Resolution of a Two-Component Mixture

Gabrillo. Shaira L.
BS-Chemistry, University of San Carlos – Talamban Campus

Two-component mixture containing cobalt(II) and chromium(III) is analyzed using ultravioletvisible spectrophotometry to determine concentration of individual components. Wavelength
as to which each component absorbs most light is manually determined using Shimadzu UV1601 double beam spectrophotometer. Absorbance of both components at the two
maximum wavelengths were plotted to obtain equation of the line which was used to
calculate the concentration of cobalt(II) in the cobalt solution and of Cr(III) in the chromium
solution. Experimental total absorption was observed as roughly the sum of the absorbance
of each component at a given wavelength. Concentration of Co(II) and Cr(III) in the mixture
was calculated based on the additive property of absorbance in solutions.
I. Introduction
In the field of analytical chemistry,
various graphical and numerical spectral
methods have been applied to the routine
preparations containing complex mixtures.
extensively for binary mixture analyses.
theoretical basis and applications of
laboratory, it is sometimes necessary to
determine individually the concentrations
of two or more substances in the same
solution. There are instances when the
presence of one species in a sample does
not influence the measurement of another
species in the same sample; i.e., they do
not interfere. Thus, the absorption of light
by the components of the sample solution
is additive; that is, the total absorption of
light at any given wavelength is just the
sum of the absorbances the two
substances would show if measured
individually under the same conditions [2].
If both substances do not absorb light at
each other's absorption maximum, then
the measurements can be done by
reading the solution absorbance at both
substances, and determining each from
separate beer law plots. However, though
this is ideal and very probable in theory, it
seldom happens that two absorption
spectra are independent of each other.
Usually at the wavelength maximum of
component 1, component 2 almost always
has at least some light absorbing
character. Given this, the absorbance at

each of the two wavelengths is composed
of contributions from both components,
and the problem arises of how to separate
them [3].
Ultraviolet–visible spectroscopy or
ultraviolet-visible spectrophotometry (UVVis or UV/Vis) is routinely used in
analytical chemistry for the quantitative
determination of different analytes. UV/Vis
spectroscopy uses light in the visible and
adjacent (near-UV and near-infrared)
ranges to measure the wavelength of the
energy absorbed by specific chemical
species. The absorption or reflectance in
the visible range directly affects the
perceived color of the chemicals involved
To solve the problem of how to
concentration separately from a mixture, a
possible case would be to read the
absorbance using UV-Vis. At a certain
wavelength, analyte 1 barely absorbs any
energy and analyte 2 strongly absorbs,
and at another wavelength the converse
was true. Even though the analytes
absorb, albeit weakly, across the spectra,
by choosing wavelengths where the
absorption of 1 is strong and 2 is weak,
and vice versa, it is still possible to
determine their concentrations because
the absorbances are additive [2].
In this experiment, a mixture of
Co2+ and Cr3+ will be resolved by
measuring the absorbance of the mixture
at the measured λmax of each component
using UV-Vis spectroscopy. A calibration
curve is then plotted using the absorbance
of both chemical species at both the λ max.
By using these two different wavelengths,
it is possible to generate two equations for

