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1.
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)
Electrophoresis, Capillary , Humans , Infant, Newborn , Mass Screening , Mutation , Thalassemia , beta-Thalassemia/genetics
2.
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 , Biomarkers , Leukemia, Myeloid, Acute/genetics , Polymerase Chain Reaction , DNA, Complementary , Reverse Transcription , Electrophoresis, Capillary
3.
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
4.
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
5.
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)
Calreticulin/genetics , Electrophoresis, Capillary , Humans , Janus Kinase 2/genetics , Mutation , Myeloproliferative Disorders/genetics , Neoplasms , Patients , Polymerase Chain Reaction
6.
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)
Electrophoresis, Capillary , Genotype , Hematopoietic Stem Cell Transplantation , Humans , Microsatellite Repeats , Tissue Donors
7.
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 , China , Electrophoresis, Capillary , Hemoglobinopathies , Hemoglobins, Abnormal , Humans , Thailand , Thalassemia
8.
Bol. latinoam. Caribe plantas med. aromát ; 36(1): 5-10, 2020. ilus, Graf
Article in Spanish | MTYCI, LILACS, MTYCI | 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
9.
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
10.
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)
Blood Protein Electrophoresis , Electrophoresis, Capillary , Erythrocyte Inclusions , Pathology , Erythrocyte Indices , Female , Genetic Testing , Hemoglobin H , Humans , Male , Mass Screening , Pregnancy , Pregnancy Complications, Hematologic , Blood , Diagnosis , Retrospective Studies , Sensitivity and Specificity , Singapore , alpha-Thalassemia , Blood , Diagnosis
11.
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 , alpha-Thalassemia , Anemia , Blood Cell Count , Child , Chromatography, Liquid , Electrophoresis, Capillary , Erythrocyte Indices , Ferritins , Hematologic Diseases , Humans , Iraq , Iron , Male , Sickle Cell Trait , Transferrin
12.
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
13.
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
14.
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
15.
Frontiers of Medicine ; (4): 196-205, 2018.
Article in English | WPRIM | ID: wpr-772750

ABSTRACT

We employed a multiplex polymerase chain reaction (PCR) coupled with capillary electrophoresis (mPCR-CE) targeting six Clostridium difficile genes, including tpi, tcdA, tcdB, cdtA, cdtB, and a deletion in tcdC for simultaneous detection and characterization of toxigenic C. difficile directly from fecal specimens. The mPCR-CE had a limit of detection of 10 colony-forming units per reaction with no cross-reactions with other related bacterial genes. Clinical validation was performed on 354 consecutively collected stool specimens from patients with suspected C. difficile infection and 45 isolates. The results were compared with a reference standard combined with BD MAX Cdiff, real-time cell analysis assay (RTCA), and mPCR-CE. The toxigenic C. difficile species were detected in 36 isolates and 45 stool specimens by the mPCR-CE, which provided a positive rate of 20.3% (81/399). The mPCR-CE had a specificity of 97.2% and a sensitivity of 96.0%, which was higher than RTCA (x = 5.67, P = 0.017) but lower than BD MAX Cdiff (P = 0.245). Among the 45 strains, 44 (97.8%) were determined as nonribotype 027 by the mPCR-CE, which was fully agreed with PCR ribotyping. Even though ribotypes 017 (n = 8, 17.8%), 001 (n = 6, 13.3%), and 012 (n = 7, 15.6%) were predominant in this region, ribotype 027 was an important genotype monitored routinely. The mPCR-CE provided an alternative diagnosis tool for the simultaneous detection of toxigenic C. difficile in stool and potentially differentiated between RT027 and non-RT027.


Subject(s)
Clostridium Infections , Diagnosis , Clostridioides difficile , Genetics , Electrophoresis, Capillary , Feces , Microbiology , Genes, Bacterial , Humans , Polymerase Chain Reaction , Ribotyping , Sensitivity and Specificity
16.
Journal of Experimental Hematology ; (6): 1663-1667, 2018.
Article in Chinese | WPRIM | ID: wpr-773039

ABSTRACT

OBJECTIVE@#To investigate the feasibility of sensitive and quantitative detection of MYD88 gene L265P mutation in lymphoma patients by using ARMS-PCR combined with capillary electrophoresis.@*METHODS@#ARMS-PCR amplified MYD88 gene was analyzed by capillary electrophoresis in ABI 3730 sequencer; Exon 5 of the same gene was sequenced bi-directionally as reported.@*RESULTS@#The sensitivity of detection L265P mutations by the ARMS-PCR combined with capillary electrophoresis and direct sequencing was 0.2% and 5%, respectively, according to the detection of the gradient-diluted plasmid standards. The detection rate of 184 patients was 13.59% and 8.28%, respectively (p<0.001). Moreover, the former method can successfully detect the mutation ratio(R=0.979), and the repeatabilities (CV=2.86%, 1.94%, 5.49%) are acceptable.@*CONCLUSION@#ARMS-PCR combined with capillary electrophoresis can quantitatively detect the MYD88 gene L265P mutation, and the detection sensitivity is significantly higher than sanger sequencing. As a supplement to the latter, it can effectively lead to the earlier diagnose and monitoring of minimal residual disease.


