G1314f vwd 1260

For the detection of biogenic amine (BA) content, samples were subjected to an extraction with trichloroacetic acid followed by derivatization with dansyl chloride, as described by Pasini et al. [10 (link)]. The biogenic amines content was analysed using a HPLC Agilent 1260 Infinity (Agilent Technologies Inc., Santa Clara, CA, USA) with a UV detector (G1314F VWD 1260) at 254 nm. The amounts of amines were expressed as mg/L by reference to a calibration curve obtained with aqueous biogenic amine standards derivatized as described for the samples.

The samples were extracted with trichloroacetic acid, according to Pasini et al. [16 (link)]. The extracts were subjected to a dansyl chloride derivatization (Sigma Aldrich, Gallarate, Italy), according to Martuscelli et al. [17 (link)]. An HPLC Agilent Technologies 1260 Infinity with the automatic injector (G1329B ALS 1260, loop of 20 µL), equipped with a UV detector (G1314F VWD 1260) set at 254 nm, was used. For the chromatographic separation a C18 Waters Spherisorb ODS-2 (150 × 4.6 mm, 3 µm) column was used with the following gradient elution: 0–1 min acetonitrile/water 35:65, 1–6 min acetonitrile/water 55:45, 6–16 min acetonitrile/water 60:40, 16–24 min acetonitrile/water 90:10, 24–35 min acetonitrile/water 90:10, 35–40 min acetonitrile/water 35:65, 40–45 min acetonitrile/water 35:65, all at a flow rate 0.6 mL/min.
The amounts of amines were expressed as mg/kg with reference to a calibration curve obtained through aqueous dansyl-chloride-derivatized amine standards of concentrations ranging from 10 to 200 mg/L (Sigma-Aldrich, Milano, Italy). The detection limit for all the amines was 3 mg/kg of the sample under the adopted conditions. All the analyses were performed in triplicate.

Twenty colonies of each isolated species were tested for biogenic amine production on agar media, according to the Bover-Cid and Holzapfel [45 (link)] method. Three spoiled and unspoiled meat homogenates were randomly sampled to detect the biogenic amines using the following method: The samples were extracted with trichloroacetic acid (5%), following the method described by Pasini et al. [46 (link)]. After dansyl-chloride derivatisation (Sigma Aldrich, Gallarate, Italy) [47 (link)], the extracts were analysed in an Agilent Technologies 1260 Infinity HPLC equipped with an automatic injector (G1329B ALS 1260, loop of 20 µL) and with a UV detector (G1314F VWD 1260) set at 254 nm, according to the method reported by Barbieri et al. [48 (link)]. The amounts of amines were expressed as mg/kg with reference to a calibration curve obtained through aqueous dansyl-chloride-derivatised amine standards of concentrations ranging from 10 to 200 mg/L (Sigma-Aldrich, Milano, Italy). The detection limit for all the amines was 3 mg/kg sample under the adopted conditions.