Table 1. Absorbance readings of solutions at each of the wavelengths listed were recorded for later usage. Results and Discussions Absorbance is an additive property of solutions. 570. λmax obtained for each component was then used for absorbance measurements of the aliquots of the solution mixture. Wavelengths at which cobalt and chromium show maximum absorbance values (λmax) were recorded for analysis. Solutions for a Beer’s law plot of Cr3+ were prepared by diluting 5ml. Approximately 2.C. The total absorbance of several components at a single wavelength is equal to the sum of the individual absorbance values that would be measured if each component were separate in its own cuvette. For each solution. 480. Clean cuvettes were pointedly used throughout the whole wavelength determination procedure. and 580-nm. Preparation of Sample Solution Approximately 5. λ determination data. λmax of the individual component solutions was manually determined. II. . and 20ml of the cobalt solution separately into three 25ml volumetric flasks.4700g of solid Co(NO3)2∙6H2O was weighed in an analytical balanced and was transferred into a 100-mL beaker where it was dissolved in minimal distilled water.B. 500.0500 M Cr3+ solution was prepared from solid Cr(NO3)3∙9H2O. Absorbances for the corresponding λmax of each component were then then measured and plotted to obtain an equation relative to each component’s concentration.A. λmax determination for Co2+. A wavelength vs absorbance plot was generated for both cobalt and chromium solutions from the absorbance values recorded using the UV-Vis spectrophotometer. absorbance values of the unknown solution mixture were measured. Using the both λmax obtained from the two component solutions. concentration of Co2+ and Cr3+ were calculated.0000g of solid Cr(NO3)3∙9H2O was weighed in an analytical balanced and was transferred into a 100-mL beaker where it was dissolved in minimal distilled water. molar absorptivity of Co2+ and Cr3+ at both λmax was calculated. From the molar absorptivity and absorbance of each component. Starting at 375nm. 490. the absorbance values of both component solutions were measured at every 25nm interval including wavelengths 470. The resulting solution was quantitatively transferred into a 100-ml volumetric flask where it was diluted up to the mark.1880 M Co2+ solution was prepared from solid Co(NO3)2∙6H2O while 0. Beer’s law Plot for Cobalt and Chromium Solutions and Data Analysis Absorbance values were plotted on MS Excel. 575. 0. 10ml. II.both components. 10ml. Solutions for a Beer’s law plot of Co2+ were prepared by diluting 5ml. Spectrophotometric Determination Using Shimadzu UV-1601 double beam spectrophotometer. Using the equation of the line generated in the plots. and 20ml of the cobalt solution separately into three 25ml volumetric flasks. λmax was determined manually using a UV-Vis spectrophotometer. Compound mixture containing unknown ratio of cobalt(II) and chromium(III) ions was prepared by the instructor in a 25ml volumetric flask. R2 value and equation of the line was automatically determined along with the generated graph. Solving them simultaneously will give the concentrations of the two components. Materials and methods This experiment was performed in the instrumentation room and organic laboratory of the University of San Carlos – Talamban Campus. was recorded to observe λ max of each component. II. Shimadzu UV1601 double beam spectrophotometer was used to measure λmax of each solution and of the mixture. II. Mixture was diluted to 25 mL prior to absorbance determination. as shown in tables 1 and 2 below. III. The resulting solution was quantitatively transferred into a 100-ml volumetric flask where it was diluted up to the mark.

173 0. λmax determination for Cr3+.54 8 0.54 8 0.069 0.443 0.03 1 0.126 0.54 8 0.108 0. Absorption of Cr3+ at λmax of Cr3+ and Co2+ Cr(NO3)3 volum e ∙ aliquo 9H2O t T1 10m L 20m L Absorbance T2 T3 T4 Ave . 0. Data recorded.332 0.303 0.05 4 0.54 8 0.03 1 0.045 0.213 0.063 0. shown in graphs 1 to 4: Table 3.124 0. and 5.2 0 0.15 0 0.159 0.199 0.032 0. were plotted in the absorbance vs concentration scattered graph.25 2 0.236 0.144 0.15 1 0.246 0.037 0.303 0.458 0.302 0.140 0.304 0.16 9 0.276 0.050 0.25 3 0.075 0.12 7 0.017 375 400 425 450 470 475 480 490 500 525 550 570 575 580 600 20mL 0.331 0.383 0.248 0.49 3 Ave .09 1 0.119 0.218 10m L 20m L 5 mL 0.03 0.54 8 0.04 9 0.092 0.332 Table 4.11x + 0.28 9 0.28 9 0.01 8 0.49 3 From the measured absorbance of the individual components of the mixture.403 0.033 0.303 0.28 9 0.331 Avera ge 0.084 0.082 0. calibration curves showing concentration vs absorbance were plotted (graph 1 to 4) using excel to show the precision of data and the equation of the line which can be used to solve for each component’s concentration: .054 0. as shown in tables 3.095 0.054 0.081 0.6 Table 2.519 0. 0.153 0.489 0.01 0.02 0.028 5mL 0.055 0.09 1 0.λ(nm) Absorbance 10mL 0.026 0.16 9 f(x) = 11.545 0.01 8 0. Absorbance 10mL 0.12 8 0.04 9 0.098 0.119 0.03 1 0.491 0.419 0.05 3 0.16 9 0.03 1 0.05 4 0.128 0.148 0.248 0.228 0.112 375 400 425 450 470 475 480 490 500 525 550 570 575 580 600 0.4 Absorbance 20mL 0.170 0.260 0.25 3 0. Absorption of Co2+ at λmax of Cr3+ and Co2+ Co(NO3) volum e 2∙ aliquo 6H2O T1 Absorbance T2 T3 T4 t @400 nm @500 nm 5 mL 10m L 20m L 5 mL 10m L 20m L 0.16 9 0.087 0.109 0.106 0.023 0.047 5 mL @400 nm @500 nm λmax λ(nm) 5mL 0.049 0.090 0.49 3 0.065 0. M Table 5.09 1 0.16 9 0.142 0.478 0.333 T2 Absorbance T3 T4 0.25 2 0.12 8 0.252 0.49 3 0.28 9 0.023 0.186 0.09 1 0.05 4 0.127 0.05 λmax Concentration.04 0. 4.488 0.15 1 0.131 0.15 1 0.01 8 0. absorbance values of individual components and the mixture were read.03 1 0.02 R² = 1 0.094 0.050 0. Absorption of mixture at λmax of Cr3+ and Co2+ λmax T1 (nm) 400 500 0.227 0.04 9 0.080 0.148 0.04 9 0.021 0.032 0.030 0.282 0.360 0.05 4 0.208 0.28 9 0.046 0.048 0.49 4 0.426 0.15 1 0.01 8 0.12 8 0.12 8 0.09 1 0.128 0.280 0.04 9 0.148 0.026 0.01 8 0.098 0.25 3 0.249 0.283 Based on the λmax obtained.

