1. Albumin Analysis by Potentiometric Method

2. Experimental

3. Materials

4. Monomer Dimethylaminopropyl methacrylamide ( DMAPMA, Rohm GmbH, Chemische Fabrik, Germany, 600-750 ppm MEHQ ) Crosslinking agent Tetraethylene glycol dimethacrylate ( TEGDMA, Fluka, 60 ppm hydroquinone ) Initiator Cumene hydroperoxide ( CHP, Fluka, 80% purity ) Tetramethyl ethylenediamine ( TMEDA, TCI, GR grade )

5. Modification agent 2-Mercapto ethanol ( Acros, 99% purity ) Electrolyte Potassium chloride ( Merck, GR grade ) Washing solvent Methanol ( Lab-Scan, HPLC grade, ) water ( deionized grade )

6. Procedure

7. Cleaning of Al 2 O 3 substrate Acetone ultrasonification for 5 mins Ethanol ultrasonification for 5 mins Deionizied water ultrasonification for 3 mins Drying in oven

8. Preparation of Au electrode Imprinting Au gel on substrate by halftone Annealing Au thick film by programming Etching by 1.00 N NaOH and HCl solutions for 10 mins, respectively, then, washing by H 2 O thoroughly Drying Au electrode by N 2 gas at room temperature

9. Modification of Au electrode Preparation of 1.00 mM thiol ethanol/ water (4/1 v/v ) solution Modification of Au electrode in thiol solution 12 hrs at least under room temperature Washing electrode by deionizied water and ethanol thoroughly Drying modified electrode by N 2 at room temperature

10. Preparation of Au MIP sensing film Spin-coating albumin containing reactive solution on Au electrode Photo-polymerization by 365 nm UV lightening for 36 hrs under N 2 circumstance Washing out albumin molecule by 20 wt % methanol aqueous solution and detected by UV/Vis No albumin was detected in the eluted solution by the UV/Vis

11. Experimental Sequence of Potentiometric Method Preparation of electrode Stabilization of electrode potential in open circuit Determine  V of electrode after albumin addition Linear regression of  V and albumin concn.