141
8. Triethylamine (Et 3 N >99%) (see Note 4).
9. Purification of tetrahydrofuran (THF; dried, max 0.005%
water) was performed using standard high-vacuum techniques
and dried over sodium potassium alloy.
10. Ethyl acetate (>99.5%) was fractionally distilled over phosphorous pentoxide.
11. Hexane (>99%, Merck) was fractionally distilled over sodium
and subsequently distilled over n-BuLi.
12. Solid Dox (98%) and Pacl (98%) were used as purchased (see
Note 5).
3 Methods
1. Use size-exclusion chromatography (SEC) to determine the M n
and M w /M n values. Use two SEC systems, one in order to
obtain the molecular weights of the protected polymers and the
other for the determination of the amount of Pacl contained in
the vesicles. The first system is composed of a 600 high-pressure
liquid chromatographic pump, columns for high temperatures,
a differential refractometer detector, and a two-angle (15°, 19°)
light scattering detector at 60 °C. A 0.1 N LiBr solution in
DMF is used as an eluent at a rate of 1 mL
.
min
−1
. The second
system is composed of a 600 high-pressure liquid chromatographic pump, columns, and a differential refractometer detector at 25 °C. The system runs using a mixture of CHCl 3 /Et 3 N
as a mobile phase in a ratio of 95/5 (v/v).
2. Achieve the
1
H NMR spectra of the polymers and NCAs in
CDCl 3 at room temperature, and that of the triblock polymers
in dimethyl sulfoxide (DMSO).
3. Carry out Fourier transform infrared measurements (FT-IR) in
KBr pellets at room temperature, in the range of 450–4000 cm
−1
.
Use less than 1 mg of samples for each spectrum.
4. Perform UV spectroscopy from 190 to 500 nm, at room temperature with cells requiring 120 mL.
5. Conduct dynamic light scattering measurements with a system
composed of a goniometer with a stepper motor controller, a
variable power Ar
+
laser operating at 690 nm and with 10 mW
power, a temperature control unit, and a pump/filtering unit.
Analyze correlation functions by the cumulant method and the
Contin software. The correlation function is collected at 90°.
Perform all measurements in isotonic Tris buffer (0.010 M) at
pH = 7.4. Measure in the concentration range between 5 × 10
−5
and 1 × 10
−6
g
.
mL
−1
.
3.1 Instrumentation
Nanostructured Polymersomes for Controlled Drug Delivery
8. Triethylamine (Et 3 N >99%) (see Note 4).
9. Purification of tetrahydrofuran (THF; dried, max 0.005%
water) was performed using standard high-vacuum techniques
and dried over sodium potassium alloy.
10. Ethyl acetate (>99.5%) was fractionally distilled over phosphorous pentoxide.
11. Hexane (>99%, Merck) was fractionally distilled over sodium
and subsequently distilled over n-BuLi.
12. Solid Dox (98%) and Pacl (98%) were used as purchased (see
Note 5).
3 Methods
1. Use size-exclusion chromatography (SEC) to determine the M n
and M w /M n values. Use two SEC systems, one in order to
obtain the molecular weights of the protected polymers and the
other for the determination of the amount of Pacl contained in
the vesicles. The first system is composed of a 600 high-pressure
liquid chromatographic pump, columns for high temperatures,
a differential refractometer detector, and a two-angle (15°, 19°)
light scattering detector at 60 °C. A 0.1 N LiBr solution in
DMF is used as an eluent at a rate of 1 mL
.
min
−1
. The second
system is composed of a 600 high-pressure liquid chromatographic pump, columns, and a differential refractometer detector at 25 °C. The system runs using a mixture of CHCl 3 /Et 3 N
as a mobile phase in a ratio of 95/5 (v/v).
2. Achieve the
1
H NMR spectra of the polymers and NCAs in
CDCl 3 at room temperature, and that of the triblock polymers
in dimethyl sulfoxide (DMSO).
3. Carry out Fourier transform infrared measurements (FT-IR) in
KBr pellets at room temperature, in the range of 450–4000 cm
−1
.
Use less than 1 mg of samples for each spectrum.
4. Perform UV spectroscopy from 190 to 500 nm, at room temperature with cells requiring 120 mL.
5. Conduct dynamic light scattering measurements with a system
composed of a goniometer with a stepper motor controller, a
variable power Ar
+
laser operating at 690 nm and with 10 mW
power, a temperature control unit, and a pump/filtering unit.
Analyze correlation functions by the cumulant method and the
Contin software. The correlation function is collected at 90°.
Perform all measurements in isotonic Tris buffer (0.010 M) at
pH = 7.4. Measure in the concentration range between 5 × 10
−5
and 1 × 10
−6
g
.
mL
−1
.
3.1 Instrumentation
Nanostructured Polymersomes for Controlled Drug Delivery