Laborota 4000 hb efficient

Three varieties of Capsicum annuum with different levels of pungency were analysed. These included the non-pungent variety Friariello (GP: green pepper), two different samples of the pungent Cayenne variety (RP1: red pepper 1; RP2: red pepper 2) and one pungent Dzuljunska Sipka variety (RP3: red pepper 3). GP and RP1 were purchased at the local markets of Avellino (AV), Italy (40°54.8964′ N and 14°47.4618′ E); RP2 was purchased at the local market of Grottaminarda (AV), Italy (41°4.1832′ N and 15°3.5364′ E); and RP3 was kindly provided by a farm located in Sicily (Southern Italy). Fresh pepper fruits were rapidly washed in distilled water, freeze-dried and kept at −20 °C until use. Once the peduncles were removed, the freeze-dried peppers were ground in a kitchen grinder and, for each variety, 0.5 g aliquots of ground sample were subjected to extraction with 5 mL of 80% aqueous methanol for 30 min in an ultrasonic bath (Astrason 10E, Farmingdale, NY, USA). After centrifugation (4000 rpm, 4 °C, for 10 min), the supernatant was removed, the pellet was suspended in 5 mL of 80% aqueous methanol and the extraction was repeated under the same conditions. The two supernatants were pooled and dried first under nitrogen flow and then in a rotary evaporator (LaboRota 4000/HB Efficient, Heidolph, Schwabach, Germany). Samples were stored at −20 °C until use.

Three varieties of Capsicum annuum with different levels of pungency were analysed. These included the non-pungent variety Friariello (GP: green pepper), two different samples of the pungent Cayenne variety (RP1: red pepper 1; RP2: red pepper 2) and one pungent Dzuljunska Sipka variety (RP3: red pepper 3). GP and RP1 were purchased at the local markets of Avellino (AV), Italy (40°54.8964′ N and 14°47.4618′ E); RP2 was purchased at the local market of Grottaminarda (AV), Italy (41°4.1832′ N and 15°3.5364′ E); and RP3 was kindly provided by a farm located in Sicily (Southern Italy). Fresh pepper fruits were rapidly washed in distilled water, freeze-dried and kept at −20 °C until use. Once the peduncles were removed, the freeze-dried peppers were ground in a kitchen grinder and, for each variety, 0.5 g aliquots of ground sample were subjected to extraction with 5 mL of 80% aqueous methanol for 30 min in an ultrasonic bath (Astrason 10E, Farmingdale, NY, USA). After centrifugation (4000 rpm, 4 °C, for 10 min), the supernatant was removed, the pellet was suspended in 5 mL of 80% aqueous methanol and the extraction was repeated under the same conditions. The two supernatants were pooled and dried first under nitrogen flow and then in a rotary evaporator (LaboRota 4000/HB Efficient, Heidolph, Schwabach, Germany). Samples were stored at −20 °C until use.

Five apples of the three varieties Annurca (Malus pumila Miller cv Annurca), Red Delicious (M. pumila Miller cv Red Delicious), and Golden Delicious (M. pumila Miller cv Golden Delicious), were randomly selected from batches purchased on different days in a local supermarket in Avellino (Italy). All fruits were quickly washed in distilled water, cut into quarters, and seeds and core were removed. Fruit parts were finely chopped and, for each variety, aliquots of 1 g were treated with 5 ml of 80% aqueous acetone to extract phenolic compounds. Each extraction was carried out for 24 h on a horizontal shaker in a refrigerated chamber at 4 °C. After centrifugation, the supernatant was removed, the pellet was suspended in 5 ml of 80% aqueous acetone and the extraction was carried out for a further 24 h under the same conditions. The two supernatants were pooled and dried in a rotary evaporator (LaboRota 4000/HB Efficient, Heidolph, Schwabach, Germany). Dry extracts were suspended in a mixture of water/1-butanol (1/1; v/v) and subjected to liquid-liquid extraction to remove water-soluble ingredients such as sugars, inorganic salts. This operation was repeated three times, and n-butanol phases were recovered, pooled, dried in a rotary evaporator and stored at −20 °C until used.