ABSTRACT In this experiment, spectrophotometry is used to quantitatively determine the concentration of Copper (II) stock sample solution and Beer-Lambert’s Law was used to analyze the obtained data. Spectrophotometry measures the transmission properties of materials as a function of wavelength, analytically. It is a technique that basically uses light to determine chemical concentrations and is useful in locating equivalence points of titration. Spectrophotometry has been used to discover the Antartic “ozone hole” in 1985, which led to awareness on the vulnerability of earth’s atmosphere to radiations coming from the sun. The experiment started in the preparation of a 250 mL standard 2500 ppm Cu(II) stock solution, then the determination of the maximum analytical wavelength (λ max) was done by measuring the absorbance of the concentrated Cu(II) against reagent blank from 700 nm to 200 nm. The analytical wavelength was known to be 632.0 After this was the preparation of the calibration curve by the use of solutions with 2.00 mL, 4.00 mL, 6.00 mL, 8.00 mL, and 10.00 mL Cu(II) with absorbance recorded at 0.091, 0.182, 0.281, 0.372, and 0.491 respectively. These values were treated by linear regression to get the equation of the line which is y= 9.9 X 10-4 ― 0.0136 and linearity R2 equal to 0.9975 implying precision and accurate collected data. Henceforth, Cu (II) concentration of the stock sample was determined to be 502.36 ppm. Furthermore, the low standard deviation which is equal to 1.54, the RSD equal to 2.90 ppt and the confidence interval equal to 502.36± 3.84 means that the data are highly precise therefore making the experiment successful.

INTRODUCTION This experiment aims to quantitatively determine the concentration of Copper (II) in solutions containing different amount of Cu (II) standard stock solutions and added with ammonia (NH3) then diluted with distilled water to obtain a 50 mL solution, thru a method called spectrophotometry. Spectrophotometry is an absorption measurement wherein more of the reactants or products absorb radiation or an absorbing indicator will be added in an analyte solution [1]. Moreover, the UV-Vis spectrophotometer measures transmission properties of materials as a function of wavelength. It can measure the absorbance of the solution by a radiant energy with a certain intensity (I0) directed at a sample solution in a transparent sell or cuvette and then the solution unabsorbed is transmitted with the intensity I [3]. It was used in 1986 to measure the concentration of the Antartic atmosphere which concluded that the ozone depletion occurred after polar sunrise and that the concentration of the chemically active chlorine in the stratosphere is approximately 100 times greater than the predicted gas-phase [2] leading to awareness in the consequences that this problem might lead. Furthermore, Beer-Lambert’s Law or absorption law is used to analyze the data gathered to determine the unknown Cu (II) concentration. As shown in (1), a, accounts for how well a substance absorbs light or its

8.9 X 10-4x ― 0. and 10. 0. Cu2+ + NH3→ Cu(NH3)42+ (2) To be able to find the Cu(II) concentration of the unknown solution. 6. Calibration Curve The analytical wavelength is used because to determine the absorbance values of all the samples that will be measured at and is the basis for creating the calibration curve.00. a 2.00. 4. Then 0. and c. After this. Using a UV-Vis spectrophotometer.00. the calibration curve was prepared by measuring the absorbance of the 0. with units of ppm.00.00. 6. Then the standard stock solutions 2.00. and 10.00. 6.00 mL standard stock solution with 10 mL NH3 and diluted to mark in a 50 mL volumetric flask at the analytical wavelength maximum 632. the Cu(II) concentration of the unknown solution was determined. 4. 2. 8. making the line more accurate. Figure 1 shows the plot of absorbance against different concentrations of the standard Cu(II) with the equation of the line y= 9. the analytical wavelength was first determined by measuring the absorbance of the most concentrated working standard Cu(II) which is the solution containing 10 mL Cu(II) against a reagent blank from 700 nm to 200 nm.0.185 and 0. . The solution with no Cu (II) served as reagent blank. Subsequently. and 10.00. 2.00 mL was measured for absorbance using the analytical wavelength obtained previously. And lastly. the given solution with an unknown Cu (II) concentration was also added with 10 mL NH3 and diluted to a 50 mL volumetric flask with distilled water was subjected into three trials and its absorbance is known to be 0.2 0 0 200 400 600 Concentration of Standard Cu(II).9975 which implies a precise and accurate data. The wavelength with the maximum point implies that it is the highest level light can be absorbed in the solution thus.6 Absorbance 0.00 mL standard stock solution was placed in a 50 ml volumetric flask and to each a 10 mL concentrated NH3 was added then diluted to mark. The absorbance near peak absorption is broad and fairly constant unlike other wavelength regions where absorbance varies greatly[4] while the measure of absorbance against a reagent blank is done to lessen the choices of what analytical wavelength should be chosen. 6. This experiment needed a 250 mL standard 2500 ppm Cu(II) stock solution.4 0. the absorbance of Cu (II) was measured against 700 nm to 200 nm reagent blank by the use of a UV-Vis spectrophotometer.00 mL amount of Cu(II) was pipetted out of the Cu(NO3)2∙3H2O then was added with NH3 and diluted in a 50 mL volumetric flask with distilled water to make a standard stock solution. Ammonia (NH3) was added to convert the Cu(II) solutions into Cu(NH3)42+ species as shown in the reaction (2). ppm Figure 1. 0.00.absorptity with units of ⁄ . and 10.00.00. Monochromatic radiation still contains light of different wavelengths however narrow. 2. 4.00. absorbance of the Cu(II) solution was known to be 632.00. RESULTS AND DISCUSSION Solutions with 0.[3] A=abc (1) This method gives a quantitative measurement on how the amount of decrease of absorption depends on the concentration of the absorbing molecules and also the path length which absorption occurs [2].185.00. The analytical wavelength was obtained and plotted against absorbance by the apparatus. concentration of the component of interest.0136 with the linearity R2 equal to 0.87 g of Cu(NO3)2∙ 3 H2O crystals was then dissolved to form this solution.185.00. is the path length or the width of the cell or cuvette in which the sample is contained with units of cm .00.0 obtained in the first step. Controlling the wavelength from 700 nm to 200 nm is significant to increase the accuracy of the calibration curve. 8. 8. in this case the Cu(II). After this.186 which gives an average absorbance of 0. b.

