The amount of RG7388 and Entinostat within NPs
was assessed using
high performance liquid chromatography (HPLC) and absorbance spectrometry,
respectively. The NP pellet was lysed using a mixture of 1:1 acetonitrile
and dimethyl sulfoxide (DMSO) to release any entrapped drug. RG7388
entrapment was detected by HPLC using a C18 reverse phase column (Phenomenex,
150 × 4.6 mm, 5 μM). The flow rate was set to be constant
at 1 mL/min at 25 °C. 30 μL of 1 mg/mL of sample was injected
per run, and the absorbance was detected at 273 nm and compared to
a series of standards prepared by spiking known amounts of free RG7388
into blank NPs (BNPs) in 1:1 acetonitrile:DMSO. Entinostat entrapment
was quantified by measurement of absorbance at 330 nm using a plate
reader (Biotek) and again compared to a series of standards prepared
by spiking known amounts of free Entinostat into BNPs in 1:1 acetonitrile:DMSO
(Supplementary Figure 1 ). Drug loading
was calculated using the formula below.
was assessed using
high performance liquid chromatography (HPLC) and absorbance spectrometry,
respectively. The NP pellet was lysed using a mixture of 1:1 acetonitrile
and dimethyl sulfoxide (DMSO) to release any entrapped drug. RG7388
entrapment was detected by HPLC using a C18 reverse phase column (Phenomenex,
150 × 4.6 mm, 5 μM). The flow rate was set to be constant
at 1 mL/min at 25 °C. 30 μL of 1 mg/mL of sample was injected
per run, and the absorbance was detected at 273 nm and compared to
a series of standards prepared by spiking known amounts of free RG7388
into blank NPs (BNPs) in 1:1 acetonitrile:DMSO. Entinostat entrapment
was quantified by measurement of absorbance at 330 nm using a plate
reader (Biotek) and again compared to a series of standards prepared
by spiking known amounts of free Entinostat into BNPs in 1:1 acetonitrile:DMSO
(
was calculated using the formula below.
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