高效液相色谱法测定提纯结核菌素效价的研究  

Potency determination of tuberculin purified protein derivative by HPLC

作　　者：张秀英[1] 关孚时[1] 王在时[1] 蒋卉[1] 陆连寿[1] 戴志红[1] 李翠[1] 

机构地区：[1]中国兽医药品监察所,北京100081

出　　处：《中国预防兽医学报》2011年第2期137-140,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家十一五国家科技支撑计划重大项目"疫苗免疫效果评价与标准物质研制"(2006BAD06A10)

摘　　要：为探讨高效液相色谱法测定牛型提纯结核菌素(PPD)效价的可行性,本研究根据PPD中蛋白成分的紫外光吸收波长,采用专用聚苯乙烯/二乙烯苯树脂色谱柱(150 mm×4.6 mm,5μm,300 nm),流动相为乙腈-水-三氟乙酸(25∶75∶0.1),检测波长为271 nm,建立了灵敏、快速的PPD效价的高效液相色谱测定法。在0.4×104 IU/mL～1.1×104 IU/mL的浓度范围内,效价与峰面积呈良好的线性关系,线性回归方程为y=2×106x-6783.6,相关系数R=1.000。将PPD样品用灭菌水稀释后,注入液相色谱仪,测得效价为2.59×104 IU/mg,与传统豚鼠标定法的测定结果(2.91×104 IU/mg)基本一致。该方法简便,灵敏度高,检测速度快,可以较好地控制PPD的质量。According to the characteristics of UV absorption for purified protein derivative（PPD）,a sensitive and quick method of HPLC for determination of PPD was developed.The liquid chromatography separation was carried on polystyrene-divinyl benzene column（150 mm×4.6 mm,5 μm,300 nm） with acetonitrile-water-trifluoroacetate（25∶75∶0.1） as mobile phase with the detection wavelength 271 nm.The equation was y=2×106x-6783.6,r=1.000,the calibration curve was linear in the range of 0.4-11,000 IU/mL.PPD was diluted by sterile water and determined on HPLC.The potency of PPD sample was 25,900 IU/mg by HPLC,which was identify with that by method of guinea pig calibration（29,100 IU/mg）.Results shouwed that the method was rapid and high sensitivity for the quantification of PPD.

关 键 词：牛型提纯结核菌素 效价 高效液相色谱法 

分 类 号：S852.61[农业科学—基础兽医学]