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AP Lab 4

AP Lab 4

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Published by Shawn Joiner
AP Lab 4, done in HL Biology IB style. ;P
AP Lab 4, done in HL Biology IB style. ;P

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Published by: Shawn Joiner on May 03, 2010
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AP BIOLOGY LAB 4Study of Photosynthesis Rate in DifferentChloroplasts Suspensions under Different Light Conditions
During the process of this lab, the researcher and his lab partners determined therate of photosynthesis in isolated chloroplasts using the dye DPIP; learned how toseparate pigments and calculate their R values; described a technique used to determine photosynthetic rates; compared photosynthetic rates at different wavelengths of lightusing controlled experiments and explained why the rate of photosynthesis varies under different environmental conditions.The researcher and his lab mates did this by labeling 5 cuvettes 1-5, then coveredthe walls and bottom of cuvette 2 with foil and made a foil cap to cover the top. 1ml of  phosphate buffer was added to each cuvette; 4ml of distilled water was added to c1, 3mlof the water in 2, 3, 4 along with 1ml of DPIP. In c5, 3ml plus 3 drops of water and 3mlof DPIP was added. The researcher and his lab mates then followed the instructionswithin the lab booklet to obtain suspensions for the chloroplasts.The results from our experiment indicated a major connection between lightexposed to live chloroplasts and light transparency through a cuvette (amount of DPIPreduced). All mixtures except the unboiled chloroplasts exposed to light remainedrelatively constant in light transmittance. This includes unboiled/dark, boiled light, andno chloroplast/light mixtures. The dark unboiled suspension did increase a small amountin light transmittance, but as cited as error, this was probably due to unwanted lightemission.
 Experiment using the scientific method.
:In this lab you will:1. separate plant pigments using chromatography and2. measure the rate of photosynthesis in isolated chloroplasts using the dye DPIP.The transfer of electrons during the light-dependent reactions of photosynthesis reducesDPIP, changing it from blue to colorless.Before doing this lab you should understand:
how chromatography separates two or more compounds that are initially presentin a mixture;
the process of photosynthesis;
the function of plant pigments;
the relationship between light wavelength and photosynthetic rate; and
the relationship between light intensity and photosynthetic rate.
After doing this lab you should be able to:
separate pigments and calculate their R values;
describe a technique to determine photosynthetic rates;
compare photosynthetic rates at different light intensities or different wavelengthsof light using controlled experiments; and
explain why the rate of photosynthesis varies under different environmentalconditions.
Table 4.4: Transmittance (%)Time (minutes)Cuvette0510152 Unboiled/Dark3027.830.533.23 Unboiled/ Light549899.998.24 Boiled/Light4747.24746.85 No Chloroplasts/Light42.842.842.842.7Table 4.1Distance Moved by Pigment Band (mm)Band NumberDistance (mm)Band Colou1.0Yellow- Brown2.5Light Green3.30Green4.48YellowDistant Solvent Front Moved _60_ mmAnalysis of ResultsThe relationship of the distance moved by a pigment to the distance moved by the solventis a constant called R. It can be calculated for each of the four pigments using thefollowing formula:R= distance pigment migrated (mm) /distance solvent front migrated (mm)Record your R values in Table 4.2Table 4.2.8 = R for Xanthophyll (yellow).5 = R for Chlorophyll a (bright green to blue green)
0 = R for Chlorophyll b (yellow green to olive green)Topics for Discussion1. What factors are involved in the separation of the pigments? - The solubility, size of  particles, and their attractiveness to the paper are all involved in the separation.2. Would you expect the R value of a pigment to be the same if a different solvent wereused? Explain. - No, the different solubilities of the pigments would change the Rf values. For example chlorophyll b is only soluble to fat solutions.3. What type of chlorophyll does the reaction center contain? What are the roles of theother pigments? - The reaction center contains chlorophyll a. The other pigments collectdifferent light waves and transfer the energy to chlorophyll a.Analysis of ResultsPlot the percentage of transmittance from the four cuvettes on Graph 4.1.Label each plotted line.For this graph you will need to determine the following:a. The independent variable: __________ Use this to label the horizontal (x) axis. b. The dependent variable: ________ Use this to label the vertical (y) axis.

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