functions of STR locus customization, STR genotype determination, automatic chimeric analysis, and detection information database management, which can automatically generate an integrated report including multiple sequential chimeric results and trend graphs for the same patient and can be accessed and used simultaneously by different users through different browser interfaces. The results of automated analysis by the platform are completely consistent with that of manual analysis by experienced technicians, and the possibility of manual analysis error is reduced through automation. The time required for automatic analysis using this platform is approximately 1/6-1/5 of manual analysis.@*CONCLUSION@#The automatic analysis platform built in this study is operation stable and reliable in analysis results, which can improve work efficiency and report connotation, thus worthing popularized and applicable.


Subject(s)
Humans , Electrophoresis, Capillary , Genotype , Hematopoietic Stem Cell Transplantation , Microsatellite Repeats , Tissue Donors
13.
Journal of Experimental Hematology ; (6): 1998-2003, 2020.
Article in Chinese | WPRIM | ID: wpr-880005

ABSTRACT

OBJECTIVE@#To evaluate the proformance of multiplex PCR and capillary electrophoresis(MPCE) in the detection of JAK2V617F and CALR mutation in myeloproliferative neoplasms(MPN).@*METHODS@#The specificity primers of JAK2617F gene mutation and the primers of CALR gene were designed at the same time. The JAK2V617F and CALR gene primers were labeled with Cy5 fluorescence, all the primers were mixed in one tube for multiplex PCR and the PCR prodcuts were analysised by capillary electrophoresis. Then detection limit and sensitivity of MPCE were evaluated, and compared with comercial diagnostic kit.@*RESULTS@#JAK2V617F and CALR gene mutations could be detect by MPCE in one PCR test. JAK2V617F mutation could be detected at 0.01 ng genomic DNA, double positive JAK2V617F and CLAR gene mutations could be detected at 0.1 ng genomic DNA, at least 0.1% JAK2V617F positive mutation could be detected. The consistency between MPCE and commercial diagnostic gene mutation kit was 100%.@*CONCLUSION@#It is developed that a new gene mutation detection method of JAK2 V617F and CLAR gene based on MPCE in our experiment and it can be used as a new reagent for molecular diagnosis of MPN patients.


Subject(s)
Humans , Calreticulin/genetics , Electrophoresis, Capillary , Janus Kinase 2/genetics , Mutation , Myeloproliferative Disorders/genetics , Neoplasms , Patients , Polymerase Chain Reaction
14.
Bol. latinoam. Caribe plantas med. aromát ; 36(1): 5-10, 2020. ilus, Graf
Article in Spanish | MTYCI, LILACS | ID: biblio-1146008

ABSTRACT

Ligaria cuneifolia (R. et P.) Tiegh. ­Loranthaceae­ es una especie hemiparásita que se desarrolla sobre diferentes hospedantes. Es conocida con el nombre vulgar de "liga" o "liguilla". Debido a su similitud morfológica, constituye el sustituto natural del "muérdago europeo", por lo cual es denominado "muérdago criollo". Las drogas vegetales son matrices complejas en las cuales múltiples componentes actúan en forma sinérgica y son responsables de la acción farmacológica. Con el fin de dar sustento científico al uso folclórico de L. cuneifolia se estudiaron distintas formas de obtención de los extractos, se evaluaron diferentes hospedantes y regiones fitogeográficas. Se desarrolló y validó un método de electroforesis capilar para construir fingerprints o perfiles cromatográficos característicos que permitan evaluar los distintos componentes con el fin de estandarizar los extractos. Se efectuó la comparación con otras técnicas cromatográficas, tales como en cromatografía en capa delgada (TLC) y líquida de alta resolución (HPLC). A su vez, se procedió al aislamiento, purificación y análisis estructural de los compuestos de interés por técnicas espectroscópicas y cromatográficas. Se identificaron diez compuestos, de los cuales cuatro son reportados por primera vez en esta especie. La electroforesis capilar probó ser una técnica adecuada para el control de calidad de los extractos y una alternativa atractiva a las técnicas cromatográficas tradicionales.


