Exp 6 - Fluorescence copy

Exp 6 - Fluorescence copy - CHE 226-001 Experiment 6...

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] CHE 226-001 Experiment 6 Optical Spectroscopy of Porphyrins Pre-Lab Questions 1. The analyte of interest is an aromatic, tetrapyrrolic macrocycle named porphyrin. Structure of Porphyrin 2. The organic phase is made up of porphyrin and methanol, while the aqueous layer is made up of HCl and ethyl acetate. 3. UV-vis spectroscopy is an optical spectroscopy technique that uses UV and other visible portions on the electromagnetic radiation scale to interact with pi-electrons. The wavelengths used in this technique are the following (in nm): 407, 505, 541, 575, and 630. Porphyrin can be analyzed using this technique because it contains 11 pi-bonds. 4. Fluorescence is different from UV-vis in that it relies on the fact that once an electron is excited by light, it emits photons. The fluorescence instrument can then detect these photons, and the data can be used for analysis of the analyte. The detector is positioned at a 90° angle from the light source to prevent light from the source from getting to the detector. Procedures The purpose of this experiment is to determine the amount of porphyrin in the average household brown egg. The completion of this experiment will also result in the understanding of optical spectroscopy techniques such as UV-vis and Fluorescence. The complete procedures can be found in the CHE 226 blackboard handouts section, named “Experiment 6 - fluorescence”. Theory As described in the pre-lab questions, the UV-vis and fluorescence are optical spectroscopy techniques where a sample is manipulated using UV light. The analyte for this
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