Most of the chemicals were obtained from MERCK, Germany, and used as received without further purifications. Acetonitrile was purchased from SAMCHUN Pure Chemical Co., South Korea, and Cation Exchange Resin, Dowex-50W-x8, was obtained from Sigma Aldrich, USA. The Preyssler-type HPA was prepared in our laboratory. Aqueous solutions were prepared from double distilled deionized water. All the chemical reactions were carried out at room temperature.
Dowex 50wx8
Manufactured by Merck Group
Sourced in United States, China
Dowex 50WX8 is a strong acid cation exchange resin with a polystyrene-divinylbenzene (PS-DVB) matrix. It has a high exchange capacity and is suitable for a variety of applications, including water treatment, ion exchange chromatography, and other industrial processes.
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Lab products found in correlation
Triethylamine tea, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Acetonitrile, by Samchun (1 mentions)
Amberlite ira 67, by Merck Group (1 mentions)
Amberlite ira 743, by Merck Group (1 mentions)
Vista mpx, by Agilent Technologies (1 mentions)
Magnesium sulphate, by Merck Group (1 mentions)
Ethyl chloroformate, by Merck Group (1 mentions)
Amberlyst 15, by Merck Group (1 mentions)
Monomuls 90 o18, by BASF (1 mentions)
Resveratrol, by Merck Group (1 mentions)
Hydrazine sulfate, by Merck Group (1 mentions)
Nhs lc lc biotin, by Thermo Fisher Scientific (1 mentions)
Pyridine, by Merck Group (1 mentions)
Sodium dodecyl sulphate (sds), by Merck Group (1 mentions)
Tetrabutylammonium acetate, by Merck Group (1 mentions)
Pluronic f68, by Merck Group (1 mentions)
Sorafenib free base, by Active Motif (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Glacial acetic acid, by Merck Group (1 mentions)
9 protocols using dowex 50wx8
1
Synthesis of Preyssler-type HPA
2
Ultrapure Boron Isotopic Analysis
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Hydrochloric acid (Guaranteed Reagent) was redistilled in a sealed vessel to remove the exogenous B. The cesium carbonate (spectroscopic pure) was of 99.994% purity. High-purity graphite was added to a mixture of ethanol solution (80%) to obtain the final solution corresponding to 13 mg/g graphite. The isotopic reference standard used in this study was NIST SRM 951 boric acid (Gaithersburg, MD, USA). A solution of mannitol of 1.82% (w/v) and Cs2CO3 solution containing 12.3 mg/mL of Cs+ was also prepared. Sodium carbonate, ammonia hydroxide, and sodium chloride were of the analytical grade reagent. Borax and boric acid were of Guaranteed grade Reagent.
The resins, B specific resin Amberlite IRA 743, strong cation exchange resin Dowex 50W X8, and weakly anion exchange resin Amberlite IRA 67, were purchased from Sigma-Aldrich Co. LLC, China.
High purity water with a B blank less than 0.008 μg was redistilled by subboiling distillation and passed through a resin column filled with B specific resin (Amberlite IRA 743), which was used to prepare the standard solution and working solution.
An inductively coupled plasma optical emission spectrometer (ICP-OES, Vista MPX, Varian, USA) with a 40 MHz radio frequency generator and a charge coupled device detector (Vista Chip) was used to detect B.
The resins, B specific resin Amberlite IRA 743, strong cation exchange resin Dowex 50W X8, and weakly anion exchange resin Amberlite IRA 67, were purchased from Sigma-Aldrich Co. LLC, China.
High purity water with a B blank less than 0.008 μg was redistilled by subboiling distillation and passed through a resin column filled with B specific resin (Amberlite IRA 743), which was used to prepare the standard solution and working solution.
An inductively coupled plasma optical emission spectrometer (ICP-OES, Vista MPX, Varian, USA) with a 40 MHz radio frequency generator and a charge coupled device detector (Vista Chip) was used to detect B.
3
Determination of Carboxyl Modification in CMHA
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To determine the degree of carboxyl modification of CMHA, a modified titration assay was used [21 ]. Briefly, CMHA was dissolved at 10 mg/ml in DI water. Cationic exchange resin (Dowex 50WX8, 100–200 mesh, Sigma-Aldrich) was added to the solution and was mixed for 2 hrs at 37°C. The solution with resin was then filtered first through a 0.8μm filter and then through a 0.22μm filter to remove the resin. The resulting solution of acidified CMHA was then lyophilized. Dry acidified CMHA (125 mg) was dissolved in 10ml of 0.1N NaOH and diluted with 15ml of DI water. This solution was then titrated with 0.05N HCl, using phenolphthalein as an indicator. Blank titrations contained no CMHA. The degree of substitution was calculated using the following equations:
where: nCOOH = mol. carboxyl grps
Vb = vol. HCl needed to titrate blank
V = vol. HCl needed to titrate sample
CHCl = conc. of HCl
where: DS = degree of substitution
MWDSU = MW of an unsubstituted disaccharide unit
mdry = mass of dry CMHA
MWI = increase in MW due to carboxyl grp substitution
The percent carboxyl modification was then calculated DS × 100%. Each lot of CMHA was acidified twice separately, and the percent modification of the two runs averaged.
where: nCOOH = mol. carboxyl grps
Vb = vol. HCl needed to titrate blank
V = vol. HCl needed to titrate sample
CHCl = conc. of HCl
where: DS = degree of substitution
MWDSU = MW of an unsubstituted disaccharide unit
mdry = mass of dry CMHA
MWI = increase in MW due to carboxyl grp substitution
The percent carboxyl modification was then calculated DS × 100%. Each lot of CMHA was acidified twice separately, and the percent modification of the two runs averaged.