Ligaria cuneifolia (R. et P.) Tiegh. ­Loranthaceae­ is a hemiparasite plant which grows on different host trees. It is popularly referred to as "liga" or "liguilla". Due to its morphological similarity, it is considered as the natural substitute for the European mistletoe, for which is known as the "Argentine mistletoe". Herbal drugs are complex matrices in which multiple components acting synergistically are responsible for the pharmacological activity. In order to provide scientific support to the popular use of L. cuneifolia, a capillary electrophoretic method was developed and validated to build a chromatographic profile or fingerprint that allows the evaluation of different components for extract standardization. A comparison was made with other chromatographic techniques such as TLC and HPLC. Isolation, purification and structural analysis of compounds were performed by chromatographic and spectroscopic methods. Ten analytes were identified, four of which are reported for the first time in L. cuneifolia. Capillary electrophoresis proved to be an appropriate tool for the quality control of herbal drugs, as well as an attractive alternative to traditional chromatographic techniques.


Subject(s)
Electrophoresis, Capillary , Loranthaceae , Mistletoe , Chromatography, Thin Layer , Flavonols
15.
Rev. nefrol. diál. traspl ; 39(2): 93-100, jun. 2019. ilus.; gráf.
Article in Spanish | LILACS | ID: biblio-1352684

ABSTRACT

Introducción: Las interferencias en el proteinograma electroforético por electroforesis capilar incluyen la aparición de picos con concentraciones y movilidades electroforéticas, que podrían simular la presencia de un componente monoclonal. Objetivos: Ante la aparición de un pico adicional con movilidad intera2-ß por electroforesis capilar (Minicap®-Sebia), el objetivo fue identificar el interferente y evaluar su relación con la funcionalidad renal. Material y métodos: Se estudiaron muestras de suero que presentaron dicha interferencia en un período de un año mediante proteinograma en soporte sólido, electroinmunofijación e inmunoelectroforesis. Se adicionó in vitro el probable interferente para confirmar su movilidad electroforética. Se evaluó el impacto de la corrección de la interferencia con la herramienta "eliminación de artefactos" (Phoresis®-Sebia) y la correlación de la concentración del pico a línea de base del interferente con la estimación de la tasa de filtrado glomerular (CKD- EPI). Resultados: La integración a la línea de base de los picos fue de 0,07-0,36 g/dL. No se observaron particularidades al realizar los estudios complementarios. Se evidenció, en todos los casos, la administración de iopamidol como medio de contraste, confirmándose su movilidad electroforética por su adición in vitro. Mediante la herramienta "eliminación de artefactos" se recuperaron los niveles basales de las fracciones. Se demostró la existencia de una correlación entre la concentración del pico a línea de base del interferente y la estimación de la tasa de filtración glomerular por CKD-EPI (r=-0.534, p<0.0001). Conclusiones: Se identificó al interferente como Iopamidol y se demostró su relación con la disminución de la tasa de filtración glomerular


Introduction: Interferences in the electrophoretic proteinogram by capillary electrophoresis include the appearance of peaks with concentrations and electrophoretic mobilities, which could simulate the presence of a monoclonal component. Objectives: In the light of an additional peak with interα2-ß mobility by capillary electrophoresis (MINICAP®-Sebia), the aim was to identify the interferent and evaluate its connection to renal functionality. Methods: Serum samples that presented this interference over a period of one year were studied by proteinogram on solid support, electroimmunofixation and immunoelectrophoresis. The probable interferent was added in vitro to confirm its electrophoretic mobility. The impact of the interference correction with the "artifact removal" tool (Phoresis®-Sebia) and the correlation of the baseline peak concentration of the interferent with the estimation of the glomerular filtration rate (CKD-EPI) were evaluated. Results: The integration to the baseline of the peaks was 0.07-0.36 g/dL. No particularities were observed when performing the complementary studies. In all cases, the administration of Iopamidol as a contrast medium was demonstrated, confirming its electrophoretic mobility due to its in vitro addition. Using the "artifact removal" tool, the basal levels of the fractions were recovered. The existence of a correlation between the concentration of the baseline peak of the interferent and the estimation of the glomerular filtration rate by CKD-EPI was shown (r=-0.534, p <0.0001). Conclusions: The interferent was identified as Iopamidol and its connection to the decrease in the glomerular filtration rate was demonstrated


