Assays were performed according to manufacturer’s instructions. The BACE1 assay kit was purchased from DiscoveRx (Fremont, CA, USA). ACS grade solvents (Fisher Scientific, Waltham, MA, USA) were distilled and filtered before use. HP-20 resin, flash column stationary phases, and HPLC columns were purchased from Supelco (St. Louis, MO, USA), Sorbent Technologies (Atlanta, GA, USA), and Phenomenex (Torrance, CA, USA), respectively.
Hp 20 resin
Manufactured by Merck Group
Sourced in United States
The HP-20 resin is a chromatographic material used for the separation and purification of various compounds. It is a porous polymer-based resin with a high surface area, allowing for efficient adsorption and desorption of target molecules. The HP-20 resin can be utilized in a range of applications, including the isolation of natural products, the purification of pharmaceuticals, and the cleanup of environmental samples.
Automatically generated - may contain errors
Lab products found in correlation
Acs grade solvents, by Thermo Fisher Scientific (2 mentions)
Flash column stationary phases, by Merck Group (2 mentions)
Ethyl acetate, by Thermo Fisher Scientific (1 mentions)
Tsb medium, by BD (1 mentions)
Ferrozine, by Sinopharm (1 mentions)
Fdu 2100, by Eyela (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Sinopharm (1 mentions)
Methanol, by Sinopharm (1 mentions)
Ethanol, by Sinopharm (1 mentions)
N hexane, by Sinopharm (1 mentions)
Ferric chloride, by Sinopharm (1 mentions)
Potassium ferricyanide, by Sinopharm (1 mentions)
Ferrous chloride, by Sinopharm (1 mentions)
Quillaja saponin, by Merck Group (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid (abts), by Merck Group (1 mentions)
N butanol, by Merck Group (1 mentions)
Rna isolation kit, by Roche (1 mentions)
Asi 100, by Thermo Fisher Scientific (1 mentions)
9 protocols using hp 20 resin
1
BACE1 Enzyme Activity Assay
2
Purification of Organic Solvents
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ACS grade solvents (Fisher Scientific, Waltham, MA, USA) were distilled and filtered before use. HP-20 resin, flash column stationary phases, and HPLC columns were purchased from Supelco (St. Louis, MO, USA), Sorbent Technologies (Atlanta, GA, USA), and Phenomenex (Torrance, CA, USA), respectively.
3
Bioactive Compound Extraction from Diverse Actinobacteria
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All twenty-five strains were pre-cultured (5 mL, 28 °C, 160 rpm for 7 days) in ISP2 medium [82 (link)], ISP3 medium [82 (link)] A1M1 medium [27 (link)] and 10-fold diluted TSB medium (BD™) prepared in distilled water. Instant ocean (18 g/L) was added in each of them. Each culture [ISP2/ISP3/A1M1/10-fold dil. TSB medium (50 mL) with activated HP-20 resin (Sigma) (2.5 g)] was inoculated (5% v/v pre-culture) and fermented (14 days, 28 °C, 160 rpm). The culture was then centrifuged (4000 rpm, 20 min), the supernatant removed, and the cell/resin pellet lyophilised until dry (Thermo Savant MicroModulyo, Thermo Fisher Scientific, Waltham, MA, USA). The lyophilised cell/resin pellet was extracted twice with ethyl acetate (Fisher Scientific, Loughborough, UK, reagent grade) (20 mL, 100 rpm, 25 °C). The extracts were combined, dried (under N2), the weight recorded and stored (4 °C).
4
Purification of Rb3 and Rd Ginsenosides
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The reaction mixture consisted of the Rb2 and Rb3 mixture at the final concentration of 50 mg/mL in 200 mL of crude recombinant Bgp2 (pH 7.0). After incubation for 12 h at 37°C, the mixture was centrifuged at 4000 × g for 15 min, and then the supernatant was loaded to a column packed with HP20 resin (340 g) (Sigma, St. Louis, MO). Two liters of water were used to remove unbound hydrophilic compounds and free sugar molecules, and the absorbed ginsenosides were eluted using three bed volumes of 95% ethanol. The eluted ethanol solution with Rb3 and Rd was evaporated in vacuo to remove the ethanol.
