Spinach

Have you ever really wondered how different variables can affect how plants go through photosynthesis? Well, in this experiment, the purpose was to see how various environmental conditions can affect the overall photosynthetic capacity of a specific plant. The factors, light, darkness, cold, and heat were applied to see how the different components would affect the photosynthesis on spinach plants. Each group was given a different factor to test. Out group was given the light factor. The hypothesis for this experiment is that when adding light as a factor, the light will affect the overall plant photosynthesis.

There are two main types of chlorophyll, chlorophyll a which absorbs wavelengths of 430nm (blue) and 662 (red) and is the main photosynthetic pigment, and chlorophyll b, which doesn’t directly participate in the photosynthetic process, but is capable of donating its energy to chlorophyll a

The concentrations and absorbances of the red and blue dyes were used to find the concentration of the purple dyes. From the graph of the blue dye, the linear equation for absorbance was y = mx + b. From that formula came the equation y = 7.915 x 104 (x) + 0.02489, where y represents absorbance, m is slope, x is concentration/molarity, and b is the constant/y-intercept. The same set up was performed for the red dye, but the equation produced was y = 1.045 x 104 (x) +.001298. The equations found when graphing absorbance vs. concentration were used to find the concentration of the purple dyes. The absorbance for purple dye 3 on the red wavelength of 470 nm equaled 0.149 and 0.818 for the blue wavelength of 635 nm. For purple dye 1

The purpose of this experiment is to determine the maximum absorbance of fast green, and the chlorophylls, also in the case of fast green create a concentration curve to determine an unknown substance. Each test will use the spectrophotometer.

The procedures for experiment A, B, and C all start the same. The first step is to put on goggles and get the data collection device set properly. The labquest needs to be plugged into the colorimeter accurately so that a click is heard when putting it in. The labquest needs to be reading digitally and the colorimeter needs to be set to 635 nm. Then shake the chloroplast solution and take a clean cuvette and fill it with 3 mL of distilled water, 3 drops of the chloroplast solution, and cap it. This is used as a blank to calibrate your labquest. Double check that the labquest is reading absorbance, this assures that the colorimeter is plugged into the labquest accurately. Insert the blank into the colorimeter and hit the calibration button. Take out the blank and empty it. The labquest is now set to experiment with. Make sure that the heat bank is set in front of the lamp and that the lamp is on. The cuvette must be placed on the opposite side of the heat bank in the path of light in the box so that no other light can interfere with the experiment.

Scientists use an instrument called a spectrometer to quantitatively determine the amount of light absorbed by a solution. The primary inner parts of a typical spectrometer are described below. The spectrometer has a light source that emits white light containing a vast mixture of different wavelengths of electromagnetic radiation. The wavelength of interest is then selected using a monochromator (“mono” meaning one and “chromate” meaning color) and an additional exit slit. The separation of white light into different colors (wavelengths) is known as diffraction. The selected light then reaches the sample and depending on how the light interacts with the chemical compound of interest, some of the light is absorbed and some passes straight through. By comparing the amount of light entering the sample (P0) with the amount of light reaching the detector (P), the spectrometer is able to tell how much light is absorbed by the sample.

In the beginning of this experiment, our TA added water, salt, and 75/25 hexane/acetone to spinach leaves to a blender and blended the mixture to assume equal amounts for each group and to avoid erros if each student had to do the blending. The addition of the water to the mixture allowed the it to separate into a distinct organic layer after being run in a centrifuge, which was available to be collected at the top of the centrifuge. Salt reduces solubility, which will force the organic parts of the mixture (the desired pigments for example) to separate into the organic layer at the top. Lastly, 75/25 hexane/acetone is added because this is a moderately polar solvent and will useful for both the non-polar and polar pigments present within the spinach leaves. A mixed solution of hexanes and acetone must be used because acetone is very polar, while hexane in very non-polar, and the spinach leaves contain both non-polar and polar pigments in them that are important in the extraction and for further analysis. The mixture was placed in the centrifuge so the solids in the mixture (mostly cellulose) could be separated from the liquids into separate distinct layers for further extraction and testing. In the tube, the organic substances separated into the top layer, whereas the water layer remains at the bottom of the tube below the solid layer made up of mainly cellulose.

