Lowry法测定b型流感嗜血杆菌结合疫苗蛋白含量干扰现象的排除

目的探讨Lowry法测定b型流感嗜血杆菌结合疫苗蛋白含量干扰现象的排除方法。方法分别应用《中国药典》三部(2005版)中Lowry法与第5版《欧洲药典》2.5.16法测定b型流感嗜血杆菌结合疫苗原液的蛋白含量,并以碳二亚胺(EDAC)为干扰性物质,验证两方法排除干扰的能力。结果同一批结合疫苗原液用两种方法测定的结果不一致,应用《中国药典》方法测定的结果明显高于第5版《欧洲药典》的方法。在多糖的活化衍生及多糖蛋白结合过程中加入EDAC、己二酰二肼(ADH)等,可干扰测定结果,使结果偏高。结论应用Lowry法测定多糖蛋白结合疫苗中蛋白含量时,应排除干扰性物质的影响。

Removal of Interference in Lowry Method for Determination of Protein Content in Haemophilus influenza Type b Conjngate Vaccine

Objective To investigate a route to remove the interference in Lowry method for determination of protein content in Haemophilus influenza type b conjugate vaccine.Methods The protein content in the bulk of Haemophilus influenza type b conjugate vaccine was determined by Lowry method described in Chinese Pharmacopeia(VolumeⅢ,2005 edition)and the method described in Section 2.5.16 of European Pharmacopeia(The fifth edition)respectively,and the abilities in removing interference of the two methods were verified using EDAC as an indicator.Results The determination results of protein content by the method described in Chinese Pharmacopeia was significantly higher than that by the method described in European Pharmacopeia,which was due to the introduction of DEAC and ADH during activation and derivation of polysaccharide and conjugation of polysaccharide protein.Conclusion An procedure for removing the interference substance shall be introduced to Lowry method for determination of protein content in polysaccharide-protein conjugate vaccine.