For sample extraction, 1 g of leaves (corresponding to 5 leaves) was weighed and crushed in liquid nitrogen. The resulting powder was resuspended in 3 mL of 70% acetone, homogenized for 5 min by an Ultra-Turrax® T-25 basic (IKA®-Werke GmbH & Co. KG, Staufen im Breisgau, Germany) and sonicated for about 20 min with Elma Transsonic T 460/H. A second step of homogenization was added to ensure the total lysis of plant material. The final mixture was centrifuged for 5 min at 4000 r.p.m. (1500 r.c.f.) (Eppendorf® Microcentrifuge 5415D, Hamburg, Germany). Sample extraction was performed for colorimetric analysis using 3 plants. The experiment was conducted in triplicate. Results are expressed as the average of the three replicas ± the standard deviation.
Transsonic t 460 h
Manufactured by Elma Schmidbauer
Sourced in Germany, Switzerland
The Transsonic T 460/H is a laboratory equipment product. It is a high-performance ultrasonic homogenizer designed for efficient sample preparation and cell disruption in a variety of applications.
Automatically generated - may contain errors
4 protocols using transsonic t 460 h
1
Leaf Pigment Extraction Protocol
2
Acetone Extraction of Flour Compounds
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For each triplicate sample, 9 mL of 70% acetone was added immediately to 0.5 g of flour [27 (link)]. The tube was then rapidly homogenized by an Ultra-Turrax® T-25 basic (IKA®-Werke GmbH & Co. KG, Staufen im Breisgau, Germany) and sonicated for 20 min with an Elma Transsonic T 460/H. Finally, the sample was centrifuged for 5 min at 1500 RCF (Eppendorf® Microcentrifuge 5415D, Hamburg, Germany).
3
Tomato Phytochemical Extraction Protocol
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For each tomato cultivar, five fruits were selected randomly and chopped. Then, 1 g of peel and 1 g of pulp were weighed and 6 mL (for peel) and 3 mL (for pulp) of 70% acetone were immediately added. Samples were homogenized by Turrax (Ultra-Turrax® T25 based IKA, Saint Louis, MO, USA) for 5 min, then placed in a sonicator (Elma Transsonic T 460/H, Wezikon, Switzerland) for 15 min and then homogenized again by Turrax. Samples were then centrifuged at 4000× g r.p.m. for 5 min (Eppendorf® 5415D centrifuge, Hamburg, Germany). Finally, supernatants (i.e., the extract) were transferred to 2 mL Eppendorf tubes.
4
Preparation of Lyophilized Tomato Peel Extracts
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Lyophilized tomato peel sample was prepared according to Beconcini et al. [34 (link)], with some modifications. Briefly, frozen tomatoes were hand peeled and weighed to obtain 2 g of tomato peel and then 6 mL of water was added. The mixture was homogenized (Ultra-Turrax® T25 based IKA, Saint Louis, MS, USA) for about 3 min and sonicated (Elma Transsonic T 460/H, Wetzikon, Switzerland) for 20 min to guarantee complete cellular decomposition. The mix was again homogenized one minute and centrifuged 5 min at 13,000 rpm (Eppendorf® 5415D centrifuge, Hamburg, Germany) to separate the biomolecules from the pellet. The supernatant was filtered through a 0.45 µm cellulose acetate membrane filter (Sartorius, Göttingen, Germany). Finally, the tomato peel extracts were freeze-dried 48 h (freeze dryer LIO 5P, 5pascal, Italy). The freeze-dried tomato peel extracts were transferred into airtight containers and stored at −20 °C. The freeze-dried tomato peel extracts were diluted in water for further analysis.
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