A Shimadzu RF-5301 PC spectrofluorophotometer (Tokyo, Japan) was used for fluorimetric measurements. The slit width of both excitation and emission monochromator was set at 5 nm. The pH values of solutions were measured using Hanna pH meter (Hanna Instruments Brazil, São Paulo, Brazil) with a combined electrode. The solutions were sonicated using ultrasonic cleaner, Branson Transonics Corporation, Eagle Road, Danbury, USA. Surface morphology studies were carried out using scanning electron microscope (SEM), JEOL JSM-5400 LV instrument (Oxford, USA), A Nicolet 6700 FTIR Advanced Gold Spectrometer, supported with OMNIC 8 software (Thermo Electron Scientific instruments Corp., Madison, WI USA) was used to record FTIR spectra and Elemental analysis was done using OXFORD INA Energy Dispersive X-ray Spectrometer (EDX). Raman spectra were recorded with a Bruker Senterra Raman microscope (Bruker Optics Inc., Germany) with 785 nm excitation, 1200 rulings mm−1 holographic grating, and a charge-coupled device detector. The average particle size of the prepared nanoparticles were analyzed by photon correlation spectroscopy using a Zeta sizer 3000 HAS (Malvern, Instruments GmbH, Germany).
Zetasizer 3000 ha
Manufactured by Malvern Panalytical
Sourced in United Kingdom
The Zetasizer 3000 HAS is a dynamic light scattering instrument used for the measurement of particle size and zeta potential in liquid samples. It is capable of measuring particle sizes in the range of 0.6 nanometers to 6 micrometers.
Automatically generated - may contain errors
Lab products found in correlation
Hanna ph meter, by Hanna Instruments (1 mentions)
Jsm 5400 lv instrument, by JEOL (1 mentions)
Rf 5301pc spectrofluorophotometer, by Shimadzu (1 mentions)
Senterra raman microscope, by Bruker (1 mentions)
Hpps 5001, by Malvern Panalytical (1 mentions)
Nano zs zetasizer, by Malvern Panalytical (1 mentions)
Avance 400 mhz nmr spectrometer, by Bruker (1 mentions)
Agilent 1200 series, by Agilent Technologies (1 mentions)
Scanning electron microscopy, by Zeiss (1 mentions)
Transmission electron microscopy, by JEOL (1 mentions)
Maldi tof mass spectrometer, by Shimadzu (1 mentions)
Ellman s reagent, by Merck Group (1 mentions)
100cx transmission electron microscope, by JEOL (1 mentions)
Iris intrepid 2 xsp, by Thermo Fisher Scientific (1 mentions)
Jem 3200fs, by JEOL (1 mentions)
Zetasizer nano s90, by Malvern Panalytical (1 mentions)
Cary 5000, by Agilent Technologies (1 mentions)
H 7650, by Hitachi (1 mentions)
Dnase rnase free water, by Thermo Fisher Scientific (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
14 protocols using zetasizer 3000 ha
1
Comprehensive Characterization of Nanoparticles
2
Chitosan-TPP Nanoparticle Characterization
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The z-average particle size, distribution and poly dispersity index (PDI) of the chitosan-TPP nano-particles were measured at 25°C by dynamic light scattering (DLS) on a high performance particle sizer (HPPS-5001, Malvern, UK). The zeta potential of nanoparticles was measured by Zetasizer 3000 HAS (Malvern, UK). Mean values were obtained from the analysis of three different batches, each measured three times. Encapsulation efficiency (EE) was calculated.
3
Comprehensive Characterization of Nanoformulations
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Brüker Avance 400 MHz NMR spectrometer (Germany) was used for NMR analysis. For NF characterization, a microscope with camera Leica, model: DM1000 (US listed microscope) with (LEICA EC3 digital camera), a UV spectrophotometer (Jasco, Japan), a Nano Zeta Sizer (ZS) (Malvern Instruments, Malvern, UK), a particle sizer (Zeta sizer 3000 HAS; Malvern Instruments Ltd, Worcestershire, UK), a rotation viscometer (Brookfield-DVBT) and a DSC 131 evo (SETARAM Inc., France) were utilized. For biological study, a confocal laser scanning microscope (CLSM) (model SP2; Leica, Mannheim, Germany) equipped with an environmental chamber was used.
4
Characterization of Drug-Loaded Nanoparticles
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Mean size and surface zeta potential of prepared nanoparticles were determined using dynamic light scattering (DLS) analyzer (Zetasizer 3000 HAS; Malvern Instruments, Malvern, UK). Each measurement of samples diluted appropriately was made in triplicate at 25°C.
