Simultaneous Determination of Seven Residual Solvents in Nilethylenol Raw Material by GC / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of residual methanol， ethanol， acetonitrile， isopropanol， methylbenzene， tetrahydrofuran and ethyl acetate in Nilethylenol raw material. METHODS： GC was performed. The determination was performed on DB-WAX capillary column. The detector was hydrogen flame ionization detector with split ratio of 5 ∶ 1. The carrier gas was nitrogen （purity： 99.999％） at the flow rate of 1.0 mL/min. The sample size was 1 μL， directly sampling with bonded crosslinked polyethylene glycol as stationary phase. The initial temperature was 40 ℃ and was maintained for 5 min， increased to 90 ℃ at 10 ℃/min， and then increased to 200 ℃ at 5 ℃/min. The temperature of injector was 220 ℃， and detector temperature was 230 ℃. RESULTS： The linear range was 0.24-12.00 μg/mL for methanol （r＝0.999 7）， 0.40-20.00 μg/mL for ethanol （r＝0.999 5）， 0.033-1.64 μg/mL for acetonitrile （r＝0.999 8）， 0.40-20.00 μg/mL for isopropanol （r＝0.999 5）， 0.071-3.56 μg/mL for methylbenzene （r＝0.999 6）， 0.058-2.88 μg/mL for tetrahydrofuran （r＝0.999 8）， 0.40-20.00 μg/mL for ethyl acetate （r＝0.999 7）， respectively. The limits of quantitation were 0.24， 0.40， 0.033， 0.40， 0.071， 0.058， 0.40 μg/mL， respectively. The limits of detection were 0.08， 0.10， 0.01， 0.13， 0.02， 0.02， 0.13 μg/mL， respectively. RSDs of precision， stability and reproducibility tests were all lower than 2％. The recoveries were 98.17％-100.48％ （RSD＝0.92％，n＝9）， 97.77％-101.30％（RSD＝1.32％，n＝9）， 97.56％-100.85％（RSD＝1.20％，n＝9）， 98.64％-100.92％（RSD＝0.87％，n＝9）， 98.54％-100.62％（RSD＝0.76％，n＝9），98.26％-100.00％（RSD＝0.74％，n＝9）， 98.30％-100.59％（RSD＝0.76％，n＝9）， respectively. CONCLUSIONS： The method has high sensitivity and good accuracy， and can be used for the simultaneous determination of residual methanol， ethanol， acetonitrile， isopropane， methylbenzene， tetrahydrofuran and ethyl acetate in Nilethylenol raw material.

Simultaneous Determination of 4 Residual Organic Solvents in Lacidipine Raw Material by GC / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of 4 residual organic solvents such as ethanol,dichloromethane,isopropanol and ethyl acetate in lacidipine raw material.METHODS:GC method was adopted.The determination was performed on DB-624 capillary column at the rate of 2.0 mL/min using nitrogen gas as carrier and FID as the detector by temperature programming.The temperature of detector was set at 250 ℃.Carries gas was nitrogen.The split ratio was 10 ∶ 1.The sample size was 10 μL.RESULTS:The linear ranges of ethanol,dichloromethane,isopropanol and ethyl acetate were 0.259 5-2.076 mg/mL (r=0.999 6),0.055 3-0.316 mg/mL (r=0.999 2),0.342 3-1.956 mg/mL (r=0.999 5),0.370 6-2.118 mg/mL(r=0.999 4),respectively.The limits of quantification were 2.96,2.24,4.4,4.44 μg/mL,and the detection limits were 0.887,0.672,1.3,1.32 μg/ mL,respectively.RSDs of precision,stability and reproducibility tests were all lower than 4.0％.The recoveries were 95.00％-101.17％ (RSD=2.19％,n=9),96.22％-104.53％ (RSD=3.27％,n=9),96.20％-104.90％ (RSD=2.41％,n=9),95.60 ％-104.48 ％ (RSD=2.85 ％,n=9).CONCLUSIONS:The method is simple,sensitive and reliable,and is suitable for simultaneous determination of 4 residual organic solvents in lacidipine raw material.

