Quantification of Quantum Dots in Solution using Surface Plasmon
Resonance and Analytical Ultracentrifugation
Chinonso Opara,
Hyewon Kwon,
Xiaoge Hu,
William Atkins, and
John Sumida
University of Washington, Department of Medicinal Chemistry, Seattle, WA
University of Washington, Department of Bioengineering, Seattle, WA Introduction Results Conclusions Acknowledgements We would like to acknowledge funding from the Amgen Foundation as part of the University of Washington 2012 Amgen Scholars Program (CO) and also from the NIH National Institutes of General Medical Sciences' Grant #12 R25 GM 058501-05A1 "UW IMSD Program". This work was performed in the Analytical Biopharmacy Core, funded through the Center for the Intracellular Delivery of Biologics by the Washington State Life Sciences Discovery Fund. Transforming SPR Response Units to Refractive Index via Correction Factor Our method shows promise as a new way to measure the concentration of quantum dots in solution. We determined a dn/dc and mass extinction coefficients for all wavelengths at which absorbance is observed. Mass extinction coefficients allow for simple UV-vis concentration measurements. AUC Synthetic Boundary J (r) Scales with Concentration Mass Extinction Coefficient from Concentration and Absorbance Knowing dn/dc, we use J(r) to measure concentration. Figure 4. An AUC synthetic boundary measurement of the total fringe increment, J(r), of a given solution of CdSe TOPO-PMAT quantum dots. CdSe TOPO-PMAT Quantum Dot dn/dc Figure 6. The concentration of a quantum dot solution makes it possible to take an absorbance spectrum and determine the mass extinction coefficient for a continuous set of wavelengths. This allows the use of UV-vis to measure concentration. Figure 3. We are able to determine the refractive index increment by using the density increment, measured by AUC, to relate density to concentration. Quantum dots are nanoparticles with several applications, including medical imaging and drug delivery. However, there are many limitations to determining their concentration. It is our goal to create a new method for measuring the concentration of quantum dots in solution. We use surface plasmon resonance (SPR) to determine the refractive index increment, dn/dc, of CdSe TOPO-PMAT quantum dots. The dn/dc allows us to relate the total fringe increment, J(r), measured by analytical ultracentrifugation (AUC) to the concentration of these quantum dots. Figure 1. A conversion factor relating SPR response units to refractive index was determined by comparing the concentration dependence of NaCl response units to the NaCl literature refractive index increment. *CRC Handbook of Chemistry and Physics, 92 nd Edition, 2010-2011. Figure 2. The correction factor was verified by using it to measure the dn/dc of bovine serum albumin (BSA). Our experimental value of 0.186 agrees with the **literature value. Figure 5. Total fringe Increment scales with concentration **Vollmer et al. Protein detection by optical shift of a resonant microcavity. Applied Physics Letters. Vol 28, Num 21 (2002).
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