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Determining the k a of an acid-base indicator. Experiment 3. Indicator. HIn + H 2 O ↔ In - + H 3 O + Red blue Ka = [H 3 O + ] [In - ]/[HIn] [H 3 O + ] = Ka [HIn]/[In - ] (-log [H 3 O + ]) = (-log Ka) + (log [In - ]/[HIn]) pH = pKa + log [In - ]/[HIn]. Determining Ka.
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Determining the ka of an acid-base indicator Experiment 3
Indicator HIn + H2O ↔ In- + H3O+ Redblue Ka = [H3O+] [In-]/[HIn] [H3O+] = Ka [HIn]/[In-] (-log [H3O+]) = (-log Ka) + (log [In-]/[HIn]) pH = pKa + log [In-]/[HIn]
Determining Ka • Ka = [H3O+] [In-]/[HIn] • Adjust pH slightly and measure the concentration of all three species, independently • H3O+ (pH meter) • In- and HIn (spectroscopy)
Quantitative Spectroscopy • Beer’s Law Al1 = el1bc e is molar absorptivity (unique for a given compound at l1) b is path length c concentration
Standard Practice • Prepare standards of known concentration • Measure absorbance at lmax • Plot A vs. concentration • Obtain slope • Use slope (and intercept) to determine the concentration of the analyte in the unknown
Spectroscopy • Indicators give fairly broad visible absorption spectra • Blue solution absorbs in the red region • Red solution absorbs in the blue region • But there is usually some significant overlap
Absorption measurement Must make measurements at two wavelengths Al1 = el1HInbcHIn + el1InbcIn- Al2 = el2HInbcHIn + el2InbcIn- Must have the slopes of four Beer’s law plots. Then you have two equations with two unknowns.
The four slopes • Measure standard solutions of Indicator adjusted to high pH (three pH units above pKa) [In-]/[HIn] = 1000 • Measure standard solutions of Indicator adjusted to low pH (three pH units below pKa) [HIn]/[In-] = 1000 • These are provided for you.
In the lab • Prepare Indicator solution adjusted to the proper pH • Measure its pH and Absorbance at 430 nm and 565 nm • Adjust pH with dilute HCl or NaOH by .2 pH units and remeasure • Continue until you have five measurements above and below the starting pH
Spreadsheet • Type in pH and corresponding absorbance measurements and the spreadsheet will calculate Ka and pKa values automatically • Calculate [H+] from the pH • Solves simultaneous equation for [HIn] and [In-] • Solves for Ka, Ka = [H+][In-]/[HIn] • Solves for pKa, pKa = -logKa • Calculates a standard deviation