Dr. Du Education Lecture Notes Year 12 Chemistry Module 8: Applying Chemical Ideas Spectroscopic Analysis (II) Name: Class: Week: Term: Year: Ultraviolet-visible spectrophotometty.........scssssssssssssnesseennneesvensteYear 12 Chemistry Spectroscopic Analysis (l!) Content Analysis of Inorganic Substances Inquiry question: How are the ions present in the environment identified and measured? Students: + analyse the need for monitoring the environment + SI # + conduct qualitative investigations —using flame tests, precipitation and complexation reactions as appropriate = to test for the presence in aqueous solution of the following ions: ®™ i = cations: barium (Ba), calcium (Ca), magnesium (Mg°*), lead(II) (Pb), silver ion (Ag"), copper(II) (Cu), iron(ID) (Fe), iron(IN) (Fe) ~ anions: chloride (C1), bromide (Br), iodide (1+), hydroxide (OH?), acetate (CHsCOO ), carbonate (COs? ), sulfate (SO,*>), phosphate (PO,) + conduct investigations and/or process data involving: = gravimetric analysis — precipitation titrations + conduct investigations and/or process data to determine the concentration of coloured species and/or metal ions in aqueous solution, including but not limited to, the use of: — colourimetry = _ultraviolet-visible spectrophotometry atomic absorption spectroscopy ©Daily Progress Version 2Year 12 Chemistry Spectroscopic Analysis (Il) Ultraviolet-visible spectrophotometry Two quantitative techniques which use the Beer-Lambert Law, Colorimetry and Atomic Absorption Spectroscopy (AAS), have been previously covered. They require standard curve preparation and subsequent analysis. This is also the case with another spectrophotometric method: Ultraviolet-visible (UV-visible) spectrophotometry using a spectrophotometer or spectrometer. When the energy from UV or visible light is absorbed by a molecule, one of its electrons jumps from a lower energy to a higher energy molecular orbital, Unlike the use of a filter in colorimetry, UV-visible spectrophotometry uses radiation with a range of wavelengths, selected via a monochromator, which are directed through a sample of interest, and a detector records which wavelengths were absorbed and to what extent the absorption occurred. The range of wavelengths is: UV (200-400 nm) and visible (400-760 nm). There are two light sources to cover the entire UV/Vis range: ‘+ a deuterium discharge lamp for a continuous spectrum in the UV region between 160 and 375 nm ‘© a tungsten filament globe for the visible providing radiation over a range of 350-2500 nm (at 3000 K) ‘The solution that is to be analysed is scanned across a range of wavelengths to select a specific wavelength that shows maximal absorbance and minimum interference from other components in the same sample (see Figure 4 in later section), Absorption of electromagnetic radiation Molecular absorption of electromagnetic radiation occurs via many modes ~ these changes of energy state of a molecule include: = electronic state (electron transitions between orbitals), AE(e) = AE(electronic) = 150—600 kJ mol”? —60 k} mol" = vibrational state / = AE(vibrational) = rotational state AF(r) = AE(rotational) ~ 3 kJ mol! Notice that the energy required to shift electrons to higher orbitals is the largest absorber of radiation energy. However, the wavelength of radiation absorbed is dependent on the total energy that is absorbed. This is shown, below with some simple equations: AE \E(e) + AE(v) + AE(r) Total energy consists of different modes of absorption nis Planck’s constant = 6,626 x 10 Js OE =hx f {fis the frequency of the radiation (in Hertz, Hz) ¢ is the speed of e-m radiation = 3.00 x 10 ms! c=fxa 1. is the wavelength of the radiation (m or nm) ag =" ~ © The total energy (J) that is absorbed is inversely related to wavelength ‘The wavelengths at which a compound absorbs light depends on the energy gap between the orbitals that are ion. This energy gap is determined by the structure of the molecule. A molecule with only single bonds absorbs light at shorter wavelengths (in the high energy UV) as shown in the NESA-provided table below. More complex bonding pattems are associated with the absorption of visible light involved in the trai Daily Progress 1 Version 2Year 12 Chemistry Spectroscopic Analysis (I) Chromophores A chromophore is a group of atoms responsible for the UV/Visible absorption of a molecule, e.g. double bonds C=C, C=C-C=C, C=O, N=N and aromatic rings. The second bonding arrangement of alternating single and double bonds is referred to a conjugated double bond system, Some chromophores that absorb in