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Food Chem ; 305: 125450, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31499291


A round robin comparison was performed in order to test the performance of a recently developed LC-MS/MS method for quantification of 6 folate forms. Eighty-nine samples representing the food groups of fruits, vegetables, legumes, cereals, dairy products, meat, and offal were analyzed by two LC-MS/MS methods and a microbiological assay (MA). A plant-origin deconjugase enzyme (Arabidopsis thaliana) for deconjugation of folates (PE-LC-MS/MS), or animal-origin deconjugase (rat serum and chicken pancreas) (AE-LC-MS/MS) was used in the LC-MS/MS methods, each in a single enzymatic step. In contrast, the MA involved tri-enzyme extraction including human plasma as a deconjugase. A significant bias of 17% lower and 25% higher results was found when PE-LC-MS/MS was compared to MA and AE-LC-MS/MS, respectively. The PE-LC-MS/MS provides fast quantification of various folate vitamers and total folate content, which could be a proper substitute to the currently standardized but imprecise and time-consuming microbiological assay in the future.

J Chromatogr A ; 1594: 34-44, 2019 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-30799066


A rapid, sensitive and reproducible method for analysis of naturally-occurring folates and folic acid in food has been developed and validated. A single-enzyme extraction step, in which a pure recombinant enzyme of plant origin (Arabidopsis thaliana) was used, enabled fast and reproducible deglutamylation during folate extraction within the incubation time of 1 h. Six commonly occurring folate forms (tetrahydrofolate, 5,10-methenyltetrahydrofolate, 10-formylfolic acid, 5-formyltetrahydrofolate, folic acid and 5-methyltetrahydrofolate) were detected and quantified in 9 min using liquid chromatography-tandem mass spectrometry (LC-MS/MS). 13C5-labeled 5-formyltetrahydrofolate, 13C5-labeled folic acid and 13C5-labeled 5-methyltetrahydrofolate were used as internal standards for the quantification. The method is described by a calibration curve (R2>0.99 and trueness 85-115%), a limit of quantification at 0.1 µg/100 g, trueness at 80-120% in spiked samples and certified reference materials, and a precision <10%. However, the precision in quantification of tetrahydrofolate was not within the acceptable limits due to the lack of use of the corresponding internal standard. An interconversion study of unstable formyl forms was performed which showed that 50% of 5,10-methenyltetrahydrofolate is converted to 5-formyltetrahydrofolate during the analysis. The developed LC-MS/MS method is a candidate for a future standard method for folate analysis in food.

Cromatografia Líquida , Ácido Fólico/análise , Análise de Alimentos/métodos , Proteínas de Plantas/metabolismo , Espectrometria de Massas em Tandem , Calibragem , Ácido Fólico/análogos & derivados , Limite de Detecção , Tetra-Hidrofolatos/análise
Food Chem ; 254: 144-149, 2018 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-29548435


Information on the retention of vitamin D in food following household cooking is scarce. So far the retention of its metabolites vitamin D3, vitamin D2, and 25-hydroxyvitamin D3 has shown that the type of food and the cooking method are the essential determinants, and there is no significant difference between the metabolites. We investigated the retention of vitamin D3 and vitamin D2 in sunflower oil, vitamin D3 in rainbow trout, and vitamin D2 in button mushrooms. The investigated cooking methods were boiling at different pH, steam cooking, microwave cooking, pan-frying, and oven baking. There was no difference between the retention of vitamin D3 and vitamin D2 added to sunflower oil, which ranged from 70 to 99%. In rainbow trout, the retention of vitamin D3 at 85-114% was not significantly different from 100%, except for panfrying at 85%. However, the retention of vitamin D2 in mushrooms at 62-88% was significantly different from 100% (p ≤ 0.05).

Agaricus/química , Colecalciferol/análise , Culinária/métodos , Ergocalciferóis/análise , Oncorhynchus mykiss , Óleo de Girassol/química , Animais , Estabilidade de Medicamentos , Carne/análise