4
Synthesis and Characterization of Resveratrol-Loaded Liquid Crystalline Nanosystems
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All liquid crystalline nanosystems were prepared from glyceryl monooleate lipid Monomuls® 90-O18 (1-(cis-9-octadecenoyl)-rac-glycerol), (GMO) (BASF, Ludwigshafen, Germany) and used without further purification. All formulations were prepared in HPLC-grade water.
Resveratrol was acquired from Sigma-Aldrich Chemical Co (St. Louis, MO, USA).
Dichloromethane (POCH) was dried over CaH2 and distilled under reduced pressure prior to use. THF (POCH) was distilled over a sodium-potassium alloy. Dowex 50WX8 (Sigma-Aldrich, Darmstadt, Germany) was washed with dry THF before use. Methoxy ether poly(ethylene oxide) 5000 (mPEO123-OH, TCI, Mn(GPC) = 5 200, Mn(NMR) = 5 400) was dried by two azeotropic distillations using anhydrous toluene. L-lactide (LA) (>99.5%, Forusorb) was purified by sublimation two times before use. 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) (98%, Sigma-Aldrich) was distilled under reduced pressure over BaO. Triethylamine (TEA, >99%, Sigma-Aldrich) was distilled under reduced pressure over BaO. 2,2-Bis(hydroxymethyl)propionic acid (bis-MPA, >97%, TCI), ethanol (96%, POCH), ethyl chloroformate (97%, Sigma-Aldrich), Amberlyst 15 (Sigma-Aldrich) and magnesium sulphate (Chempur, Piekary Slaskie, Poland) were used as received.
Resveratrol was acquired from Sigma-Aldrich Chemical Co (St. Louis, MO, USA).
Dichloromethane (POCH) was dried over CaH2 and distilled under reduced pressure prior to use. THF (POCH) was distilled over a sodium-potassium alloy. Dowex 50WX8 (Sigma-Aldrich, Darmstadt, Germany) was washed with dry THF before use. Methoxy ether poly(ethylene oxide) 5000 (mPEO123-OH, TCI, Mn(GPC) = 5 200, Mn(NMR) = 5 400) was dried by two azeotropic distillations using anhydrous toluene. L-lactide (LA) (>99.5%, Forusorb) was purified by sublimation two times before use. 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) (98%, Sigma-Aldrich) was distilled under reduced pressure over BaO. Triethylamine (TEA, >99%, Sigma-Aldrich) was distilled under reduced pressure over BaO. 2,2-Bis(hydroxymethyl)propionic acid (bis-MPA, >97%, TCI), ethanol (96%, POCH), ethyl chloroformate (97%, Sigma-Aldrich), Amberlyst 15 (Sigma-Aldrich) and magnesium sulphate (Chempur, Piekary Slaskie, Poland) were used as received.
5
Characterization of Dowex 50WX8 Resin
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The chemicals used in this work were of analytical reagent grade (AR) and most of them were used without further purification. Stock solutions of Pr(III), DY(III) and Y(III) (1000 mg/L) were prepared by dissolving a known amount of the metal oxide in minimum concentrated nitric acid and evaporated to near dryness and then made up to the mark in a measuring flask with double distilled water. The desired required concentrations of test solutions were prepared by favorable dilution with a known concentrated nitric acid of the stock solutions. Dowex 50WX8 which is a strong acid cation resin containing 8% divinylbenzene (DVB) [20 (link)], Fig. 1 was purchased from sigma Aldrich. The chemical and physical specifications of Dowex 50X8 are given in table given in Table 1 .![]()
Chemical structure of hydrogenated Dowex 50WX8 resin
Specification data sheet of Dowex 50WX8
| Product specification | |
|---|---|
| Appearance (Color) | Faint Yellow to Brown |
| Appearance (Form) | Beads |
| Chemical form | Strongly acidic hydrogen form containing 8% divinylbenzene (DVB) |
| Mesh size | 100–200 mesh |
| Particle size | Distribution > 90% within 300 to 1180 UM |
| Wet capacity | > 1.7 _MEQ/ML |
| Moisture content | 50–60% |
6
PF4 Interaction with Heparin and Enoxaparin
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HP and LMWH (enoxaparin) were obtained from the United States Pharmacopoeia convention. Recombinant human PF4 was purchased from Sino Biological (Beijing, China). NHS-LC-LC-Biotin was purchased from Thermo Fisher Scientific (Shanghai, China). Deuterium oxide (99.9% D2O) was obtained from Qingdao Tenglong Weibo Technology Co., Ltd. (Qingdao, China). The 70% aqueous hydrazine was obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Hydrazine sulfate, dimethyl sulfoxide, pyridine and Dowex 50WX8 (hydrogen form, 50 to 100 mesh) were purchased from Sigma-Aldrich (Shanghai, China). N, O-bis(trimethylsilyl)acetamide (BTSA) was obtained from Jinming Biotechnology (Beijing, China). Phosphate-Buffered Saline (PBS) was purchased from Wuhan Procell Biological Technology (Wuhan, China).
