Qualitative Analysis Quantative Analysis Physical Properties

Chemical Properties

What chemical species are present in a substance or sample How much of each chemical species is present in a substance or sample Properties that can be observed or measured without changing composition of the matter ( hence sampe element/ compound is present before and after a physical change) e.g appearance, texture, density, solubility, colour, malleability Properties that become evident during a chemical reaction ( chemical properties cannot be determined by viewing or touching substance) e.g reactivity with other chemicals, flame tests, pH, heat of combustion

Key knowledge How does chromatography work? Types of chromatography What each type can do? Quantitative Qualitative What substances can each type analyse? What do the results look like? What do the results mean? How do we analyse to results?

CHROMOTOGRAPHY Q. Why use Chromotography

Its used in pharmaceutical research, detection of banned substances, forensic work eg. ( detection of explosives)

Chromatography

Form of chemical analysis Where unknown substance is compared to observations made with known substances Under same conditions In order to separate components of a mixture By using two separate phases Stationary Phase Solid or liquid substance Coated onto solid surface NOTE : should posses large surface area Mobile phase Solvent used Which is Gas or liquid That moves through the stationary pahse Origin The line slightly above the initial solvent level , on which samples are placed it is drawn with a lead pencil. Solvent Front The level reached by the mobile phase when the chromatogram is removed. Adsorbed Attached to the stationary phase ( analogy like climbing a ladder you to not absorb into you just interact with it) Desorbed Dissolved back into the mobile phase NOTE : Separation is caused by adsorption to the stationary phase by making and breaking weak bonds with the surface of stationary phase ( weaker the bonds, faster component moves over stationary phase) and desorption back into the mobile phase.

Principle of Separation Some substances are more strongly attracted to the stationary phase than others Substances that have stronger it is more strongly adsorbed to the stationary intermolecular forces phase it spends more time attached to the stationary phase - - the substance moves more slowly up the plate Substances that are more soluble in spend more time dissolved in the mobile phase the mobile phase move more rapidly up the plate Separate according to polarity number and type of functional groups/ atoms as larger molecules move slower and have slower retention times. Revision of Intermolecular Bonding Dispersion forces ( also referred to as instaneous dipole. Van der walls)

Dipole Interactions Hydrogen Bonding

Attraction between (negative) electrons in one molecule and (positive) nuclei in neighbouring molecules Most important between non-polar molecules Increases with molar mass (more electrons) Attraction between polar molecules Increases with number of polar groups Attraction molecules with -OH or -NH groups ( NOF) Strongest form of intermolecular bonding

Q. What are two things the separation of sample components based on?

Q. How does the solvent carry the sample up the plate? Uses of Chromatography Identification : to help identify components by seperature mixtures Purify substance : to purify sample containing mixture of substances Check purity checking to see sample is pure Confirming presence of component in mixture Concentration : help identify concentration of components in sample

How strongly each component adsorbs onto the stationary phase How readily each component dissolves into the mobile phase And separation is caused by the different rates of movement as they are carried through stationary phase by the mobile phse Sample molecules move up the plate by adsorbing by attaching to the stationary phase and desorbing by dissolving back into the mobile phase.

Types of Chromatography Techniques Qualitative only Paper Chromatography only used to identify the component and is not quantitative as Thin Layer it doesnt indicate the concentration of sample. Chromatography(TLC) Qualitative and Quantative High Performance Liquid Chromatography(HPLC) Qualitative retention time helps identify component ( will have same retention time if run under same conditions Gas- Liquid Quantative concentration of component can be worked out Chromotography ( GLC) by peak area ( given)

Paper Chromatography & Thin layer and (TLC)

Qualitative : because it is only used to identify the component and is not quantitative as it doesnt indicate the concentration of sample. Based on how far components move up the plate (Rf) Used to separate small amounts of substances Components undergo repeated transfer of particles back and forth between the stationary phase and mobile phase and separate according to the differences in attraction between mobile and stationary phase( The greater the attraction to mobile phase, faster move up) Components made visible if not visible ( e.g fluorescing under UV light, or under certain chemicals) Stationary Phase Mobile Phase Paper High quality absorbent paper Appropriate solvent that will Chromatography easily dissolve the sample Thin Layer Fine absorbent powder coated Chromatography ( onto plastic or glass sheet. TLC) e.g silica gel ( SiO2) , alumina( aluminium oxide (Al2O3), cellouse

