Satish Pradhan Dnyanasadhana College, Thane. Department of Chemistry M Satish Pradhan Dnyanasadhana College, Thane. Department of Chemistry M.Sc. Analytical Chemistry Sem-I Language of Analytical Chemistry by Dr.G.R.Bhagure

Language of Analytical Chemistry (8L) 1.1.1 Analytical Perspective, Common analytical Problems, terms involved in Analytical Chemistry(analysis, determination, measurement, techniques, methods, procedures and protocol) 1.1.2 An overview of analytical methods of analytical methods, types of instrumental methods, instruments for analysis, data domains, electrical and non electrical domains, detectors, transducers and sensors, selection of an analytical method, accuracy, precision, selectivity, sensitivity, detection limit and dynamic range

1.1.3 Errors,determinate errors,and Indeterminate errors,types of determinte errors,takaling of errors. 1.1.4 Quntitive methods of analysis: calibration curve, standard addition and internal standard methods

1.2 Quality in Analytical Chemistry (7L) 1.2.1 Quality Management Systems (QMS) Evolution and significance of Quality Management Systems , Types of quality standards for Laboratories, Total quality management(TQM) Philosophy implementation of TQM(Reference of Kaizen,six sigma approch & 5S) Quality Audits &reviews, Responsibility of Laboratory staff for quality and problems

1.2.2 Safety in Laboratories Basic concepts of safety in Laboratories, Personal Protection equipment (PPE) OSHA , Toxic Hazard(TH) classifications, Hazardous Chemical Processes including (processs calorimetry/thermal build up concepts)

1.2.3 Accreditations Accreditation of Laboratories, Introduction to ISO series, Indian Government Standard (ISI, Hallmark, Agmark) 1.2.4 Good Laboratory Practices (GLP) Principle,Objective,OECD Guidelines, The USFDA 21 CFR 58, Klimisch score

Definition of Analytical Chemistry Analytical Chemistry is branch of chemistry which deals with the study of theory and practice of methods used to determine composition of matter.

The Analytical Perspective What is the “analytical perspective”? Many analytical chemists describe this perspective as an analytical approach to solving problems. Although there are probably as many descriptions of the analytical approach as there are analytical chemists, it is convenient for our purposes to treat it as a five-step process:

1. Identify and define the problem. The Analytical Perspective 1. Identify and define the problem. 2. Design the experimental procedure. 3. Conduct an experiment, and gather data. 4. Analyze the experimental data. 5. Propose a solution to the problem.

1 Determine the type of information nedded Qualitative, Quantitative ,characterisation or fundamental 2 Identify interferent Select method Establish validation data Establish samplinng startagy 3 Calibrate Instruments and equipments Standerdise reagent Gather data 4 Reduce or transform data Analyse statstics verify results Interpretate results 5 Conduct external evaluation

Common Analytical Modules ❖ Qualitative Analysis – Identification of components from a sample ❖ Quantitative Analysis - Estimation of ‘how much’ of a component is present ❖ Characterization Analysis ❖ Fundamental Analysis

Term Involved in Analytical Chemistry : Analysis: Analyte's: Matrix: Determination: Measurement: Technique: Method: Procedure: Protocol:

The Language of Analytical Chemistry 1 Analysis: A process that provides chemical or physical information about the constituents in the sample or the sample itself. 2 Analyte's: The constituents of interest in a sample. 3 Matrix: All other constituents in a sample except for the analyte's. 4 Determination: An analysis of a sample to find the identity, concentration, or properties of the analyte. 5 Measurement: An experimental determination of an analyte’s chemical or physical properties.

The Language of Analytical Chemistry 6 Technique: A chemical or physical principle that can be used to analyze a sample. 7 Method: A method is the application of a technique for the determination of a specific analyte in a specific matrix. 8 Procedure: Written directions outlining how to analyze a sample. 9 Protocol: a protocol is a set of stringent written guidelines detailing the procedure that must be followed if the agency specifying the protocol is to accept the results of the analysis. Protocols are commonly encountered when analytical chemistry is used to support or define public policy.

