Experiment No.

6 Atomic Absorption Spectroscopy (AAS)

CHEM 127.1 - MAD Duro, Marlon Espiritu, Kevin Sotelo, Tiffany

INTRODUCTION

Basics
One of the most common methods used for analyzing metals in a sample Uses absorption of emitted light from free atoms Used for qualitative and quantitative analysis
Requires standards of known concentration for quantitative analysis

Basics
Involves atomization at very high temperatures The light emitted from sample give a line spectrum
AAS detects the emission from first half of excitation process (during absorption and before transition to excited state)

Notes
Very selective elements have different sets of energy levels Follows Beer-Lambert law
Limited by quality of monochromator Bandwidth of absorbing species must be broader than light source Fixed by narrowing radiation sources

Basic Parts
1. 2. 3. 4. 5. Lamp Atomizer Nebulizer Monochromator Photomultiplier tube (PMT)

Lamp
Hollow Cathode Lamp (HCL)
Most common lamps Provides constant yet intense light line for element of interest Cathode is made of same element analyzed and contains low pressure inert gas Light from HCL goes through glass transparent to UV-Vis region Generates very narrow spectral lines

Lamp

Lamp
Electrodeless Discharge Lamp (EDL)
Contains small amount of analyte in form of salt/metal Narrower spectral lines

Deuterium (D2)
Used for background correction Limited wavelength range (190-320nm)

Continuum sources

Atomizers
Flame Destroys analyte ions and complexes Involves the following processes:
Desolvation, vaporization, atomization, ionization

Creates elemental form of element of interest Used for liquid or dissolved samples

Flame Atomizer

Nebulizer
Controls flow rate of sample Mixes fuel and oxidant pressure generated sucks sample through tube
Fuel is usually acetylene

Creates an aerosol of the sample aerosol + fuel + oxidant

heterogeneous mixture that goes to the burner Leftover sample goes to glass waste container

Monochromator
Filters specific bands for the element of interest for entry to the PMT The line from the light source (usually HCL) is isolated Allows light not absorbed by sample to pass through

Photomultiplier Tube
The light is detected by the PMT PMT readings comes from presence of analyte in flame The elemental form of the analyte absorbs light and the corresponding decrease in the PMT reading is transformed into analytical data

Apparatus

Procedure
Fit the specific light source lamp to the lamp housing, and switch on the instrument. Light the source lamp, adjust the wavelength dial of the spectroscope to the wavelength of the analytical line specified, and set at an appropriate current value and slit-width. Using the supporting gas and combustible gas specified, ignite the mixture of these gases, adjust the gas flow rate and pressure, and make the zero adjustment after nebulizing the solvent into the flame. Nebulize the test solution or the standard solution or control solution prepared by the method prescribed elsewhere, and measure the absorbance.

Analysis
Calibration curve Method
Preparation of a standard curve, followed by measurement of the adsorbance of the unknown.

Standard Addition method

To equal volumes of more than 2 of different test solutions, add the standard solution so that the stepwise increasing amounts of the object element are contained in the solutions, and add the solvent to make a definite volume.

Standard Addition Method

Beer-Lambert Law: Unknown:

A = kC A0 = kCx

C x V x C s Vs Unknown + standard: A k V VT T

Standard Addition Method

Dividing A by A0:

C x Vx C s Vs A A0 C x VT
A A 0 VT C s C x V x C s Vs C xCs Vx 1 Vs C C s x

Working Equation:

A y A 0

VT C s

Experiment:

DETERMINATION OF TRACE LEVELS OF COPPER AND LEAD IN VEGETABLE SAMPLES USING THE ATOMIC ABSORPTION SPECTROPHOTOMETER

Copper (Cu)
A micronutrient needed for the absorption and transport of iron in the blood stream. RDA: 900 g / day Main sources are shellfish, beans and mushrooms. Also found in vegetables in trace amounts.

Experimental
Preparation of stock solutions
0.5 g of Cu(s) dissolved in 1:1 nitric acid Diluted to 250 mL in a vol. flask Take 5 mL aliquot and dilute to 100 mL Stock solution: 100 ppm Cu

Experimental
Preparation of standard solutions
5 100-mL vol. flasks (1-5) 0.50,1.25, 2.50, 5.00 mL of the stock solution added to flasks 1-4 Flask 5 is reagent blank.

Experimental
Preparation of sample
From 1 kg of vegetables, dried for two weeks 2 g of dried leaves used. Digested in conc. HNO3 for 20 min 10 mL d. H2O added and filtered while washing out the filter paper into 50-mL vol. flask

Experimental
Analysis of vegetable sample
Absorbance of standardd soln and each sample recorded using the AAS using the setting for Cu. Solution was diluted 1:5 if absorbance is too high. Three trials performed

Experimental
Standard addition
10 mL digested solution in 5 50-mL vol. flasks 0.00, 1.00, 2.50, 5.00, 10.00 ppm of added standards then diluted to the mark. Absorbance of each solution measured. Plot of A/Ao versus Vs made.

