Class: BE210
Group: R5
Members:  FACILITATOR Hobart Lee,TIME & TASK KEEPER Jehann Biggs, SCRIBE Jenea McLaughlin, PRESENTER David Kim
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Abstract:

Column chromatography with the reducing agent sodium dithionite was used to reduce methemoglobin into hemoglobin, while G-25 Sephadex gel was used to pack the column at a speed of 1.3 ml/min.  Spectral analysis of the hemoglobin indicated three peaks at 412 nm, 541 nm, and 576 nm compared to the single peak of methemoglobin at 406 nm.  Molar extinction coefficients were obtained from linear regressions of Absorbance vs. Dilution plots.  The ?(?) of all hemoglobin samples were statistically consistent with one another at p<0.05 for each of the three wavelengths.  The values of the molar extinction coefficients were 3.8 ? 0.5 x 105 M-1cm-1 95% CI at 412nm, 5.1 ? 0.9 x 104 M-1cm-1 95% CI at 541nm and 4.9 ? 0.8 x 104 M-1cm-1 95% CI at 576nm and only the value at 412nm was significantly different from literature values.  Quadratic functions were fit to the peaks at 412, 541 and 576nm for samples of hemoglobin and samples of hemoglobin with methemoglobin impurity: the peak at 576nm showed a significant difference (p<0.05) between the quadratic regression coefficients of the pure and impure hemoglobin.