Analysis

DIGESTER : PHARMACEUTICAL ANALYSIS DIGESTER
PHARMACEUTICAL ANALYSIS
DIGESTER -65
TYPES OF ERROR
Error is the difference between the true result (accepted true result) and the measured
result.
Determinate Determinate errors are caused by faults in the analytical procedure
Error or the instruments used in the analysis.
Determinate errors are systematic errors.
Indeterminate Indeterminate errors are not constant or biased. They are random
Error in nature and are the cause of slight variations in results of replicate
samples made by the same analyst under the same conditions
Gross Error Gross errors differ from indeterminate and determinate errors.
They usually occur only occasionally, are often large, and may cause
a result to be either high or low. They are often the product of
human errors.
Absolute error Absolute error = Observed value – True or most probable value
Relative error Relative error = Absolute error /True value1000 per thousand
DIGESTER -66
ACCURACY AND PRECISION
Accuracy Accuracy is the closeness of experimental value to the true value.

Precision Precision is the closeness of measurement from one another.
Reproducibility Reproducibility expresses the precision between laboratories
Repeatability Repeatability expresses the precision under the same operating
conditions over a short interval of time.
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107
DIGESTER GPAT DISCUSSION CENTER : MAKES STUDY EASY
DIGESTER -67
PRECISION MEASURE
Mean or Mean is obtained by dividing the sum of a set of measurements by the

average or number of individual results in the set.
relative mean mean,m =
 Mn Where
deviation n
M is individual measurement
n is the total number of measure
Median Median is a about which all the other value are equally distributed.
Mean deviation Mean deviation of a single measurement is the mean of the
derivations of all the individual measurements.
d -=
 Mn - m 
N
d-= mean deviation
= Absolute value of the deviation of the Mnth number
Relative mean Relative mean deviation is the mean deviation divided by the mean.
deviation Relative mean deviation = mean deviation ×100
mean
Average d -
Average deviation D =
deviation n
d-= Average deviation
n = Total number of measurements
Standard
  Mn - m 
2
deviation s=
N
DIGESTER -68
ELECTROMAGNETIC SPECTRUM

108
REGION λ WAVELENGTH ν FREQUENCY (HERTZ)

Cosmic rays 10-9 nm
Gamma rays Below 1 Å 1020 to 10 24
X rays 1 Å to 10 Å 1018
U. V. rays Far 10 nm to 200 nm 1016
Near 200 nm to 400 nm 1015
Visible rays 400 nm to 800 nm 1014
Infra-red Near 0.8 µm to 2 µm
middle 2 µm to 15 µm 1012 to 1014
Far 15 µm to 1000 µm
Microwave 0.05 cm to 25 cm 1010 to 1012
Radio frequency Above 25 cm 106 to 1010
DIGESTER -69
CHARACTERISTICS OF WAVE
CHARACTERISTICS DESCRIPTION
Frequency Number of waves per unit time.
Expressed in cycle per second (cps)/Hertz/Fresnel
Wavelength Distance between two nearest parts of wave in the same phase
Expressed in nm/ A°/μm
Wave number Number of waves per unit length
Expressed in cm-1 or Kaiser
Max-Plank E = h.ν = h c/λ
Equation Where E = energy of photon ν = frequency
h = Plank constant (6.6 × 10–27 erg-sec)
c = velocity of light , λ = wavelength
DIGESTER -70
CLASSIFICATION OF SPECTROSCOPIC TECHNIQUES
PRINCIPLE SPECTROSCOPIC METHOD

Absorption of  Ultraviolet and Visible Spectrophotometry
Radiation  Infrared Spectrophotometry
 Nuclear Magnetic Resonance (NMR)
 Electrons Spin Resonance (ESR)
 X-ray spectroscopy
 Atomic Absorption
 Colorimetry
Emission of  X-ray spectroscopy
Radiation  Atomic emission
 Fluorescence and phosphorescence
Spectrophotometry
 Ultraviolet and Visible Spectrophotometry
 X-ray spectroscopy

109
Refraction of  Refractometry and Interferometry

Radiation
Diffraction of  X ray
radiation  Electron Diffraction
Mass to charge  Mass Spectrometry
ratio
Scattering of  Raman spectroscopy
radiation  Nephelometry and Turbidimetry
Reflectance  Dynamic reflectance spectroscopy (DRS)
Rotation of  Polarimetry
Radiation  Optical rotary Dispersion
 Circular Dichroism
DIGESTER -71
ANALYTICAL TECHNIQUE AND MOLECULAR CHANGE
ANALYTICAL TECHNIQUE MOLECULAR CHANGES

NMR Spectroscopy Nuclei are oriented in magnetic field
Mass Spectroscopy Ionization of molecule/ fragmentation
IR Spectroscopy Vibration by light
Atomic emission Spectroscopy Light emission-excited electron
UV visible Spectroscopy Excitation of weakly bonded electrons
DIGESTER -72
SPECTROSCOPY AND THEIR PRINCIPLE
SPECTROSCOPIC METHODS PRINCIPLE

Absorption spectroscopy Measures the absorbance or percentage of transmittance
(ultraviolet, visible, radiation (of particular wavelength)
infrared)
Mass spectrometry Observe the position & intensity of signals in mass
spectrum causing the ionization of molecules
NMR spectrometry Observe the position & intensity line in NMR spectrum
when protons interact with EMR in radio frequency
region
Atomic absorption Measures the intensity of absorption when atoms absorb
spectrometry monochromatic light
X-ray spectroscopy Measure the position & intensity of spectral lines during
emission of X-ray spectrum by atoms under influence of
X-rays
Refractometry Measures the refractive index by causing refraction of
light by matter
Polarimetry Measure the optical relation by causing rotation of plane
polarized light