7
Synthesis and Characterization of PEGylated Polymer Nanocarriers
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AminoPEG Mw = 5000 was purchased from Nanocs Inc. (New York, NY, USA), poly(vinyl alcohol) (PVA, Mw = 30,000–70,000 g/mol, 87–90% hydrolysed), Pluronic F-68, dichloromethane (DCM), acetone, glacial acetic acid, dimethyl sulfoxide (DMSO), sodium azide, 1-ethyl-3(3-dimethylaminopropyl) carbodiimide (EDC), N-hydroxysuccinimide (NHS), sodium dodecyl sulphate (SDS), RPMI-1640 medium and triethyl amine (TEA) were obtained from Sigma Aldrich (Saint Louis, MO, USA). Sorafenib (free base) and doxorubicin hydrochloride were purchased from Active Biochem (Hong Kong, PRC). The following reagents were used for the [R,S]-β-butyrolactone oligomerization reaction: β-butyrolactone (BL, ≥98%) purified as described previously [43 (link)], tetrabutylammonium acetate (97%), Dowex® 50WX8 (hydrogen form, 200–400 mesh) obtained from Sigma Aldrich (Saint Louis, MO, USA), as well as solvents: tetrahydrofuran over molecular sieves (THF, pure), n-hexane (pure p.a.), and chloroform (98.5%) obtained from Avantor Performance Materials Poland S.A. (Gliwice, Poland); all were used without further purification. The HCT-116 human colon carcinoma cell line was a kind gift from the National Institute of Oncology (Budapest, Hungary).
8
MALDI Mass Spectrometry of Lipid A
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MALDI negative-ion mass spectrometry analyses were performed on a PerSeptive Voyager-DE STR model time-of-flight mass spectrometer of Applied Biosystem, (I2BC, Université de Paris Sud XI). The apparatus is equipped with a 337 nm nitrogen laser and spectra were recorded in the linear negative-ion mode with delayed extraction.
The ion-accelerating voltage was set at 20 kV. Dihydroxybenzoic acid (DHB) (Sigma) suspended in 0.1 M citric acid was used as a matrix. A few microliters of lipid A were dissolved in a mixture of chloroform:methanol:water (3:1.5:0.25 v/v/v) at 1 mg/mL and desalted with a few grains of ion-exchange resin (Dowex 50W-X8, Sigma, St. Louis, MO, USA) (H+), in an Eppendorf tube. A 1 µL aliquot of the solution (50 µL) was deposited on the target and covered with the same amount of the matrix suspended at 10 mg/mL in the same mixture of solvents [39 (link)]. Different ratios between the samples and DHB were tested when necessary. B. pertussis lipid A was used as an external standard.
The ion-accelerating voltage was set at 20 kV. Dihydroxybenzoic acid (DHB) (Sigma) suspended in 0.1 M citric acid was used as a matrix. A few microliters of lipid A were dissolved in a mixture of chloroform:methanol:water (3:1.5:0.25 v/v/v) at 1 mg/mL and desalted with a few grains of ion-exchange resin (Dowex 50W-X8, Sigma, St. Louis, MO, USA) (H+), in an Eppendorf tube. A 1 µL aliquot of the solution (50 µL) was deposited on the target and covered with the same amount of the matrix suspended at 10 mg/mL in the same mixture of solvents [39 (link)]. Different ratios between the samples and DHB were tested when necessary. B. pertussis lipid A was used as an external standard.
9
Synthesis and Purification of Sulfated Aglycones
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Products were synthesised by the treatment of initial aglycones (500 mg) with sulfur trioxide -pyridine. The aglycones (500 mg) and sulfur trioxide -pyridine (one equivalent for all compounds, with the exception of vanillic acid (VA) for which two equivalents were used) were dissolved in 10 ml of anhydrous pyridine and kept at 658C with constant stirring for 24 h. The reaction was stopped by the addition of water. Solvents were dried in vacuo and the residue was dissolved in water. The unreacted aglycones were separated with ethyl acetate, and the product in the water phase was purified on a Dowex 50W-X8 ion-exchange column (Sigma-Aldrich) loaded with Na þ . The purified compounds were then dried in vacuo and characterised by 1 H and 13
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