Proposition for Mechanism of Separation of Folates by HPLC-DAD: Practical Application in New Zealand Spinach (Tetragonia expansa)
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- 作者：Emmanuela Prado de Paiva Azevedo…
- 关键词：Folate ; Lewis acid and base ; HPLC ; DAD ; Isocratic ; Chromatographic parameters ; Tetrahydrofolate ; 5 ; Methyltetrahydrofolate ; 10 ; Formyltetrahydrofolate ; 5 ; Formyltetrahydrofolate ; Folic acid ; Pteroic acid ; Tetragonia expansa
- 刊名：Food Analytical Methods
- 出版年：2016
- 出版时间：May 2016
- 年：2016
- 卷：9
- 期：5
- 页码：1321-1332
- 全文大小：732 KB
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Clayton Anderson de Azevedo Filho (2)
Beate Saegesser Santos (3)
Lorena Trajano da Silva (1)
Ana Elizabeth Cavalcante Fai (5)
José Almiro da Paixão (4)
1. Rural Technology Department, Federal Rural University of Pernambuco, Rua Dom Manoel de Medeiros, s/n, Dois Irmãos, 52171-900, Recife, PE, Brazil
2. Higher Education Association of Caruaru, Av. Portugal s/n, Universitário, 55016901, Caruaru, PE, Brazil
3. Pharmacy Department, Federal University of Pernambuco, Av. Prof. Moraes Rego, s/n, Campus Universitário, 50670-901, Recife, PE, Brazil
5. Department of Basic and Experimental Nutrition, Institute of Nutrition, Rio de Janeiro State University, R. São Francisco Xavier, 524, Pavilhão João Lyra Filho, 12 andar, Sala 12.002, Bloco D, 20559-900, Rio de Janeiro, RJ, Brazil
4. Nutrition Department, Federal University of Pernambuco, Av. Prof. Moraes Rego, s/n, Campus Universitário, 50670-901, Recife, PE, Brazil - 刊物类别：Chemistry and Materials Science
- 刊物主题：Chemistry
Food Science
Chemistry
Microbiology
Analytical Chemistry - 出版者：Springer New York
- ISSN：1936-976X
文摘
A fast and simple method for determination of six folate forms was developed through a study of standards and tested in New Zealand spinach (Tetragonia expansa) samples. A systematic evaluation of the standard solubilization conditions, pH, and molar concentration of the aqueous solvent of the mobile phase was performed and evaluated by means of chromatographic parameters: retention time (t r), capacity factor (k), alpha (α), and resolution (Rs). The optimal separation was achieved using phosphate buffer pH 2.0, 100 mmol L−1 and methanol in the ratio 85:15 isocratic mode using C18 Spherisorb ODS-2, 2.0 × 100 mm, 5 μm column, and DAD detection 200–400 nm. The following k values were obtained: 0.58, 1.17, 1.06, 1.98, 3.03, and 6.05, respectively, to tetrahydrofolate, 5-methyltetrahydrofolate, 10-formyltetrahydrofolate, 5-formyltetrahydrofolate, folic, and pteroic acid and α values of 1.81, 1.10, 1.69, 1.53, and 1.99. The elution order of the six folate forms is explained using the Lewis theory of acids and bases. This method was applied to samples of New Zealand spinach and monitored through chromatographic and spectral parameters obtained in the region of maximum absorption of λ. Moreover, the similarity and the purity of the peaks in standards and samples were investigated. For all folate forms, α and Rs values were above 1, except tetrahydrofolate. The similarity index was above 75 % and a purity above 90 % in samples, except for 10-formytetrahydrofolate, indicating that the method has good selectivity for complex matrices.