Subject(s)
Humans , Iopamidol , Blood Proteins , Electrophoresis, Capillary , Contrast Media , Immunoelectrophoresis , Serum/drug effects , Glomerular Filtration Barrier
16.
Blood Research ; : 38-44, 2019.
Article in English | WPRIM | ID: wpr-739436

ABSTRACT

BACKGROUND: Microcytic anemia, the most common form of anemia in children and adolescents, is a heterogeneous group of diseases that is acquired or inherited. We assessed the frequency and causes of microcytosis in children and adolescents with the sickle cell trait (SCT). METHODS: This descriptive study included 95 subjects (49 males and 46 females) with SCT who attended Basra Center for Hereditary Blood Diseases for evaluation. Investigations included complete blood count, high performance liquid chromatography, capillary electrophoresis, and measurement of serum ferritin and transferrin levels. RESULTS: SCT subjects had a low hemoglobin (Hb) concentration (9.79±1.75 g/dL), low mean corpuscular volume (MCV, 67.43±9.22), low mean corpuscular Hb (21.15±3.64), and a normal red cell distribution width (RDW, 14.00±2.30). Among 95 SCT subjects, 81 (85.26%) had microcytosis, 12 (12.63%) had normal MCV, and 2 (2.11%) exhibited macrocytosis. Sixty-three (77.78%) SCT subjects with microcytosis were iron deficient, and 18 (22.22%) had normal iron levels. The mean serum ferritin and HbA2 levels were significantly lower, while the RDW, sickle Hb, and serum transferrin levels were significantly higher in patients with microcytosis and iron deficiency compared to non-iron deficient subjects (P0.05). CONCLUSION: Despite the frequent occurrence of iron deficiency in SCT subjects, co-inheritance of alpha-thalassemia seemed to be the cause of low MCV in non-iron deficient individuals with microcytosis. Genetic analysis is required to understand the genetic basis of this phenomenon.


Subject(s)
Adolescent , Child , Humans , Male , alpha-Thalassemia , Anemia , Blood Cell Count , Chromatography, Liquid , Electrophoresis, Capillary , Erythrocyte Indices , Ferritins , Hematologic Diseases , Iraq , Iron , Sickle Cell Trait , Transferrin
17.
Article in English | WPRIM | ID: wpr-777408

ABSTRACT

INTRODUCTION@#Haemoglobinopathy testing is performed for carrier screening and evaluation of microcytic anaemia. We evaluated the effectiveness of thalassaemia screening tests at our institution and suggest ways of improving the testing algorithm.@*MATERIALS AND METHODS@#A total of 10,084 non-antenatal and 11,364 antenatal samples with alkaline gel electrophoresis (AGE), capillary electrophoresis (CE), haemoglobin H (HbH) inclusion test, mean corpuscular haemoglobin (MCH) and mean corpuscular volume (MCV) were retrospectively reviewed. A subgroup of 187 samples with genetic testing was correlated with HbH inclusions and MCH/ MCV. The effect of iron deficiency on percentage hemoglobin A2 (HbA2) was studied.@*RESULTS@#HbH inclusion test showed low sensitivity of 21.43% for α-thalassaemia mutations but higher sensitivity of 78.95% for deletion. By receiver operating characteristic (ROC) analysis, MCH ≤28 pg or MCV ≤80 fl for non-antenatal samples and MCH ≤27 pg or MCV ≤81 fl for antenatal samples had >98% sensitivity for HbH inclusions. Above these thresholds, the probability that HbH inclusions would be absent was 99%). MCH ≥28 pg had 100% sensitivity (95% CI 95.63%-100%) for α-thalassaemia mutations and 97.68% calculated NPV in the antenatal population. Detection of haemoglobin variants by CE correlated highly with AGE (99.89% sensitivity, 100% specificity). Severe iron deficiency reduced HbA2 in hemoglobin ( <0.001) and α-thalassaemia ( = 0.0035), but not in β-thalassaemia.@*CONCLUSION@#MCH/MCV thresholds have adequate sensitivity for α-thalassaemia in the antenatal population, and genotyping plays an important role as HbH inclusion test shows low sensitivity. CE without AGE, may be used as initial screening for haemoglobin variants. Our study provides contemporary data to guide thalassaemia screening algorithms in Singapore.