Simultaneous Determination of 6 Residual Organic Solvents in Xingnaojing Injection by Headspace GC / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 6 residual organic solvents in Xingnaojing injection,such as methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile.METHODS:Headspace GC method was adopted.The determination was performed on DB-624 capillary column by temperature programming with the injector temperature of 200 ℃;flame ionization detector was adopted with the temperature of 250 ℃;carrier gas was nitrogen with flow rate of 25 mL/min and split ratio of 35 ∶ 1;headspace sampling size was 1 mL,and heating temperature of headspace sampling was 80 ℃;equilibrium time was 15 min.RESULTS:The linear ranges of methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile were 15.00-240.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-399.99 μg/mL(r=0.999 9),25.00-399.99 μg/mL(r=0.999 8) and 5.00-80.00 μg/mL(r=0.999 9).The LOQ were 5.98,3.94,2.05,2.13,1.39,1.24 μg/mL,and the LOD were 2.01,2.11,1.18,1.56,1.15,0.01 μg/mL,respectively.RSDs of precision tests were all less than 2.0％,stability and repetitive tests only ethyl acetate was detected,RSD＜2.0％;the recoveries were 93.59％-99.02％ (RSD=2.62％,n=6),92.42％-98.40％ (RSD=2.43％,n=6),94.81％-104.64％ (RSD=3.47 ％,n=6),94.56％-106.73％ (RSD=4.21％,n=6),97.04％-106.33％(RSD=3.50％,n=6)and 98.40％-107.97％ (RSD=3.37％,n=6).CONCLUSIONS:The method is specific,rapid,simple and accurate,and can be used for simultaneous determination of 6 residual organic solvents in Xingnaojing injection.

Simultaneous Determination of 6 Residual Organic Solvents in Xingnaojing Injection by Headspace GC / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 6 residual organic solvents in Xingnaojing injection,such as methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile.METHODS:Headspace GC method was adopted.The determination was performed on DB-624 capillary column by temperature programming with the injector temperature of 200 ℃;flame ionization detector was adopted with the temperature of 250 ℃;carrier gas was nitrogen with flow rate of 25 mL/min and split ratio of 35 ∶ 1;headspace sampling size was 1 mL,and heating temperature of headspace sampling was 80 ℃;equilibrium time was 15 min.RESULTS:The linear ranges of methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile were 15.00-240.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-399.99 μg/mL(r=0.999 9),25.00-399.99 μg/mL(r=0.999 8) and 5.00-80.00 μg/mL(r=0.999 9).The LOQ were 5.98,3.94,2.05,2.13,1.39,1.24 μg/mL,and the LOD were 2.01,2.11,1.18,1.56,1.15,0.01 μg/mL,respectively.RSDs of precision tests were all less than 2.0％,stability and repetitive tests only ethyl acetate was detected,RSD＜2.0％;the recoveries were 93.59％-99.02％ (RSD=2.62％,n=6),92.42％-98.40％ (RSD=2.43％,n=6),94.81％-104.64％ (RSD=3.47 ％,n=6),94.56％-106.73％ (RSD=4.21％,n=6),97.04％-106.33％(RSD=3.50％,n=6)and 98.40％-107.97％ (RSD=3.37％,n=6).CONCLUSIONS:The method is specific,rapid,simple and accurate,and can be used for simultaneous determination of 6 residual organic solvents in Xingnaojing injection.

Simultaneous Determination of 6 Residual Organic Solvents in Omeprazole Magnesium by Headspace GC / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of 6 kinds of residual organic solvents in Omeprazole magnesium,such as methanol,isopropanol,acetonitrile,dichloromethane,ethyl acetate and toluenein. METHODS:Headspace GC was adopted. The determination was performed on the column of DB-624 column by temperature programming. The inlet temperature was 200 ℃,and hydrogen flame ionization detector was adopted with the temperature of 250 ℃;nitrogen gas was used as carrier gas with flow rate of 2.0 mL/min;the splitting-radio was 5 : 1,and sample size was 1 mL;the headspace tem-perature was 80 ℃ and the equilibrium time was 20 min. RESULTS:The linear range were 12.56-628.00 μg/mL for methanol(r=0.9997),20.22-1011.20 μg/mL for isopropanol(r=0.9999),1.96-97.76 μg/mL for acetonitrile(r=0.9997),3.10-154.88 μg/mL for dichloromethane(r=0.9998),20.69-1034.56 μg/mL for ethyl acetate(r=0.9998),and 3.53-176.72 μg/mL for toluene(r=0.9998);the limits of quantitation were 1.00,0.91,0.47,0.93,0.41 and 0.35 μg/mL respectively;the limits of detection were 0.31,0.30,0.14,0.31,0.12 and 0.11 μg/mL respectively;RSDs of precision,stability and reproducibility tests were lower than 3%;the recoveries were 94.53%-101.29%(RSD=2.15%,n=9),97.78%-103.42%(RSD=1.77%,n=9),96.99%-105.76%(RSD=2.59%,n=9),96.83%-102.05%(RSD=1.86%,n=9),97.98%-101.13%(RSD=0.88%,n=9)and 97.80%-102.40%(RSD=1.41%,n=9). CONCLUSIONS:The method is sensitive and accurate,can be used for the simultaneous determination of 6 kinds of residual organic solvents in Omeprazole magnesium,such as methanol,isopropanol,acetonitrile,dichloromethane,eth-yl acetate and toluenein.