Analytical Methods Chemical methods Physical Methods

Physical Change

Chemical Change Combustion of Fuel

Chemical Methods of Analysis Classification of Chemical Methods of Analysis Gravimetry or Gravimetric analysis Volumetry Volumetric analysis

Chemical reaction / change Precipitation Neutralization Oxidation Reduction Complexation

Analytical Technique developed on Precipitation Reaction Gravimetry Metal ion Non Metal ion Chemical Methods

Volumetry Chemical Methods Analytical Technique based on Chemical Reaction Volumetry Acid Base Titration Precipitation Titration Complexometric Titrations Redox Titration Chemical Methods

Technique based on Oxidation Reaction Elemental Analyzer Carbon Hydrogen Nitrogen Sulphur

Classification of Instrumental methods Optical methods Electroanlytical methods Separation methods Miscellaneous methods

Optical Property Optical Methods Absorption Emission Scattering Fluorescence Phosphorescence Optical Activity Optical Methods

Optical methods Absorption spectroscopy Emission spectroscopy. Atomic absorption Spectroscopy. Infrared Absorption Spectroscopy Fluorophotometry Turbidimetry and Nephelometry Raman Spectroscopy

Technique based on the Principles of Absorption of Light UV-Visible Spectroscopy IR Spectroscopy Atomic Absorption Spectroscopy

Single Beam COLRIMETER Radiation Source Collimating Lens Filter Sample Cell Photocell Read Out Device

SINGLE BEAM SPECTROPHOTOMETER Radiation Source Collimating Lense O.2 Read out Meter Amplifier PMT Detector Sample Cuvette Grating SINGLE BEAM SPECTROPHOTOMETER

DOUBLE BEAM SPECTROPHOTOMETER PMT Detector Blank Cuvette Mirror Grating Tungsten Lamp Read Out Meter Mirror Sample Cuvette Mirror Deuterium Lamp PMT Detector DOUBLE BEAM SPECTROPHOTOMETER

Atomic Absorption Spectrophotometer 62 different metals Rotating Chopper Hollow Cathode Lamp P.M.T.Detector Flame Amplifier Read Out Grating  Robert Wilhelm Bunsen and Gustav Robert Kirchhoff, both professors at the University of Heidelberg, Germany. The modern form of AAS 1950s by a team of Australian chemists. They were led by Sir Alan Walsh  Power Supply Sample Solution

Technique based on the Principles of atomic Emission of Light Flame Emission Spectroscopy Photoemission or Photoelectron spectroscopy (ESCA ) Inductively Coupled plasma Atomic Emission Spectroscopy

Flame Photometer Na, Mg, K, Li Slit Collimating Mirror P.M.T.Detector Amplifier Read Out Fuel Oxidant Prism Monochromator Sample Solution

ICPAES SPCTROMETER Amplifier 60 metals can be determined up to PMT Detector Plasma Monochromator Amplifier Computer System Read out device system ICPAES SPCTROMETER Nebulizer 60 metals can be determined up to

Phosphorence Spectroscopy Technique developed on the Principles of Molecular Emission of Light Fluorescence Spectroscopy Phosphorence Spectroscopy

Instrument: Single Beam Fluorimeter Sample Cell U.V. Light & visible light Primary filter Secondary filter Photocell /PMT detector Read Out Device

Phosphorimeter PMT detector Read Out Device

Technique based on the Principles of Scattering of Light Turbidimetry Suitable for Large Particle size Drinking water Nephelometry Suitable for Small Particle size Medicinal preparation like saline, eye drops

Turbidimeter visible light Filter Photocell Detector Read Out Device Sample Cell visible light Filter

Nephelometer visible light Light Trap Photocell Detector Sample Cell Graduated Disc Collimating Lens Light Trap visible light Photocell Detector Sample Cell Read Out Device

Polarimeter Polarizer Monochromatic light Analyzer Sample Cell Monochromatic light nonlinear

Electrical Conductivity Electrical Property Electrical Conductivity Emf Of Cell Diffusion Current Electric Charge Resistivity Electrical Impedance

Electrochemical Methods Electrical Conductivity Electrical Conductivity Meter Emf Of Cell Potentiometer pH pH meter Diffusion Current Polorography

Technique based on the Principles of Electrolysis Voltammetry Electrogrvimetry Polorography Amperometry

Separation techniques Why separations is essential ?