After removal of unentrapped drug through ultrafiltration method using Amicon Ultra-4 centrifugal filter tubes (10 kDa cutoff; Millipore), the entrapment efficiency (EE) and drug-loading (DL) of various ST-loaded nanoparticles were calculated using the following equations, where W0 and W are the total amount of ST in each nanoparticle suspension and ultrafiltrate (free ST), respectively, and Q0 is the total amount of the feeding materials:
The amount of ST was quantified by high-performance liquid chromatography (HPLC; Agilent 1200 series; Agilent Technologies, Palo Alto, CA, USA) equipped with a ultraviolet detector at 238 nm using a shim-pack VP-ODS column (150×4.6 mm, 5 µm) at 30°C. The mobile phase consisted of acetonitrile and water (80:20, v/v) with a flow rate of 1.0 mL/min.
After removal of unentrapped drug through ultrafiltration method using Amicon Ultra-4 centrifugal filter tubes (10 kDa cutoff; Millipore), the entrapment efficiency (EE) and drug-loading (DL) of various ST-loaded nanoparticles were calculated using the following equations, where W0 and W are the total amount of ST in each nanoparticle suspension and ultrafiltrate (free ST), respectively, and Q0 is the total amount of the feeding materials:
The amount of ST was quantified by high-performance liquid chromatography (HPLC; Agilent 1200 series; Agilent Technologies, Palo Alto, CA, USA) equipped with a ultraviolet detector at 238 nm using a shim-pack VP-ODS column (150×4.6 mm, 5 µm) at 30°C. The mobile phase consisted of acetonitrile and water (80:20, v/v) with a flow rate of 1.0 mL/min.
5
Nanoparticle Characterization and Stability
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Characterization of the nanoparticles was carried out with scanning electron microscopy (Zeiss, Germany) and transmission electron microscopy (JEOL, Japan). The three-dimensional structure was measured by atomic force microscopy (ATM, CU, China). The size, polydispersity index (PDI) and zeta potential were determined by a Nano ZS (Zetasizer 3000 HAS, Malvern Instrument, England). A MALDI-TOF mass spectrometer (Shimadzu, Japan) was used to evaluate the stability of the proteins in the nanoparticles. The entrapment efficiency was calculated according to the following formulas: Entrapment efficiency (%) = (Amounttotal Vo − Amountloading Vo)/Amounttotal Vo × 100.
6
Characterization of PEGylated Gold Nanorods
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The characterization of AuNRs was carried out using a Cary 500 UV–Vis Spectrometer (Agilent Technologies, Santa Clara, CA, USA) for the spectroscopic measurements, and a JEOL 100 CX transmission electron microscope (TEM) (JEOL Ltd., Tokyo, Japan) was used to image the samples. Zeta potentials were measured using a ZetaSizer 3000 HAS (Malvern Instruments, Worcestershire, UK). Zeta potential was tested to characterize the surface conjugation with PEG. Ellman’s reagents (react with free-SH group; Sigma-Aldrich) were used to quantify the number of PEG molecules bound to the surface of the AuNRs.29 (link) Ellman’s reagent reacts with free-SH group and can be measured calorimetrically at 412 nm. By subtracting the absorbance of the residual PEG, in the supernatant solution after PEG conjugation and centrifugation, from the original absorbance of the solution before adding to AuNRs, we can calculate the number of PEG molecules on the surface of AuNRs by Beer’s law. The results of characterization are described below (see “Formulation and characterization of the AuNRs” section).
7
Liquid Exfoliation of Black Phosphorus
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The BPs were prepared by a modified liquid exfoliation technique established by our group. Briefly, 25 mg of the BP crystal powder were added to 25 mL of NMP and sonicated with a sonic probe in an ice bath for 10 h at 1080 W (2 s on and 5 s off), followed by bath sonication for another 10 h at 300 W. The dispersion was centrifuged at 9000 rpm for 10 min to remove oversized BP sheets and the supernatant containing BPs was centrifuged at 15000 rpm for 10 min. The precipitate was collected and re-suspended in NMP for storage. The BPs were rinsed with ethanol and water prior to experiments.
The TEM images were obtained from JEM-3200FS (JEOL, Japan) at an acceleration voltage of 200 kV. The size distribution and zeta potential of the BPs were determined by DLS using the Zetasizer 3000 HAS (Malvern Instruments Ltd., UK). XPS was carried out on the Thermo Fisher ESCALAB 250Xi XPS and Raman scattering was performed on the Jobin-Yvon LabRam HR-VIS high-resolution confocal Raman microscope (Horiba, Japan) with the 633 nm laser as the excitation source. The concentration of BPs was determined by inductively-coupled plasma atomic emission spectroscopy (IRIS Intrepid II XSP, Thermo Electron Corporation).
