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Experiment_1.docx

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Department
Chemistry
Course
CHM2354
Professor
Don't Know
Semester
Winter

Description
Advanced InstrumentalAnalysis Laboratory CHM4116 Laboratory#1 Separation andAnalysis of ChlorophyllA& B from Spinach leaves Abstract: Molecular Absorption spectroscopy can be employed in the lab to deduce concentration of a compound in a sample based on its absorption in comparison to a reference standard. Here we attempt to undergo an analytical analysis of the chlorophyll A/B content of a representative sample of spinach leaves. The analysis was only performed on the spinach leaves themselves, 3 and excluded veins and stems. ChlorophyllsA & B were extracted using aqueous acetone and their absorbance measured at two wavelengths, 645nm & 663nm. The uniformity of the sampling technique was to be confirmed usingANOVAstatistical methods. Chlorophyll Aconstituted 67.5% of the spinach samples on average, and Chlorophyll B constituted 32.5% of the spinach samples on average. Experimental: For method and procedure please refer CHM4116: Advanced Instrumental Analysis Laboratory manual. Pages 11-16 A) Instruments:  Cary 100 Spectrometer  Stir plate  Samples were prepared using analytical balances for weight  Mortar & Pistil Results: Table.1: Raw absorbance Readings SampleAbsorbance λ A1 A2 A3 B1 B2 B3 C1 C2 C 3 645n 0.26180 0.071993 0.175262 0.161997 0.14256 0.152368 0.158265 0.184782 0. m 6 4 9 5 6 8 8 18 96 46 9 663n 0.49791 0.189973 0.466512 0.436706 0.38066 0.404235 0.423381 0.4871107 0. m 3 2 8 4 1 49 84 82 9 Table.2: ChlorophyllA& B concentrations in Spinach leaves Λ 645 Λ663 m ChlorophyllChlorophyllA] [Chlorophyll B] [Chlorophyll [Chlorophyll Sample (g) (ug/mL) (ug/mL) A] B] (ug/mL) (ug/mL) A1 0.4042g 16.60 7.99 10.21 4.91 A2 0.4222g 18.82 9.06 15.73 7.55 A3 0.4246g 16.58 7.96 10.19 4.79 B1 0.4060g 15.33 7.36 9.54 4.58 B2 0.4193g 13.49 6.48 8.32 3.99 B3 0.4128g 14.42 6.92 8.83 4.24 C1 0.4301g 14.97 7.19 9.25 4.44 C2 0.4152g 17.48 8.39 10.65 5.11 C3 0.4090g 17.94 8.61 10.89 5.23 Table.3: ANOVAanalysis ChlorophyllA Chlorophyll B SS df MS F SS df Ms F Within 19.15 2 0.3657 67.30 2.837 2 2.467 1.478 6 Between 221.5 8 24.61 51.57 8 3.647
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