Classical Separation Technique Classical Separation Technique Based on solubilities filtration, precipitation, crystallization Based on Volatility distillation Based on Electrical effect Electrophoresis Based on Gravity: Centrifugation Classical Separation Technique

Based on Retention capacity of stationary phase Modern methods of separation Techniques Chromatography Adsorption Chromatography Partition Chromatography Based on Retention capacity of stationary phase

Adsorption Chromatography Size Exclusion Chromatography TLC GSC ION Exchange Size Exclusion Chromatography

TLC: Application of sample and development of Chromatogram by Ascending way High Polar Less Polar

ION EXCHANGE CHROMATOGRAPHY H+ Na+ H+ R

ION EXCHANGE CHROMATOGRAPHY H+ Cl- OH- R

Partition Chromatography Paper Chromatography HPLC GLC

Separation in Paper Chromatography Solvent Front S O L V E N t F W Solute front Solute front Application of Sample Original Line

WORKING OF HPLC

Separation in Paper Chromatography Solvent Front S O L V E N t F W Solute front Solute front Application of Sample Original Line

GAS CHROMATOGRAPHIC INSTRUMENT Pressure regulator and flow control Sample Injection port Detector * * * * Recorder column Column oven Least Volatile Most Volatile Carrier Gas

New Versions of Chromatography

GC-MS LC-MS New Versions of Chromatography Fusion of Two Technique Separation and Determination of Molecular weight

GC-MS

LC-MS

Applications of GC-MS Drug detection, Fire Investigation Environmental Analysis Identification of unknown samples Trace elements At airport security to detect substances

Environmental Monitoring Biotechnology Food Processing Environmental Monitoring Pharmaceuticals Agrochemicals Cosmetic Industries Applications of LC-MS

Thermal Methods mass change versus temperature or time TGA Differential Thermal Analysis temperature difference versus temperature or time Evolved Gas Analysis analysis of gases evolved during heating of a material, usually decomposition products Differential Scanning Calorimetery heat flow changes versus temperature or time Thermal stability of a material

Thermogravimetric analysis Substance Chemical Changes Heated Physical Changes Thermogravimetric analysis

Technique based on Radioactivity NAA Archeology Forensic Science Geology Semiconductor Industry Isotope Dilution Technique Clinical Diagnosis

Selection of Analytical method: 1 Concentration of the component. 2 The complexity of the materials/presence of interfering material 3 The probable concentration of the species of interest 4 Degree of Accuracy 5 Sensitivity and detection limit 6 Duration of an analysis 7 Speed ,time and Cost of analysis 8 Availability of equipments 9 Skill person for handling the instrument All the above factors should be taken into account combinedly, to select the proper method.

1.1.3 Errors,determinate errors,and Indeterminate errors,types of determinte errors,takaling of errors. 1.1.4 Quntitive methods of analysis: calibration curve, standard addition and internal standard methods

Quantitative Analysis 1. Calibration Curve method: 2. Method of Standard Addition 3. Internal Standard Method:

Quantitative Analysis: Calibration Curve method: A calibration curve is used to determine the unknown concentration of an element in a solution. The instrument is calibrated using several solutions of known concentrations. The property to be measured of each known solution is measured and then a calibration curve of property measured v/s concentration is plotted. The property of a sample solution is measured .The unknown concentration of the element is then calculated from the calibration curve

Calibration Curve method Sr. No. Concentration of KMnO4 O.D. 1 5 0.02 2 10 0.04 3 15 0.06 4 20 0.08 25 0.1 6 Unknown 0.05

Calibration Curve method: Measurable Property is directly proportional to concentration Measurable Property

Method of Standard Addition: When Standard Addition Method is useful Sample composition is unknown Sample composition affect on analysis To nullify the effect of Matrix

Method of Standard Addition: In this method the property of unknown (X) is first find out against blank. Then a series of standards having definite amount of unknown (X) plus varying amount of standard are prepared and diluted to same volume in each case .Their property is then measured. A graph of measured property against concentrations of standard (S) gives a linear curve. The concentration of the unknown can be determined by extrapolation of line which cuts to X axis.

Method of Standard Addition Sr. No. Volume Of Sample Solution Concentration of KMnO4 O.D. 1 5 0.02 2 0.06 3 10 0.08 4 15 0.1 20 0.12 6 25 0.14

. . . . . Standard addition method Property Absorbance--- . . . . . Absorbance--- X X+S X+2S X+3S X+4S Concentration----- Concentration of Unknown.

