Determination the Calibration Curve of Cobalt Nitrate by Spectrophotometer

1. INSTRUMENTAL ANALYSIS LAB. Mr Haydar A.M.S BSc, MSc Faculty of Science Chemistry Department Email: [email protected]

2. Exp. 3 Determination the Calibration Curve of Cobalt Nitrate by Spectrophotometer Objectives In this ­­experiment, you will Prepare a set of Co +2 standard solutions of known concentrations, Measure the A or %T of the known solutions and construct a calibration curve, and Measure the A or %T for unknown solution and determine its concentration from the calibration curve.

3. Ultraviolet/Visible Molecular Absorption Spectrometry The instrument used to measure the amount of light absorbed by the sample is a spectrophotometer .

4. Components of a Spectrophotometer 4

5. 5 Wavelength Selectors

6. Light source Grating Rotating the grating changes the wavelength going through the sample slits slits Sample filter Phototube Wavelength Selectors When blank is the sample I o is determined otherwise I is measured Separates white light into various colors detects light & measures intensity - white light of constant intensity 6

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8. 8 Sample Containers

9. Range

10. Transmittance (T) is defined as the amount of light passing through the sample solution (I) divided by the amount of incident radiation (I o ). T = I/ I o Transmittance is often expressed as a percentage or %T = I/I o × 100% The Absorbance (A) of a medium is defined by the equation: A = log I o /I = -log T

11. As the radiation is absorbed in the sample, the total intensity of radiation is reduced as it travels through the sample. This results in a non-linear relationship between transmittance and concentration Absorbance (A) is based on the amount of light absorbed by the solution and is defined as the log of the inverse of the transmittance. A = log 10 (1/T) Within limits, the relationship between absorbance and concentration is linear.

12. The blank The blank contains all substances except the analyte . Is used to set the absorbance to zero: A blank = 0 This removes any absorption of light due to these substances and the cell. All measured absorbance is due to analyte . Analyte is a substance undergoing analysis.

13. Beer’s law Beer’s law states that the amount of light absorbed by a solution ( colored ) is directly proportional to the path length (b) through the medium and the concentration (c) of the absorbing species, A (absorbance) = log I o /I = a b c Where (a) is the proportionality constant called the absorptivity b is the path length of the sample i.e. the inside cross section of the sample cuvette (cm ). When the concentration is expressed in ( mol /L) , the absorptivity is called the molar absorptivity and is given the symbol (є) A = є b c (units of є = L.mole -1 .cm -1 )

14. Determination the calibration curve of cobalt nitrate by spectrophotometer

15. Procedure 1. Obtain TWO matched cuvettes, the first cuvette is for distilled water and the second one is for the cobalt solution. 2. Prepare the following solutions From stock solution (0.2M) Co(NO­­ 3 ) 2­ .6H 2 O (M) Absorbance (A) 0.02   0.04   0.06   0.08     Part A Calibration Curve

16. Procedure Continued 3. Set the wavelength dial 505 nm 4. Measure the absorbance of the blank (water). 5. Measure the absorbance of the cobalt solutions and record the absorbance 6. Empty and rinse the cuvettes thoroughly with water. 7. plot the absorbance versus concentration 8. Measure LOD, LOQ and sensitivity

17. Procedure Continued 1. Measure the absorbance of the unknown concentration 2. use a calibration curve that you plot it in part A and determine the concentration of the unknown solution. Part B Determination the absorbance of the unknown

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