RESUMO
@#Objective To develop and verify a method for determination of cyanide residues in polysaccharide-protein conjugate vaccines based on flow injection analysis(FIA)with amperometric detector.Methods After removing macromolecular substances in the samples by ultrafiltration,the cyanide residues were determined by a 3700 automatic chemical analyzer with the injection time of 35 s,injection volume of 200 μL,pump speed of 40%,sample cycle time of 140 s,ultraviolet wavelength of 312 nm and an Ampere detector.The developed method was verified for the specificity,matrix effect,linear range,limit of detection(LOD),limit of quantitation(LOQ),accuracy,presicion and stability.The cyanide residues in the polysaccharide derivative bulk(13 batches)and conjugate bulk(21 batches)of Haemophilus influenzae type b conjugate vaccine and group A and C meningococcal conjugate vaccine produced by 5 manufacturers were determined by the developed method.Results Blank samples showed no interference to detection;The recoveries of the matrix effector solution of Haemophilus influenzae type b conjugate vaccine polysaccharide derivative and conjugate bulk,group A and C meningococcal conjugate vaccine polysaccharide derivative and conjugate bulk were 97.4%,102.4%,96.8% and 99.8% respectively,with all CV values less than 15%;In the range of 0.312 5 ～ 80 ng/mL,cyanogroup concentration showed a good linear relationship with peak height with a regression equation of y = 133.13 x + 57.556,R2= 0.999 1;The LOD was 0.2 ng/mL,and the LOQ was 0.6 ng/mL.The average recoveries of the control solution were 108.9%,106.5%,103.5% respectively with RSD value of 6.4% in the groups added with 5,10 and 20 ng/mL of cyanogroup and the CV values of precision verification were all less than 15%;The average concentrations of the control solution injected 20 times continuously were 76,38,18and 5 ng/mL with all CV values less than 15%,when cyanogroup concentration was 80,40,20 and 5 ng/mL,respectively.Cyanide residues were detected in 13 batches of derivative bulk samples,while not in 2 batches of 21 batches of polysaccharide conjugate bulk samples,and were detected in the others.Conclusion The developed method had good accuracy,precision and stability,which might be applied to the quantitative determination of cyanide residues in polysaccharideprotein conjugate vaccines.
RESUMO
El análisis químico día a día se acerca más a la automatización, buscando satisfacer las necesidades actuales de resultados rápidos y confiables. Los sistemas de análisis en flujo (FIA - Flow Injection Analysis) son una de las formas de aproximarse a la automatización. En este artículo se presentan los pasos necesarios para implementar una metodología FIA, para la determinación de Pb(II) en agua, partiendo de la revisión de los procedimientos clásicos y describiendo detalladamente los pasos necesarios para implementar la técnica de análisis en flujo. El trabajo produjo un método de análisis de Pb en agua que usa ditizona disuelta en isopropanol (agente cromogénico), en presencia de bromuro de cetiltrimetil amonio (CTAB), para solubilizar en agua el complejo, cuyas características más sobresalientes fueron: volumen de inyección de muestra de 81,7 µL, velocidad de flujo de 8,0 mL/min, tiempo de toma de espectros 1,4 s e intervalo lineal de 1,0 a 40 mg L-1.
Chemical analysis has evolved towards automation to satisfy the current requirements: fast analysis and certainty in the results. Flow injection analysis (FIA) is a way to reach automation. This work presents the necessary steps to obtain an optimized FIA methodology for the determination of Pb(II) in water by classic methods. The result was a FIA method to determinate Pb with dithizone (chromogenic agent) dissolved in iso-propyl alcohol, using cethyltrimethylammonium bromide (CTAB) to solubilize the complex. The main characteristics of the method were: injection sample volume 81.7 µL, flow 8.0 mL/min, spectra acquisition time 1.4 s and linear range 1 to 40 mg L-1.
Cada dia, a análise química é mais cerca da automatização com o fim de satisfazer as necessidades atuais de resultados rápidos e confiáveis. Os sistemas de análise em fluxo (FIA - Flow Inyection Analysis) são uma das formas de aproximação à automatização. Este artigo apresenta os passos necessários para implementar uma metodologia FIA para a determinação de Pb(II) em água, partindo da revisão dos procedimentos clássicos e descrevendo detalhadamente os passos necessários para implementar a técnica de análise em fluxo. Os resultado são um método de análise de Pb em água que usa ditizona dissolvida em isopropanol (agente cromogénico) na presença de bromuro de cetiltrimetil amônio (CTAB), usado para solubilizar o complexo em água. As características principais do método foram: volume de injeção de amostra de 81,7 µL, velocidade de fluxo de 8,0 mL min-1, tempo de aquisição de espectros de 1,4 s e intervalo linear de 0.9 a 40 mg L-1.