Internal Standard Method: When Internal Standard Method is useful It is useful for analysis when quantity of sample analyzed or the instruments responses varies slightly from run to run. Such response are difficult to control When sample loss occur during sample preparation

Internal Standard Method: A series of standard solution containing the same elements as that present in sample solution is prepared. A fixed quantity of suitable internal standard is then added to each of standard solutions, blank and sample solutions alike. The measurable property of each of above standard solutions and sample solutions are measured. The measurable property for each of above standard solution (Is) & (Ii) and sample solution (Ix&Ii) are measured at different wavelength one corresponds to element and other corresponds to the internal standard. These measurements are made against blank .The ratio of measured property of the standard solutions to that of internal standard (Is/Ii) are plotted against the concentration of standard solutions. This gives a straight line from this curve concentration of sample solution can be read by finding where the ratio (Ix/Ii) falls on concentration scale.

Internal Standard Method: Sr. No. Volume Of Sample Solution Concentration Internal Standard (ppm) O.D. 1 Blank 5 0.02 2 sample 0.06 3 0.08 4 10 0.1 15 0.12 6 20 0.14

Property of Internal standard Internal Standard Method Property of Sample Property of Internal standard Concentration of Unknown. Concentration-----

Property to be measured Types of Instruments

Signal/Characteristic property Instrumental Methods Emission of radiation Emission spectroscopy (X-ray, UV, visible, electron, Auger); fluorescence, phosphorescence, and luminescence (X-ray, UV, and visible) Absorption of radiation Spectrophotometry and photometry (X-ray, UV, visible, IR); photoacoustic spectroscopy; nuclear magnetic resonance and electron spin resonance spectroscopy Scattering of radiation Turbidimetry; nephelometry; Raman spectroscopy Refraction of radiation Refractometry; interferometry Diffraction of radiation X-Ray and electron diffraction methods

Polarimetry; optical rotary dispersion; circular dichroism Rotation of radiation Polarimetry; optical rotary dispersion; circular dichroism Electrical potential Potentiometry; chronopotentiometry Electrical charge Coulometry Electrical current Polarography; amperometry Electrical resistance Conductometry Mass-to-charge ratio Mass spectrometry Rate of reaction Kinetic methods Thermal properties Thermal conductivity and enthalpy Radioactivity Neutron Activation and isotope dilution methods

DATA DOMAINS Electrical and Non Electrical Domains, Detectors, Transducers and Sensors, Sensors

Data domains Information contained in Chemical and physical Characteristics of property of interest is called as data domain.

Non-Electrical domain Current, voltage, frequency, charge, pulse width, are called as electrical domain Physical and chemical property , scale, position and number are called as non electrical domain

Instruments converts Information for human Interpretation in terms of property interest and manipulation Nonelectrical domain Information contained in Chemical and physical Characteristics of property of interest Nonelectrical domain Transduction processing Electric Domain

CLASSIFICATION OF TRANSDUCERS The input transducer The output transducer The input transducer is called the sensor, because it senses the desired physical quantity and converts it into another energy form. It converts information in electrical domain to information in non-electrical domain (Voltmeters, computer screens, ADC)

Types OF TRANSDUCERS PHOTO TRANSDUCERS THERMAL TTRANSDUCERS BIOSENSOR

PHOTO TRANSDUCER Photo transducer is device that converts light energy into electrical energy are called Photo- transducer

PHOTOCELL DETECTOR (--) D D G C E- G B A +

PHOTO TUBE DETECTOR Photo Cathode Collector Anode - AMPLIFIER RECORDER

A SENSOR A sensor some time also called as detector that measures a physical quantity and converts it into signal which can read by an observer

Thermal Transducer Thermocouple Thermocouple sensor used for high temperature measurement. Thermocouple converts temperature to an out put voltage which can read by a voltammeter. Hg Mercury in glass thermometer converts measured temperature. Uses : Cars, Aero plane, Chemical plant ,Elevators, Doors , Manufacturing machine ,robotics.

Biological Sensors: A biosensor is an analytical device used for detection of analyte that combines a biological component with a physiochemical detector.