6 0. which in turn would be used to calculate the concentration of the individual components of the mixture. M Graph 4. Concentration values obtained were to be used in calculation for the molar absorptivity (equation 1) of each component. Calibration curve of Cr3+ at 500nm 0.15 0. Calculation of concentrations of Co2+ and Cr3+ in the unknown mixture makes use of the equations of the line obtained from each calibration curve of the individual components.1 f(x) = 3.15 Concentration. M NOTE: For concentration calculation. Molar absorptivity formula Equation 2.2 0 0 0.02 0. Calibration curve of Cr3+ at 400nm 0. Calibration curve of Co2+ at 500nm Graph 3.05 0.05 0.04 0.02 0 0 0 0.32x + 0. and then solving for x (see Calculations section). Calibration curve of Co2+ at 400nm 0.01 R² = 1 0. Formulafor concentration using the additive property of absorbance Making use of the additive property of absorbance in solutions.03 0.2 Absorbance 0.01 R² = 1 0. Equation 1. M Concentration.Graph 1.2 Precision test (R2) results were greater than 0. Concentration of Co2+ and Cr3+ were calculated by substituting the maximum absorbance at the corresponding wavelength of each component into y of the corresponding equation of the line.1 0.15 f(x) = 0.06 0.05 0.01 R² = 1 0. refer to Calculation section of the article.1 0.36x + 0.2 Concentration. concentration .4 Absorbance f(x) = 3.05 0. Graph 2.04 Absorbance 0.998 indicating that precision of data acquired is within acceptable range.01 0.22x + 0.

Conclusion Ideal two-component mixture analysis contains components that absorb light at a specific wavelength and does not interfere with light absorption of the other species at a different wavelength. John Wiley & Sons. [4] . 1984. concentration of individual components can be calculated using equation 2 by substituting the appropriate values into the equation (See Calculation section).02442M Cr3+. however. VI. Heineman.Skoog. [2] Chemistry Experiments for Instrumental Methods. Experimental total absorption can still be observed as more or less the sum of the absorbance of each component at a given wavelength. ERDAL DINÇ. James. Calculations VIII.. Douglas A. F. VI. References [1] ALTERNATIVE APPROACHES TO THE SPECTRAL QUANTITATIVE RESOLUTION of TWO-COMPONENT MIXTURE BY WAVELET FAMILIES.02004M Co2+ and 0.of each component of the mixture can be calculated. Holler. Stanley R.). AND DUMITRU BALEANU. Sawyer. (2007). Crouch. Since it is established that absorbance reading at 400 nm is the sum of the absorbance of Cr3+ and Co2+ at 400 nm (the same is true for absorbance at 500 nm). Using the additive property of absorbance and calculating for concentration of cobalt(II) and chromium(III) in the unknown mixture gave the concentration values of 0. [3] Principles of Instrumental Analysis (6th ed. albeit the small deviations from the theoretical value. each component almost always absorbs some light from the λmax of the other. Beebe. In application. FAHRETTIN ARSLAN. 2009.