5
Mulberry Fruit Antioxidant and Anti-inflammatory Potential
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Mulberry fruit, M. alba of J33 variety, was harvested at ripe stage from the Plantation of National Mulberry Orchard (Zhenjiang, Jiangsu province, PR China). Fruits were dried by freeze dryer (EYELA FDU-2100, Japan) and milled into powder for further use. The Experimental Animal Center of Jiangsu University (Zhenjiang, China) provided clean grade female ICR mice weighing 30 g with an age of 6 weeks. Moreover, the experiments were performed in accordance with regional legal regulations (Ethical issue number UJS-LAER-2015080101). HP-20 resin was purchased from Sigma Chemical Co. (St. Louis, MO, USA), whereas sea sand, n-hexane, ethanol, methanol, 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferrozine, potassium ferricyanide, ferrous chloride, and ferric chloride were purchased from Sino pharm Chemical Reagent Co. Ltd. (Shanghai, China). All other reagents used in this study were of analytical grade.
6
Biochemicals and Cell Culture Reagents
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All of the chemicals (HP-20 resin, Quillaja saponin, MTT, ABTS, DPPH, …) were purchased from Sigma- Aldrich Co. (St. Louis, MO, USA) and all solvents (ethanol, n-butanol, dichloromethane, ascetic acid) were purchased from Merck (Darmstadt, Germany). Cell cultured reagents were obtained from Gibco-USA. RNA isolation kit from Roche Company was purchased. cDNA synthesis kit and RT-PCR kit were purchased from Thermo Scientific Company.
7
Isolation and Fractionation of Plum Polyphenols
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Chromatography with the use of HP-20 resin (Sigma-Aldrich) was utilized to extract semipurified polyphenolic fractions from dried plum powder. First, a crude polyphenol extract was derived from sonicating 500 g dried plum in 80% methanol under pulsated nitrogen gas 2 times. The liquid phase was subjected to column chromatography by using 300 g HP-20 resin. The column was then rinsed 5 times with 200 mL deionized water to eliminate a water-soluble carbohydrate-rich extract. After the rinses, the column was washed with 100% methanol to yield a water-insoluble total polyphenolic–rich extract. This extract was then subjected to additional column chromatography by using 200 g HP-20 resin. Six semipurified polyphenol fractions of similar solubility properties were eluted from the column with increasing concentrations of methanol (i.e., 0%, 20%, 40%, 60%, 80%, and 100% methanol). These fractions were lyophilized and the weights of each fraction, derived from 500 g dried plum powder, were as follows: dried plum fraction (DP-Fr) A, 17.85 g; DP-FrB, 4.42 g; DP-FrC, 3.27 g; DP-FrD, 4.14 g; DP-FrE, 2.16 g; and DP-FrF, 1.39 g.
8
Comprehensive Spectroscopic Analysis of Natural Products
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NMR spectra were acquired in CD3OD on Bruker Avance DRX III 600 instruments ( 1 H at 600 MHz and 13 C at 150 MHz). Standard pulse sequences and parameters were used to obtain 1D 1 H and 13 C and 2D COSY, ROESY, TOCSY, HSQC-TOCSY, and HMBC spectra. HR-ESI-MS data were gained using a Micromass Q-TOF high-resolution mass spectrometer. Optical rotations were determined in MeOH by Perkin-Elmer 241 polarimeter. TLC was performed on precoated silicagel 60 F254 Merck and compounds were visualized by spraying the dried plates with 50% H2SO4, followed by heating. CC was carried out on HP-20 resin (Sigma Aldrich).
