1. Simulated Body Fluid (SBF)
SBF is used for many experiments that involve studying bioactivity of biomaterials. Its ion concentration is similar to human plasma and gives a close understanding of how a system will behave biochemically under in vivo conditions. Its composition is given below.
Composition of SBF – 1L, pH 7.4
| Order | Reagent | Amount |
| 1 | NaCl | 7.996 g |
| 2 | NaHCO3 | 0.350 g |
| 3 | KCl | 0.224 g |
| 4 | K2HPO4･3H2O | 0.228 g |
| 5 | MgCl2･6H2O | 0.305 g |
| 6 | 1M-HCl | 40 mL |
| (About 90 % of total amount of HCl to be added) |
| 7 | CaCl2 | 0.278 g |
| 8 | Na2SO4 | 0.071 g |
Protocol:
- Wash all the glassware with 1N HCl solution, neutral detergent, and ion-exchanged- distilled water, dry them.
- Put 500 ml of ion-exchanged- distilled water into a 1-liter bottle (polyethylene bottle) and cover the bottle with a watch glass.
- Stir the water in the bottle with a magnetic stirrer, and dissolve the reagents one by one (after the previous reagent has dissolved completely) in the order as given in the table.
- Adjust the temperature of the solution in the bottle at 36.5℃ using a temperature control water bath or using a thermometer, and adjust pH of the solution at pH 7.4 by stirring the solution and titrating 1N-HCl solution (The water used for washing the electrode should be added back to the solution after removing pH electrode).
- Transfer the solution from the bottle to a volumetric glass flask. Add the water used for washing the polyethylene bottle to the solution in the flask.
- Adjust the total volume of the solution to 1 liter by adding ion-exchanged- distilled water and shaking the flask at 20℃.
Transfer the solution from the flask to a polyethylene or polystyrene bottle, and store the bottle in a refrigerator at 5-10℃ (Don’t use this solution as SBF or the container if any precipitation is seen during the storage).