The TEM images were obtained from JEM-3200FS (JEOL, Japan) at an acceleration voltage of 200 kV. The size distribution and zeta potential of the BPs were determined by DLS using the Zetasizer 3000 HAS (Malvern Instruments Ltd., UK). XPS was carried out on the Thermo Fisher ESCALAB 250Xi XPS and Raman scattering was performed on the Jobin-Yvon LabRam HR-VIS high-resolution confocal Raman microscope (Horiba, Japan) with the 633 nm laser as the excitation source. The concentration of BPs was determined by inductively-coupled plasma atomic emission spectroscopy (IRIS Intrepid II XSP, Thermo Electron Corporation).
8
Characterization of SMEDDS Containing ATR and EZT
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The mean droplet sizes and polydispersity index of the microemulsions were analyzed using dynamic light scattering (DLS) at 25°C with a scattering angle of 90° (Zetasizer nano S90, Malvern Instruments Ltd., Malvern, U.K.). In addition, zeta potential of the same sample was analyzed using laser Doppler electrophoresis (Zetasizer 3000 HAS, Malvern Instruments Ltd.) at 25°C.
Preparation of SMEDDS Containing ATR and EZT SMEDDS was prepared by mixing specific ratios of drugs, oil, surfactant, and cosurfactant. First, surfactant, and cosurfactant were accurately weighed in a glass vial and mixed by a magnetic stirrer. Then, the oil was added into the vial and continuously mixed for 5 min. ATR and EZT were slowly added to the resultant mixture followed by continuous stirring until a homogeneous, transparent mixture was obtained. The mixture was then stored in a glass vial in ambient temperature.
UV Absorption Spectra Analysis Appropriate amount of excipients were added to separate vials containing ATR or EZT dissolved in methanol. The UV absorption spectra of each solution from 190 nm to 300 nm were measured using a UV-Vis-NIR spectrophotometer (Cary 5000, Agilent Technologies, Santa Clara, CA, U.S.A.).
Preparation of SMEDDS Containing ATR and EZT SMEDDS was prepared by mixing specific ratios of drugs, oil, surfactant, and cosurfactant. First, surfactant, and cosurfactant were accurately weighed in a glass vial and mixed by a magnetic stirrer. Then, the oil was added into the vial and continuously mixed for 5 min. ATR and EZT were slowly added to the resultant mixture followed by continuous stirring until a homogeneous, transparent mixture was obtained. The mixture was then stored in a glass vial in ambient temperature.
UV Absorption Spectra Analysis Appropriate amount of excipients were added to separate vials containing ATR or EZT dissolved in methanol. The UV absorption spectra of each solution from 190 nm to 300 nm were measured using a UV-Vis-NIR spectrophotometer (Cary 5000, Agilent Technologies, Santa Clara, CA, U.S.A.).
9
Kudingcha Nanoparticles Preparation and Characterization
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Kudingcha nanoparticles were prepared using previously reported methods [17] . In summary, chitosan was dissolved in 1% acetic acid, forming a solution at the concentration of 2.0 mg/mL, and tripolyphosphate (TPP) was dissolved in distilled water at 1.0 mg/mL. A specific volume of chitosan solution was obtained and slowly added into the total flavonoids of the Kudingcha solution at room temperature with magnetic stirring (600 rpm). This mixed solution was slowly placed into the TPP solution using a 4th syringe needle for 45 min. Finally, a nanosuspension was spontaneously obtained when blue opalescence appeared [18] . Spray-drying was performed for the nanosuspension with a Lab Spray Dryer L-117 (Laiheng Scientific Co. Ltd, Beijing, China). The nozzle was 0.5 mm, and the other adjustable parameters, including the inlet and outlet temperatures, solution pump flow rate, and aspirator partial vacuum, were similar to those in our previous studies [19, 20] .
The particle size and size distributions of the nanoparticles were determined with a particle sizer (Zetasizer 3000 HAS, Malvern Instruments Ltd., Worcs, UK). The morphology of the nanoparticles was examined using transmission electron microscopy (H-7650, Japan Hitachi).
The particle size and size distributions of the nanoparticles were determined with a particle sizer (Zetasizer 3000 HAS, Malvern Instruments Ltd., Worcs, UK). The morphology of the nanoparticles was examined using transmission electron microscopy (H-7650, Japan Hitachi).
10
Zeta Potential Analysis of Exosomes and Liposomes
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ζ-Potential measurements were performed with a Zetasizer 3000-HA (Malvern Instruments, UK). Exosomes isolated via the ExoSpin kit (see above) were 1:100 in PBS + 0.05% Tween-20 for measurements. Liposomes isolated via the ExoSpin kit (see above) were diluted 1:1000 in PBS + 0.05% Tween-20 for measurements. 5 mL of each sample was applied to a flow cell, and 50 ζ-potential measurements taken. Standard settings were used (viscosity = 0.89, dielectric constant = 80, temperature = 25°C).
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