Flash chromatography was conducted on a Grace Reveleris system equipped with dual UV and ELSD detection using Grace® cartridges (Silica gel or RP-C18). HPLC was performed on a Dionex apparatus equipped with an ASI100, ultimate 3000 Pump, a diode array detector UVD 340S and a chromeleon software. A prepacked RP-C18 column (Phenomenex 250 x 15 mm, Luna 5 µ) was used for semi-preparative HPLC. The eluting mobile phase consisted of H2O with TFA (0.0025%) and CH3CN with a flow rate of 5 mL/min and the chromatogram was monitored at 205, 210, 254, and 300 nm.
Flash chromatography was conducted on a Grace Reveleris system equipped with dual UV and ELSD detection using Grace® cartridges (Silica gel or RP-C18). HPLC was performed on a Dionex apparatus equipped with an ASI100, ultimate 3000 Pump, a diode array detector UVD 340S and a chromeleon software. A prepacked RP-C18 column (Phenomenex 250 x 15 mm, Luna 5 µ) was used for semi-preparative HPLC. The eluting mobile phase consisted of H2O with TFA (0.0025%) and CH3CN with a flow rate of 5 mL/min and the chromatogram was monitored at 205, 210, 254, and 300 nm.
9
Analytical Characterization of Organic Compounds
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Optical rotations were measured on a Perkin Elmer model 341 polarimeter (589 nm, 20 °C).
NMR data were performed in CD 3 OD on Bruker Avance 500. HRESIMS data were gained using a Micromass Q-TOF high-resolution mass spectrometer. Mass spectra were recorded in the positive-ion mode in the range m/z 100-2000, with a mass resolution of 20000 and an acceleration voltage of 0.7 kV. CC was carried out on HP-20 resin (Sigma Aldrich). Flash chromatography was conducted on a Grace Reveleris system equipped with dual UV and ELSD detection using Grace® cartridges (Silica gel or RP-C 18 ). HPLC separations were performed on a Dionex apparatus equipped with an ASI-100 autosampler, an Ultimate 3000 pump, a STH 585 column oven, a diode array detector UVD 340S and a Chromeleon software. A prepacked RP-C 18 column (Phenomenex 250 x 15 mm, Luna 5 µ) was used for semi-preparative HPLC. The eluting mobile phase consisted of H 2 O with TFA (0.0025%) and CH 3 CN with a flow rate of 5 mL/min and the chromatogram was monitored at 205 and 210 nm. TLC were carried out using silica gel 60 F 254 pre-coated aluminium plates (0.2 mm, Merck). Spots were visualized through developing agent (CHCl 3 :MeOH:H 2 O, 14:6:1) and chromogenic agent (50% aq. H 2 SO 4 ) subsequent heating.
NMR data were performed in CD 3 OD on Bruker Avance 500. HRESIMS data were gained using a Micromass Q-TOF high-resolution mass spectrometer. Mass spectra were recorded in the positive-ion mode in the range m/z 100-2000, with a mass resolution of 20000 and an acceleration voltage of 0.7 kV. CC was carried out on HP-20 resin (Sigma Aldrich). Flash chromatography was conducted on a Grace Reveleris system equipped with dual UV and ELSD detection using Grace® cartridges (Silica gel or RP-C 18 ). HPLC separations were performed on a Dionex apparatus equipped with an ASI-100 autosampler, an Ultimate 3000 pump, a STH 585 column oven, a diode array detector UVD 340S and a Chromeleon software. A prepacked RP-C 18 column (Phenomenex 250 x 15 mm, Luna 5 µ) was used for semi-preparative HPLC. The eluting mobile phase consisted of H 2 O with TFA (0.0025%) and CH 3 CN with a flow rate of 5 mL/min and the chromatogram was monitored at 205 and 210 nm. TLC were carried out using silica gel 60 F 254 pre-coated aluminium plates (0.2 mm, Merck). Spots were visualized through developing agent (CHCl 3 :MeOH:H 2 O, 14:6:1) and chromogenic agent (50% aq. H 2 SO 4 ) subsequent heating.
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