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2 protocols using sea sand

1

Extraction and Analysis of Butterfat Composition

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Dichloromethane, hexane, methanol, acetonitrile, dimethylformamide (HPLC grade) and sulfuric acid (98% purity) were purchased from Labscan (Dublin, Ireland). Sodium carbonate, sea sand and sodium sulfate anhydrous were supplied by Panreac (Barcelona, Spain). Sodium methoxide (95% purity) was supplied by Sigma-Aldrich (St. Louis, MO, USA). Butterfat BCR-164 (EU Commissions; Brussels, Belgium) was supplied by Fedelco Inc. (Madrid, Spain). CO 2 (99.99% purity) was supplied by Carburos Metálicos (Madrid, Spain).
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2

Analytical Techniques for Phytochemical Analysis

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Absolute ethanol for extractions was purchased from VWR International (Leuven, Belgium). ACN with HPLC-MS quality, was purchased from Fisher (Thermo Fisher Scientific, Leicestershire, UK). Ultrapure water with a resistivity value of 18.2 MΩ was obtained from a Milli-Q system (Millipore, Bedford, MA, USA). Folin-Ciocalteu reagent, ABTS (2,2-azinobis (3-ethylbenzothiazoline-6-sulfonate)), trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), potassium persulfate, aluminum chloride and the standard compounds: rutin, p-coumaric acid, kaempferol, 3,4-dyhydroxybenzoic acid, quercetin, chlorogenic acid and caffeic acid were from Sigma-Aldrich (Steinheim, Germany). Sea sand was from Panreac (Barcelona, Spain).
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