Doxorubicin hydrochloride

The thin-lipid-film hydration method was used to prepare liposomes from the mixture. 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC, Echelon Biosciences, Salt Lake City, UT, USA) and cholesterol (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) at a weight ratio of 10:1 was used for the neutral liposome. For the cationic liposome, DPPC, 1,2-dipalmitoyl-3-trimethylammonium-propane (chloride salt, 16:0 TAP; Avanti Polar Lipids; Birmingham, AL, USA) and cholesterol were used at a weight ratio of 5:5:1 and for the anionic liposome, DPPC, 1,2-dipalmitoyl-sn-glycero-3-phosphoserine plus sodium salt (Echelon Biosciences) and cholesterol were used at a weight ratio of 5:5:1.
The lipids were dissolved in dichloromethane and the solvent was removed via evaporation under reduced pressure at 55°C. Subsequently, a thin film of lipids was dispersed in distilled water (lipid concentration of 2.2 mg/ml) with doxorubicin hydrochloride (LC Laboratories, Woburn, MA, USA) by sonication. Dialysis was then performed in order to remove the unloaded doxorubicin using distilled water for 1 h. After disrupting the liposomes completely with a detergent solution (1% Triton X-100; Samchun, Pyeongtaek-Si, South Korea), the amount of doxorubicin in the liposomes was evaluated based on the fluorescence of doxorubicin (490/570 nm).

1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-Dipalmitoyl-sn-glycero-3-phosphoserine, sodium salt (DPPS) were purchased from Echelon Biosciences (Salt Lake City, UT, USA). 1,2-dipalmitoyl-3-trimethylammonium-propane [chloride salt (16:0 TAP)] was purchased from Avanti Polar Lipids (Birmingham, AL, USA). Cholesterol was purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Dichloromethane was purchased from Daejung Chemical (Cheongwon, Korea). A dialysis membrane [pre-wetted RC Tubing (MWCO: 25 kD)] was purchased from Spectrum (Spectra/Por; Laguna Hills, CA, USA). Dimethyl sulfoxide, 99.0% (methyl sulfoxide, DMSO) and Triton X-100 were purchased from Samchun Pure Chemical (Pyeongtaek, Korea). Phosphate-buffered saline (PBS, pH 7.4) was purchased from Gibco (Thermo Fisher Scientific, Inc., Waltham, MA, USA). Doxorubicin hydrochloride was purchased from LC laboratories (Woburn, MA, USA).
Methods

S. potatorumL. seeds were purchased
from Sirigiri Venkappa
Ayurveda store, Nandyala, Andhra Pradesh, India. Dimethylaminoethyl
methacrylate (DMAEMA), acrylamide, N,N-methylene bisacrylamide (MBA), and ammonium persulphate (APS) were
purchased from Sigma-Aldrich Chemical Co. 5-Fluorouracil, silver nitrate,
sodium hydroxide, and hydrochloric acid were purchased from SD Fine
Chemicals, Mumbai, India. Doxorubicin hydrochloride (DOX) was received
from LC laboratories. All of the chemicals were used as received,
and double-distilled water (DDW) was used for all experiments.

The reagents used in this work were iron(III) chloride hexahydrate (97%), iron(II) chloride tetrahydrate (97%), ammonium solution (25%), citric acid (>99%), sodium citrate, dopamine hydrochloride (99%), PAMAM dendrimers g.5.0 (5 wt.% in methanol), phosphate buffered saline (PBS; pH = 6.8 or 7.4; 10 mM), sodium tetraphenylborate (NaB; pH = 8.5; 10 mM), Tris aminomethane buffer (Tris; pH = 8.5; 10 mM), doxorubicin hydrochloride (>99%) (Doxo·HCl), O-[N-(3-maleimidopropionyl)aminoethyl]-O′-[3-(N-succinimidyloxy)-3-oxopropyl]heptacosaethylene glycol (>90%). All of the chemical reagents were purchased from Sigma-Aldrich, except for the dopamine hydrochloride (Alfa Aesar) and doxorubicin hydrochloride (LC Laboratories). All reagents and solvents were of reagent-grade quality.