36. Moreover.01 M can be tested because as the distance between ions or molecules become smaller in concentrated solutions. et al. It arises when the blank and analyte solutions are not of equivalent optical[1]. The Beer-Lambert’s Law or the absorption law was used to determine the unknown concentration of Cu(II) which is 502.0 maximum analytical wavelength or λmax. Fundamentals of Analytical Chemistry Eight edition. the lower standard deviation which is equal to 1.0136. Modern Analytical Chemistry. they disrupt the extent of absorption of its neighbors[1].84 this means that the data are highly precise making the experiment a success.36 ppm by spectophotometry by using the 632. One of which is testing the solutions with decreasing concentrations of Cu(II) instead of increasing Cu(II) concentration which leads to a larger concentration of the Cu(II) in the sample. however some fibers might still remain. For this experiment (4) is used to account for the absorbance. United States of America. This will lead to a lower absorbance of the solution since the translucent part will also absorb light even if it can be minimal it is not negligible and the fingerprints and tissue fibers might block the path of the light that accounts for the absorbance of the solution. Daniel C. identification of iron waste content in water. SUMMARY AND CONCLUSIONS The concentration of the sample Cu(II) was determined to be 502. Analytical Chemistry Laboratory Manual. 2013. C.. Spectrophotometry can also be used in the determination of the composition of gases. [4] Harvey.. A= log = log The Beer-Lambert’s Law also has its limitations and one of which is that it can only treat data which were measured by monochromatic radiation in spectrophotometers because light here. REFERENCES [1] Skoog. Furthermore. [2] Harris. only solutions with concentration less than 0. the RSD is equal to 2. New York City: McGraw-Hill Higher Education.36± 3.90 ppt and the confidence interval is equal to 502. Quantitative Chemical Analysis Seventh edition (Electronic-book) [3] Alcantara.54. Douglas A. The λmax was determined by measuring the absorbance of the solution against the reagent blank controlling the wavelength from 700 nm to 200 nm.. The calibration curve was then obtained with the equation of the line y= 9. The linearity implies precise and accurate data. This can be minimized by wiping the cuvette with tissue. The handling of cuvette in the transparent part instead of the translucent one. has a very narrow wavelength while polychromatic light causes a negative deviation in the law [4]. Thomson Learning. Possible sources of error can be personal which includes the handling of the UV-vis spectrophotometer.The equation of the line is y= 9. 2004. Inc. T= (3) (4) Further source of error are mismatched cells. where y is the absorbance.9 X 10-4x― 0. x is the concentration of the solution and b is the y-intercept greater than zero signifies a constant factor throughout the samples. et al. . 2000. Quezon City: UP Diliman. T is accounted (3) is used. When transmittance.9975. and transferin protein found in blood and the determination of the rate of an enzyme-catalyzed reaction in a DNA in the field of biochemistry. D. A. and directed it in the laser with the transparent part and when fingerprints are left on the cuvette.9 X 10-4― 0. m is the slope of the line which accounts for the absorptity multiplied by the path length.0136 and linearity R2 equal to 0..

36 .606060 ( X= 501.0 mL Volume of aliquot from stock solution: 20.091 0.185 0.00 8.00 10.APPENDIX Raw Data Absorption Spectrum Wavelength at maximum absorption (λ max): 632.9X10-4x – 0.0136) y= absorbance (0.0136 = X= 200. ppm Trial 1 y= 9.491 Linear equation of the calibration curve: y= 9.52 ppm Trial 2 y= 9.185) 0.9x 10-4x + (-0. mL 2.185) – Absorbance 0. ppm 100 200 300 400 500 Absorbance 0.182 0.00 4.9x 10-4x + (-0.372 0.00 6.185 0.00 Concentration of Standard Cu(II).0136) y= (0.0136) Sample Analysis Volume of stock sample solution: 50.185 Concentration of Stock Sample Cu(II).0 mL Volume of diluted unknown solution: 50.0 mL Trial 1 2 3 Average Calculations Determination of the Concentration of Stock Sample Cu(II).185= 9.0 Calibration Curve Concentration of working standard Cu(II) solution: 2500 ppm Volume of Working Standard Solution.281 0.9x 10-4x + (-0.04 502.52 504.186 0.52 501. ppm 501.

606060 ( X= 501.84 .36 ppm Standard deviation s= 1.185= 9.186= 9.0136 = X= 200.9X10-4x – 0.0.90 ppt Confidence Interval at 95% level of confidence Confidence limit=X± √ – – Confidence limit= 502.545 Relative Standard Deviation (RSD) RSD= X 1000 ppt RSD= X 1000 ppt RSD= 2.9X10-4x – 0.186) 0.52 ppm Trial 3 y= 9.04 ppm Average Concentration= 502.616161 ( X= 504.36± 3.0136 = X= 201.0136) y= (0.9x 10-4x + (-0.