Abstract: The purpose of this lab is to separate and identify pigments and other molecules within plant cells by a process called chromatography. We will also be measuring the rate of photosynthesis in isolated chloroplasts. Beta carotene, the most abundant carotene in plants, is carried along near the solvent front because it is very soluble in the solvent being used and because it forms no hydrogen bonds with cellulose. Xanthophyll is found further from the solvent font because it is less soluble in the solvent and has been slowed down by hydrogen bonding to the cellulose. Chlorophylls contain oxygen and nitrogen and are bound more tightly to the paper than the other pigments.

Pigments extracted from different greens have different polarities and may be different colors. Mixed pigments can be separated using chromatography paper. Chromatography paper is able to separate mixed pigments due to their polarity and solubility. Pigments of chlorophyll a, chlorophyll b and beta carotene will be separated on chromatography paper because each has its own polarity and solubility, which results in different distance traveled up the paper. Beta carotene is non-polar so it travels the highest distance, followed by chlorophyll a. Chlorophyll b is the most polar; therefore, it travels the shortest distance. The separated pigments on the chromatography paper can be eluted in acetone and absorbance spectrum is

This experiment demonstrates the effects of pH on the rate of photosynthesis by examining the behavior of leaf disks in different pH solutions under light. In this experiment, we used five different pH levels: pH 5, pH 6, pH 7, pH 8 and pH 9. These solutions were created using a combination of hydrochloric acid and sodium hydroxide. Spinancia olcerea or spinach, leaves were used in the experiment to examine the effects of pH on the rate of photosynthesis. The rate of photosynthesis was measured by counting the number of leaf disks that rose to the surface of the solution after each minute. In acidic solutions, the rate of photosynthesis increased while in basic solutions, the rate of photosynthesis decreased.

The photosynthesis lab is comprised of three short experiments. These experiments showed how to understand and apply the absorbance spectrum and which colors and wavelengths correspond to visible light. In order to fully understand this absorbance spectrum and how to apply it, we initially prepared a substance comprised of acetone, a large spinach leaf and petroleum ether and measured its absorbance in the spectrophotometer. This showed us what wavelengths corresponded to the most absorption of the spinach leaf. It is understood that the least amount of absorbance should occur after 500 nm and the lower the number, the lower the absorbency. Thus, the 700 ranges is the very least amount of absorbency and the results showed us that the lowest amount of absorbance was around 740nm, which is an accurate, representation of this solution. Next, to understand what wavelengths of light drive the light reactions you can visibly see in photosynthesis, we used CMS in a variety of tubes and measured its absorbency under white light, no light, and colored filters such as red and green. We then used the chlorophyll extract we originally prepared and painted this on a chromatography strip. We measured the band distances to find out the number of pigments in the spinach leaf. These processes helped emphasize how the chloroplast pigment extract and chloroplast membrane suspension have different functional capabilities and how photosynthesis works.

This would have major impact on plant growth and floor (Battaglia et al. 2003; Endler 1993). This is connected with the wavelength of the light and much of this research is based on observation of the highest amount of photosynthetic activity, that is measured with samples that are exposed to dark (unfiltered) light at different wavelengths (Lee 1987). Therefore the aim of this experiment is to assess the photosynthetic activity using chloroplast isolate from spinach beet. This study will determine how wavelength affects the rate of photosynthesis. Based on past studies and research it can be hypothesised that chloroplast exposed to white light would have quicker photosynthetic activity then those exposed to red light. This is expected to occur because chlorophyll a and b consists of accessory pigments which inhibit different wavelengths of light, as a result this would raise the amount of light energy being consumed. The control of this study will be light independent reaction as chloroplasts are kept in dark to stop photosynthesizing due to inadequate development of ATP and NADPH from light reaction. While the reaction in room is hypothesised to photosynthesis at a lowest possible rate (Sadava et al. 2011)

The purpose of this experiment was to take spinach leaves and extract the chlorophyll and carotenoid pigments by using acetone as the solvent. The chlorophyll and carotenoid pigments were extracted by using column chromography and alumina was used as the solvent. Solvents of different polarities were used, starting with the least polar, to extract the certain components from the leaves. They were then analyzed by using thin- layer chromatography.

For lab 12, it is hypothesized that chlorophylls a and b are present in a plant leaf and contribute to the starch production in photosynthesis. Also, products of photosynthesis will be present in leaf tissue exposed to red and blue light wavelengths for several days, but a decreased presence in leaf tissue exposed to green and black light wavelengths. In lab 13, it is expected that since chlorophylls a and b are more polar and smaller molecules than the anthyocyanins and carotenoids, they will travel higher up the chromatography paper than the other pigments.

8) Steps 1 - 8 were repeated using the wavelengths of 360 nm to 900