Determination of Folic Acid in Serum by SPE and High Performance Liquid Chromatography with Photochemical Spectrofluorim
This paper described a combined high-performance liquid chromatography (HPLC) -photochemical spectrofluorimetry/solid-phase extraction (SPE) procedure for determining the concentration of unmetabolized folic acid in human serum. After being extracted by S
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Determination of Folic Acid in Serum by SPE and High Performance Liquid Chromatography with Photochemical Spectrofluorimetry Caiju Zhou, Haijie Hu and Tong-Cun Zhang Abstract This paper described a combined high-performance liquid chromatography (HPLC) -photochemical spectrofluorimetry/solid-phase extraction (SPE) procedure for determining the concentration of unmetabolized folic acid in human serum. After being extracted by SPE (C18-H) column, the serum samples were irradiated under the UV lamp for 11 h, separated on a C18 HPLC column with methanol/water as the mobile phases and determined by a fluorescence detector. Fluorescence excitation and emission wavelengths were at 280 and 443 nm, respectively. The linear range was 1.9 * 8.0 ng/mL. The recovery was from 88.9 to 123.6 % with RSD in the range of 4.1–6.4 %.
Keywords Folic acid High-performance liquid chromatography Photochemical spectrofluorimetry Solid-phase extraction
199.1 Introduction Folic acid (pteroylglutamate), a member of water-soluble vitamin B family, is used as a drug to treat anemia, mainly for megaloblastic anemia, and also for pernicious anemia as secondary treatment drug. It is also involved in the biosynthesis of amino acids and nucleic acids in the human body, and jointly promotes the formation of red blood cells with vitamin B12 [1]. In addition, folic acid is associated with some other diseases, including neural tube defects [2], colon cancer [3], cardiovascular disease [4], and childhood mental retardation [5]. Therefore, rapid and accurate determination of folic acid in human tissue is of great significance for C. Zhou (&) H. Hu T.-C. Zhang Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, People’s Republic of China e-mail: [email protected]
T.-C. Zhang et al. (eds.), Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012), Lecture Notes in Electrical Engineering 251, DOI: 10.1007/978-3-642-37925-3_199, Ó Springer-Verlag Berlin Heidelberg 2014
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the prevention, diagnosis, and treatment of diseases caused by folic acid. Due to its poor stability, complex composition, and very low content in natural samples, the analysis of folic acid is a great challenge. The accuracy and sensitivity of folic acid analysis highly depends on the merits of the extraction method as well as detection techniques. Currently, there are microbiological assay (MA), radioimmunoassay, capillary electrophoresis and HPLC [6] to determine folic acid in biological samples. There are, however, several limitations with MA, e.g., tedious laboratory work, and multiple interferences from the sample matrix [7, 8]. HPLC method has a great potential due to its rapid progress of separation and detection techniques, as well as the possibility of automatizing the purification procedure. The objective of this work was to develop and validate a simple and sensitive HPLC method suitable for
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