固相萃取-光化学荧光-高效液相色谱测定血浆中叶酸含量

Determination of folic acid in plasma by solid phase extraction and high performance liquid chromatography with photochemical spectrofluorimetry

目的采用固相萃取-光化学荧光-高效液相色谱法测定血浆中叶酸含量。方法叶酸血浆样品经LD605固相萃取小柱分离净化,在紫外灯下照射,经高效液相色谱分离,荧光检测。结果线性范围在0.100-100.0mg·L-1之间,方法回收率不低于79.4%,精密度(n=5)小于13.9%,检测限为0.2ng。结论本法具有准确、灵敏、选择性好、回收率高等特点,能满足临床常规血药浓度监测。

OBJECTIVE To develop a method for determining the concentration of folic acid in plasma by solid phase extraction and high performance liquid chromatography with photochemical spectrofluorimetry. METHODS After being extracted by subminiature column of LD605 macroreticular resin, the plasma samples were irradiated under the UV lamp for 50 min. And then, the samples were separated on a C18 column with methanol/Na2HPO4-NaH2PO4 buffer as the mobile phase. The fluorescence intensity was determined by a fluorescence detector. Fluorescence excitation and emission wavelengths were at 280 nm and 443 nm. RESULTS The linear range was 0.100 - 100.0 mg· L^- 1. The recovery was higher than 79.4%. RSD was lower than 13.9%. The detect limit was 0.2 ng. CONCLUSION The method is accurate, sensitive, high selectivity and effective for folic acid and can be used in routine determination.