Capillary Electrophoretic Determination of Selenium and Tellurium Oxyanions
- Slides: 20
Capillary Electrophoretic Determination of Selenium and Tellurium Oxyanions in Bacterial Cultures by Bala Krishna Pathem and Thomas G. Chasteen Department of Chemistry Sam Houston State University 1
Instrument configuration Source: ceandcec. com 2
Flow reversal n For Anions: – In order to speed up the separation, the EOF can be reversed by addition of cationic surfactants. – A negative voltage is applied. – We have used TTAB and HTAB in our work. 3
Source: ceandcec. com 4
Method development n We developed a method for the identification of selenium species in presence of TSB or LB bacterial growth medium, complex and well-defined media. n We applied the method for simultaneous estimation of selenate and selenite or tellurite in live cultures during their biological reduction by bacteria. 5
Capillary n Fused silica capillaries were used for all the studies done in this research. n Dimensions: 50 cm effective length, 75 µm i. d, 375 µm O. D. 6
Method n Buffer: 15 m. M KH 2 PO 4 + 1 m. M TTAB n p. H - 10. 5 n Injection Pressure: 0. 5 psi Time: 5 sec n Separation Voltage: – 25. 0 KV n Capillary temp: 25 o. C n Run time: 5 min n Detection: 190 nm for selenium oxyanions 220 nm for tellurium oxyanions 7
Bacterial growth conditions n Two bacterial species were used in our research. 1. Precultures of Bacillus sp. were grown in TSB (p. H 7. 0) at 30 o. C and then amended with 1. 0 m. M selenate. 2. Precultures of genetically modified clone of E. coli K 12 (1 VH) were grown in LB medium with Ampicillin (p. H 7. 0) at 37 o. C and then amended with 0. 05 m. M tellurite. 8
Figure 1 9
Bioreduction of selenate by Bacillus sp. Figure 2 10
Figure 3 11
Figure 4 12
Figure 5 13
Bioreduction of 0. 05 m. M tellurite by 1 VH m. AU Te. O 32 - 0 h 5 h 24 h after inoculation Figure 6 14
Figure 7 15
Conclusions n Once bacteria enters stationary phase (4 h): – Rate of selenate reduction decreases – Selenite reduction process begins or increases significantly, 25% drop in Se. O 32 - in 1 h n Varied bacterial response to toxic salts – Selenite is more toxic than selenate for both these organisms, based on specific growth rates 16
Conclusions n Method offers excellent LOD (3 S/N) – Selenate : 1. 0 ppm or 0. 007 m. M – Selenite : 0. 25 ppm or 0. 002 m. M n Linearity – Selenate: 0. 9996 (0. 1 m. M to 1. 0 m. M) – Selenite : 0. 9998 (0. 1 m. M to 1. 0 m. M) 17
Conclusions n This method affords high sample throughput and minimal or no sample preparation for biological samples. n Perfect tool for the Qualitative or Quantitative analysis of selenium and tellurium oxyanions in solution. n Applicable for analysis in environmental or agricultural samples. 18
Acknowledgements This work was supported by Sam Houston State University’s Faculty Enhancement Research Fund The Robert A. Welch Foundation 19
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