Lab Report on the Separation of Blue Dextran and Cobalt Chloride on Sephadex G25<\/p>\n
Name of Student<\/p>\n
Date of Submission<\/p>\n
Lab Report on the Separation of Blue Dextran and Cobalt Chloride on Sephadex G25<\/p>\n
Results<\/p>\n
The radius, height of bed and the bed volume of the column<\/p>\n
The formula for the volume of a right cylinder is 3.142*r*r*h<\/p>\n
Where r is the radius of the column and h is the height which are both expressed in cm<\/p>\n
For the Econo column which is clamped in an upright position the radius is the diameter divided by 2<\/p>\n
Which is 0.7 \/2 = 0.35<\/p>\n
The height is 20 cm<\/p>\n
Therefore, the volume is 3.142 * 0.35*0.35*20 = 7.6979 cm3<\/p>\n
2. The plots for the elution profiles is as shown below<\/p>\n
The graph below shows the elution profile of blue dextran. The absorbance versus fraction number using the wavelength data is the first loading<\/p>\n
Sample ID Description 619.6<\/p>\n
512.9<\/p>\n
Sample1920 Blue Dextran Fraction 1 0.0361 0.0356<\/p>\n
Sample1921 Fraction 2 0.0121 0.0125<\/p>\n
Sample1922 Fraction 3 0.1563 0.0389<\/p>\n
Sample1923 Fraction 4 1.1281 0.2408<\/p>\n
Sample1924 Fraction 5 0.2651 0.0570<\/p>\n
Sample1925 Fraction 6 0.0416 0.0096<\/p>\n
Sample1926 Fraction 7 0.0121 0.0051<\/p>\n
The Graph above is a graph of absorbance of blue dextran fraction against the fraction number<\/p>\n
619.6 512.9<\/p>\n
Sample1927 Cobalt Chloride Fraction 1 0.0010 0.0008<\/p>\n
Sample1928 Fraction 1 0.0013 0.0012<\/p>\n
Sample1929 Fraction 2 0.0010 0.0008<\/p>\n
Sample1930 Fraction 3 0.0029 0.0026<\/p>\n
Sample1931 Fraction 4 0.0022 0.0038<\/p>\n
Sample1932 Fraction 5 0.0095 0.1363<\/p>\n
Sample1933 Fraction 6 0.0282 0.4087<\/p>\n
Sample1934 Fraction 7 0.0155 0.2063<\/p>\n
Sample1935 Fraction 8 0.0024 0.0206<\/p>\n
Sample1936 Fraction 9 0.0014 0.0025<\/p>\n
Sample1937 Fraction 10 0.0019 0.0015<\/p>\n
The graph above is a plot of absorbance against the fraction number. <\/p>\n
The graph for absorbance versus fraction number for the mixture<\/p>\n
Sample ID Description 619.6 512.9<\/p>\n
Sample1938 Mixture Fraction 1 -0.0004 -0.0006<\/p>\n
Sample1939 Fraction 2 -0.0004 -0.0006<\/p>\n
Sample1940 Fraction 3 -0.0004 -0.0002<\/p>\n
Sample1941 Fraction 4 0.1345 0.0290<\/p>\n
Sample1942 Fraction 5 0.2255 0.0487<\/p>\n
Sample1943 Fraction 6 0.0312 0.0446<\/p>\n
Sample1944 Fraction 7 0.0447 0.5608<\/p>\n
Sample1945 Fraction 8 0.0574 0.8250<\/p>\n
Sample1946 Fraction 9 0.0230 0.3293<\/p>\n
Sample1947 Fraction 10 0.0022 0.0257<\/p>\n
Sample1948 Fraction 11 0.0007 0.0039<\/p>\n
3. The elution volume for Blue dextran<\/p>\n
Ve = Vo + KdVi<\/p>\n
Where Ve is the elution volume <\/p>\n
Vo is the volume external to the medium<\/p>\n
Vi is the volume included within the gel<\/p>\n
Ve = 0.25 + 1(.5)<\/p>\n
Ve = 0.25 + 1.5<\/p>\n
Ve = 1.75ml<\/p>\n
The elution volume for cobalt chloride<\/p>\n
Ve = Vo + KdVi<\/p>\n
Ve = 1 + 1(1.5)<\/p>\n
Ve = 1 + 1.5<\/p>\n
Ve = 2.5ml<\/p>\n
4. For blue dextran the mass is `0.0042g<\/p>\n
0.4moles\/1 * 1 mole * 1L\/1000ml = 0.004 moles<\/p>\n
Cobalt Chloride = 237.93g<\/p>\n
0.004 moles * 237.93g = 0.952g (Expected) Cobalt Chloride<\/p>\n
Actual Cobalt Chloride is 0.9683g<\/p>\n
The Cobalt peak maxima is (512.85nm, 1.5373A)<\/p>\n
The Blue dextran maxima is (619.6nm, 0.32954A)<\/p>\n
Discussion <\/p>\n
The results appear not to be consistent despite the fact that there may be some errors. This may be because the samples stick to the column. The inconsistency is due to errors in calibration. The gel filtration columns should be calibrated regularly as the elution volumes change with time. Since the lab experiment occurred under limited timeframe, cleaning was intense. My results were not consistent since I accidently mixed up the chemicals in the laboratory. The degree of separation was consistent. <\/p>\n
The main method of making the separation more effective is by ensuring the purification is done perfectly. Unlike other chromatography methods, the strength of gel chromatography comes in through the interaction of the according to shapes and sizes. Two samples which differentiate with respect to the molecular size come up due to differences in the size. If the shape counteracts the size difference the separation may be ineffective (Liu et al,.). <\/p>\n
Reference<\/p>\n
Liu, H., Nishide, D., Tanaka, T., & Kataura, H. (2011). Large-scale single-chirality separation of single-wall carbon nanotubes by simple gel chromatography.\u00a0Nature communications,\u00a02, 309.<\/p>\n","protected":false},"excerpt":{"rendered":"
Lab Report on the Separation of Blue Dextran and Cobalt Chloride on Sephadex G25 Name of Student Date of Submission<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[],"tags":[],"class_list":["post-34471","post","type-post","status-publish","format-standard","hentry"],"yoast_head":"\n