Simultaneous determination of cyanide and thiocyanate in milk by GC-MS/MS using cetyltrimethylammonium bromide as both phase transfer catalyst and protein precipitant.

A method was developed for simultaneous determination of cyanide and thiocyanate in milk by gas chromatography-tandem quadrupole mass spectrometry (GC-MS/MS). Cyanide and thiocyanate were derivatized with pentafluorobenzyl bromide (PFBBr) as PFB-CN and PFB-SCN, respectively. Cetyltrimethylammonium bromide (CTAB) was employed both as a phase transfer catalyst and a protein precipitant in the sample pretreatment, which facilitates the separation of the organic and aqueous phases, and greatly simplifies the pretreatment procedures to achieve simultaneous and rapid determination of cyanide and thiocyanate. Under the optimized conditions, the limits of detection (LODs) of cyanide and thiocyanate in milk were 0.006 mg/kg and 0.015 mg/kg, and the spiked recoveries ranged from 90.1% to 98.2% and from 91.8% to 98.9% with relative standard deviations (RSDs) less than 18.9% and 15.2%, respectively. The proposed method was validated as a simple, fast and highly sensitive method for the determination of cyanide and thiocyanate in milk.

Physicochemical Properties and Antibiofilm Activity of Tricalcium Silicate Cement and its Association with Cetrimide / Propiedades físico-químicas y actividad antibiofilm del cemento de silicato tricálcico y su asociación con cetrimida

Abstract Cetrimide (CTR) is a cationic surfactant detergent with antimicrobial and antibiofilm activity. The aim of this study was to evaluate setting time, pH, solubility and antibiofilm activity of tricalcium silicate cement (TSC) with zirconium oxide (ZrO2) and its association with 0.2 and 0.4% cetrimide. Initial and final setting times (IST and FST) were assessed based on ISO-6876. pH was evaluated at periods of 1,3,7,14 and 21 days. Solubility was analyzed by weight loss. A modified direct contact test (MDCT) on the biofilm of Enterococcus faecalis formed on bovine root dentin blocks was performed, after 6 hours of manipulation and 15 hours of contact time. The analysis was performed by UFC mL ¯¹ counting. The data were analyzed by ANOVA and Tukey's tests (α=0.05). Higher IST was observed for TSC/ZrO2+CTR in both concentrations than for TSC/ZrO2 and lower FST for TSC/ZrO2+0.4% CTR (p0.05). All materials exhibited increased mass. TSC/ZrO2+CTR 0.4% had lower mass gain than the other materials (p<0.05). The highest antibiofilm activity was observed for TSC/ZrO2+CTR in both concentrations, when compared with the positive control (p<0.05). In conclusion, CTR exhibited potential to promote greater antibiofilm activity to tricalcium silicate cement, without harming its physicochemical properties of setting time, pH and solubility.

Resumen Cetrimida (CTR) es un detergente y surfactante catiónico con actividad antimicrobiana y antibiofilm. El objetivo de este estudio fue evaluar el tiempo de fraguado, pH, solubilidad y actividad antibiofilm del cemento de silicato tricálcico (CST) con óxido de zirconio (ZrO2) y su asociación con CTR a 0.2% y 0.4%. Tiempo de fraguado inicial y final (TFI y TFF) fueron determinados con base en las normas ISO-6876. pH fue evaluado en los períodos de 1,3, 7,14 y 21 días. Solubilidad fue analizada por la pérdida de masa/peso. El test de contacto directo modificado (TCDM) fue realizado sobre biofilm de Enterococcus faecalis formado en dentina radicular bovina, después de 6 horas de manipulación de los cementos y 15 horas de contacto. El análisis fue realizado por la cuantificación de UFC mL¯¹. Los datos fueron analizados usando las pruebas de ANOVA y Tukey (α=0.05). Mayor TFI fue observado para CST/ ZrO2+CTR en las dos concentraciones que para TSC/ZrO2 y menor TFF para TSC/ ZrO2+CTR 0.4% (p0.05). Todos los materiales mostraron aumento de masa. TSC/ZrO2+CTR 0.4% tuvo menor gano de masa que los otros materiales (p<0.05). Mayor actividad antibiofilm fue observado para CST/ZrO2+CTR en las dos concentraciones, cuando comparados con el grupo control positivo (p<0.05). En conclusión, CTR demostró potencial para promover superior actividad antibiofilm al cemento de silicato tricálcico (CST), sin perjudicar sus propiedades físico-químicas de tiempo de fraguado, pH y solubilidad.

Quantification of residual cetyltrimethylammonium bromide (CTAB) and sodium deoxycholate (DOC) in Haemophilus influenzae type b (Hib) polysaccharide using NMR.

CTAB and DOC are used as reagents in the purification of Hib polysaccharide. Polysaccharide is purified by precipitation with CTAB from fermented broth followed by solvent extraction and DOC is used to remove the protein impurities. The reagents used in the purification process should be removed from the product as per regulatory requirements. These two residual reagents can be easily identified and quantified in purified Haemophilus influenzae type b polysaccharide by NMR. The LOD of these residual reagents is 0.1% (10 µg/mL) and LOQ is 0.5% (50 µg/mL) with respect to polysaccharide determined from the spectrum. The absence of the peaks corresponding to CTAB and DOC in the NMR spectrum of purified polysaccharide confirms either they are absent or present at less than 0.1%. The present study provides supporting data from the regulatory viewpoint, which can help in circumventing the time-consuming studies for the vaccine manufacturers to develop different analytical methods for identification and quantification of CTAB and DOC as per regulatory requirements.