The homogenates were then centrifuged for 30 min (50,000g) (Alresa HZ50, Orto Alresa, Aljavit, Madrid) at 5 °C. The absorbance of the filtered supernatant was read at 525, 572 and 730 nm. Percentage of MMb was determining using the formula of Krzywicki (1979):
MMb(%)=1.395–((A572–A730)/(A525–A730))∗100MMb(%)=1.395–((A572–A730)/(A525–A730))∗100
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Samples were kept on ice during the whole assay.