Subject(s)
DNA Mutational Analysis , Electrophoresis, Capillary , Humans , Lymphoma , Mutation , Myeloid Differentiation Factor 88 , Genetics , Polymerase Chain Reaction
17.
São Paulo; s.n; s.n; 2018. 88 p. graf, tab.
Thesis in Portuguese | LILACS | ID: biblio-980422

ABSTRACT

Estudos envolvendo os glicocorticoides merecem destaque devido a serem hormônios responsáveis pela transferência de informações e instruções às células, desta forma regulando o metabolismo, desenvolvimento, crescimento, função imune e também auxiliam no controle das funções tanto reprodutivas quanto tecidual. Estes também são sintetizados e amplamente utilizados com finalidade terapêutica processos alérgicos, tratamento de doenças autoimunes, em transplantes no pré-operatório e/ou pós-operatório-, devido a sua eficiente ação como imunossupressores e anti-inflamatórios. Os dois primeiros capítulos deste trabalho exibem uma revisão da literatura com foco em considerações gerais sobre os glicocorticoides, metodologias empregadas na análise destes hormônios e fundamentos da eletroforese capilar. Na sequência, o quarto capitulo, mostra a otimização da separação de 17 glicocorticoides utilizando cromatografia eletrocinética micelar devido a alto grau hidrofóbico dos analitos. Para tal, a composição do eletrólito consistiu em 20mM de tetraborato de sódio (pH=9.3) e 30 mM de dodecil sulfato de sódio (como surfactante), e a interação soluto-micela e, portanto, retenção do soluto, foi manipulada com a adição (volume/volume) de solventes orgânicos na composição de até 20% acetonitrila (ACN), 20% etanol (EtOH) e 1% tetrahidrofurano (THF), a qual se baseia num modelo de desenho de misturas (totalizando dez diferentes eletrólitos), e através desta abordagem um ótimo de separação foi obtido (13,3% EtOH, 3,3% ACN e 0,17% THF). A melhor condição de separação foi testada qualitativamente numa amostra de urina de um voluntário que faz uso contínuo de prednisona como terapia corticoidal. As misturas de solventes estudadas neste trabalho afetam a solubilidade dos hormônios na fase aquosa e a estrutura micelar também sofre grande impacto,principalmente na camada de solvatação. O quarto capítulo busca racionalizar tais efeitos através da obtenção de descritores, e as informações contidas nos descritores hidrofóbicos e hidrofílicos são sempre relevantes e contribuem nas correlações encontradas. Obteve três grupos de comportamento distinto, onde a capacidade doadora e aceptora de prótons para a realização de ligações de hidrogênios foram as interações consideradas as mais relevantes para o comportamento observado da separação. E o capítulo final, apresenta possibilidades de aproveitamento no controle de qualidade na indústria farmacêutica, métodos baseados na injeção e tensão inversas foram propostos a fim de ganho de tempo de análise (máximo de 5 minutos), estes foram validados seguindo o protocolo preconizado pela ANVISA (Agência Nacional de Vigilância Sanitária) nos parâmetros: precisão, exatidão, seletividade, linearidade, limites de detecção e quantificação e robustez; e aplicados na quantificação de quatro (diferentes formulações comerciais contendo glicocorticoides (prednisona 20 mg, betametasona 4 mg, furoato de mometasona 200 mcg e dipropionato de beclometasona 200 mcg)


