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Simultaneous determination of 4‐hydroxyphenyl lactic acid, 4‐hydroxyphenyl acetic acid, and 3,4‐hydroxyphenyl propionic acid in human urine by ultra‐high performance liquid chromatography with fluorescence detection
Author(s) -
Yang Yongli,
Liu Fan,
Wan Yiqun
Publication year - 2017
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201700014
Subject(s) - acetic acid , lactic acid , chromatography , chemistry , urine , ammonium acetate , detection limit , high performance liquid chromatography , ammonium bromide , bromide , extraction (chemistry) , ammonium , pulmonary surfactant , organic chemistry , biochemistry , bacteria , biology , genetics
A simple and reliable method was established for simultaneous determination of 4‐hydroxyphenyl acetic acid, 4‐hydroxyphenyl lactic acid, and 3,4‐hydroxyphenyl propionic acid in human urine by high‐performance liquid chromatography with fluorescence detection. Solid‐phase extraction was used to eliminate the interferences in urine. The separation of three analytes was achieved using a C 18 column and a mobile phase formed by a 95:5 v/v mixture of 50 mmol/L ammonium acetate buffer at pH 6.8 that contained 5 mmol/L tetrabutyl ammonium bromide and acetonitrile. Under the optimized conditions, the detection limits of 4‐hydroxyphenyl acetic acid, 4‐hydroxyphenyl lactic acid, and 3,4‐hydroxyphenyl propionic acid were 4.8 × 10 −3 , 8.80 × 10 −3 , and 9.00 × 10 −3 mg/L, respectively, and the recoveries were in the range of 85.0–120.0% with relative standard deviations of 1.5–3.1%. This method was used to analyze urine samples from breast cancer patients, healthy people and post‐surgery breast cancer patients. Significant differences in urinary levels of 4‐hydroxyphenyl acetic acid and 4‐hydroxyphenyl lactic acid could be found between the breast cancer patients group and other two groups. No effect of age and sex was observed on the urinary levels of 4‐hydroxyphenyl acetic acid and 4‐hydroxyphenyl lactic acid. This method might be helpful for cancer biomarkers discovery in urine.