1. BEHAVIOR OF CISPLATIN INVESTIGATED BY SPECTROPHOTOMETRY AND CAPILLARY ELECTROPHORESIS Hoai Viet Nguyen Reg.č.projektu: CZ.1.07/2.3.00/20.0148 Název projektu: Mezinárodní spolupráce v oblasti "in vivo" zobrazovacích technik Název: Školitel: Datum: 22.02.2013

2. Cisplatin Platinum inorganic complex Used in the management of non-small lung cancer, malignant melanoma, endometric cancer and adrenocorticol carcinoma. The therapeutic is drastically narrowed. 2

3. Spectrophotometric analysis Measurements preformed by SPECORD 210 (Analytik Jena, Germany) -Absorbtion spectra was form 200 to 700 nm -Cuvettes : 1cm, Hellma, Essex, UK -BGE : - 0.2 M sodium borate (pH 7, 7.5, 8, 8.5,9) - 0.2 M sodium phosphate (pH 6, 6.5, 7, 7.5, 8) -Sample : CisPt (500, 400, 300, 200, 100 and 50 µM) 3

4. Capillary electrophoresis analysis Measurements performed by 3D Hwelett Pacard with DAD - Detection at 200, 272, 293 and 360 nm -Uncoated fused silica capillary was used with l tot = 64.5cm, l eff = 56cm with ID = 50 µm -Separation condition: voltage 20kV, injection 20 sec by 3.4 kPa -BGE: 0.2 M sodium borate (pH 7, 7.5, 8) -Sample: CisPt 500, 400, 300, 200, 100 and 50 µM 4

5. Fig. 1: Spectra of 500 µM CisPt with borate buffer at different pH (7, 7.5, 8, 8.5, 9) Fig. 2: Absorbance of CisPt with different concentrations (500, 400, 300, 200, 100 and 50 µM) in borate buffer at different pH (7, 7.5, 8, 8.5 and 9) 272nm 293nm 360nm Borate buffer 5

6. Phosphate buffer Fig. 3: Spectra of 500µM CisPt with phosphate buffer at different pH (6, 6.5 7, 7.5 and 8) Fig. 4: Absorbance of CisPt with different concentrations (500, 400, 300, 200, 100 and 50 µM) in borate buffer at different pH (6, 6.5, 7, 7.5 and 8) 272nm 293nm 360nm 6

7. Result and discussion Fig. 5: Spectra of 500 µM CisPt with borate and phosphate buffer at different pH (7, 7.5 and 8) 7

8. µ e = µ a - µ EOF µ e : Effective electrophoretic mobility µ a : Apparent mobility µ EOF : Electroosmotic flow mobility I : Effective capillary length (m) L : Total capillary length (m) t : Migration time (s) V : Electric field (V) Effective electrophoretic mobility 8 Cation +µ Anion - µ

9. Result and discussion Fig. 6: CE-UV (λ=200 nm) of Spectra of 500µM CisPt with borate buffer at different pH (7, 7.5, 8, 8.5 and 9) 9

10. Effective electrophoretic mobility Fig 6. Effect of pH on effective electrophoretic mobility. Conditions: 500µM CisPt with borate buffer at different pH (8, 8.5 and 9) 10

11. Conclusion CE-UV detection is powerful method for cisPt analysis. Absorbance of cisplatin in borate buffer at different pH (9, 8.5, 8, 7.5 and 7) and phosphate buffer at different pH (8, 7.5, 7, 6.5 and 6) was investigated by UV-vis spectrometry. Behavior of cisplatin in borate buffer at pH 7.5 is different from cisplatin in borate buffer at pH 9, 8.5 and 8. Therefore, there are complicated processes happening at pH 7.5. 11

12. Thanks to Ing. Maja Stanisavljević Mgr. Marketa Vaculovicova PhD Mgr. Lukas Nejdl Prof. Ing.Rene Kizek PhD Financial support from following projects PGS16_2012, Liga proti rahovine Praha 2012/2013,NANOLABSYSCZ.1.07/2.3.00/20.0148 and CEITEC CZ.1.05/1.1.00/02.0068. 12

13. Thank you for your attention 13