Portacubetas Cubeta:disolucindemedida

Refrigerante:baotermost8co, Pel$ercooler

Propertyofluminescencespectrum
Fluorescencevsphosphorescence 1. Phosphorescenceisalwaysatlongerwavelengthcomparedwithuorescence 2. Phosphorescenceisnarrowercomparedwithuorescence 3. Phosphorescenceisweakercomparedwithuorescence Why? Absorp8onvsemission 1. absorp8onismirroredrela8vetoemission 2. Absorp8onisalwaysontheshorterwavelengthcomparedtoemission 3. Absorp8onvibra8onalprogressionreectsvibra8onallevelintheelectronicexcited states,whiletheemissionvibra8onalprogressionreectsvibra8onallevelinthe electronicgroundstates 4. 0transi8onofabsorp8onisnotoverlapwiththe0ofemission Why?

Fluorescencespectroscopy

Fluorescencespectroscopy
Lightsource
Excita$on monochromator

Beam spliNer sample

Q:whytheemissionis measuredat90rela$veto theexcita$on?

Emission Monochromator Reference diode

PMT

Amplier

Computer

Emissionspectrum:holdtheexcita$onwavelengthsteadyandmeasuretheemissionatvarious wavelengths Excita$onspectrum:varytheexcita$onwavelengthandvarythewavelengthmeasuredforthe emi<edlight

Fluorescencespectroscopy:welldenedmolecules

Summaryofspectrophotometrictechniques
Describethemaincomponentsofaspectrophotometeranddis$nguishbetweensingle doublebeaminstruments Describesuitablesourcesforultraviolet(UV)/visible(vis),infrared(IR)andatomic absorp$on(AA)instruments Describeandassessadvantagesanddisadvantagesofvariousmonochromatorse.g.Prism, dirac$ongra$ngs Explainhowtoassesthequalityofgra$ng Explainhowphotomul$pliersanddiodedetectorswork Explaintheadvantageofmul$plexdetec$ng Describetheluminescencespectroscopyandenergytransferprocess Comparetheemissionandabsorp$onspectrum

TOMIC
BSORPTION PECTROSCOPY

ATOMS
Bohrs shell model: SODIUM atom Electrons

Nucleus Shells

Nucleusprotons (+ve)andneutrons (neutral). Electrons(ve) chargedpar8cle. Shellsconsistsof subshells.

Shell,Subshells,Electrons
Electrons are arranged according to their energy levels. They are arrange in subshells, the subshells are arrange in shells and shells arearrangearoundthenucleus.
Atom (around the nucleus) !Note: Electronsthatarenearthenucleushas a lower energy level than the electrons that are much far away. However, they experience stronger aNrac8on in the nucleus than those ones that are furtheraway. Shells

Subshells Electrons

Exci8ngatoms
groundstate:isastatuswheretheatoms electronsareintheirlowestpossibleenergy level.(stable) excitedstate:anotherstatuswherethe atomselectronsabsorbenoughenergyto bepromotedtoahigherlevel.Therefore, theyarenotintheirlowestenergylevel. (unstable)

Excitedatom
Generally, atoms are in their ground state but when an atom receives enough input of energy that their electrons requires to be promoted to a higher energy level. They will then turn to their excited state.

Since, an atoms excited state is very unstable it rapidly jump back down to its ground state. This jump then causes the atom to release the energy it absorbed in the form of photons of light.

Ground state: Excited state: sodium atom

!Note: Take note that the electron can go back down to its original place in more than one jump.

WhatisAAS?
(AAS)AtomicAbsorp8onSpectroscopyis:
Quan8ta8vetechnique Typicalsamples:lowviscositysamples Generallyusestodeterminetheamountofseveral metals(e.g.Cu,Fe,Zn,Mg)inthesoil,blood,air, water,andfood. Occasionallycanbeuseasaqualita8vetechnique (caninden8fychemicalsthatarepresent)68 elements

 In the electromagne8c spectrum, it uses thevisibleparttodetectthepresenceof metals(p.77chembook) Founda8on of the technique: the absorp8onofthelightenergythathasthe right wavelength causes the electrons from the sample to be promoted from a lower energy level to a higher energy level.