Calibration Curve
Standard blank ppm Cu 0.0
0.8

A'
0.7 R = 0.9941

0.0123 0.0000

1
2 3 4*

1.0
2.5 5.0 10.0

0.1937 0.1814
0.6

0.3685 0.3562 0.6789 0.6666 1.1751 1.1628

A' 0.4 0.5

0.3

0.2

0.1

0
0 1 2 ppm Cu 3 4 5

Analysis of Vegetables
kangkong
Sample 1 A 0.0498 A' 0.0375 ppm Cu soln 0.09945 sample 2.486

Sample computations: A = 0.0375 Vs = 50.0 mL wsample = 2.00 g Solution: ppm Cu =

2
3

0.0475
0.0475

0.0352
0.0352

0.08177
0.08177 0.08767

2.044
2.044 2.192

0.0375 - 0.02456 0.130113

Average

= 0.099449 ppm Cu
Sample:

kamote tops
Sample 1 2 3 A 0.1219 0.1278 0.1134 A' 0.1096 0.1155 0.1011 ppm Cu soln 0.6536 0.6989 0.5883 0.6469 sample 16.34 17.47 14.71 16.17

50.0 mL 0.09945 ppm ppm Cu = 2.00 g

= 2.486 ppm Cu

Average

Standard Addition
kangkong
Sample 1 Solution Vs (mL) A y A y A y Sample 2 Sample 3

1
2 3 4 5

0.00
0.50 1.25 2.50 5.00

0.0119
0.1619 0.3350 0.6165 1.0865

0.5000
6.803 14.08 25.90 45.65

0.0127
0.1632 0.3311 0.6147 1.0820

0.5000
6.425 13.04 24.20 42.60

0.0133
0.1612 0.3282 0.6148 1.0820

0.5000
6.060 12.34 23.11 40.68

slope
y-int r

8.908
2.106 0.997

8.307
1.984 0.997

7.940
1.848 0.998

Solution
ppm Cu Sample

0.1123
14.03

0.1204
15.05

0.1259
15.74

Standard Addition
kamote tops
Sample 1 Solution Vs (mL) A 0.0272 0.1696 0.3308 0.6200 1.1073 y 0.5000 3.118 6.081 11.40 20.35 3.934 1.013 0.999 Solution Sample 0.2542 31.78 A 0.0261 0.1679 0.3315 0.6191 1.1078 y 0.5000 3.216 6.3506 11.86 21.22 4.105 1.035 0.999 0.2436 30.45 A 0.0276 0.1733 0.3317 0.6202 1.1088 y 0.5000 3.139 6.009 11.26 20.09 3.875 1.025 0.999 0.2581 32.26 Sample 2 Sample 3

1
2

0.00 0.50 1.25 2.50 5.00

3
4

5
slope y-int r ppm Cu

Standard Addition
Sample computations: Slope = 8.908 ppm-1 Solution: Cx = 1/(8.908 ppm-1) = 0.1123 ppm Sample:

50.0mL ppm Cu = 0.1123ppm 10.0mL

=14.03 ppm Cu

50.0mL 2.00g

Average ppm Cu in kangkong: 14.94 ppm (1.494 mg per 100 g) Average ppm Cu in kamote tops: 31.49 ppm (3.149 mg per 100 g)

Experiment No.11 Nuclear Magnetic Resonance (NMR) Spectroscopy

CHEM 127.1 - MAD Groups 5 and 6

INSTRUMENTATION

Wide-Line NMR
Wide-line NMR
1. 2. 3. 4. Electromagnet Oscillator Modulation and lock-in detection Signal acquisition

Pulse NMR
1. 2. 3. 4. Programmable Pulse Generator Transmitter Probe circuit Reciever

NMR Spectrometer

NMR Spectrophotometer

Video Clip

Sample Quality Affects Spectra

Low NMR sample quality increases every peaks width, making it hard to resolve small couplings and frequency differences.

Factors Affecting Sample Quality

NMR spectra are strongly affected by both the sample contents and the NMR tube. These factors include:

Boundaries Between Materials Cause Problems

We shim to make the field more uniform across the sample, but sample factors can limit shimmings effectiveness. Every material gets magnetized in a magnetic field, and the strength of its response is its magnetic susceptibility (). To ensure field uniformity, one must minimize such interfaces.

Sample Height
Sample should at least exceed the detection region. Three fingers rule

 Solid particles must be absent in the sample.

NMR Tube Straightness (Camber) and Concentricity

A tube is held at the top, but the sample must be aligned precisely in the center of the probe for maximum performance.

 The centers of the inner and outer surfaces of the tube may not coincide well. Poor positioning of the sample in the coil and nonuniformity of the glass wall thickness create shimming problems.

NMR Tube Cleaning

Rinse 1x with samples solvent Rinse 5-10x with non- chlorinated solvent, e.g. acteone &/or H2O Rinse 1x with D2O Store inverted on a lab wipe Dry with stream of N2 If absolutely necessary, dry flat in oven 125 C, 45 min NEVER STORE IN AN OVEN!

Processing of NMR

RESULTS

Compound 1

Compound 1
O S O HO
Methyl 4-(2-hydroxyethyl)benzenesulfonate (1). C9H12O4S, 1H NMR (500 MHz, CHLOROFORM-d) ppm 2.42 (s, 3 H) 2.58 (br. s, 1 H) 3.77 (t, J=4.76 Hz, 2 H) 4.09 (t, J=4.40 Hz, 15 H) 7.33 (d, J=8.06 Hz, 15 H) 7.77 (d, J=8.43 Hz, 15 H).

O CH3

Compound 2

Compound 2
O S O CH3 O

O S O

O CH3

Biphenyl-4,4'-diyldimethanediyl dimethanesulfonate (2). C16H18O6S2, 1H NMR (500 MHz, CHLOROFORM-d) ppm 2.45 (s, 6 H) 4.17 (s, 4 H) 7.33 (d, J=7.69 Hz, 4 H) 7.72 (d, J=8.06 Hz, 4 H).