Determination of copper (II) concentration by colorimetric method

Institute of Biology, College of Science University of the Philippines, Diliman, Quezon City, Philippines
May 15, 2013

ABSTRACT The importance of colors were displayed in this experiment. This shows how different concentrations of a substance can affect the absorbance. The linear relationship of concentration and absorbance is summarized in Beers law. In this experiment, it was shown how Beers law was used in order to obtain the concentration of a substance. This was done by analyzing the absorbance of the same substance but of different unknown concentration using a spectrophotometer that has wavelength of maximum absorbance of 626nm. From the obtained absorbance values, the concentration was then calculated. From these data, a linear equation was obtained using linear regression. This equation was then used to calculate the concentration of an unknown substance. The unknown substances concentration has an average of 2371 ppm. Overall, this experiment proved that indeed, the concentration and absorbance of a substance has a linear relationship.

INTRODUCTION
Colors play an important role in the field of analytical chemistry. This does not only apply for detecting color indicator end points but can also be used in colorimetry. Generally, calorimetry is the science of measuring color (cg.tuwien.ac.at). To define further, it is the method of trace analysis that involves absorption of radiations having frequencies in the visible spectrum (Pickering,1966). Hence, it is the variation of the color of a system with change in concentration of some component forms the basis of what the chemist commonly terms calorimetric analysis (Jeffery et. al. 1989). Simply speaking, it can determine the concentration of a substance by measuring the light absorbed with respect to a known concentration of a substance. The color being measured may be innate to the substance or it can be due to the formation of a colored compound by the addition of an appropriate reagent. Hence, it can be said that the most important idea in colorimetry is that color intensity is proportional to the concentration (depts.clackamas.edu). However, to get the concentration, a spectrophotometer is needed, as in the case of the experiment. This is done

so as to understand the concepts of colorimetry in chemical analysis. When light (monochromatic or heterogeneous) falls upon a homogeneous medium, a portion of the incident light is reflected, a portion is absorbed within the medium, and the remainder is transmitted. The reflected light is what we can perceive using our eyes and is usually the complementary color of the one that is absorbed while the absorbed and transmitted light is what we cannot see and what is being measured using the spectrophotometer. Below is a table showing the different colors that are absorbed at different wavelengths and their complementary color as perceived by our eyes.

Table 1. Complementary colors

reaction took place upon the addition of ammonia (Klingenberg,1965):

Cu 2 Cu ( NH 3 ) 4 2 ( Dark blue)
The blank reagent is the flask containing the 0.00 mL stock solution and 10 mL ammonia.
Taken from Fundamentals of Analytical Chemistry, 8th Edition

The relationship between the absorbed light and the concentration of the solution can be seen using Beers law. Beer studied the effect of concentration of the colored constituent in solution upon the light transmission or absorption and came up with this equation: A = a b c (Equation 1) In the equation, A, refers to the absorbance by the solution while a is the molar absorptivity with a unit of L mol-1 cm-1. This can also be symbolized by . Additionally, b is the path length of light while c is the concentration of solution with a unit of mol L-1. For convenience, absorbance can also be calculated from transmittance, which is the fraction of the incident radiant power transmitted by the sample (Pecksok, 1976). Transmittance is expressed as: T=(I/Io) (Equation 1) From equation 2, absorbance can be calculated as: A= -log T (Equation 3) To apply the concepts of colorimentry, this experiment measured the absorbance of a standard sample of different unknown concentrations. Also, an unknown sample was analyzed by measuring its absorbance to obtain its concentration.

The absorbance of the most concentrated working standard solution was then measured against the blank reagent. The blank reagent was used to take into account the transmitted radiation lost due to scattering, reflection, and etc (Pecsok, 1976). From the data, the wavelength of maximum absorption (max) was then obtained. The value of max in this experiment is 626nm. From this obtained max, the absorption of the remaining working standard solutions was then measured. A graph was made by plotting the absorbance of the working standard solutions against their concentration. The slope, y-intercept, and the regression coefficient was also determined using linear regression. The shape of the calibration curve often depends on the bandwidth. That was why the wavelength range needed to be scanned to determine the best analytical wavelength for analysis. At this wavelength, change in absorbance with concentration is at a maximum, thus yielding greater sensitivity and higher accuracy (Pecsok, 1976). The unknown sample taken from the class instructor was also analyzed in the same way the working standard samples were analyzed only that, three absorbance readings were taken. The concentration of the unknown sample was calculated using the equation obtained from the linear regression done to the working standard samples. RESULTS AND DISCUSSION From the experiment, the wavelength of maximum absorption (max) obtained has a value 626nm. Below is the table showing the results of the values of absorbance for each working standard, obtained from the spectrophotometer reading. The table also shows the calculated Cu (II) concentration for each working standard. The resulting concentration was calculated using the equation found in Appendix 1. Table 2. Corresponding Cu (II) concentration from the obtained absorbance values for each working standard