Q. Why is a lid place on top of containing holding chromatogram

Q. Why must the stationary phase and origin be perfectly horizontal?

Reduce rate of evaporation of mobile phase Protects chromatogram from temperature changes, contamination Which may alter Rf values Solvent front travels evenly up the stationary phase

Mark the solvent straight away and centres as quickly as possible after chromatogram is removed NOTE: solvent can still move after removed from the beaker Measure to middle of the spot NOTE : better quality will have finer dots ( high
resolution

Must be above solvent so sample does not dissolve in the solvent Draw in grey lead pencil as ink can run and move the origin hence wont know where the substance started incorrect Rf value calculated

Rf values: identifying samples Enables us to compare between No two compounds chromatograms have identically during chromatography Sample will different Rf values when conditions are changed e.g length of coloumn, temperature, solvent, stationary phase Rf = Distance Moved by particular dot as measured FROM THE ORIGIN Distance moved by solvent front as measured FROM THE ORIGIN NOTE : must write FROM THE ORIGIN Example : Calculate the Rf value for sample A and use the table to identify the sample

Distance moved: - Sample A : 35mm - Solvent from : 60mm Rf = 35/60 = 0.58 Rf 0 0-1 1

Component remains at origin No affinity for mobile phase Some affinity for both the mobile phase and stationary phase Component located at solvent front Low affinity to stationary phase To achieve better separation When the mobile and stationary phase are the same

Q. Why would you want the solvent front to run for longer hence be closer to the top of the paper ? Q. When can Rf values be compared?

For Rf values to be the same between chromatograms consistency of Size of sample Choice of solvent Complete saturation of vapor in which paper is developed

Qualitative/ Quantative
Paper chromatography

Stationary phase

Mobile phase

Advantages

Limitations

Qualitativ e

High Quality Paper

Water based or polar -

Thin Layer Chromatography ( TLC)

fine powder (e.g. Al2O3 or silca gel) on a glass or plastic plate

usually an organic (nonpolar) solvent

Large samples Cheap Easy & simple to use can be stored for future better resolution of polar mixtures Faster & more sensitive ( used on smaller samples) Corrosive materials can be used as absorbent layer made up of inorganic substances Better on non polar mixtures more choice of stationary phases

Corrosive materials cannot be used not as sensitive as TLC - components can streak delicate plates hard to handle colorless mixtures have to be colored cannot be stored for future examination as colored is not permanent

both cannot separate complex mixtures containing similar compouds

Two- way Chromatograms

Uses two different solvents on the same sample Allows for better separation of complex mixtures Method 1. Place sample spot on corner of chromatogram 2. Run the chromatogram as normal in the first solvent 3. Turn the chromatogram on its side and run in the second solvent Example The amino acids present in a sample of fruit juice can be detected by thin-layer chromatography. To achieve better separation of the complex mixture of substances present in the juice, a two-way chromatogram was prepared.

1.a) Calculate the Rf value for each component in solvent 1. b) Attempt to identify each component based on solvent A data.

2.a) Calculate the Rf value for each component in solvent B. b) Use this data to further identify each component.

Q. Why has the chromatogram been run in two solvents?

Two-way chromatogram produces better separation of components of complex mixtures. No single solvent can separate all amino acids. 4. Draw the chromatogram as it would have appeared after being run in solvent A only.

COLUMN CHROMOTOGRAPHY
Qualitative : can only identify substance glass or metal column packed with alumina or silca gel sample carried by mobile liquid phase under influence of gravity or pressure eluent goes to spectrometer and retention time is recorded Column Used to separate mixtures into individual components Chromatography By separating via differing rates of adsoption and desorption

NOTE Rt value does not depend on concentration of sample Rt is characteristic of component under conditions it is performed in Longer columns produce better resolution ( more separation occurs) Polar vs non polar will determine rate Retention time dependence factor Type of bonding (e.g can be hydrogen bonding, inic, dispersion forces ) with stationary phase Affinity for mobile phase Normal Phase Chromatography : With a Polar Stationary phase and non polar mobile phase ( ASSUME Same length compound , different Least polar chains eluted first homolgous series Polar compounds are in column for longer time Same homologous series, different Shorter chain eluted first ( less dispersion forces) chain length Reverse Phase Chromatography : With Non polar stationary phase and polar mobile phase Same length compound , different Most polar chains eluted first homolgous series Non- polar compounds are in column for longer time Same homologous series, different Shorter chain eluted first ( less dispersion forces) chain length