Subject(s)
Female , Humans , Male , Pregnancy , Blood Protein Electrophoresis , Electrophoresis, Capillary , Erythrocyte Inclusions , Pathology , Erythrocyte Indices , Genetic Testing , Hemoglobin H , Mass Screening , Pregnancy Complications, Hematologic , Blood , Diagnosis , Retrospective Studies , Sensitivity and Specificity , Singapore , alpha-Thalassemia , Blood , Diagnosis
18.
São Paulo; s.n; s.n; 2019. 59 p. graf, tab, ilus.
Thesis in Portuguese | LILACS | ID: biblio-1008520

ABSTRACT

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


Subject(s)
Nanoparticles , Drug Liberation , In Vitro Techniques , Chromatography, High Pressure Liquid/methods , Electrophoresis, Capillary/methods , Imatinib Mesylate/analysis
19.
Rev. chil. pediatr ; 90(1): 26-35, 2019. tab
Article in Spanish | LILACS | ID: biblio-990883

ABSTRACT

OBJETIVO: Determinar la factibilidad de la identificación genética a un grupo de recién nacidos prove nientes de un hospital público de Lima-Perú. MATERIAL Y MÉTODO: Estudio descriptivo de corte trans versal, realizado por Registro de Identificación y Estado Civil de Perú, en recién nacidos vivos y sus respectivas madres, provenientes del Hospital Carlos Lanfranco La Hoz (Puente Piedra-Lima) du rante el mes de enero del 2015. Las muestras fueron colectadas en tarjetas FTA (Fast Technology for Analysis of nucleic acids) que permitieron un análisis directo por PCR (Polymerase Chain Reaction) y electroforesis capilar de 21 marcadores genéticos de tipo STR (Short Tandem Repeats), incluyendo el marcador amelogenina para la determinación del sexo. RESULTADOS: Se incluyeron un total de 44 madres y 45 recién nacidos (existió un parto gemelar). La probabilidad de maternidad fue mayor al 99.9% en todos los casos. No se encontraron dificultades en la toma de muestra, ni en el transporte del material. El material biológico obtenido fue suficiente para la obtención de ADN para realizar la identificación del recién nacido. CONCLUSIONES: El procedimiento de identificación genética fue factible de realizar en este hospital. Se identificaron etapas del proceso que podrían mejorarse para la posible aplicación de este procedimiento a una mayor escala en el Perú.


OBJECTIVE: To determine the feasibility of genetic identification in a group of newborns from a public hospital in Lima, Peru. MATERIAL AND METHOD: Descriptive cross-sectional study, carried out by the National Registry of Identification and Civil Status of Peru, on live newborns and their mothers, from the Carlos Lanfranco La Hoz Hospital (Puente Piedra, Lima) during January. 2015. The samples were collected in FTA (Fast Technology for Analysis of nucleic acids) cards that allowed a direct analysis by PCR (Polymerase Chain Reaction) and capillary electrophoresis of 21 STR markers (Short Tandem Repeats), including the amelogenin marker for gender determination. RESULTS: 44 mothers and 45 newborns were included (there was a twin birth). The probability of maternity was higher than 99.9% in all cases. There were no difficulties in the sampling or in transporting the material. The obtained biological material was enough to collect DNA to identify the newborn. CONCLUSIONS: The genetic identification procedure was possible to perform in this hospital. Stages of the process that could be improved were identified for the eventual application of this procedure on a larger scale in Peru.


Subject(s)
Humans , Male , Female , Infant, Newborn , Pedigree , Genetic Testing/methods , Neonatal Screening/methods , Peru , Genetic Markers , Pilot Projects , Feasibility Studies , Polymerase Chain Reaction , Cross-Sectional Studies , Microsatellite Repeats , Electrophoresis, Capillary , Medical Errors/prevention & control
20.
Braz. J. Pharm. Sci. (Online) ; 55: e17786, 2019. tab, graf
Article in English | LILACS | ID: biblio-1039051

ABSTRACT

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.


Subject(s)
Research Design , Electrophoresis, Capillary/standards , Proton Pump Inhibitors/analysis
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