摘要
研究了直接用于液相色谱测定甲醛与2,4-二硝基苯肼的反应条件,使反应在菌疫苗类生物制品自身的pH范围内即弱酸性条件下,无需酸碱度调节,反应产物甲醛衍生物不需要有机溶剂萃取,避免损失,可直接用于液相色谱分析.在检出限及精确度方面更是优越于分光光度法.将样品用2,4-二硝基苯肼溶液衍生后,经C18化学键合硅胶为固定相,以乙睛：水（60：40,体积比）为流动相.紫外检测波长 360 nm进行测定.方法的平均回收率为95.4%（n=5）,两种生物制品 6 次独立测定的相对标准偏差分别为重组乙型肝炎疫苗（CHO）RSD=0.92%,吸附白喉破伤风联合疫苗RSD=3.92%.甲醛（0.6～3.0 μg/mL）范围内,浓度与吸收面积值呈良好的线性关系.结果显示,该研究方法的色谱条件能准确定量、灵敏、准确、重复性好,优于药典的分光光度法,可用于实际检测分析.
Reaction conditions of formaldehyde with 2,4-dinitrophenylhydrazine that could be directly used in liquid chromatography were studied. Under these conditions bio-products of microbial vaccines could react in their own pH range i.e. weak acid condition. It did not need to adjust their pH values, and the reactive products, derivatives of formaldehyde, need not be extracted with organic solvent to avoid the loss, therefore could be directly used in liquid chromatography analysis. Furthermore, it is more superior to spectrophotometer in the aspects of the examining limit and the measure precision. After the sample was derived with 2,4-dinitrophenylhydrazine then the determination was carried out with C18 chemical bonded silica gel as a partitioner phase and catonitrile-water （60：40） as mobile phase, and UV wavelength at 360 nm. The average recovery of the added sample was 95.4 % （n= 5） RSD got by 6 determinations in bio-products. Hepatitis B vaccine made by recombinant DNA technique in CHO cell was 0.92 %, diphtheria and tetanus combined vaccine, adsorbed was 3,92 % ; There was a good linear relationship between the concentration of formaldehyde and absorption area value in range of 0.6 - 3.0μg/mL. The results showed that this chromatographic condition of the studied method was highly reproducible and accurate, sensitive, and more superior to spectrophotometer described in pharmacopoeia. Therefore, it can be used in practical examination analysis.
出处
《微生物学杂志》
CAS
CSCD
2006年第5期104-106,共3页
Journal of Microbiology
