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4 protocols using simplicity ultrapure water system

1

Plum Distillate Storage Stability Study

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The plum distillates were diluted with deionized water (Simplicity® Ultrapure Water System, 18.2 MΩ∙cm resistivity (25 °C) at 0.5 L/min, Merck Millipore) from initial ASVs of 76.77% v/v and 81.92% v/v to 37.5, 40 and 50% v/v. Next, the samples were placed in glass bottles and stored under the following conditions, for 64 days:

ambient temperature (+ 20 ± 2 °C), daylight,

ambient temperature (+ 20 ± 2 °C), darkness,

cold storage (+ 8 °C), darkness,

temperature close to freezing (− 16 °C for ASVs of 37.5% v/v and 40% v/v, − 25 °C for ASV of 50% v/v), darkness.

After 10, 20, 30, 34, 42, 50, 57 and 64 days the turbidity of the samples was measured. After 64 days, the samples were filtered through filter sheets and then subjected to turbidity measurements, as well as to chromatographic and organoleptic analysis. The control samples were the distillates before treatment. Prior to filtration, the used filter sheets were rinsed with deionized water for 10 min in the direction of flow.
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2

Industrial-Scale Distillate Production

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The raw materials used were distillates produced on an industrial scale by the Polish companies. Rye distillate (ABV of 93.26% v/v) was sourced from the agricultural distillery Zbig-Rol (Prusinowice, Poland). Plum distillate (ABV of 82.03% v/v) was provided by the company +H2O (Chociszew, Poland). Deionized water was produced using the Simplicity® Ultrapure Water System, 18.2 MΏ·cm resistivity (25 °C) at 0.5 L/min, Merck Millipore (Burlington, MA, USA).
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3

Synthesis and Characterization of P1 Compound

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The synthesis of P1 is reported
in the literature.54 (link) All samples were prepared
in water that was distilled and then purified by a Millipore purification
system (Millipore Simplicity Ultrapure Water System). Buffer solution
(N-(2-hydroxyethyl)piperazine-N′-ethanesulfonic
acid (HEPES), 10 mM, pH = 7.4) was prepared using 4-(2-hydroxyethyl)-1-piperazine
ethanesulfonic acid and sodium hydroxide from Sigma-Aldrich. Tris(3-aminoethyl)amine
and potassium pyrophosphate were also purchased from Sigma-Aldrich.
All chemicals were used as received, unless otherwise noted.
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4

Protein Sample Preparation Workflow

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Trifluoroacetic acid (TFA), ammonium bicarbonate (ABC), formic acid (FA), dithiothreitol (DTT), iodoacetamide (IAM), and Tween 20 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium hydroxide, DMSO, iodomethane and trichloromethane were purchased from Merck Millipore (Darmstadt, Germany). Trypsin was purchased from Promega (15,664 unit/mg, Sequence Grade, WI, USA). Acrylamide, SDS and TEMED were obtained from Bio-Rad (Hercules, CA, USA). Acetonitrile (ACN) was purchased from J.T. Baker (Phillipsburg, NJ, USA). Deionized water was generated with a Simplicity Ultrapure Water System (Millipore, Burlington, MA, USA) with a measured value of 18 MΩ.
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