The average hydrodynamic diameter (Dh) and polydispersity index (PI) of LE-MixNano were determined using dynamic light scattering (DLS) analysis (Beckman Coulter® N4 Plus, Beckman Coulter s.r.l, Milan, Italy). Five minutes before the DLS measurements, each sample was adequately diluted with ultrapure water, previously filtered through a 0.45 μm pore size filter (Phenex® RC Syringe filter, Phenomenex, Torrance, CA, USA), choosing the final concentration to reach a measurement intensity ranging from 5 × 104 to 1 × 106 counts per second (cps). Dh and PI for each LE-MixNano preparation were measured at 20 °C with 6 runs of three different samples at an angle of 90° [35 (link)] and a run time of 200 s.
Phenex rc syringe filter
Manufactured by Phenomenex
Sourced in Italy, United States, France
Phenex-RC Syringe Filters are disposable filtration devices designed for sample preparation in analytical laboratories. They feature a regenerated cellulose (RC) membrane that is compatible with a variety of solvents and solutions. These filters are used to remove particulates and clarify samples prior to analysis or further processing.
Automatically generated - may contain errors
Lab products found in correlation
N4 plus, by Beckman Coulter (1 mentions)
Ultimate 3000, by Thermo Fisher Scientific (1 mentions)
Corona veo charged aerosol detector, by Thermo Fisher Scientific (1 mentions)
Propionylcarnitine, by Merck Group (1 mentions)
Acetylcarnitine, by Merck Group (1 mentions)
Butyrylcarnitine, by Merck Group (1 mentions)
Carnitine, by Merck Group (1 mentions)
Uric acid, by Merck Group (1 mentions)
1 methylhistidine, by Merck Group (1 mentions)
15 f2t isop, by Cayman Chemical (1 mentions)
15 e 2t isop, by Cayman Chemical (1 mentions)
Salivette roll shaped polyester swabs, by Sarstedt (1 mentions)
7 protocols using phenex rc syringe filter
1
DLS Analysis of LE-MixNano Particles
2
Quantitative Analysis of CyA Nanomicelles
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The percentage of CyA solubilised by using the different mixtures of surfactants and mucoadhesive polymer was determined by RP-HPLC analysis after dilution of the nanomicellar formulations with organic solvent. An aliquot (1.0 mL) of each ASMP-Nano formulation (Table 1 ) was filtered through 0.2 μm pore size filter (Phenex RC Syringe filter, Phenomenex®, BO, Italy) and added with acetonitrile (ACN, 20.0 mL) to favour the dissolution of CyA into organic solvent. The amount of CyA was detected by HPLC analysis and each analysis was repeated on three different samples.
3
Chemical Stability of Cyclosporine A
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To evaluate the chemical stability of CyA, three different batches of the selected Nano1HAB-CyA formulation were prepared following the method described in the paragraph 2.2.1. After preparation, the samples were filtered in sterile condition (0.2 μm, Phenex RC Syringe filter, Phenomenex®, BO, Italy) and packaged into different crimp vials. Then, they were incubated at 4 °C and 20 °C up to 90 days and the concentration of CyA in the samples was analysed over time by HPLC method after appropriate dilution with acetonitrile. The CyA detected in the samples was reported as percentage of initial amount and the coefficient of variation (CV%) was calculated according to the equation: (SD/mean) * 100, where SD represents standard deviation.
4
Reversed-phase HPLC for Mono-rhamnolipid Quantification
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Reversed-phase chromatography was performed for analyzing mono-rhamnolipid concentrations based on a method developed earlier (Behrens et al., 2016 (link); Tiso et al., 2016 (link)). For sample preparation, the supernatant was mixed 1:1 with acetonitrile and stored at 4°C overnight. Subsequently, the mixture was centrifuged at 16,500 g for 2 min. All samples were filtered with Phenex RC syringe filters (0.2 μm, Ø 4 mm, Phenomenex, Torrance, USA). The HPLC system Ultimate 3000 with a Corona Veo Charged Aerosol Detector (Thermo Fisher Scientific, Waltham, MA, USA) was used. For separation, a NUCLEODUR C18 Gravity 150 × 4.6 mm column (particle size: 3 mm, Macherey-Nagel GmbH & Co. KG, Düren, Germany) was used. The flow rate was set to 1 ml min−1 and the column oven temperature was set to 40°C. Acetonitrile (A) and 0.2% (v/v) formic acid in ultra-pure water (B) were used as running buffers. The method started with a ratio of 70% buffer A: 30% buffer B and a linear gradient was applied to reach a ratio of 80:20% in 8 min. The acetonitrile fraction was increased linearly from 80 to 100% between 9 and 10 min and decreased linearly to 70% between 11 and 12.5 min. The measurement was stopped after 15 min.