the UV region are provided in your data sheets (from NESA) and have been reproduced below, Tuble / gives a broad grouping of the types of absorbing compounds. Table 2 provides a list of wavelengths that are observed and complementary colours (for visible part of the spectrum). UY absorption (This is not a definitive list and is approximate.) Chromophare By, (Hm) Chromophore [Aas Cam) 173178 cH 122 c 196 22 cc 135 coal 13 c=c 162 c—Br 208 Spectral regions Region 2 (am) Absorbing compounds | Far ultraviolet <[Link] Saturated (Single bonds) and mono-unsaturated (one double ot (vacuum UV region) triple bond) y-uns iny double bonds) and aromatic (ring (Near) ultraviolet 190—[Link] Polyunsaturated (many doubl ) Hic (ring structure, benzene-type) Visible light region 380—780 nm coloured Table I: Main types of absorbing compounds Visible light absorption 2.(am) Colour of light Colour of absorbing substance 400-435 | viotet yellow-green 435—480 blue yellow 480—490 ‘green-blue orange + 490-500 blue-green red-orange 500—S60 ‘green red | 560 —sa0 sreen-yellow violet 580—595 yellow-orange blue - 595—620 red-orange sgreen-blue 620760 red blue-green Table 2: A list of absorption wavelengths and complementary colours Obaily Progress 2 Version 2Year 12 Chemistry Spectroscopic Analysis (I) Question 1 Calculate the frequency of the absorbing radiation if a solution is observed to be red-coloured. Question 2 ‘An orange solution absorbs at 485 nm, Calculate the energy absorbed by this radiation. Absorbing species Organic compounds Double and triple bonds and non-bonded electrons on atoms such as N, O and the halogens are the structural features that cause absorption in the UV/Visible region. ‘Compounds without the multiple bonds and non-bonded electrons absorb below 200 nm. However, measurements in the far UV (< 200 nm) require a vacuum, since the components of air absorb strongly (double (O-O) or triple (NaN) bond and lots of non-bonded electrons: 4 on each oxygen, 2 on each nitrogen). ‘This makes for a less convenient, more complex spectrometer and too much interference for quantitative analysis. So, when we talk about UV spectra, we are actually referring to near UV. Fora compound to absorb strongly above 200 nm, it requires a number of multiple bonds in conjugation (alternating single-multiple) and/or a number of atoms with non-bonding electrons. The following compounds (in Figure 1) show the effect of the presence or ¥ absence of absorbing groups. The wavelength in nm of maximum absorption ¥. we a is also shown. Abbreviated structural formulae are given. The abbreviated jf ‘i 2 O structural formula for benzene is compared to its (full) structural formula. am v a Q/0|/0|e*|co <200 210 256 282 312 Oo oOo Co co on ‘OH 2H 260 265 270 | 280 300 Figure 1: The effect of conjugation and non-bonding electron pairs Daily Progress 3 Version 2Year 12 Chemistry Spectroscopic Analysis (i) For an organic compound to absorb in the visible region (even if the peak is not past 400 nm), it will need numerous conjugated double bonds and non-bonding electrons. Figure 2 shows the structure of methyl orange (the yellow form in a neutral solution) to illustrate this. wos) nen) WOH, Figure 2: Methyl orange Inorganic compounds Some simple metal ions absorb in the ultraviolet or visible region, because they have valence electrons, e.g. Cu®* and Ni*, but usually what absorption there is, is not particularly strong, and therefore, not useful for quantitative analysis. In the majority of cases, complexes of metal ions and ligands are needed for intense absorption, Polyatomic ions, such as permanganate and dichromate, get their much stronger absorbance from a combination of factors: multiple bonds and non-bonding electrons, Question 3 Compare the structures of cyclohexane and phenol to explain the difference in the wavelengths of maximum absorption (given in nm). OH <200 nm 270 nm cyclohexane phenol Question 4 ‘Compare the structures of toluene and phenol to explain the difference in the wavelengths of maximum absorption (given in nm). OH | 260 nm 270 nm toluene phenol ©baily Progress 4 Version 2Year 12 Chemistry Spectroscopic Analysis (I) Question 5 Identify the colour and wavelength of absorption of methyl orange and list the absorbing groups that allow for this to occur. UV-visible spectra Figure 3 below is a simplified absorbance spectrum of a molecule which absorbs light in the UV range. (It is uncommon to have only one peak.) Peaks in UV spectra tend to be quite broad. Typically, there are two features Yamax= 260 nm that we look for: ‘© The peak