High Performance (or pressure) Liquid Chromatography (HPLC)

Components : Stationary phase Mobile phase Sample Detector ( eyes or device ) From the graph in recorder you want to know 1. Retention time - the number of compounds, identify the component 2. Peak height concentration
- solid or viscous liquid coating - chosen to give good separation - small particles: large surface area (allows frequent adsorption and desorption, giving better separation) A solvent pumped through under high pressure. - UV is absorbed by any molecules in the eluent - results are presented as a chromatogram In Gas liquid chromatography the sample will need to get into gaseous form and we do this by heating usually lower molecular weight and smaller of 300 of molar units or less , hence boiling points is important and also if it the sample is heat sensitive it will decompose in GLC

Components Stationary Phase

Mobile Phase Detector

Q. Compare HPLC to GlC

Lowest retention time: A Lowest concentration : lowest peak area If this was HPLC : will have a non polar coloumn and polar solvent Most polar :A Q. Why do larger Stronger dispersion forces give the sample a greater affinity for the molecules have a stationary phase: stronger adsorption if a solid stationary phase, or longer Rt? greater solubility if a liquid stationary phase. NOTE : affinity to stationary phase means strength of adsoption to the solid stationary phase or solubility in a liquid stationary phase

Qualitative Analysis Rt (retention time) Quantative Analysis Area under peak

Time taken for the substance to pass through the machine which is used to identify molecules by comparing its Rt value to that of a known standard To calculate the amount of a substance: 1. Measure the peak area (or peak height) on the chromatogram. 2. Compare to the area produced by a series of standard solutions using a calibration graph.

Uses HPLC is generally used to identify organic compounds, especially those with a large MR: hormones proteins pharmaceuticals food components Units measured in Ppm parts per million Ppb parts per bilion 1ppm = 1mg/L 1ppm = 1 microgram/L Example Question A herbal tea extract was analysed using HPLC, The chromatogram is shown below.

Q. How many components are evident? Q. What was the Rt of the component that was bound least strongly to the stationary phase. Explain your answer.

Q.Which component was present in the highest amounts? Explain your answer.

Four components, corresponding to the four peaks. The component with an Rt of about 1.4 minutes was bound least strongly to the stationary phase. The less strongly a component is bound the more time it spends in solution with the mobile phase and the more rapidly it will move through the chromatograph. The peak with an Rt of about 2.9 minutes was present in the highest concentration. The area under this peak is the largest.

Gas Chromatography

NOTE : nitrogen is the common carrier gas Mobile gas ( usually nitrogen gas ) carrier gas phase Sample Vapourised as it is injected into the GC. Why? Injection point very close to oven or inside the oven want it in quickly Flame Current produced as sample burns producing ions. Ionisation Detector Stationary Gas solid Chromatography: phase porous beads Gas liquid Chromatography: beads coated with a high boiling point liquid hence is very thick and VISCOUS ( e.g. like oil and honey) Factors affecting retention time High Temperature decreases Rt

High pressure / flow rate More polar stationary phase with a nonpolar sample Longer column

decreases Rt

Particles have greater energy less affinity for the mobile phase NOTE : - Peaks can be shorter and wider but important that PEAK AREA IS THE SAME Mobile phase carries sample through more rapidly Particles have less affinity for the mobile phase hence will move quicker

decreases Rt

increases Rt

Increased solubility of the sample in the stationary phase:

increases Rt

Further to travel BONUS more distinct separation between two components which are short distance apart MINUS takes more time Greater affinity: more time spent dissolved in liquid stationary phase so less time mobile phase

Uses

GC can be used for organic substances which can be vaporized without decomposing: with MR up to about 300 molar mass. Larger molecules must be analysed using HPLC. GC is the most sensitive technique ( more sensitive than HPLC however GLC cannot be used for many substances ) : detects as little as 10-12 g. Limitations Is not sutiable for some substances which are vaporised or degraded when heated ( even caramalized in the case of honey) In gas chromatography Q.Why is the column The column is packed with very fine particles to increase the surface packed with very fine area increases the rate of adsorption and desorption in each of the particles? stationary phase. This, increasing the degree of separation. Q. Why is the injection port heated? Q. Discus the relative advantages of HPLC and GC. What types of substance is each used for? The injection port is heated to vapourise the sample. The sample is carried through the GC as a gas. GC and HPLC are both used for organic compounds. GC is the most sensitive, but can only be used for substances which can be vapourised without decomposing: MR < 300. HPLC is used for larger or less stable molecules.

