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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
[Determination of phenylalanine and tyrosine in serum by high-performance liquid chromatography with fluorescent
1Department of Clinical Laboratory, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Objective:
To determine phenylalanine and tyrosine in serum by high-performance liquid chromatography (HPLC) method.
Methods:
Serum precipitated with perchloric acid was isocratically eluted using a base-deactivated C8 column with 10% acetonitrile in water as the mobile phase. The flow rate was 1.0 ml/min. Fluorescent detector worked at an excitation wavelength of 210 nm and emission wavelength of 303 nm.
Results:
Good linearity was observed in the range of 12.2 approximately 1 220.0 micromol/L for phenylalanine, the intra- and inter-assay coefficients of variation (CV) were 4.07% and 7.17% respectively, and the average recovery was 102.5%. There also was a good linearity in the range of 5.5 approximately 550.0 micromoL/L for tyrosine. The intra- and inter-assay coefficients of variation were 2.99% and 5.23% respectively, and the average recovery was 102.2%. The total assay time for the serum sample was within 30 min. We found a good correlation compared with HPLC ultraviolet detection.
Conclusion:
This method is simple, rapid, accurate in measuring phenylalanine and tyrosine in serum quantitatively.
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