METHODS
Five working standard solution were prepared by pipetting out 0.00, 2.00, 4.00, 6.00, 8.00, and 10.00 mL standard Cu (II) solution with a concentration of 2500 ppm. Each working standard solution was placed in a 50 mL volumetric flask. To each flask, 10 mL ammonia solution was added and the solution was then diluted to the mark. The purpose of adding ammonia was to intensify the color of Cu (II) as it has a light blue color before the addition of ammonia. The following

Volume of working standard solution, mL 2 4 6 8 10

Concentration of standard Cu (II), ppm 100 200 300 400 500

Absorbance 0.088 0.177 0.269 0.358 0.447

water first and followed by the sample to be analyzed to make sure that it is indeed he sample alone that is being analyzed. However, even if all the necessary instructions are followed, there are limitations in using the Beers law. The following are the limitations of the linearity of Beers law taken from the University of Adelides Department of Chemistry: Deviations in absorptivity coefficients at high concentrations (>0.01M) due to electrostatic interactions between molecules in close proximity Scattering of light due to particulates in the sample Fluoresecence or phosphorescence of the sample Changes in refractive index at high analyte concentration Shifts in chemical equilibria as a function of concentration Non-monochromatic radiation, deviations can be minimized by using a relatively flat part of the absorption spectrum such as the maximum of an absorption band Stray light

From the values in table 2, the graph below was generated showing the linear relationship between absorbance and concentration. This is the same reason why it was necessary to use absorbance rather than transmittance since absorption peaks appear as deep valleys in a transmittance plot whereas an absorbance plot would produce a linear result (Pecsok,1976). The graph also contains the linear equation needed to calculate the Cu (II) concentration of the unknown sample. The resulting R2 with a value of 0.9971 means that the values that we obtained are good since it is close to 1.

Figure 1. Concetration (in x-axis) v.s. absorbance (in yaxis) The derived formula in order to calculate Cu (II) concentration of the unknown sample is shown in Appendix 2, where a detailed calculation is also placed in order show how concentration in each trial was obtained. The table below shows the calculated concentration of the unknown sample. Table 3. Cu (II) Concentration Trial 1 2 3 Absorbance 0.320 0.319 0.319 0.319 Conccentration, ppm 2376 2369 2368 2371 Cu(II)

CONCLUSION
From the results gathered from the experiment, the linear relationship of concentration and absorbance as indicated by the Beers law was observed. However, the Beers Law dont apply all e time s there are limitations to it be it chemically or instrumentally.

REFERENCES
Beer-Lmbert Law. University of Adelide, Department of Chemistry. chemistry.adelaide.edu.au. Web. 15 May 2013 Colorimetry. Clackamas Community College. depts.clackamas.edu. Web. 14 May 2013 Jeffery, G.H., Bassett, J. , Mendham, J., and Denney, R.C. 1989. Vogels Textbook of Quantitative Chemical Analysis 5th Edition. The School of Chemistry,Thames Polytechnic, London

Average

Using the average concentration, when compared to the original value of the solution which is 2500 ppm, the value obtained in the experiment only has a 5.16 % error. These errors may be due to the way the sample was analyzed. When placing he sample inside the cuvette, it is important that there are no traces of fingerprints and etc. at the sides of the cavity. Also, it is important that the cuvette is washed with distilled

Klingenberg, Joseph. 1965.Introduction to Quantitative Chemistry. Reinhold Publishing Group

Kovic, M. Colorimetry. Institute of Computer Graphics and Algorithms. cg.tuwien.ac.at. Web. 14 May 2013 Pecsok, Robert, et al. 1976.Modern Method of Chemical Analysis, 2nd edition. John Wiley & Sons Inc. Pickering, William F. 1966.Fundamental Principles of ChemicalAnalysis. Elsevier Publishing Company. Skoog, West, Holler, and Crouch. 2004. Fundamentals of Analytical Chemistry, 8th edition

APPENDIX

Appendix 1. Formula to get Cu (II) concentration

ppm working standard ppm working standard ppm working standard ppm working standard ppm working standard

(2500 ppm)(10 mL) 500 ppm 50mL (2500 ppm)(8 mL) 400 ppm 50mL (2500 ppm)(6 mL) 300 ppm 50mL (2500 ppm)(4 mL) 200 ppm 50mL (2500 ppm)(2 mL) 100 ppm 50mL

Appendix 2. Formula to get the Cu (II) concentration in the unknown sample

Ab ( Dilution factor ) m

0.320 (0.0127) 50.0 ( ) 2376.43 2376 ppm 0.01 7.00 0.319 (0.0127) 50.0 C2 andC3 ( ) 2369.29 2369 ppm 0.01 7.00 C1