The organic acid and sugar content was determined using the method proposed previously by Wojdyło et al.30 (link) by HPLC–PDA (Waters Co.; Milford, CT, USA) and HPLC-ELSD (PL-ELS 1000; Merck; Hitachi, Japan), respectively. The sample (approx. 3 g of fruits and 1 g of leaves) mixed with distilled water, sonicated (Sonic 6D; Polsonic, Warsaw, Poland) for 15 min and boiled for 30 min, finally sample was centrifuged (MPW-55; Warsaw, Poland) at 12,000xg for 10 min at 4 °C. The supernatant (2.5 mL) was applied onto the Sep-Pak C-18 (1 g, Millipore Waters, Milford, MA, USA) and finally eluted by water to Eppendorf tubes. The extract before analysis was filtered through 0.20 μm hydrophilic PTFE membrane (Millex Simplicity Filter; Merck, Germany). All samples were assayed in triplicate repetition. Results expressed as g per 100 g dry weight (dw).
Millex simplicity filter
Manufactured by Merck Group
Sourced in Germany
The Millex Simplicity Filter is a sterile, disposable filter designed for various laboratory applications. It features a 0.22 μm membrane and a simple, easy-to-use design. The filter is intended to remove particulates and microorganisms from liquids, ensuring sample integrity and purity.
Automatically generated - may contain errors
Lab products found in correlation
Sonic 6d, by Polsonic (3 mentions)
G2 q tof micro mass spectrometer, by Waters Corporation (2 mentions)
Mpw 55, by Polsonic (1 mentions)
Hplc pda, by Waters Corporation (1 mentions)
Pl els 1000, by Merck Group (1 mentions)
Acquity ultra performance lc system, by Waters Corporation (1 mentions)
Acquity uplc system, by Waters Corporation (1 mentions)
Pda detector, by Waters Corporation (1 mentions)
Acquity uplc beh c18 rp column, by Waters Corporation (1 mentions)
8 protocols using millex simplicity filter
1
Determination of Organic Acids and Sugars
2
Polyphenol Identification in Coffee Extracts
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Analysis was started from the centrifugation of all samples at 19,000× g for 10 min, and the supernatant was filtered through a hydrophilic PTFE 0.20 µm membrane (MillexSimplicity Filter, Merck, Rahway, NJ, USA) and used for analysis.
Identification of polyphenols of coffee extracts was carried out using an ACQUITY Ultra Performance LC system equipped with a PDA detector (Waters Corporation, Milford, MA, USA) with a mass detector G2 Q-TOF micro mass spectrometer (Waters, Manchester, UK) equipped with an ESI source operating in negative mode. A UPLC BEH C18 column (1.7 µm, 2.1 mm × 100 mm, Waters Corporation, Milford, MA, USA) was used. Samples (10 µL) were eluted according to the linear gradient described previously by Oziembłowski et al. [12 (link)]. Each compound was optimized to its estimated molecular mass in the negative mode, before and after fragmentation, and monitored at 320 nm. All experiments were conducted in triplicate. The results were expressed as milligrams per 1 L of extract.
Identification of polyphenols of coffee extracts was carried out using an ACQUITY Ultra Performance LC system equipped with a PDA detector (Waters Corporation, Milford, MA, USA) with a mass detector G2 Q-TOF micro mass spectrometer (Waters, Manchester, UK) equipped with an ESI source operating in negative mode. A UPLC BEH C18 column (1.7 µm, 2.1 mm × 100 mm, Waters Corporation, Milford, MA, USA) was used. Samples (10 µL) were eluted according to the linear gradient described previously by Oziembłowski et al. [12 (link)]. Each compound was optimized to its estimated molecular mass in the negative mode, before and after fragmentation, and monitored at 320 nm. All experiments were conducted in triplicate. The results were expressed as milligrams per 1 L of extract.
3
Polyphenol Extraction and Analysis
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Polyphenols were extracted according to the procedure reported by Wojdyło et al. [21 (link)]. Briefly, in the first step, products were weighed and thoroughly mixed with 30% methanol solution, followed by the addition of acetic (1%) and ascorbic (1%) acids. The mixtures were sonicated for 15 min, stored at 4 °C for 20 h, and sonicated again for 15 min. Subsequently, all the supernatant was collected after centrifugation and the ready extracts were filtered through a 0.20 μm hydrophilic PTFE membrane (Millex Simplicity Filter; Merck, Germany). Qualitative and quantitative analyses of polyphenols were performed by LC-PDA/MS QTof and UPLC-PDA, respectively, following Wojdyło et al. [22 (link)]. For the quantitative analysis, the phenolic compounds were quantified at 280 nm (dihydrochalcones and flavan-3-ols), 320 nm (phenolic acids), 360 nm (flavonols), and 520 nm (anthocyanins). The polymeric procyanidins were analysed by the phloroglucinol method according to Kennedy and Jones [23 (link)] and the results were expressed as mg/100 g fw.
4
Polyphenol Extraction and Analysis in Fruit Samples
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Methanol extracts for determination of polyphenolic compounds were prepared as follows: The freeze-dried powder of fruits (~1 g) was vortexed for 1 min with 6 mL methanol/water/acetic acid/ascorbic acid (30:68:1:1, v/v/v/m), sonicated for 20 min (Sonic 6D; Polsonic, Warsaw, Poland) and left for 24 hours at 4 °C. Then, the extract was sonicated again for 20 min, and centrifuged at 19.000 × g for 10 min at 4 °C. Finally, the extract was filtred by 0.20 μm hydrophilic PTFE membrane (Millex Simplicity Filter; Merck, Germany) and used for phenolic compounds identification by LC-PDA-QTOF-ESI-MS and quantification by UPLC-PDA. For the determination of antioxidant and in vitro biological activities, the same protocol as that described above was used, but a methanol/water (80:20, v/v) with 1% hydrochloric acid mixture was used for extraction.
