21. 70% cold ethanol: Mix 70 mL of absolute ethanol and 30 mL
of distilled water, pre-cool at À20
C.
22. Lewis lung cancer cells.
2.4 Anticancer
Efficiency In Vivo
1. 1 mL syringe.
2. Electronic scale (ten thousandth balance).
3. Surgical
Instruments
(surgical
scissors,
tweezers
ophthalmology).
4. C57BL/6J mice (20 Æ 2 g).
3 Methods
3.1 Preparation
Process of CC@CHLNV
1. Operate all steps in darkness (see Note 6). Weigh 10.00 mg CC,
70.24 mg glycerin monostearate, 54.07 mg isopropyl palmitate, 117.35 mg Lipoid S 75 (see Note 8). Transfer them to the
50 mL round-bottomed flask.
2. Add 20 mL of absolute ethanol to the same round-bottomed
flask and sonicate for 2 min to obtain a yellowish solution.
3. Evaporate solution at 30
C under hypobaric conditions for
40 min to remove ethanol and obtain a yellow film (see Note
9).
4. Take out the yellow powder and store them in 4
C.
5. Add 241.66 mg of the yellow powder to 10 mL of distilled
water containing CTAB and SDS (56.26 μmol:112.52 μmol)
and sonicate for 10 min to obtain CC@CHLNV.
3.2 Pharmacokinetic
Experiment
1. Randomly divide rats into four groups, fast for 12 h before
administration (free drinking water), and orally administer with
0.9% NaCl (negative group), blank CC@CHLNV (Blank control group), 0.9 mg/mL CC suspension (treatment group 1),
and CC@CHLNV (15 mg/kg, treatment group 2), respectively. Using a 5 mL syringe with 16-gauge straight needle,
intravenously inject with 0.16 mg/mL CC suspension and
CC@CHLNV (2.5 mg/kg) for the latter two group,
respectively.
2. Analyze plasma CC concentration by HPLC. Under ether
anesthesia, collect ophthalmic blood samples at 0, 5, 10, 15,
30, 45, 60, 120, 180 min, centrifuge at 6000 rpm for 10 min.
Obtain plasma samples from the upper layers of supernatants,
fix with 100 μL nitrendipine (5 μg/mL) and 1 mL of ethyl
acetate. Vortex the mixture for 3 min and then centrifuge at
12,000 rpm for 10 min. Remove the organic layer and evaporate under nitrogen flow. Reconstitute the residue with 100 μL
of the acetonitrile and 5% glacial acetic acid.
Catanionic Hybrid Lipid Nanovesicles
61
of distilled water, pre-cool at À20
C.
22. Lewis lung cancer cells.
2.4 Anticancer
Efficiency In Vivo
1. 1 mL syringe.
2. Electronic scale (ten thousandth balance).
3. Surgical
Instruments
(surgical
scissors,
tweezers
ophthalmology).
4. C57BL/6J mice (20 Æ 2 g).
3 Methods
3.1 Preparation
Process of CC@CHLNV
1. Operate all steps in darkness (see Note 6). Weigh 10.00 mg CC,
70.24 mg glycerin monostearate, 54.07 mg isopropyl palmitate, 117.35 mg Lipoid S 75 (see Note 8). Transfer them to the
50 mL round-bottomed flask.
2. Add 20 mL of absolute ethanol to the same round-bottomed
flask and sonicate for 2 min to obtain a yellowish solution.
3. Evaporate solution at 30
C under hypobaric conditions for
40 min to remove ethanol and obtain a yellow film (see Note
9).
4. Take out the yellow powder and store them in 4
C.
5. Add 241.66 mg of the yellow powder to 10 mL of distilled
water containing CTAB and SDS (56.26 μmol:112.52 μmol)
and sonicate for 10 min to obtain CC@CHLNV.
3.2 Pharmacokinetic
Experiment
1. Randomly divide rats into four groups, fast for 12 h before
administration (free drinking water), and orally administer with
0.9% NaCl (negative group), blank CC@CHLNV (Blank control group), 0.9 mg/mL CC suspension (treatment group 1),
and CC@CHLNV (15 mg/kg, treatment group 2), respectively. Using a 5 mL syringe with 16-gauge straight needle,
intravenously inject with 0.16 mg/mL CC suspension and
CC@CHLNV (2.5 mg/kg) for the latter two group,
respectively.
2. Analyze plasma CC concentration by HPLC. Under ether
anesthesia, collect ophthalmic blood samples at 0, 5, 10, 15,
30, 45, 60, 120, 180 min, centrifuge at 6000 rpm for 10 min.
Obtain plasma samples from the upper layers of supernatants,
fix with 100 μL nitrendipine (5 μg/mL) and 1 mL of ethyl
acetate. Vortex the mixture for 3 min and then centrifuge at
12,000 rpm for 10 min. Remove the organic layer and evaporate under nitrogen flow. Reconstitute the residue with 100 μL
of the acetonitrile and 5% glacial acetic acid.
Catanionic Hybrid Lipid Nanovesicles
61