目的建立测定生物制品保护剂中蔗糖和乳糖含量的高效液相色谱(HPLC)法。方法样品用磺基水杨酸沉淀、离心除去蛋白质,上清液采用氨基柱分离,示差折光检测器进行监测,依外标法建立校正曲线,并对建立的方法进行验证。采用建立的方法测定4种疫苗成品中蔗糖和乳糖的含量。结果蔗糖和乳糖的标准曲线的相关系数r2均达0.999 9以上。该方法能将两种糖完全分离;测定两种糖的特异性较好;该法检测4个浓度的蔗糖和乳糖的平均回收率分别为97.9%和100.5%,变异系数分别为1.28%和0.48%;最低检测限及定量限分别为0.01 mg/ml和0.1 mg/ml;采用该法测定的4种疫苗成品中蔗糖和乳糖的变异系数分别为2.59%和1.44%。结论已建立了测定生物制品保护剂中蔗糖和乳糖含量的HPLC法,该方法定量准确,重复性好,可作为生物制品保护剂中蔗糖和乳糖含量的常规检测方法。 Objective To establish a high performance liquid chromatography (HPLC) method for the determination of sucrose and lactose in biological protective agents. Methods The samples were precipitated with sulfosalicylic acid and centrifuged to remove protein. The supernatant was separated by amino column and monitored by refractive index detector. The calibration curve was established by external standard method and the established method was validated. The established method was used to determine the contents of sucrose and lactose in the four vaccines. Results The correlation coefficient r2 of standard curve of sucrose and lactose reached above 0.999 9. The method can completely separate the two kinds of sugars; the specificity of the two kinds of sugars is good; the average recoveries of the four concentrations of sucrose and lactose measured by the method are respectively 97.9% and 100.5%; the coefficient of variation is 1.28% and 0.48 %; The lowest limit of detection and limit of quantification were 0.01 mg / ml and 0.1 mg / ml, respectively. The coefficients of variation of sucrose and lactose in the four vaccines were 2.59% and 1.44%, respectively. Conclusion HPLC method has been established for the determination of sucrose and lactose in biological protective agent. The method is accurate and reproducible. It can be used as a routine method for the determination of sucrose and lactose in biological protective agents.