A mixture of liquid alkanes is analysed using a GC. Identify the peaks using your knowledge of inter-molecular forces. Explain your answer The four alkanes analysed are: decane: C10H22 hexane: C6H14 octane: C8H18 pentane: C5H12 Pentane comes out first less molar mass less dispersion forces less interaction travels faster slower retention time A: pentane, B: hexane, C: octane, D: decane. The larger molecules have stronger dispersion forces so will spend more time bound to the stationary phase therefore will take longer to pass through the GLC and gave a longer R t. Exercise: Concentration of Ethanol in Wine The ethanol (b.p. 78C) sample of old cask wine is analysed using GC. Four standards and the test sample were prepared as shown ion the table below. Propanol (b.p. 97C) was used as a reference to enable comparison where different volumes of solution may be injected into the GC.

Talk about set up of standards & dilution factor for test sample 50 100 dilution factor of two. Q. Why do we need to know about boiling Want the coloumn slightly higher than boiling point? point so substance stays in gaseous form

Five chromatograms

Need propanol as variable amount is injected into machine: obvious by inspection. Q. Why are the putting in 20ml ? As a reference standard - Non linear results due to sensitivity of the dector changing Allows for comparison of ratio of peak heights 1. Complete the table by calculating the peak area ratio: ethanol propanol

Q. Calculate the concentration of the original wine sample.

Original wine sample was 50 mL, final diluted sample was 100 mL. Dilution factor = 100/50 = 2 Concentration of original wine = 2 x 5.05 = 10.1 % Ethanol is oxidised into ethanoic acid (to produce vinegar).

Q. The label on the wine cask stated the concentration to be 11.5%. Give a reason for the difference (apart from a simple mistake performing the analysis). Q. The analysis was conducted at 110C. Why was this temperature chosen rather than 60C? Q. Why is it bad to store wine upright

All components of the samples must be gases: the temperature must be above their boiling points.

The cork will dry out hence causing the cork to shrink , then bacteria and yeast and fungi go into the win they multiply after drinking the wine. So CH3CH20H ( ethanol ) due to drunken party yeast as it is oxidised CH3COOH ( ethanoic acid )

terms: mobile phase, stationary phase, adsorption, desorption, Rf values, retention time Gas chromatography samples are analysed by converting them into gases. These gases are then passed through a column packed with special materials to achieve separation. HPLC works in a similar fashion to GC, except that the mixture to be analysed remains as a liquid. GC is used when substances are easy to vaporise and HPLC for mixtures harder to vaporise or for mixtures that might be damaged by high temperatures. Retention time is the time from injection until the component is detected in the column. Components are identified by their retention times. GC and HPLC can be calibrated for quantitative analysis by obtaining readings from a number of standards and then plotting a calibration curve. GC is used for substances with relative molecular mass of up to 300 that are volatile and organic. HPLC can be used for substances with relative molecular mass over 300, including substances that are unsuitable for GC because they decompose when heated. HPLC can also be used to isolate and purify substances.

SPECTROSCOPY
Key Knowledge Principles and applications of spectroscopic techniques and interpretation of qualitative and quantitative data from atomic absorption spectroscopy (AAS), infrared spectroscopy (IR), mass spectroscopy, nuclear magnetic resonance spectroscopy (NMR), and visible and ultraviolet spectroscopy (visible-UV); Matching analytical technique/s to a particular task. Basic Principles 1. Atoms or molecules absorb and emit electromagnetic radiation of specific energies. 2. Atoms or molecules undergo a change when they absorb electromagnetic radiation. 3. Different parts of the electromagnetic spectrum affect different parts of the atom or molecule. Spectroscopy use of light or electromagnetic radiation for analysis Electromagnetic Form of energy radiation Consists of electric and magnetic fields That travel at the speed of light e.g light, radiowaves, xrays ( difference is that have different amount of energy found in the photons) Photon Single unit of electromagnetic radiation Consisting of mass- less participle travelling In a wave like motion NOTE : each photo contains a certain amount ( quantam ) of energy Q. What allows the atoms or molecule move to a higher energy level in spectroscopy techniques? Q. What is the difference between the movements of atoms to a higher energy level compared with molecules? In each technique the atom absorbs a specific quantam of energy which allows the atom to move to a higher energy level - In atoms the movement is of the electrons to a higher energy level(electronic energy levels, E = hv) - where as in molecules the electorns move to a higher energy level but the movement of molecules also increases to a higher vbrational, rotational and nucleur spin energy ( these energy levels are quantified fixed values)