DiscoveryofAAS
1952,theAustralianscien8stAlanWalshwasworking onthemeasurementofsmallconcentra8onsofmetals attheCSIROusingatomicemissionspectroscopy. TheideaofAAScameintohismindashewasgardening athisMelbournehome.OnthenormalSundaymorning hehadtheideaaboutlookingatthelightabsorbedby theatomsexceptthanlookingatthelighttheyemits. AlanWalshdidnotjustdiscoveraprocessthathasthe abilitytosavelivesbutalsoproventhatatomswillonly absorblightthathastheEXACTvaluerequiresto promotetheirelectrontoahigherlevel.

HowdoesAASworks?

Lamp monochromator Burner slit detector meter

Howcanweobtainthedata?
The intensity of the light coming through the cathode lampismeasured Theintensityofthelightisthen again measured and compared totherstresult.=absorbance

The higher the concentration of the metal that is being observe in the sample the greater the absorbance.

The light can then be absorbed by the atoms from the sample that has been vaporized in the flame. This wavelength can then promote the electrons to a higher energy level = excited state

Howdoweanalyzethedata?
Bycomparingthelightintensitythathaspassedthroughthe sample(refertopreviousdiagram)withthatofthesamelight aoerithaspassedthroughablank,theabsorbanceismeasured. Theabsorbanceofdierentstandardsolu8onsofacompoundof theelementarealsomeasuredandacalibra8oncurveis constructed. AbsorbanceisploNedagainstconcentra8on.Wethenusethe calibra8oncurvetodeterminetheunknownconcentra8on.

ExampleofaCalibra8oncurve
AnAASwasusedtodeterminetheconcentra8onofleadions(inppm) in sh. The AAS was set up with a lamp that emiNed light with a wavelength that is absorbed by lead atoms. The AAS was calibrated usingdierentsolu8onscontainingknownconcentra8onsofleadions. Thegraphonthenextslideshowsthevaria8onofabsorbancewiththe concentra8onoflead. A2.0gsampleoftheshwasgroundupandheatedonahotplatewith 10mlofnitricacid.Thismixturewaslteredandthensprayedintothe ame of the AAS. The absorbance reading was 6.0. Determine the concentra8onofleadionsinthesh.

Graph:

GC/MS

Sample in

Sample out

(solid or heavy liquid coated onto a solid or support system)

GCMethods
ParameterGroup SDW05.13000s SDW05.24000s WPP05.04000s WPP06.03000s SHW06.21000s Method EPA502.2 EPA551.1 EPA604 EPA625 SW8468141A Compounds VOCs ClVOCs/Pests Phenols SVOCs OrgPPests

Instrumentation
Injection Port - Sample introduction

ManualDirectInjec8on AutomatedAutosampler PurgeandTrap

Instrumenta8onOven
TemperatureControl
Isothermal Gradient

Columns
Packed

Capillary

Polarity

Nonpolar

Polar

Phases

Instrumentation - Detectors
Destruc8ve
MassSpectral(CI/EI)[625] FlameIoniza8on(FID)[604] NitrogenPhosphorus(NPD)[8141A] FlamePhotometric(FPD)[8141A] Electroly8cConduc8vity(Hall/ELCD)[502.2]

Instrumentation - Detectors
NonDestruc8ve

ThermalConduc8vity(TCD) ElectronCapture(ECD)[551.1] PhotoIoniza8on(PID)[502.2]

Chromatograms551.1

Chromatograms604

SupelcoPTE5

SupelcoSPB50

HPLC
HighPerformanceLiquidChromatography FourTypes
Par88onChromatography Adsorp8on(LiquidSolid)Chromatography IonExchangeChromatography SizeExclusion(Gel)Chromatography

PlanarChromatography
Simple,CheapModelingforHPLCCondi8ons

HPLCSystem

h<p://hplc.chem.shu.edu/BOOK/intro/int_inst.html

ScopeofHPLC

Sensi8ve Quan8ta8ve Nonvola8leCompounds ThermallyFragile Compounds BroadApplicability

BiochemicalSpecies Pharmaceu8cals Pes8cides InorganicandOrganometallics IndustrialChemicals