【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。更多还原

【Abstract】 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.更多还原