Callistemon Citrinus Lab Report

Materials and Methods:
PLGA (50:50 copolymer compositions; MW 30,000 Da), berberine chloride hydrate (MW 371.81 Da), poly vinyl alcohol (MW 89,000 Da) and 15 and 50 mL Corning centrifuge tubes were purchased from Aldrich (St. Louis, MO, USA). Acetone was purchased from Fischer Scientific Laboratory (Fair Lawn, NJ, USA). Phosphate buffer pH 7.0 was purchased from EMD Chemicals Inc. (Gibbstown, NJ, USA). 0.2 micron syringe filters were obtained from Millipore Corporation (Carrigtwohill, Ireland).
1.1. Collection and preparation of crude methanolic plant extract (CMPE) of Callistemon citrinus (CC)
Sample collection: The plant used in current study was Callistemon citrinus and it was collected from various parts of District Chakwal located in Punjab Province of Pakistan. The branches and leaves of Callistemon citrinus was selected for sample preparation.
Extract preparation:
The extracts of Callistemon citrinus was prepared by grinding the sample into fine powder. Methanol was used as solvent for preparation of extracts. 20 grams of sample was dissolved in 200ml of methanol. After proper mixing, they were kept in
Briefly, 10 mg of berberine chloride was dissolved in 1 ml of methanol and 7 mg of PLGA was dissolved in 1 ml of acetone. Later on, berberine chloride solution was added into PLGA solution and stirred at 500 rpm for 45 minutes. Various concentrations of PVA ranging from 0.25 to 1% were prepared by heating on hot plate at 90C and stirring around 500rpm in order to get complete dissolution of PVA. PLGA berberine solution (organic phase) was then added to PVA (aqueous phase) in a drop wise manner. This process resulted in aggregation of PLGA NPs which was prevented by adding 2ml dH2O. The solvents both acetone and methanol were removed in rotary evaporator. PLGA NPs were collected by centrifugation at 14,000 rpm for 15 minutes and lyophilized for