5
Polyphenol Identification in Alder Extracts
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The stock solutions were centrifuged at 19,000× g for 10 min, and the supernatant was filtered through a hydrophilic PTFE 0.20 µm membrane (Millex Simplicity Filter, Merck, Darmstadt, Germany) and used for analysis. Identification of polyphenols of A. glutinosa L. extracts was carried out using an ACQUITY UPLC system equipped with a PDA detector (Waters Corporation, Milford, MA, USA) and G2 Q-TOF micro mass spectrometer (Waters, Manchester, UK) equipped with an electrospray ionization source operating in negative mode. Identification and quantification of phenolic compounds were performed according to the method described by Kolniak-Ostek and Oszmiański [54 (link)]. The compounds were monitored at 254 nm (ellagitannins and ellagic acid derivatives), 320 nm (phenolic acids) and 360 nm (flavonols). Calibration curves were determined experimentally for caffeic, ferulic and ellagic acid and isorhamnetin 3-O-glucoside. Ferulic acid derivatives were expressed as ferulic acid, methyl-caffeoyl-quinate was expressed as caffeic acid, isorhamnetin derivatives were expressed as isorhamnetin 3-O-glucoside and ellagitannins and ellagic acid derivatives were expressed as ellagic acid All experiments were performed in triplicate. The results are expressed as milligrams per 100 g of dry matter (DM).
6
Antioxidant Extraction from Dry Plant Materials
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Ground dry plant materials (1 g) were weighed into a test tube and slightly stirred with 20 mL of 80% aqueous methanol with 1% of HCl. Then, the samples were sonicated for 15 min and left at 4 °C in darkness for 24 h. After this period, the samples were again sonicated, then centrifuged for 10 min (20,878× g), and the supernatants were collected at 4 °C to be used within 24 h. Before the analysis by LC–MS and HPLC, the extract was filtered by a 0.20 μm hydrophilic polytetrafluoroethene (PTFE) membrane (Millex Simplicity Filter; Merck, Darmstadt, Germany). For the determination of antioxidant activities, the same protocol as that described above was used, but a methanol/water (80:20, v/v) with 1% hydrochloric acid mixture was used for extraction.
7
Methanol-based Antioxidant and Polyphenol Extraction
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The extract was prepared following methods described in previous studies (Tkacz et al., 2021) (link). The extracts were provided using methanol 80% in distilled water used for antioxidant analysis and methanol 30% (methanol/water/acetic acid/ascorbic acid: 30/68/1/1, v/v/v/m) for polyphenolic measurement. Antioxidant assessment and polyphenolic measurement were performed in methanol solution. Therefore, the extracts were prepared in a methanol solution. The solution was added into 1 g of sample, and it was mixed using a vortex for 1 min. The mixture was sonicated (Sonic 6D, Polsonic, Warsaw, Poland) for 20 min and left at 4 • C. After 24 h, it was again sonicated for 20 min followed by centrifugation at 19000×g at 4 • C for 10 min. The extract was separated using a hydrophilic PTFE membrane (0.20 μm) (Millex Simplicity Filter, Merck, Germany).
8
Extraction and Analysis of Garlic Carotenoids
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Sample extraction was performed as described by Lachowicz et al. [13] . Freeze-dried different parts of wild garlic (~ 0.35 g) were mixed with 5 mL of hexane:acetone:methanol (2:1:1), shaken at 20 °C for 30 min and 40 kHz. Next, the slurry was centrifuged at 19,000g for 10 min, and the supernatant was evaporated to dryness. The pellet was re-extracted using 2 mL of 100% methanol, filtered through a hydrophilic PTFE 0.20 µm membrane (Millex Simplicity Filter, Merck, Darmstadt, Germany), and used for analysis.
For the extraction of carotenoids, a protocol similar to that described earlier was followed [14] . Compounds were separated with an ACQUITY UPLC BEH RP C18 column (1.7 µm, 2.1 mm × 100 mm, Waters Corp., Milford, MA, USA) at 32 °C. The elution solvents were ACN:MeOH (7:3, v/v) (A) and 0.1% formic acid (B). Samples (10 µL) were eluted according to the linear gradient described by Delphino-Rius et al. [15] . The runs were monitored at 450 and 650 nm. The PDA spectra were measured over the wavelength range of 200-700 nm in steps of 2 nm. The retention times and spectra were compared to those of the authentic standards. All incubations were done in triplicate. The results were expressed as milligram per kilogram of dry matter (mg/kg dm).
For the extraction of carotenoids, a protocol similar to that described earlier was followed [14] . Compounds were separated with an ACQUITY UPLC BEH RP C18 column (1.7 µm, 2.1 mm × 100 mm, Waters Corp., Milford, MA, USA) at 32 °C. The elution solvents were ACN:MeOH (7:3, v/v) (A) and 0.1% formic acid (B). Samples (10 µL) were eluted according to the linear gradient described by Delphino-Rius et al. [15] . The runs were monitored at 450 and 650 nm. The PDA spectra were measured over the wavelength range of 200-700 nm in steps of 2 nm. The retention times and spectra were compared to those of the authentic standards. All incubations were done in triplicate. The results were expressed as milligram per kilogram of dry matter (mg/kg dm).
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