The average particle diameter, particle size distribution, and surface charge were determined by dynamic light scattering (DLS) technique using a Zetasizer Nano ZS (Malvern, UK).23 (link) The morphology and size of the particles were observed using JEM-1010 transmission electron microscopy (JEOL, Tokyo, Japan). The encapsulation capacity of LPNs was determined by measuring the concentrations of CBP and PTX using a C-18 reverse-phase high-performance liquid chromatography (RP-HPLC) (Beckman Coulter, Brea, CA) at a flow rate of 1 mL/min.24 (link) Tert-butyl methyl ether (5 mL) was added to the LPNs suspension and vortex for 1 min to extract the drugs and then redissolved in acetonitrile:water (10:90) solution. The solution (1 mL) was injected into the C-18 column, and CPT and PTX were detected at 227 nm after different retention times. The amounts of CBP and PTX were quantified by HPLC. The encapsulation efficiency (EE) was determined as: (Amount of entrapped drug/amount of total drug) × 100%.
C 18 reverse phase high performance liquid chromatography rp hplc
Manufactured by Beckman Coulter
The C-18 reverse-phase high-performance liquid chromatography (RP-HPLC) is a laboratory instrument used for the separation and analysis of a wide range of chemical compounds. It utilizes a stationary phase consisting of chemically modified silica particles with C-18 alkyl chains, and a mobile phase of solvents, to separate and detect analytes based on their interactions with the stationary phase.
Automatically generated - may contain errors
2 protocols using c 18 reverse phase high performance liquid chromatography rp hplc
1
Characterizing Lipoprotein Nanoparticle Formulations
2
Quantifying Drug Loading and Release in Liposomes
Check if the same lab product or an alternative is used in the 5 most similar protocols
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To study the
loading capacity of Dox, cMLV (Dox) and cMLV (Dox+PTX)
were collected and washed twice with PBS, followed by lipid extraction
of vesicles with 1% Triton X-100 treatment. Dox fluorescence (excitation
480 nm, emission 590 nm) was then measured by a Shimadzu RF-5301PC
spectrofluorometer (Japan. The amount of paclitaxel incorporated in
the cMLV(PTX) and cMLV(Dox+PTX) was determined by C-18 reverse-phase
high-performance liquid chromatography (RP-HPLC) (Beckman Coulter,
Brea, CA). The cMLV(PTX) and cMLV(Dox+PTX) suspensions were diluted
by adding water and acetonitrile to a total volume of 0.5 mL. Extraction
of paclitaxel was accomplished by adding 5 mL of tert-butyl methyl ether and votex-mixing the sample for 1 min. The mixtures
were centrifuged, and the organic layer was transferred into a glass
tube and evaporated to dryness under argon. Buffer A (95% water, 5%
acetonitrile) was used to rehydrate the glass tube. To test PTX concentration,
1 mL of the solution was injected into a C18 column, and the paclitaxel
was detected at 227 nm (flow rate 1 mL/min). To obtain the release
kinetics of Dox and PTX from liposomes, the releasing media was removed
from cMLVs incubated in 10% FBS-containing media at 37 °C and
replaced with fresh media daily. The removed media was quantified
for Dox fluorescence (by spectrofluorometer) and PTX fluorescence
(by HPLC) every day.
loading capacity of Dox, cMLV (Dox) and cMLV (Dox+PTX)
were collected and washed twice with PBS, followed by lipid extraction
of vesicles with 1% Triton X-100 treatment. Dox fluorescence (excitation
480 nm, emission 590 nm) was then measured by a Shimadzu RF-5301PC
spectrofluorometer (Japan. The amount of paclitaxel incorporated in
the cMLV(PTX) and cMLV(Dox+PTX) was determined by C-18 reverse-phase
high-performance liquid chromatography (RP-HPLC) (Beckman Coulter,
Brea, CA). The cMLV(PTX) and cMLV(Dox+PTX) suspensions were diluted
by adding water and acetonitrile to a total volume of 0.5 mL. Extraction
of paclitaxel was accomplished by adding 5 mL of tert-butyl methyl ether and votex-mixing the sample for 1 min. The mixtures
were centrifuged, and the organic layer was transferred into a glass
tube and evaporated to dryness under argon. Buffer A (95% water, 5%
acetonitrile) was used to rehydrate the glass tube. To test PTX concentration,
1 mL of the solution was injected into a C18 column, and the paclitaxel
was detected at 227 nm (flow rate 1 mL/min). To obtain the release
kinetics of Dox and PTX from liposomes, the releasing media was removed
from cMLVs incubated in 10% FBS-containing media at 37 °C and
replaced with fresh media daily. The removed media was quantified
for Dox fluorescence (by spectrofluorometer) and PTX fluorescence
(by HPLC) every day.
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