Properties of Electromagnetic Radiation

Wavelength Frequency (Hz)

Describes distance between any two consecutive identical points on a wave the number of complete cycles of the wave that pass a given point in a second

NOTE : frequency (v) of a photon is inversely proportional to wavelength ( hence when frequency increases , wavelength decreases energy of photon (E) is inversely proportional to wavelength ( shorter the wavelength more energy) the energy of photon (E) is directly proportional to its frequency The Electromagnetic Spectrum radiation from each part has a specific frequency and wavelength coloured light is in the visible spectrum (note : different colors consist of different energies or wavelengths) Using radiation in spectroscopy atoms and molecules absorb and emit specific wavelengths of electromagnetic radiaton radiation interacts with atoms and molecules in different ways the part of the atom or molecule that is affected depends upon the wavelength or energies it is subjected to Example Visible / UV radiation Valence electrons move to higher energy levels Infrared Radiation ( not enough to move valence electrons ) - Molecules themselves move to higher energy levels - By causing changes to covalent bonds in molecules Depending on strength stretch, rock, bend, twist like a spring Radiowave radiation - can change direction of spin - moving to higher nucleur spin energy In summary - different parts of electromagnetic radiation affect different parts of an atom in different ways Technique Part of Spectrum Used Part of Structure Affected by supplied radiation Flame Tests Visible Valence electrons in metal atoms Atomic Emissions Spectroscopy ( AES) Atomic Absorption Ultraviolet and Visible Spectroscopy (AAS) Colorimetry Visible Electrons in molecules (Visible Spectroscopy) UV Visible Spectroscopy Ultraviolet and Visible Infrared Spectroscopy ( IR) Infrared Bending and stretching of bonds in molecules Nuclear Magnetic Resonance Radiowaves Nuclear spin states ( Spectroscopy ( NMR) nucleons in molecules)

TYPES OF SPECTSCOPIC ANALYSES 1. Analyse energies emitted by a substance ( e.g flame tests,AES) 2. Analyse energies absorbed by a substance ( e.g AAS)
Qualitative Measuring absorbance as function of wavelength to identify substance e.g Measuring how much energy of specific wavelength is being absorbed by substance Used to determine amount of substance Infrared UV Visible Spectroscopy Colorimetry AAS UV visible spectroscopy

Quantative

Flame Test
Heat is used to excite valence electrons so they jump to higher energy levels hence electrons are in excited state (absorbed energy is used to overcome the forces of attraction between valence electrons and the positively charged nucleus) they return to lower energy levels they release specific quantam of energy in the form of photons which have discrete wavelengths that are characteristic of the element hence allowing us to identify the metal cation

note : ionisation occurs when electron leaves atom and electrons cant go half way between shells Flame Test Spectroscopy technique Allowing for identification of certain metal cations in compound Qualitative can help identify an element but is not accurate as many elements have same colour eg. both sodium and calcium emit a yellow/orange colour in the flame test.

Limitations Can only be used to identify some metal species, many metal atoms may not produce coloured flame because o Insufficient heat to excite electrons and cause them to move to higher energy level o Not enough sample was vaporised for distinct flame colour to be observed Many metals produce similar coloured flames cant identify definitively Sample destroyed in process Some metals may not produce radiation in the wavelength of visible spectrum Not quantative hence amount of metal cannot b determined Most samples not pure , flame colour will be a combination of more than one species Q. How can these disadvantages be overcome Use a hotter flame so sufficient energy is available to excite electrons in wider range of elements Use a spectroscope so light passes through prism and proper up into discrete wavelengths ( atomic emission spectrum)

