摘要
目的比较不同方法测定疫苗中游离甲醛残留量的差异。方法采用不同方法测定0.25～100μg/ml标准甲醛溶液,绘制反应曲线,初步确定各种方法的线性范围,通过重复测定,计算相关系数,对标准曲线的线性范围进行验证;分析制品中铝佐剂、离心及添加物对检测结果的影响,并对方法的适用性进行验证。结果品红亚硫酸法(《中国药典》)的线性范围为25～100μg/ml,改良乙酰丙酮法与乙酰丙酮法(Sinovac)的线性范围为0.25～100μg/ml,乙酰丙酮法(Crucell)、乙酰丙酮法(Pasteur)和盐酸苯肼法(MSD)的线性范围为0.25～50μg/ml,各方法标准曲线的相关系数为0.994～1.000。品红亚硫酸法测定未离心的含铝佐剂样品的回收率在18.06%～76.00%之间,测定离心后的含铝佐剂样品的回收率在17.19%～75.95%之间;乙酰丙酮法测定未离心的含铝佐剂样品的回收率在88.28%～103.21%之间,测定离心后的含铝佐剂样品的回收率在95.43%～102.78%之间;盐酸苯肼法测定未离心的含铝佐剂样品的回收率在99.27%～117.74%之间,测定离心后的含铝佐剂样品的回收率在100.55%～119.40%之间。各方法测定含添加物样品的回收率在94.68%～101.86%之间。改良乙酰丙酮法(412 nm)测定含甲醛疫苗的回收率在100.84%～105.88%之间,变异系数在0.28%～3.08%之间。结论改良乙酰丙酮法操作简便,灵敏度高,线性范围广,结果准确可靠,适用于疫苗中游离甲醛残留量的测定。
Objective To compare the difference in determination of residual free formaldehyde content in vaccine by various methods.Methods Standard formaldehyde solutions at concentrations of 0.25 ～ 100 μg / ml were tested by various methods to plot a reaction curve,based on which the liner range of each method was determined and verified by repeat tests and the calculation of correlation coefficients.The effects of aluminum adjuvant,centrifugation and additives on determination results were analyzed,and the methods were verified for suitability.Results The liner range of sulfurous acid-magenta method（Chinese Pharmacopoeia） was 25 ～ 100 μg / ml,while those of improved acetylacetone method and acetylacetone（Sinovac） method were 0.25 ～ 100 μg / ml,and those of acetylacetone（Crucell） method,acetylacetone（Pasteur） method and phenylhydrazine hydrochloride（MSD） method were 0.25 ～ 50 μg / ml.The correlation coefficients of standard curves of various methods ranged from 0.994 to 1.000.The recovery rates of aluminum adjuvant-containing samples before and after centrifugation were 18.06% ～ 76.00% and 17.19% ～ 75.95% by sulfurous acid-magenta method,88.28% ～ 103.21% and 95.43% ～ 102.78% by acetylacetone methods,and 99.27% ～ 117.74% and 100.55% ～ 119.40% by phenylhydrazine hydrochloride method,respectively.The recovery rates of samples containing additives determined by various methods were 94.68% ～ 101.86%.However,the recovery rates of samples containing formaldehyde determined by improved acetylacetone method（412 nm） was 100.84% ～ 105.88%,with a coefficient of variation of 0.28% ～ 3.08%.Conclusion Improved acetylacetone method is simple,sensitive,accurate,reliable and of a wide linear range,which is suitable for the determination of free formaldehyde in vaccine.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第6期749-753,共5页
Chinese Journal of Biologicals
关键词
分光光度法
乙酰丙酮
甲醛
疫苗
灵敏度
Spectrophotometry
Acetylacetone
Formaldehyde
Vaccine
Sensitivity
