摘要
目的 采用高效液相色谱法（high performance liquid chromatography,HPLC）测定钩端螺旋体疫苗中的苯酚含量。方法 色谱条件：色谱柱：Novo-Pak C-18层析柱（3.9 mm×150 mm,4μm）,流动相：甲醇-超纯水（40：60,v：v）,流速：1.0 ml/min,检测波长：270 nm,柱温：30℃,进样量：10μl。对建立的方法进行线性范围、精密性、重现性、准确性验证及初步应用。结果 苯酚浓度在18.552～129.864μg/ml范围内,色谱峰面积与苯酚浓度呈良好的线性关系（r=0.999 7）;苯酚对照品溶液连续进样6次,主峰面积的相对标准偏差（RSD）为0.25%,保留时间的RSD为0.10%;用建立的方法检测6份钩端螺旋体疫苗样品中苯酚含量的RSD为0.5%;准确性试验的总平均回收率为100.6%,RSD为1.0%;HPLC法和溴量滴定法测定4批钩端螺旋体疫苗样品中苯酚含量的结果基本一致。结论HPLC法可以更准确地测定苯酚含量,且该方法操作简便、快速,精密性高,重现性好,无干扰,适用于钩端螺旋体疫苗成品中苯酚含量的检测。
Objective To determine the phenol content in leptospira vaccine by high performance liquid chromatography （HPLC）. Methods Novo-Pak C-18 chromatographic column （3.9 mm× 3.9 mm, 4 μm） was used. Methanolultrapure water （40 ： 60, v ： v） was served as mobile phase at a flow rate of 1.0 ml/min. The detection wavelength was 270 nm, while the column temperature was 30℃, and the sample loading was 10 μl. Results The peak area showed good linear relationship to phenol concentration at a range of 18. 552 - 129. 861 μg/ml （r = 0. 999 7）. The relative standard （RSD） deviation of main peak area of phenol control solution tested for 6 times was 0. 25%, while that of retention time was 0. 10%. The RSD of determination results of phenol contents in six samples of leptospira vaccine by the developed method was 0. 5%. The total average recovery rate in accuracy test was 100. 6%, with a RSD of 1.0%. The determination results of phenol contents in samples of four batches of leptospira vaccine by HPLC were basically consistent with those by bromine content titration method. Conclusion HPLC may be used for more accurate determination of phenol content, which is simple, rapid, precise and of good reproducibility without interference. The method is suitable for the determina- tion of phenol content in final product of leptospira vaccine.
出处
《中国生物制品学杂志》
CAS
CSCD
2013年第12期1823-1826,共4页
Chinese Journal of Biologicals
基金
国家高技术研究发展计划(863计划
2012AA02A402)
关键词
高效液相色谱法
钩端螺旋体疫苗
苯酚
含量测定
High performance liquid chromatography （HPLC）
Leptospira vaccine
Phenol
Content determination
