柱前衍生化-高效液相色谱法检测疫苗中游离甲醛含量

Determination of Free Formaldehyde Content in Vaccine by Pre-column Derivatization HPLC

关键词:  高效液相色谱  柱前衍生化  甲醛  疫苗

Key words:  HPLC  pre-column derivatization  free formaldehyde  vaccine

摘要:

建立了测定疫苗中游离甲醛含量的柱前衍生化-高效液相色谱法。采用2,4-二硝基苯肼衍生疫苗中游离甲醛，乙腈︰水（65︰35）为流动相，流速为1.0 mL/min，色谱柱为Waters Atlantis T3柱（4.6 mm×250 mm，5 μm），检测波长为365 nm。在此方法下，3个不同甲醛浓度样品回收率为98.28%~105.24%，相对标准偏差（RSD）为 2.3%。本方法检测限为0.4 mg/L，定量限为1.6 mg/L。游离甲醛浓度在0.0040~1.1930 mg/mL范围内，与甲醛衍生物峰面积呈良好的线性关系（r2=0.9991）。结果表明，该方法分离度高，专属性强，操作简单且不易受疫苗基质和颜色影响，可用于测定疫苗中游离甲醛含量。

Abstract:

Pre-column derivatization HPLC method was established in order to determine the free formaldehyde content in vaccine with 2,4-dinitrophenylhydrazine (DNPH) as the derivatization reagent. The Waters Atlantis T3 (4.6 mm×250 mm, 5 μm) was used in the analysis. The mobile phase was a mixture of acetonitrile and water with a ratio of 65 to 35 by volume. The flow rate was 1.0 mL/min. UV detection was made at the wavelength of 365 nm. The recovery of samples was in a range of 98.28%~105.24%, with relative standard deviation (RSD) of 2.3%. The detection limit was 0.4 mg/L and the quantitation limit was 1.6 mg/L. The free formaldehyde content in vaccine showed good liner relationship to the peak area within a content range of 0.0040~1.1930 mg/mL (r2=0.9991). This method is separable, specific, simple and not easy to be affected by vaccine’s ground substance and color, and might be used for determination of formaldehyde content in vaccine.