Análise comparativa dos cimentos MTA cinza e branco associados ao álcool isopropílico / Comparative analysis of gray and white MTA associated with isopropyl alcohol

O objetivo deste estudo ex vivo foi investigar e comparar os efeitos antibacterianos, a citotoxicidade, a adaptação marginal e a capacidade seladora do agregado trióxido mineral (MTA), branco e cinza (Angelus, Londrina, Brasil), quando combinados com água destilada ou álcool isopropílico. Os efeitos antibacterianos de ambas as formulações foram avaliados através do teste de difusão em agar utilizando Staphylococcus aureus (ATCC25923) e Pseudomonas aeruginosa (ATCC27853) para a seleção da concentração de escolha. A geração das espécies reativas do oxigênio (ERO) foi analisada, primeiramente, utilizando cepas de Escherichia coli proficiente (AB1157) e triplo mutante (BW535), deficiente nos mecanismos de reparo. A geração de ERO também foi analisada através da degradação plasmidial (método de eletroforese) e com a utilização da sonda DCF. A geração das espécies reativas do nitrogênio (ERN) foi avaliada através da sonda DAF. A citotoxicidade dos eluatos dos cimentos foi avaliada pelo MTT, com células L929. A adaptação marginal foi feita através da microscopia eletrônica de varredura (MEV). A capacidade seladora foi realizada através de ensaios de infiltração bacteriana. Os resultados mostraram que a incorporação do isopropanol ao MTA aumentou os halos de inibição no modelo Gram-positivo (S. aureus-ATCC25923) (P <0.05). A adição de isopropanol 70% aos cimentos de MTA branco e cinza aumentou a atividade antimicrobiana para E. coli selvagem (AB1157) e triplo mutante (BW535) (P <0.0001). Não ocorreu a total degradação do DNA plasmidial para as amostras de MTA branco e cinza manipuladas com isopropanol 70%. As amostras de MTA branco e cinza manipuladas de acordo com o fabricante geraram significativamente mais espécies reativas do oxigênio (sonda DCF) (P <0.0001 e P =0.0002) e do nitrogênio (sonda DAF) (P =0.0002 e P =0.0003) do que as manipuladas com isopropanol 70%. Os cimentos manipulados com água destilada foram mais citotóxicos do que a associação com solução de isopropanol 70% (P <0.05). A análise ao MEV mostrou melhor adaptação ao preparo nas amostras com o isopropanol 70%, entretanto o ensaio de infiltração bacteriana ex vivo com Enterococcus faecalis (ATCC29212), não mostrou diferença estatística para as amostras manipuladas com isopropanol ou com água (P >0.05). Podemos concluir que a manipulação com isopropanol melhorou a atividade antimicrobiana do MTA tornando-o menos citotóxico, sem alterar a capacidade seladora.

