LABORATORY REPORT

CHM475
Experiment 8: Spectrophotometric Analysis for transition metal cations

Name: Aishah Binti Azmi Matric No. : 2012816034 Lecturers name: Pn. Kamariah Muda

Introduction In chemistry, spectrophotometry is the quantitative measurement of the reflection or transmission properties of a material as a function of wavelength. It is more specific than the general term electromagnetic spectroscopy in that spectrophotometry deals with visible light, near-ultraviolet, and near-infrared, but does not cover time-resolved spectroscopic techniques. Spectrophotometry involves the use of a spectrophotometer. A spectrophotometer is a photometer that can measure intensity as a function of the light source wavelength. Important features of spectrophotometers are spectral bandwidth and linear range of absorption or reflectance measurement. In this experiment, In order to determine the concentration of unknown solutions a standard plot of the concentration of copper vs. the absorbance reading from the spectrophotometer of solutions having known copper solution concentrations is plotted. According to Beer's Law, absorbance is directly proportional to concentration and so the resulting plot should be a straight line. This graph will be used to determine the concentrations of solutions containing known amounts of the unknown copper compound and from this we can determine the weight percent of copper. Typically, we determine the concentration of some solute of interest by react it with another substance to produce a substance that absorbs light at a particular wavelength, and comparing it to a second solution with a known concentration. When we do so, we are trusting that we have conducted the experiment correctly, and that the results accurately reflect the actual concentration of the substance.

Objective The objective is to determine the concentration of an unknown copper(II) solution by spectrophotometric analysis.

Apparatus and equipments 1. Spectrophotometer 2. Test tubes 3. Test tube rack 4. Cuvettes 5. Beakers 6. Measuring cylinders

Method 1. The spectrophotometer is switched on to warmed up and the l is set up to the max at 645 nm. Six of the test tubes are cleaned and dried and is number from 1 to 6. The first four test tubes is used as the reference solution to calibrate the spectronic 20 spectrophotometer. 2. Calibrations standard is prepared using 0.2M CuSO4.5H2O and samples of the coordination compound prepared earlier. The contents of each of test tubes is mixed thoroughly. According to the direction in table 8.1. 3. The contents is then transferred into cuvettes . the absorbance is measured at 654nm. Data is entered at the table prepared.

Questions 1. What is the general relationship between concentration and absorbance? Essentially if the path length of the light is constant there is a direct linear relationship between concentration and absorbance for a particular absorbing compound. 2. Determine the concentration of the unknown from the calibration curve. The concentration of the unknown sample prepare in test tube 5 is 0.15M and test tube 6 is 0.19M according to the calibration curve.

3. Explain how the concentration of the unknown is determined? The concentration of the unknown is determine by only knowing the absorbance tested earlier during the experiment is conducted. Then when a graph of absorbance against concentration is plotted. The concentration of both test tubes containing unknown solution concentration can be determined.

4. Supposedly you were given an unknown with a greater concentration than any of your known solutions. Explain how you would determine the concentration of the unknown. The concentration will have o be calculated and cannot be found by using only the calibration curve.

Discussion From the data that have been gathered, the concentration of the unknown in test tube 5 is 0.15M while in test tube 6 is 0.19M. This amount showed that by using spectrophotometer the amount of unknown concentration can be measured. The solution with known concentration prepared earlier is used as standard to plot the calibration curve so that we can predict the concentration of the unknown solution. The higher the concentration of certain substances the higher the

absorbance reading because it can absorb more visible light. While the experiments have been conducted there are many errors that can happen that will affect the reading or the results. The following are the most common causes of artifacts in spectrophotometric measurements: 1. Air bubbles in the cuvette. Microcuvettes are particularly susceptible to collection of air bubbles in the sample beam Particulates in the sample. Particulate matter in the sample will scatter light and result in absorbance readings that are significantly higher than the true reading. 2. Contaminating substances in diluents.. 3. Incomplete mixing of samples can yield improper results. 4. Dirty cuvettes. The cuvettes should be clean on both the outside and the inside

5. Residual liquid in the cuvette before addition of the sample. This will cause either a) dilution of the sample, resulting in an artifactually low reading; or b) contamination of the sample with a solution containing a "high absorbing" compound, resulting in an artifactually high reading. Conclusion By using spectrophotometer, the concentration of unknown solution can be made known by using standard solution with known concentration. The higher the absorbance reading the higher its concentration. References 1. http://www.ruf.rice.edu/~bioslabs/methods/protein/spectrophotometer.html 2. http://www2.bren.ucsb.edu/~keller/courses/esm223/Spectrometer_analysis.pdf 3. http://www.chm.davidson.edu/vce/spectrophotometry/Spectrophotometry.html 4. http://www.rrcap.ait.asia/male/manual/national/11Chapter11.pdf