Lab 1 Carbohydrates Bch2333

Lab BCH 2333

Section:

Lab 1 Carbohydrates: Separation Techniques Based on Molecular Size

TA:

Wednesday, January 16th, 2013

Team #4

By:

Partner:

Purpose

The purpose of this experiment is to exemplify how differences in molecular weight allow separation of polymers from their monomers. Methods of dialysis and gel filtration chromatography will be used to separate a glucose monomer from a starch polymer. Colorimetric glucose oxidase assay will be used to monitor the presence of glucose and a colorimetric iodine assay will be used to monitor the presence of starch in prepared solutions after separation

Results and Discussion

Table 1: Glucose oxidase assays
|Reagents |Volume (mL) |Positive Response |A420 |
| |Sample |Water |Total |
| |Sample |Water |
| |A420 Internal Solution |C1 (g/L) |

# |Elution Vol. (mL) |A420 |C (g/L) |Mass (mg) |Positive
Response |A600 |C (g/L) |Mass (mg) |Positive
Response | |1 |3.86 |-0.0140 |-0.0512 |-0.198 |- |0.0240 |0.00704 |0.0272 |- | |2 |7.72 |0.0210 |0.0768 |0.296 |- |0.0370 |0.0109 |0.0421 |- | |3 |11.6 |0.0610 |0.223 |0.861 |- |0.0300 |0.00880 |0.0340 |- | |4 |15.4 |0.0280 |0.102 |0.394 |- |0.314 |0.0921 |0.356 |+ | |5 |19.3 |-0.0610 |-0.223 |-0.861 |- |0.0800 |0.0235 |0.0907 |- | |6 |23.2 |0.0470 |0.172 |0.664 |+ |0.0480 |0.0141 |0.0544 |- | |7 |27.0 |0.459 |1.68 |6.48 |+ |0.000 |0.000 |0.000 |- | |8 |30.9 |0.727 |2.66 |10.3 |+ |0.0240 |0.00704 |0.0272 |- | |9

References: 1. Voet, D. and Voet, J. (2004). Biochemistry, 3rd Ed. John Wiley & Sons, 6.3, p 133. 2. Voet, D. and Voet, J. (2004). Biochemistry 3rd Ed. John Wiley & Sons, 6.3Bc p 139.