Advantages Quick & can confirm presence of a metal in sample

Atomic Emissions Spectroscopy and Spectroscopes

Very similar to flame test BUT uses the slit to focus a narrow beam of emitted light and a prism to separate the this light into its individual wavelengths. Qualitative Analysis : can be used to identify elements in sample when the emission spectra is compared to spectra of KNOWN samples PRODUCED UNDER SAME EXPERIMENTAL CONDITIONS. Different elements possess different energy levels, hence the wavelengths of radiation emitted as electrons move to lower energy level will differ. They represent the energies released as excited electrons in metal atoms movefrom excited states to lower energy levels. Different elements have different numbers of protons hence will have different energy levels. Every atom has different electron energy levels, and so has a unique emission spectrum. Prism produces spectrum: identifies elements more accurately

Q. Why do different elements produce different wavelengths of light?

Q. What do spectral lines on emission spectra represent? Q. What is the cause for the arrangement of lines on an emission spectrum characteristic of element? Q. Why can they be used to identify different atoms?

Q. What improvement makes AES a more useful technique than flame tests?

Spectroscope

Optical instrument with a slit (to allow passage of light to instrument ) and optical lens. Used to produce and examine spectra of light Radiation from sample source passes through slits and then passed through prism Different wavelengths are bent or refracted through different angles causing incoming light to split into components

Above : The emission spectra of four elements are below, these spectra are individual to each element. Limitations of atomic emission spectroscopy Only few atoms have electrons excited by heat of flame ( most remain in ground state) cant detect presence of small quantities

Atomic Absorption Spectroscopy

Q, What is the difference between emission and absorption spectroscopy?

In emission sample is heated and light from excited state emits light when it returns to ground state Where as in absorption light is put through a sample and discrete units of energy are removed these units of energy removed correspond to the amount of energy needing to excite the valence electrons.

Qualitative : can determine identify of element as the spectrum produce is the same Quantative : can determine concentration of element ( the amount absorbed determines on concentration then unabsorbed light is compared with original ongoing light Black lines( represent wavelengths absorbed) on a coloured background ( wavelengths remaining ) Many applications Mercury levels in fish: fish must be purated, added liquid diluted to test levels Iron levels in blood : dilute blood Copper in a sample of ore from a mine: copper must be dissolved it

NOTE : emission and absorption spectra are not mirror images this is to do with the number of pathways in emission spectra compared to absorbing once. Q. What is the difference between Atomic Absorption spectroscopy and Atomic Emission Spectroscopy AAS analyses light absorbed where as AES analyses light emitted

Process Atoms will ABSORB light if the energy of light is exactly that required to promote an electron from its normal energy level to a higher energy level. As each element has a unique absorption spectrum, each element to be analyzed requires its own light source that will emit light of the correct wavelength. The light used is composed of the element and when vaporized it produces light of the correct wavelength for the analyze of the particular element. Unabsorbed light read by spectrometer ( black lines represent absorbed wavelengths)

Operation of an Atomic Absorption Spectrometer - solid line emits light constantly used a reference Q. Why do we want to pulse Can be used to compared to solid line light?

When light is pulsed

Both emitted and transmitted light are measured.

Only emitted light is measured NOTE : Transmitted light is the difference between light ON and light OFF. Interpreting the data standard solutions, calibration graphs A calibration (standard) curve for an A.A.S. analysis

Q. Why did the student have to perform the dilution?

The absorbance of Sample A did not fall on the calibration graph. A dilution was required to produce a solution with a lower absorbance. Advantages of Atomic Absorption Spectroscopy Very sensitive :can detect up to ppm/ ppb More elements : Wide range of elements can be detected More accurate than AES Qualitative and Quantative

QUANTATIVE SPECTROSCOPY ANALYSIS

Radiation of specific frequency is passed through a saple and some of this radiation is absorbed or transmitted by species e.g AAS and UV Visible spectroscopy - Radiation is absorbed by ground state electrons in atoms or molecules - That radition NOT absorbed reaches detector Spectrometer Measures amount of radiation That is absorbed or transmitted by chemical species ABSORBANCE = Initial Radiation Intensity Final Radiation Intensity NOTE : - The amount of radiation absorbed is directly proportional to concentration of solution ( ONLY AT LOW CONCENTRATIONS more concentrated substances will absorb more, hence higher absorbance reading)

