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In Vitro Release

The in vitro release of PTX-lip-gel and PTX-gel (1.125 mg/mL PTX for each
formulation) was investigated by tube method. Then, 3 parallel drug release experiments
were conducted in accordance with our previous established process. Samples were carefully
distributed into tube (diameter 10 mm) and kept warm in water bath at 37oC for 30 min so
that they were transformed into gel sufficiently. Five milliliter pH 7.4 PBS (0.1% Tween80)
was added into the dissolution tubes, which were placed in a shaking incubator at 37oC and
50 rpm. At predetermined time, 3mL supernatant was collected for HPLC analysis and
replaced by the same amount of fresh PBS. Every group had three parallel samples. The in
vitro release of PTX-lip (1.5 mg/mL PTX for each formulation) was investigated by a
dialysis method. PTX-lip formulation were sealed in a preswollen dialysis bag (molecular
weight cutoff 12,000-14,000 Da) and immersed into 20 mL of sodium salicylate solution (1.0
M) at 37oC, followed by immediate shaking at 40 rpm. Sodium salicylate was used because
of its strong solubilization capacity for PTX. At each time points, the whole volume of
medium was withdrawn and replaced with an equal volume of then medium. All of the
experiments met the sink conditions and were repeated in triplicate. The analysis conditions
were the same as in section Determination of Drug Content in Docetaxel Liposome Gel.
The accumulated release was calculated according to the below equation:

where E is the accumulated release (%), VE is the sampling volume (3 mL), V0 is the initial
volume (5 mL), Ci and Cn are the drug concentrations (𝜇g/mL), i and n are the sampling
times, and m0 is the total mass of drug in the gel (𝜇g).
Pelepasan In Vitro
Pelepasan in vitro gel lip PTX dan lip PTX (1.125 mg/mL PTX untuk setiap
formulasi) diteliti dengan metode tabung. Kemudian, percobaan pelepasan 3 obat paralel
dilakukan sesuai dengan proses yang telah dilakukan sebelumnya. Sampel dimasukkan secara
hati-hati ke dalam tabung (diameter 10 mm) dan diletakkan dalam waterbath agar tetap
hangat pada suhu 37oC selama 30 menit hingga sampel berubah menjadi setengah gel. 5 mL
PBS pH 7,4 ( Tween80 0,1%) ditambahkan ke dalam tabung, yang ditempatkan dalam
inkubator getar pada suhu 37oC dan 50 rpm. Pada waktu yang telah ditentukan, supernatan
3mL dikumpulkan untuk analisis HPLC dan diganti dengan jumlah PBS baru yang sama.
Setiap kelompok memiliki tiga sampel paralel. Pelepasan in vitro lip PTX (1,5 mg/mL PTX
untuk setiap formulasi) diteliti dengan metode dialisis. Formulasi lip PTX disegel dalam
kantong dialisis preswollen (berat molekul antara 12.000-14.000 Da) dan direndam ke dalam
20 mL larutan natrium salisilat (1,0 M) pada suhu 37oC, diikuti dengan pengocokan pada 40
rpm. Sodium salicylate digunakan karena membantu kelarutan untuk PTX. Pada setiap waktu
tertentu, seluruh volume medium diambil dan diganti dengan volume yang sama. Semua
percobaan memenuhi kondisi tenggelam dan diulang dalam rangkap tiga. Kondisi analisis
sama seperti dibagian Determination of Drug Content in Docetaxel Liposome Gel.

di mana E adalah pelepasan terakumulasi (%), VE adalah volume sampling (3 mL), V0


adalah volume awal (5 mL), Ci dan Cn adalah konsentrasi obat (μg / mL), i dan n adalah
waktu sampling , dan m0 adalah total massa obat dalam gel (μg).