Studies involving glucocorticoids deserve to be highlighted because they are hormones responsible for the transfer of information and instructions to cells, thus regulating metabolism, development, growth, immune function and also assist in the control of both reproductive and tissue functions. These are also synthesized and widely used for therapeutic purposes - allergic processes, treatment of autoimmune diseases, in preoperative and/or postoperative transplants - due to their efficient action as immunosuppressants and anti-inflammatories. The first two chapters of this paper present a review of the literature focusing on general considerations about glucocorticoids, methodologies used in the analysis of these hormones and fundamentals of capillary electrophoresis. Subsequently, the fourth chapter shows the optimization of the separation of 17 glucocorticoids using micellar electrokinetic chromatography due to the high hydrophobic degree of the analytes. To this end, the electrolyte composition consisted of 20 mM sodium tetraborate (pH = 9.3) and 30 mM sodium dodecyl sulfate (as a surfactant), and the solute-micelle interaction and therefore solute retention was manipulated with organic solvent in the composition of up to 20% acetonitrile (ACN), 20% ethanol (EtOH) and 1% tetrahydrofuran (THF), which is based on a mixture design model (totaling ten different electrolytes), and through this approach an optimal separation was obtained (13.3% EtOH, 3.3% ACN and 0.17% THF). The best separation condition was qualitatively tested in a urine sample from a volunteer who makes continuous use of prednisone as corticosteroid therapy. The solvent mixtures studied in this work affect the solubility of the hormones in the aqueous phase and the micellar structure also has a great impact, especially on the solvation layer. The fourth chapter seeks to rationalize these effects by obtainingdescriptors, and the information contained in the hydrophobic and hydrophilic descriptors is always relevant and contributes to the correlations found. It obtained three groups of distinct behavior, where the donor and acceptor capacity of protons for the realization of hydrogen bonds were the interactions considered the most relevant for the observed behavior of the separation. And the final chapter presents possibilities of use in quality control in the pharmaceutical industry, methods based on injection and reverse voltage were proposed in order to gain analysis time (maximum of 5 minutes), these were validated following the protocol recommended by ANVISA (Brazilian National Agency of Sanitary Surveillance) in the parameters: precision, accuracy, selectivity, linearity, limits of detection and quantification and robustness; and applied in the quantification of four different commercial formulations containing glucocorticoids (prednisone 20 mg, betamethasone 4 mg, mometasone furoate 200 mcg and beclomethasone dipropionate 200 mcg)


Subject(s)
Electrophoresis, Capillary , Drug Compounding/statistics & numerical data , Glucocorticoids/analysis , Steroids , Chromatography, Micellar Electrokinetic Capillary/methods
18.
Braz. J. Pharm. Sci. (Online) ; 54(4): e17585, 2018. tab, graf
Article in English | LILACS | ID: biblio-1001581

ABSTRACT

A simple and fast alternative methodology using capillary zone electrophoresis (CZE) to analyze linezolid and its cationic photodegradation products in tablets has been developed. The separation was carried out on fused silica capillary and conducted using 100 mM formic acid (pH 3.0) and by applying 30 KV voltage. Detection was performed at UV 254 nm. The optimized method was validated in terms of linearity, limits of detection and quantification, precision (repeatability), stability studies (selectivity) and accuracy. Good linearity (8-20 mg L-1) was obtained and the limit of detection was 0.95 mg L-1. The greatest advantages of the CZE method were the rapid set-up of instrumentation and capillary equilibration, short analysis time (12 min), low running cost and low waste generation. The method showed good stability in determining linezolid submitted to degradation by light and to a climatic chamber and can be used as an alternative for evaluation in stability studies of linezolid in tablets, as well as for the analysis of the drug in raw materials and finished products.


Subject(s)
Tablets/classification , Electrophoresis, Capillary/instrumentation , Linezolid/analysis , Photobleaching , Drugs from the Specialized Component of Pharmaceutical Care
20.
Article in Chinese | WPRIM | ID: wpr-344157

ABSTRACT

<p><b>OBJECTIVE</b>To explore the feasibility of using PCR-based capillary electrophoresis method to analysis mutation of the TOR1A gene in a family affected with primary torsion dystonia (PTD).</p><p><b>METHODS</b>Peripheral blood sample was collected from proband and amnionic fluid from her fetus for the extraction of DNA. The 5th exon of the TOR1A gene and its flanking sequences were amplified with PCR and analyzed with agarose electrophoresis, fluorescence labeled fragment analysis and Sanger sequencing.</p><p><b>RESULTS</b>Fluorescence labeled fragment analysis was performed through capillary electrophoresis, which showed that the proband carried a c.907_909delGAG (p.Glu303del) deletional mutation of the TOR1A gene. The result was verified by Sanger sequencing. The fetus DNA was also found with the same mutation by capillary electrophoresis, inferring that the fetus was probably affected with the disease.</p><p><b>CONCLUSION</b>The mutation of c.907_909delGAG of the TOR1A gene was speculated as pathologic cause of proband in this family. Fragment analysis by capillary electrophoresis combined with DNA sequencing is an efficient test for small deletional mutations and feasible for its prenatal diagnosis.</p>


Subject(s)
Adult , Dystonia , Diagnosis , Genetics , Electrophoresis, Capillary , Female , Humans , Molecular Chaperones , Genetics , Mutation , Prenatal Diagnosis , Sequence Analysis, DNA
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