User:Moira M. Esson/Notebook/CHEM-571/2013/11/06

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Objectives Continue biomineralization. Prepare protein-AuNPs for Hemoglobin and Lysozyme. Analyze protein-AuNP solutions prepared on 10/30/2013 using UV/vis spectroscopy. UV/vis Analysis General Protocol: Place 1mL of each ratio in an 1.5mL eppendorf tube. Centrifuge at maximum speed for 30 minutes. Take a UV-Vis spectrum of each sample from 200nm to 800nm. After taking the spectrum, store the sample in a fresh eppy tube in an upright position. Figure 1. Corrected Absorbance Spectra of Lysozyme-AuNP solutions Only the 30:1 ratio exhibited an absorbance peak at 526nm. This was expected as this was the only colored solution or only solution that did not form fibers shown on 11/05/2013 Figure 2. Corrected Absorbance Spectra of Myoglobin Au-NP solutions 15:1 ratio is the only exhibited an aborbance peak at 732nm. This may indicate the optimal concentration for the preparation of fibers. Although this solution produced fibers 11/05/2013 this solution is still easily monitored using UV/vis spectroscopy. Protein-AuNP solution preparation The general protocol for the synthesis of Hemoglobin-AuNP and Lysozyme-AuNP follows the general protocol for the preparation of protein wrapped Au-NPs described on 08/28/2013. 5mL of each solution was prepared. All solutions were prepared using deionized H2O Tri-hydrate gold was used for the preparation of the gold standard solution. All stock solutions were prepared in a 10mL volumetric flask. Table 1. Preparation of Hemoglobin Stock solution Molecular weight (g/mol) 64500 mass myoglobin (g) 0.0106 volume solution (L) 0.01 Concentration (M) 1.64341E-05 Table 2. Preparation of Gold stock solution Molecular weight Au (g/mol) 393.83 mass Au(g) 0.032 volume stock(L) 0.025 Concentration (M) 0.003250133 Table 3. Preparation of Lysozyme stock solution Molecular weight (g/mol) 14307 mass lysozyme (g) 0.0717 volume solution (L) 0.05 Concentration (M) 0.00010028 Note: This stock solution was prepared by Jame's Schwabacher Table 4. Preparation of Hemoglobin-AuNP solutions Ratio Amount of gold added concentration hemoglobin (M) volume Hemoglobin (mL) Volume Water (mL) 30 0.384599609 8.33333E-06 2.535377358 2.080023032 50 0.384599609 0.000005 1.521226415 3.094173976 70 0.384599609 3.57143E-06 1.086590296 3.528810094 90 0.384599609 2.77778E-06 0.845125786 3.770274604 110 0.384599609 2.27273E-06 0.691466552 3.923933838 130 0.384599609 1.92308E-06 0.585087083 4.030313308 150 0.384599609 1.66667E-06 0.507075472 4.108324919 170 0.384599609 1.47059E-06 0.447419534 4.167980857 190 0.384599609 1.31579E-06 0.400322741 4.21507765 210 0.384599609 1.19048E-06 0.362196765 4.253203625 Table 5. Preparation of Lysozyme Au-NP solutions Ratio Amount gold added (mL) Concentration Lysozyme (M) Volume lysozyme (mL) Volume water (mL) 5 0.384599609 0.00005 2.493019545 2.122380845 10 0.384599609 0.000025 1.246509773 3.368890618 15 0.384599609 1.66667E-05 0.831006515 3.784393876 20 0.384599609 0.0000125 0.623254886 3.992145504 25 0.384599609 0.00001 0.498603909 4.116796482 30 0.384599609 8.33333E-06 0.415503258 4.199897133 35 0.384599609 7.14286E-06 0.356145649 4.259254741 40 0.384599609 0.00000625 0.311627443 4.303772947 45 0.384599609 5.55556E-06 0.277002172 4.338398219 50 0.384599609 0.000005 0.249301955 4.366098436 New ratios of lysozyme were prepared in order to determine the optimal concentration ratios for the formation of fibers.