大鼠体内三聚氰胺及其代谢物双波长HPLC检测方法的建立

Establishment of Dual Wavelength HPLC for Detection of Melamine and Its Metabolites in Rats

建立双波长高效液相色谱法同时检测不同给药剂量组大鼠体内三聚氰胺、三聚氰酸和尿酸含量的方法。采用C18色谱柱,流动相为0.5mmol/L磷酸盐(pH 7.4)-甲醇(95∶5,V/V),流速1.0mL/min,检测波长三聚氰胺和三聚氰酸为213nm,尿酸为285nm,进样量10μL,柱温25℃。样品用0.6mol/L高氯酸沉淀蛋白质。结果表明,三聚氰胺、三聚氰酸、尿酸分别在0.10~1.00×10~2、0.50~1.00×10~2、0.20~1.00×10~2μg/mL范围内线性关系良好(r=0.999 9),最低检出限分别为2.50×10^(-2)、7.50×10^(-2)、3.00×10^(-2)μg/mL,平均回收率在90.99%~96.09%之间,相对标准偏差在0.84%~1.40%之间。该方法操作简便、灵敏度高、结果准确可靠,可用于动物体内三聚氰胺、三聚氰酸及尿酸同时检测和分析。

A dual wavelength HPLC method was developed to determine melamine,cyanuric acid and uric acid in rats from different dosage groups.The determination was performed on a C18 with mobile phase con- sisted of 0.5 mmol/L phosphate buffer solution（pH 7.4）-methanol （95 ： 5, v/v） ,the flow rate was 1.0 mL/ rain.Melamine, cyanuric acid and uric acid were determined by dual wavelength,the detected wavelength of melamine and cyanuric acid were 213nm,it＇s of uric acid was 285nm.The injection volumn was 10 μL,the column temperature was 25℃.The sample was deproteinized by 0.60 mol/L perchloric acid.The results in- dicated：the good linearity ranges of melamine, cyanuric acid, uric acid were 0.10-1.00 X 102 ,0.50-1.00,0.20- 1.00 ;4 102 μg/mL （r= 0.999 9）, the lowest detection limits were 2.50 X 10 2,7.50 ）〈 10-2 , 3.00 ）4 10.2 μg/ mL,the average recoveries were between 90.99%-96.09%, the relative standard deviations （RSDs） were between 0.84%-1.40%.The method was simple,sensitive,accurate.It can be used to determine and analyze melamine, cyanuric acid and uric acid in animals.