摘要
提出了反相高效液相色谱同时测定新生儿尿液中4-羟基苯乙酸、2-羟基苯乙酸、苯乙酸和苯丙酮酸等4种苯丙酮尿症特征酸的方法。取尿液样品5.00mL,乙腈5mL,混合振荡1min,离心10min,取上清液进行液相色谱分离。用Waters Symmetry C18色谱柱作为固定相,用不同比例的乙腈和0.15%(体积分数)磷酸溶液进行梯度洗脱,检测波长为210,289nm。在上述条件下,4种特征酸分离完全,并且不受尿液中内源性物质的干扰。4种特征酸的质量浓度在一定范围内呈线性,检出限(3S/N)分别为0.43,0.22,0.62,0.42mg·L-1。按标准加入法在3个浓度水平上进行加标回收试验,测得回收率在89.1%~108%之间,测定值的相对标准偏差(n=6)小于4%。
Reversed-phase high performance liquid chromatography （RP-HPLC） was applied to the determination of 4 phenylketonuric characteristic acids, including 4-hydroxyphenylacetic acid, 2-hydroxyphenylacetic acid, phenylacetic acid and phenylpyruvic acid in newborn urine. The urine sample （5. 00 mL） was shaked with acetonitrile （5.0 mL） by swirling for lmin and centrifuged for 10 min. 4 analytes in the supernatant were separated by LC using Waters Symmetry C18 column as stationary phase, and mixtures of acetonitrile and 0. 15% （9） phosphoric acid solution in various ratios as mobile phase in gradient elution. Detections were taken at the wavelengths of 210, 289 nm. Under the above conditions, the 4 characteristic acids were separated completely, and no interference by the endogenous substances in the urine was observed. Linearity for the 4 characteristic acids were kept between definite mass concentration ranges. Values of detection limits （3S/N） for the 4 characteristic acids were 0. 43, 0.22, 0. 62, 0. 42 mg · L-1 respectively. Tests for recovery were made by standard addition method at 3 concentration levels, giving results in the range of 89.1%-108%, with RSDs （n=6） less than 4%.
出处
《理化检验（化学分册）》
CAS
CSCD
北大核心
2017年第1期12-16,共5页
Physical Testing and Chemical Analysis(Part B:Chemical Analysis)
基金
浙江省自然科学基金资助项目(M203030
Y4110536)