Thermal Transducers Infrared radiation generally does not have sufficient energy to produce a measurable current when using a photon transducer. A thermal transducer, therefore, is used for infrared spectroscopy. The absorption of infrared photons by a thermal transducer increases its temperature, changing one or more of its characteristic properties. The pneumatic transducer, for example, consists of a small tube filled with xenon gas equipped with an IR-transparent

SIGNAL PROCESSOR : A device, such as a meter or computer, that displays the signal from the transducer in a form that is easily interpreted by the analyst.

Amplifier: An Electronic device that increases the power of signal

Amplifier Weak Signal Strong Signal

Selection of Analytical method: 1 Concentration of the component. 2 The complexity of the materials/presence of interfering material 3 The probable concentration of the species of interest 4 Degree of Accuracy 5 Sensitivity and detection limit 6 Duration of an analysis 7 Speed ,time and Cost of analysis 8 Availability of equipments 9 Skill person for handling the instrument All the above factors should be taken into account combinedly, to select the proper method.

Performance characteristics of analytical Method : 1 Precision: It is the agreement between Individual observations of same set. 2 Accuracy: It is defined as closeness of the observed value with the true value. 3 Limit of detection(LOD): Minimum amount of concentration of a component that can be detected with a given degree of confidence. 4 Limit of Quantification (LOQ): Minimum amount of concentration of a component that can be estimated with a given degree of confidence is termed as LOQ.

Performance characteristics of analytical Method : 5 Limit of Linearity (LOL) : It is defined as maximum concentration range up to which instrument is produces linear response. 6 Sensitivity: It is a measure of ability of method to discriminate between two small concentration differences in the analyte. 7 Measurement of Sensitivity: It is is measured in terms of slope of the calibration curve. If the slope is greater sensitivity of the method is high and vise-versa.

Performance characteristics of analytical Method : 8 Selectivity: It is defined as degree to which the method is free from interferences from other components present in the matrix. 9 Dynamic range: It is the concentration range from limit of Quantification (LOQ) to Limit of Linearity (LOL).

1.1.3 Errors, determinate errors ,and Indeterminate errors, types of determinate errors, takaling of errors.

Any measurement involves the interaction of following three components Analyst Sample Method ERROR 11/9/2018

True Value Observed Value Difference between Error 11/9/2018

Types of Error Determinate error Indeterminate error Determinate error are those for which source can be observed or detected Indeterminate error The error are for which source can not be observed or not detected or can be pinpointed. 11/9/2018

No Source can be observed Sr. No. Characteristics Determinate error Indeterminate Error 1 Origin Source can be observed No Source can be observed 2 Magnitude Large Small 3 Direction Unidirectional No direction 4 Reproducibility Reproducible Not Reproducible 5 Effect Affect the measurement No Affect on measurement 6 Remedy Minimization possible, elimination in some cases possible No elimination 11/9/2018

Absolute error Relative error The difference between the measured value and True value Absolute error= xi -T Absolute error Absolute error / True value = xi –T/T Relative error 11/9/2018

Constant errors Proportionate errors The error in which the absolute error remains constant and the relative error changes with the change in sample size Constant errors The error in which the magnitude of the absolute error changes with change in sample size but relative error remains constant Proportionate errors 11/9/2018

Instrumental errors Methodic errors Types of Determinant error Operational errors Personal error 11/9/2018

Instrumental errors: Uncertainty in the last digit of the measurement due to least count of the instrument or volumetric glass ware. Ex. Counting /noting burette reading Improper response: Optimum condition for the working of the Instrument. Instrument works in that condition only. Ex. Working of glass electrode to measure pH using pH meter. pH of solution 1-10 can be recorded properly. If the solution is having pH greater than this range electrode system will give Improper response 11/9/2018

These types of errors obtained due to classical methods 2 Addition of excess amount of titrant 3 Incomplete reaction 4 Incomplete decomposition 5 Co-precipitation and post Co-precipitation 1 Solubility of salt These types of errors obtained due to classical methods as these methods involves no. of steps. Methodic Errors MnNH4PO4 ---- Mn2P2O7 +2NH3+H2O 11/9/2018

Operational errors Weighing of the hot crucible Loss of precipitate during filtration Blowing of last drop of in the nozzle of the precipitate Improper recording of the instrument Under washing or over washing of the ppt. Ignorance of temp. 11/9/2018