STANDARD CURVES / CALIBRATION CURVES

At low concentrations the absorbance of substance is directly proportional to the concentration of substance in solution Method 1. Dilute the sample so that is able to transmit some light / radiation 2. Colorimetry only : solutions that are not coloured may need to be coloured by mixing chemical under investigation with another substance 3. Select appropriate wavelength of radiation for analysis 4. Measure the amount of radiation absorbed by sample 5. Measure the amount of radiation that is absorbed by known concentration so component being analysed under same conditions as the sample 6. Plot absorbance of each standard solutions against concentration ( standard curve/ calibration curve) 7. Use curve to determine the concentraton of sample, NOTE : dilution factors NOTE : - Line of best fit > need to pass origin - At higher levels of concentration the absorbance- concentration relationship becomes non linear - Do not extrapolate graph at high concentrations ( but you can at low ) - Sample and known concentrations must be under same conditions Examples of spectroscopy techniques which use standard curves Colorimetry Atomic Absorption Spectroscopy UV- Visible Spectroscopy Q. 25 Why do we calibrate spectrometers each time they are sued to analyse a sample?

COLORIMETRY
Q.How do you know by looking at two glasses of cordial which one is more concentrated? Q. Why is it darker? The colour in B is darker in more concentrated than A

B looks darker It contains more pigment molecules and they absorb more light. the colour is darker as white light shines through it is absorbing lights which are not red and since there is more pigment molecules more light is absorbed.

Q. Why is it red?

Objective we can see results with eyes, but using instruments is more accurate Uses a colorimeter Basic Principles 1. The greater the concentration of the solution, the greater the intensity of the colour. 2. The colours absorbed are the complement of the colours we observe.( opposite side of the colour wheel) Solution must be coloured : To measure copper ion concentration in CuCl2 + NaSO4= CuSO4 ( coloured ) Colorimetry the process of comparing the intensity of a coloured solution with a set of standards of known concentration NOTE : SOLUTION MUST BE COLOURED ! concentration of unknown solution can be determined NOT QUALITATIVE cannot identify the solution.

Quantative

Basic Principles

NOTE : substance canot absorb radiation of the same colour only complementary colours

Colorimetry Instruments

Purpose of lamp Purpose of slit Purpose of color filter Purpose of cell Purpose of detector Method

the source of light to focus a beam of light to pre select the colour we want to go through our solution ( usually the complementary colour ) to contain test sample reads how much light has been absorbed

Make standard solutions graph a calibration curve Use it to work out unknown concentration. Q. Would you try make up 0.05 M solution from powder? Q. Why do graphs tail off at high concentrations?

Q. What is the difference between AES and AAS compared with colorimetry and UV visible spectroscopy Q. What determines what colour filter is chosen?

Weigh out too small an amount hence high chance of error. Hence we use multiple dilutions The graph is less accruate after it stops going linear ( higher concentrations) - Should aim to get diluted sample in the linear section - If the sample is in non linear section dilute it to get into linear area AES and AAS measure absorbance of discrete wavelegnths Colorimetry measures absorbance of a range of wavelengths The filter selects the band of wavelengths which are most strongly absorbed by coloured solution/

Ways to work out dilution 1. 25 ml and put into 250 ml flask for 1/10 dilution. ( example) 2. Can also use c1v1=c2v2 Revison of concentrations Concentration (n = c x V) Molarity (M) = mol/L ppm (parts per million), ie 1 g per million g, it also similar to mg/L for aqueous solutions ppb (parts per billion), ie similar to g/L %w/v, ie gram per 100mL %w/w, ie gram per 100g %v/v, ie mL per 100mL To convert M (mol/L) to g/L, it is converting mol to g, therefore X by molar mass To convert g/L to M (mol/L), by molar mass Density (d = m/V ie g/mL) for aqueous solutions density is 1 g/mL, ie 1 mL = 1 g Dilution (moles are constant) C1V1 = C2V2 Limitations Low concentrations : Not capable of testing very low concentrations Accuracy of Results : Results are not as accurate as AAS Visible Light Only : Restricted to substances that can absorb visible light, as not all substances can absorb visible light Coloured Solutions : Solutions must be coloured or be able to be coloured More than one substance : More than one substances may absorb chosen wavelength hence only suitable for pure substances. Advantages of Colorimetry Simple Equipment and reagents are inexpensive Can be used to test variet of chemical species ( most metal cations and some simple anions) Same is not destroyed