According to Analytica—EBC 7.7 method [47 ] HPLC was employed to determine the α- and β-acids in hop extracts. Extracts were filtered through a disposable syringe filter, Chromafil Xtra PET-45/25 (Macherey-Nagel, Dueren, Germany) and a 10 µL injection loop on the HPLC injector was used. The separation was achieved on the Nucleodur 5–100 C18, 125 × 4 mm HPLC analysis column (Macherey-Nagel, Dueren, Germany). The isocratic mobile phase consisting of distilled water, methanol (J.T.Baker, USA) and 85% aqueous solution of ortophosphoric acid (MERCK, Germany, Taufkirchen) in a ratio of 775/210/9 (v/v/v) was used, and the detection was carried out with a Diode array detector (DAD) set at 314 nm for α-and β-acids and 370 nm for detection of xantohumole, respectively. The quantification was performed by the external standard ICE4 (NATECO2, Wolnzach, Mainburg, Germany) for α-and β-acids and by XH 90% (Steiner Hopfen GmBH, Germany). All solvents were of analytical grade purity.
Ortophosphoric acid
Manufactured by Merck Group
Sourced in Germany
Orthophosphoric acid is a colorless, odorless, and viscous liquid chemical compound. It is the simplest phosphoric acid and is commonly used in various industrial and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Chromafil xtra pet 45 25, by Macherey-Nagel (2 mentions)
Methanol, by Merck Group (2 mentions)
Pvp k90, by Merck Group (1 mentions)
Potassium dihydrogen phosphate, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Methanol, by Avantor (1 mentions)
Pd98059, by New England Biolabs (1 mentions)
Sb203580, by Merck Group (1 mentions)
Swine anti rabbit fitc antibody, by Agilent Technologies (1 mentions)
Boron trifluoride methanol complex, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Ly294002, by Merck Group (1 mentions)
Amarogentin, by ChromaDex (1 mentions)
Gentiopicroside, by ChromaDex (1 mentions)
Low molecular weight chitosan, by Merck Group (1 mentions)
Tryptone soya broth (tsb), by Thermo Fisher Scientific (1 mentions)
Trehalose, by Merck Group (1 mentions)
Tryptone soya agar tsa, by Thermo Fisher Scientific (1 mentions)
Kolliphor p407, by BASF (1 mentions)
5 protocols using ortophosphoric acid
1
HPLC Analysis of Hop Extracts
2
Aceclofenac Solubility Enhancement
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Aceclofenac (Figure 2 ) was provided by Richter Gedeon Plc. (Budapest, Hungary), poly (vinyl-pyrrolidone) (PVP K90, Kollidon K90) was obtained from Sigma Aldrich (Merck, Darmstadt, Germany), and ethanol (EtOH, 96%) was from Chemical Company (Iasi, Romania). Methanol, acetonitrile, and orto-phosphoric acid (85%) used during the liquid chromatographic measurements were HPLC grade, obtained from Merck (Darmstadt, Germany). Ultrapure distilled water was prepared in-house by a MilliQ water purification system. Potassium dihydrogen phosphate and sodium hydroxide used throughout the in vitro dissolution studies were obtained from Merck (Darmstadt, Germany).
3
Hops Alpha-Beta Acids Quantification by HPLC
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According to Analytica-EBC 7.7 method [16] , high performance liquid chromatography (HPLC) was employed to determine the alpha-and beta-acids in hops. Ground hops (5 g) was added to 80 mL of the mixture of diethyl ether/methanol and 0.1 M aqueous hydrochloric acid in a ratio of 10/50/20 (v/v/v). Extracts were filtered through a disposable syringe filter, Chromafil Xtra PET-45/25 (Macherey-Nagel, Düren, Germany) and 10 µL injection loop on HPLC injector was used. The separation was achieved on Nucleodur 5-100 C18, 125 × 4 mm HPLC analysis column (Macherey-Nagel, Düren, Germany). Isocratic mobile phase constituted from distilled water, methanol (J.T.Baker, Phillipsburg, New Jersey, USA) and 85% aqueous solution of ortophosphoric acid (MERCK, Kenilworth, NJ, USA) in a ratio of 775/210/9 (v/v/v) was used and the detection was carried out with Diode array detector (DAD) set at 314 nm for alpha-and beta-acids and 370 nm for xanthohumol. The quantification was done by the external standard ICE4 (NATECO2, Wolnzach, Germany). All solvents were of analytical grade purity.
4
Immunohistochemical Analysis of CerS3 Expression
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The polyclonal rabbit-anti-human CerS3 antibody was from Antikörper-online (Aachen, Germany; 1:50). The secondary antibodies were the swine anti-rabbit-FITC antibody (Dako, Hamburg, Germany; 1:30) and the donkey anti-rabbit-Alexa-Fluor 555 antibody (Thermo Fisher Scientific, Darmstadt, Germany). DAPI was from Sigma-Aldrich GmbH (Taufkirchen, Germany). The following specific inhibitors were used: PPARγ antagonist (GW9962, Enzo, Lörrach, Germany), PI3K inhibitor (LY294002; Sigma-Aldrich GmbH), ERK inhibitor (PD98059, New England Biolabs GmbH, Frankfurt, Germany), p38 MAPK inhibitor (SB203580, Sigma-Aldrich GmbH) and NFκB inhibitor (GIV 3727, Merck Millipore, Darmstadt, Germany). Methanolic sodium hydroxide solution, hexane standards, loganic acid, and loganin were from Carl Roth GmbH (Karlsruhe, Germany). Boron trifluoride–methanol complex was from Merck KGaA (Darmstadt, Germany), and standard methyl heptadecanoate was from Sigma-Aldrich GmbH. Orto-phosphoric acid, 85%, Ph. Eur. p.a. grade was supplied by VWR. Amarogentin and gentiopicroside were purchased from Chromadex Inc. (Santa Ana, CA, USA).
5
Pseudomonas aeruginosa Antimicrobial Formulation
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Pseudomonas aeruginosa (ATCC 17334), Tryptone Soya Broth (TSB) and Tryptone Soya Agar (TSA) from Oxoid, Basingstoke, UK; N-acetylcysteine (Farmalabor, Assago, Milano, Italy); caprylic/capric tryglicerides (Labrafac cc, Gattefossé sas, Milano, Italy) and glycerol dibehenate (Compritol 888 pastilles, Gattefossé sas, Milano, Italy); Kolliphor p 407, a mix of polyoxyethylene (73%) and polyoxypropylene (POE-POP) with gelling and thickening action was obtained from BASF (Germany) and polyglicery l-4 sorbitan olivate phosphate (Caldic, Milano, Italy); acetonitril HPLC grade, hydrochloric acid and orto-phosphoric acid (Sigma-Aldrich, Milano, Italy), chitosan, low molecular weight (Sigma-Aldrich, Milano, Italy), trehalose (Sigma-Aldrich, Milano, Italy).
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