The aim of this study was to investigate and compare the antibacterial effects, cytotoxicity, marginal adaptation and sealing ability of mineral trioxide aggregate (MTA), white and gray (Angelus, Londrina, Brasil), when combined with distilled water or isopropyl alcohol. The antibacterial effects of both formulations were evaluated by agar diffusion method using Staphylococcus aureus (ATCC25923) and Pseudomonas aeruginosa (ATCC27853) for selecting the suitable concentration of isopropyl alcohol. The antimicrobial mechanism by the generation of reactive oxygen species (ROS) was first analyzed using Escherichia coli strains: proficient (AB1157) and triple mutant (BW535), deficient in the repair mechanisms. The ROS generation was also analyzed by degradation of DNA plasmid (electrophoresis method) and with the DCF probe. The generation of reactive nitrogen species (RNS) was assessed by DAF probe. The cytotoxicity of the eluates was assessed by MTT test, with L929 cells. The marginal adaptation was evaluated by scanning electron microscopy (SEM). The sealing ability was performed using bacterial leakage assay. The results showed that the incorporation of isopropanol to MTA increased the inhibitory haloes in Gram-positive model (S. aureus-ATCC25923) (P <0.05). Adding 70% isopropanol to white and gray MTA cements increased antimicrobial activity against both E. coli wild (AB1157) and triple-mutant (BW535) strains (P <0.0001). The DNA plasmid was not completely degraded for the samples of white and gray MTA manipulated with 70% isopropanol. The white and gray MTA samples, handled according to the manufacturer, generated significantly more ROS (DCF probe) (P <0.0001 e P =0.0002) and RNS (DAF probe) (P =0.0002 e P =0.0003) than that manipulated with 70% isopropanol. The cements handled with distilled water were more cytotoxic than when associated with isopropanol solution 70% (P <0.05). The SEM analysis showed better adaptation to the cavity prepares when the samples were filled with MTA with 70% isopropanol, however the ex vivo bacterial infiltration test with Enterococcus faecalis (ATCC29212), was not statistically different from samples manipulated with isopropanol or water (P >0.05). We can conclude that the manipulation with isopropanol improved the antimicrobial activity of the MTA rendering less citotoxicity, without altering the sealing ability.

Histo-architechtural Evaluation of Conventional Versus Two Rapid Microwave Processing Techniques.

Aims: This work aims to reduce the time of tissue processing, exclude xylene from tissue processing as well as to reduce the total quantity of reagents used per cycle of paraffin wax processing technique. Study Design: Harvesting and fixation of tissues. Grossing into triplicates, processing using three different techniques. Staining and grading of sections. Place and Duration of Study: Apparently healthy rabbits from the animal house, National Veterinary research Institute, Vom, Jos, Nigeria, between August and December, 2014. Methodology: Two apparently healthy rabbits were sacrificed and the Trachea, Lungs, Heart, Liver, Kidney, Stomach, Skin, Brain, and the Spleen were harvested and fixed in 10% buffered formalin for three days. They were grossed into triplicates, labeled and processed using the conventional, microwave without vacuum and microwave with vacuum respectively. They were sectioned and stained simultaneously using the haematoxylin and eosin staining technique as well as the Gordon and Sweet’s method. They were graded as excellent if they permitted microscopy, fair if not very good but can permit microscopy and poor if they cannot permit microscopy at all. Results: There is a drastic reduction in the duration of processing as well as the quantity of reagents used in the microwave techniques as compared to the conventional method. Xylene was completely eliminated in the microwave techniques. Tissue histo-architechture, special features as well as silver impregnation were clearly demonstrated without significant differences. Dye uptake as well as section thickness were comparable among the three techniques. Conclusion: Same-day turn-around is possible in histology, with reduced reagent consumption and elimination of xylene, without compromising section quality, dye uptake or ability to reduce silver to its metallic form. This will result to quick diagnosis hence quick intervention at a cheaper rate to both laboratories and clients. It is a good innovation in forensic and diagnostic histopathology and should be encouraged. Its compatibility with histochemical, immunohistochemical and molecular techniques should be evaluated to give a wider application.

Etanol e isopropanol: genotoxicidad y teratogénesis evaluada aplicando el modelo de Drosophila melanogaster / Ethanol and isopropanol: genotoxicity and teratogenicity evaluated using Drosophila melanogaster as a model system

El etanol y el isopropanol son, de los alcoholes alifáticos de cadena corta, los más frecuentemente asociados a la actividad humana tanto a nivel industrial como en el entorno doméstico. En este trabajo se presentan los principales hallazgos reportados en la literatura para ensayos de genotoxicidad y teratogénesis en modelos experimentales de distinto nivel de complejidad, con especial énfasis en Drosophila melanogaster. El metabolismo de estos alcoholes es semejante en Drosophila y en humanos por lo cual la mosca es un buen modelo in vivo para la evaluación de sus potenciales efectos tóxicos, genotóxicos y teratogénicos.

Ethanol and isopropanol are two of the short chain aliphatic alcohols more frequently associated to the human environment, both in the industrial and domestic conditions. The aim of this work was to present the main findings reported in the literature about their genotoxicity and teratogenicity in experimental models of different level of complexity, with special emphasis in Drosophila melanogaster. Taking into account that the metabolism of both alcohols in Drosophila and humans is similar, the fly is a good model for the evaluation of their potentially toxic, genotoxic and teratogenic effects.