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IV. Resultados y discusión. Capítulo III 259
method. The separations were performed at 30 °C by isocratic elution at a
flow rate of 1 mL/min. The injection volume was 10 µL. Detection was at
214 nm. The mobile phase was 100 mmol/L phosphate buffer at pH 6.8.
The second SEC-HPLC screening method has a greater affinity for
smaller Mw peptides (roughly 700 - 17,000 Da). The standards
thyroglobulin, aprotinin, cyctochrome C, insulin, angiotensin I, angiotensin
II, uridine and sodium azide were used to prepare a calibration curve for this
method. The separations were performed at 30 °C by isocratic elution at a
flow rate of 1 mL/min. The injection volume was 10 µL. Detection was at
214 nm. The mobile phase was acetonitrile (ACN)-H 2 O (ratio 45:100)
containing 0.1 mL/100 mL trifluoroacetic acid (TFA).
2.8 Evaluation of the anti-inflammatory bioactivity of fermented
almond milks
The anti-inflammatory bioactivity of fermented and non-fermented
almond milks were evaluated in human intestinal epithelial-like (Caco-2)
cells. Caco-2 cells were purchased from the American Type Culture
Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco's
Modified Eagle's Medium (DMEM) (Invitrogen Corp., San Diego, CA,
USA) supplemented with 10 mL/100 mL fetal bovine serum (Invitrogen
Co., Carlsbad, USA), 1 mL/100 mL non-essential amino acids, 1g/100 mL
sodium pyruvate and penicillin (100 U)/streptomycin (100 g/mL) (all lasts
purchased from Sigma–Aldrich Corp., St. Louis, MO, USA). Cells were
maintained in vented 75 cm 2 flasks in a humidified cell culture incubator
with 5% CO 2 at 37 ºC.