5
Targeted Metabolomic Analysis of Carnitines and Related Compounds
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One hundred micro-litres of each thawed 24 h urine sample were diluted with 900 μL of milliQ water/acetonitrile (90/10, v/v). For plasma deproteinization, 200 µL of fasting samples were mixed with 200 µL of cold methanol and stored at −20 °C overnight. The samples were then centrifuged and 100 µL of the supernatant was diluted with 900 μL of milliQ water/acetonitrile (90/10, v/v). All plasma and urine samples were then filtered using 0.2 nm Phenex-RC Syringe Filters with a cellulose membrane (Phenomenex, Le Pecq, France), transferred to a LC vial and either kept at −20 °C or stored at 4 °C if used within 1–2 days until analysis. Blank and quality control (QC) samples were injected every 10 analytical samples in order to control the possible instrument deviation. The blank samples contained acetonitrile and water (10/90, v/v) and QC samples contained the aliquots of all analyzed samples. All solvents used were liquid chromatography mass spectrometry (LC-MS) grade (Sigma-Aldrich; Saint Quentin Fallavier, France) and the aqueous solutions were prepared using purified Milli-Q water. The authentic standards of carnitine, acetyl-carnitine, propionyl-carnitine, butyryl-carnitine, 2-methylbutyroyl-carnitine, uric acid and 1-methylhistidine were purchased from Sigma-Aldrich.
6
Salivary Biomarkers of Oxidative Stress
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Commercially available prostaglandin E 2 (PGE 2 ), F 2 -and E 2 -isoprostanes (IsoPs), 15-F 2t -IsoP and 15-E 2t -IsoP (purity ≥99%) were from Cayman Chemical (Michigan, USA).
Not commercially available F 4 -NeuroPs (20-F 4t -NeuroP, 20-epi-F 4t -NeuroP), F 2 -dihomo-IsoPs (ent-7(R,S)-7-F 2t -dihomo-IsoP, 17-F 2t -dihomo-IsoP) and F 2 -dihomo-IsoF (7(R,S)-ST-Δ 8 -11-dihomo-IsoF) were synthesized at the Institut des Biomolecules Max Mousseron (IBMM) (Montpellier, France) according to procedures reported elsewhere [41] (link)[42] (link)[43] (link)[44] (link).
All the solutions and saliva samples were stored in sterile polypropylene containers from Eppendorf (Milan, Italy).
Phenex™-RC syringe filters (0.2 μm regenerate cellulose, 4 and 15 mm of diameter) were from Phenomenex (California, USA).
The eVol® XCHANGE analytical syringe (20-500 μL) and microextraction by packed sorbent silica-C18 Barrel Insert and Needles (BINs) were purchased from SGE Analytical Science (Melbourne, Australia).
Salivette roll-shaped polyester swabs were purchased from Sarstedt (Nümbrecht, Germany).
A Macherey Nagel Pehanon narrow range (5.2 < pH < 8.1) pH paper strips (Düren, Germany) with a resolution of 0.3 pH units was used to estimate the salivary pH.
An Amicon Ultra-0.5 mL Centrifugal Filters (Darmstadt, Germany) with a cut-off of 3 kDa was used to purify saliva samples.
Not commercially available F 4 -NeuroPs (20-F 4t -NeuroP, 20-epi-F 4t -NeuroP), F 2 -dihomo-IsoPs (ent-7(R,S)-7-F 2t -dihomo-IsoP, 17-F 2t -dihomo-IsoP) and F 2 -dihomo-IsoF (7(R,S)-ST-Δ 8 -11-dihomo-IsoF) were synthesized at the Institut des Biomolecules Max Mousseron (IBMM) (Montpellier, France) according to procedures reported elsewhere [41] (link)[42] (link)[43] (link)[44] (link).
All the solutions and saliva samples were stored in sterile polypropylene containers from Eppendorf (Milan, Italy).
Phenex™-RC syringe filters (0.2 μm regenerate cellulose, 4 and 15 mm of diameter) were from Phenomenex (California, USA).
The eVol® XCHANGE analytical syringe (20-500 μL) and microextraction by packed sorbent silica-C18 Barrel Insert and Needles (BINs) were purchased from SGE Analytical Science (Melbourne, Australia).
Salivette roll-shaped polyester swabs were purchased from Sarstedt (Nümbrecht, Germany).
A Macherey Nagel Pehanon narrow range (5.2 < pH < 8.1) pH paper strips (Düren, Germany) with a resolution of 0.3 pH units was used to estimate the salivary pH.
An Amicon Ultra-0.5 mL Centrifugal Filters (Darmstadt, Germany) with a cut-off of 3 kDa was used to purify saliva samples.
7
Urine Sample Preparation for LC-MS
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One hundred micro-litres of each thawed 24h-urine sample were diluted with 900 μl of milliQ water/acetonitrile (90/10, v/v), filtered using 0.2 nm Phenex-RC Syringe Filters with a cellulose membrane (Phenomenex, Le Pecq, France), transferred to a sealed glass autosampler vial and either kept at -20°C or stored at 4°C if used within one to two days for LC-MS analysis. Quality control (QC) samples were prepared by combining small aliquots of all samples. Analytical samples were analysed in random order, interspersed after every 10 injections with QC sample and blank (acetonitrile/water 10/90, v/v) injections to monitor instrument performance and sample stability.
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