Personal error: The error due to physical limitation of the analyst and some time bias during measurement are called as Personal error. Ex. Colourblindnees of the person unable to detect end point . 11/9/2018

Minimization of errors Calibration of apparatus and Instruments Running Blank determination Use of Independent method of analysis Running control determination Running Parallel determination standard addition method Internal standard method Amplification method 11/9/2018

Calibration of apparatus and Instruments Operational and instrumental error can be minimized Calibration of apparatus and Instruments Methodic and operational errors can be minimized Running Blank determination Standard sub., Analysed and its result compared with the true value Deviation of the obtained result from the true or expected value will be measure of Methodic and operational errors Running of control determination 11/9/2018

Use of Independent method of analysis Analysis of same sample by two method of analysis ,one which will be chosen & results obtained can be compared. Methodic and operational errors can be different Use of Independent method of analysis Analysis of same sample by two different method by same analyst, or different . Methodic error will differ in two cases ,if same analyst Methodic and personal error will be differ in two cases if different analyst. Running Parallel determination Sample is analysed alone then sample + standard substance analysed Methodic and operational errors will be same for two measurements standard addition method 11/9/2018

Internal standard method Fixed amount of reference material is added to all standard solutions ,blank and sample . Ex. Na is added in the analysis of soil while determining lithium Internal standard method detector singles are amplified to rectify the improper response of detector. With the knowledge of type of error analyst can modify existing method, type of error and magnitude. Amplification method 11/9/2018

1.1.4 Quantitative methods of analysis

Quantitative Analysis 1. Calibration Curve method: 2. Method of Standard Addition 3. Internal Standard Method:

1.1.4 Quantitative Analysis: Calibration Curve method: A calibration curve is used to determine the unknown concentration of an element in a solution. The instrument is calibrated using several solutions of known concentrations. The property to be measured of each known solution is measured and then a calibration curve of property measured v/s concentration is plotted. The property of a sample solution is measured .The unknown concentration of the element is then calculated from the calibration curve

Calibration Curve method Sr. No. Concentration of KMnO4 O.D. 1 5 0.02 2 10 0.04 3 15 0.06 4 20 0.08 25 0.1 6 Unknown 0.05

Calibration Curve method: Measurable Property is directly proportional to concentration Measurable Property

Method of Standard Addition: In this method the property of unknown (X) is first find out against blank. Then a series of standards having definite amount of unknown (X) plus varying amount of standard are prepared and diluted to same volume in each case .Their property is then measured. A graph of measured property against concentrations of standard (S) gives a linear curve. The concentration of the unknown can be determined by extrapolation of line which cuts to X axis.

Method of Standard Addition Sr. No. Volume Of Sample Solution Concentration of KMnO4 O.D. 1 5 0.02 2 0.06 3 10 0.08 4 15 0.1 20 0.12 6 25 0.14

. . . . . Standard addition method Property Absorbance--- . . . . . Absorbance--- X X+S X+2S X+3S X+4S Concentration----- Concentration of Unknown.

Internal Standard Method: When Internal Standard Method is useful It is useful for analysis when quantity of sample analyzed or the instruments responses varies slightly from run to run. Such response are difficult to control When sample loss occur during sample preparation

Internal Standard Method: A series of standard solution containing the same elements as that present in sample solution is prepared. A fixed quantity of suitable internal standard is then added to each of standard solutions, blank and sample solutions alike. The measurable property of each of above standard solutions and sample solutions are measured. The measurable property for each of above standard solution (Is) & (Ii) and sample solution (Ix&Ii) are measured at different wavelength one corresponds to element and other corresponds to the internal standard. These measurements are made against blank .The ratio of measured property of the standard solutions to that of internal standard (Is/Ii) are plotted against the concentration of standard solutions. This gives a straight line from this curve concentration of sample solution can be read by finding where the ratio (Ix/Ii) falls on concentration scale.

Internal Standard Method: Sr. No. Volume Of Sample Solution Concentration Internal Standard (ppm) O.D. 1 Blank 5 0.02 2 sample 0.06 3 0.08 4 10 0.1 15 0.12 6 20 0.14

Property of Internal standard Internal Standard Method Property of Sample Property of Internal standard Concentration of Unknown. Concentration-----

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