wavelength Ass, which is the wavelength at maximal light absorbance. In this case hos = 260 nm. ‘* How much light is absorbed at ws? The absorbance value at 260 nm (Aaco) is about 1.0 in this spectrum, Absorbance Wavelength (nm) Figure 3: dows for maximum absorbance Figure 4 Shows how one component of a mixture UV-visible spectrum can be selectively analysed by comparing the absorbance spectra, Solution A Solution B The wavelength to measure the absorbance of Solution B in this mixture cannot be 400 nm because there is also significant absorbance by solution A at this wavelength, A better wavelength to use is about 630 nm as there is negligible interference from solution A. ao se” oe ‘Wavelength (nm) Absorbance Figure 4: The suitability of hoax for analysis ©baily Progress 5 Version 2Year 12 Chemistry Spectroscopic Analysis (I) Question 6 UV-visible spectroscopy was used to measure the spectra of two solutions, A and B, Solution A was a pink colour, while Solution B was a green colour. ‘An analyst recorded the absorbance of each solution over a range of wavelengths on the same axes, ‘The resultant absorbance spectrum is shown below UV-visible spectrum Solution A Solution B 04 Absorbance 300 400 500 600 700 Wavelength (nm) (a) If 10.00 mL of Solution A was mixed with 10.00 mL of Solution B, which wavelength should be used to measure the absorbance of Solution B in this mixture? Justify your answer. (b) The analyst used two sets of standard solutions and blanks to determine the calibration curves for the two solutions, The absorbances were plotted on the same axes. The graph is shown below. Os 04 absorbance 0.3 ot {_ ° 4 8 mR 16 20 concentration (mM) 00 ‘The analyst found that Solution A had an absorbance of 0.2 when it was measured at the appropriate wavelength. If Solution A was cobalt(II) nitrate, Co(NOs)2, determine its concentration in mg L” M(Co(NOs):) = 182.9 g mol ©baily Progress 6 Version 2Year 12 Chemistry Spectroscopic Analysis (tl) (©) In another mixture, the pink compound in Solution A and the green compound in Solution B each have a concentration of approximately 1.5 * 10? M. Could the analyst reliably use both of the calibration curves to determine the concentrations for Solution A and Solution B by UV-visible spectroscopy? Justify your answer. Question 7 A green dye used to colour tinned peas is made by mixing the yellow chemical, curcumin, with another colouring agent called bright blue. ‘The individual spectra of curcumin, bright blue and a typical green dye are represented below. bright blue typical green . dye mixture Absorbance Xs, Wavelength (nm) 400 500 600 700 (a) UV-visible spectroscopy is used to analyse the curcumin content of tinned peas. Which wavelength below would be best used for absorbance measurements to determine the curcumin content of the peas? Justify your choice. 400 nm, 450 nm, 500 nm, 550 nm, 600 nm, 650 nm {8} 400 nm ‘The absorbance used for the analysis of the curcumin content should be one at which curcumin absorbs strongly but the other colouring agent present, bright blue, does not. Curcumin absorbs most strongly between 400 nm and 500 nm. Of the wavelengths suggested within this span, bright blue would absorb least, and have less impact, at 400 nm. Tn one analysis, a stock solution of curcumin was prepared by dissolving 0.100 g of curcumin (CalHas06; molar mass = 368 g mol) in 250.0 mL of water. ‘The stock solution was then diluted to make the following three standard solutions. standard solution curcumin concentration in g Lt standard | 1.00 = 109 standard 2 5.00 « 10° standard 3 1.00 « 107 ©Daily Progress 7 Version 2Year 12 Chemistry Spectroscopic Analysis (i) ‘The absorbances of the standard solutions at the chosen wavelength were measured and the following calibration curve was obtained. 020 absorbance 0.10 standard 1 standard 2 standard 3 curcumin concentration in g L-) (b) Calculate the concentration of curcumin in the stock solution in mol L-', (©) What volume of water must be added to 10.0 mL. of the stock solution to make standard 3? (@) The curcumin in a 9.780 g sample of peas was extracted into solution. The solution was filtered and made up to 100 mL in a volumetric flask. The absorbance of the sofution at the wavelength used in the above calibration was found to be 0.170. Calculate the curcumin content of the peas in milligram per gram of peas. ©baily Progress. 8 Version 2LAs Level 8, 11-15 Deane St, Burwood Phone: 9715 2622 Level 2, 182 Forest Rd, Hurstville Phone: 9586 3969 Email: office@[Link] [Link] Office Hours: Monday to Sunday: 10am — 6.30pm