Abstract
Analytical method for an accurate determination of folic acid in human serum has been established using isotope dilution ultra‐high‐pressure liquid chromatography/mass spectrometry (ID‐UPLC/MS). Sample extract was prepared from 0.5 mL of human serum spiked with 13C5‐folic acid solution and then LC–MS/MS experiment was performed on multiple reaction monitoring mode for 20 min. The method was tested for repeatability using homogeneous human serum and it showed good repeatability with relative standard deviation of 1.8%. Further evaluation was carried out through NIST SRM 1955 samples (level I, II, and III) and the measurement results were in good agreement with the certified values with good reproducibility over two periods. It was also revealed that the obtained relative expanded uncertainty was 12% which is much smaller than that of previously reported methods, indicating that this method is very reliable and accurate for determining the content of folic acid in human serum.
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