preparation were performed in accordance with EC regulation no.
333/2007.27 Moisture of squids was evaluated using 10 g of
homogenized sample according to Portuguese Standard NP 2282-
1991 and the official AOAC method.28
Samples were kept frozen, in polycarbonate containers, at .20 °C,
until analysis.
Sample Extraction, Hydrolysis, and Solid-Phase Extraction.
After defrosting, samples were minced and homogenized, and 1 g was
added of 1 mL of ascorbic acid (1 mg/mL) and 6 mL of acetonitrile.
Once vortex stirred, the sample was ultrasonicated for 5 min and
extracted by centrifugation at 4 °C and 10000g for 10 min. The
supernatant was collected and concentrated to dryness at 37 °C under
a gentle N2 stream.
For the enzymatic hydrolysis, the extract was redissolved in 4 mL of
sodium acetate buffer. Then, 20 ìL of â-glucuronidase/arylsulfatase
was added and the oxygen atmosphere replaced by nitrogen. After
incubation at 37 °C for 45 min, the enzymatic reaction was stopped by
preparation were performed in accordance with EC regulation no.
333/2007.27 Moisture of squids was evaluated using 10 g of
homogenized sample according to Portuguese Standard NP 2282-
1991 and the official AOAC method.28
Samples were kept frozen, in polycarbonate containers, at .20 °C,
until analysis.
Sample Extraction, Hydrolysis, and Solid-Phase Extraction.
After defrosting, samples were minced and homogenized, and 1 g was
added of 1 mL of ascorbic acid (1 mg/mL) and 6 mL of acetonitrile.
Once vortex stirred, the sample was ultrasonicated for 5 min and
extracted by centrifugation at 4 °C and 10000g for 10 min. The
supernatant was collected and concentrated to dryness at 37 °C under
a gentle N2 stream.
For the enzymatic hydrolysis, the extract was redissolved in 4 mL of
sodium acetate buffer. Then, 20 ìL of â-glucuronidase/arylsulfatase
was added and the oxygen atmosphere replaced by nitrogen. After
incubation at 37 °C for 45 min, the enzymatic reaction was stopped by
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