110
Fluorimetry Measures the intensity of fluorescence caused by

emission of electromagnetic radiation due to absorption
of UV radiation
Turbidimetry Measure the turbidity of a system by passing light beam
in a turbid media
Nephelometry Measure the opalescence of the media by reflection of
light by a colloidal solution
DIGESTER -73
SPECTROSCOPIC TECHNIQUES AND THEIR SOURCE OF RADIATION SOUCE
SPECTROSCOPY SOURCE OF RADIATION

Uv - visible  Deuterium lamp
spectroscopy  Hydrogen lamp
 Xenon arc lamp,
 Tungsten filament lamp,
IR spectroscopy  Nernst glower (Made up of with oxide of Zirconium oxide,
Yttrium oxide, Erbium, Thorium)
 Nernst glober
(Made up with Sintered silicon carbide)
 Mercury arc lamp
 Tungsten filament lamp
NMR spectroscopy  Radiofrequency source transmitter
Fluorescence  Low-pressure mercury vapour lamp
spectroscopy  High-pressure xenon arc lamp
ESR  Klystron oscillator
DIGESTER -74
SPECTROSCOPIC TECHNIQUES AND THEIR SOLVENTS
TECHNIQUES SOLVENTS
UV spectroscopy Ethanol, Methanol, Cyclohexane, Ether, Water
IR spectroscopy Cyclohexane, Xylene, Chloroform, Acetone, CCl4
Mass spectroscopy Methanol, Acetonitrile
NMR spectroscopy CDCL3, CCl4, DMSO4 , CS2 , D2O
ESR Methyl cyclohexane, Isooctane, Toluene

111
DIGESTER -75
SPECTROSCOPIC TECHNIQUES AND THEIR DETECTOR
SPECTROSCOPY DETECTOR MADE UP OF WITH

Barrier layer cell (photo Fe, Al, Se, Au, Ag metal
voltic cell)
Cathode: Photo-sensitive material like
Photo emissive cell sodium potassium or cesium
UV-Visible Anode: Straight wire made of nickel or
Spectroscopy platinum
Photo multiplier tube Consists of a photo emissive cathode
Thermo couple Bismuth and Antimony
Bolometer Platinum black
IR Spectroscopy Golay detector Xenon gas
Thermistor Oxide of Mn, Co, Ni
Pyroelectric Triglycerine sulphate used as medium
NMR Phase sensitive detector
ESR Silicon crystal
Geiger muller counter Argon gas, Xenon and Crypton gas
X-ray Proportionality counter NaF, p-cresol in xylene
Scintillation counter Gallium lithium
Semiconductors Silicon lithium
Faraday cup Metal cup
Mass Array detector
spectroscopy Electron multiplier
Charge detection
Fluorescence and Mercury vapour lamp
phosphorescence Xenon arc lamp
DIGESTER -76
ANALYTICAL TECHNIQUE AND THEIR REFRENCE STANDARD
ANALYTICAL TECHNIQUE REFERENCE STANDARD GRAPH

UV Spectroscopy Potassium chromate Absorbance v/s concentration
IR Spectroscopy Polystyrene film %transmittance v/s wave
number
NMR Spectroscopy Tetramethylsilane (for Radiofrequency absorption v/s
organic solution) Field strength
Silapentane ( for aqueous
solution)
Mass Spectroscopy Per fluoro kerosene Abundance v/s m/e ratio
ESR Spectroscopy DDPH (1,1diphenyl picryl Intensity v/s magnetic field
hydrazyl)

112
DIGESTER -77
UV -VISIBLE SPECTROSCOPY
Beer’s law Find relationship between transmittance and concentration of

medium
Lambert’s law Find relationship between transmittance and thickness of
medium
Beer Lambert law A = .Cl
Where,  = Molar absorption coefficient
C = Concentration
l = Thickness
Molar absorption  = A/ Cl
coefficient
Transmittance T = IT/ Io
Absorbance A = -log T
Specific absorbance Є = (A1cm1% × mol.wt)/10
Bathochromic lmax Towards to longer Wavelength
(Red shift)
Hypsochromic lmax Towards to shorter Wavelength
(Blue shift)
Hyperchromic Increases in the intensity
Hypochromic Decrease in the intensity
Isobestic point Every absorption curve which is contained in spectrum of
compound taken at different pH.
Chromophore Molecular group that absorb visible or UV light and imparts
colour to the compound. Example: Nitro group, Azo group
Auxochrome They do not have any characteristic absorption on their own but
can modify the absorption of chromphore.
Example: -OH, –OR, –NH2 , -NHR
K-Bands Originates due to ᴫ⟶ᴫ* transition in a compound with
conjugated ᴫ system
Very intense band with high εmax
Present in conjugated dienes like butadienes
R-Bands These are forbidden transitions, originates due to n⟶ᴫ*
Transition of electron of atleast one lone pair of electron on
hetero atom
Have very low εmax value , below 100
B-Bands Originates due to ᴫ⟶ᴫ* transition in aromatic or hetero-
aromatic molecules
E-Bands Originates due to electronic transitions in the benzenoid system
In cyclic conjugation, they shows two absorption bands in UV
spectra.
Energy value order σ → σ* ˃ n → σ* ˃ π → π * ˃ n → π*
for transition

113
DIGESTER -78
WOODWARD FIESER RULE
Homoannular diene: It is a cyclic diene having conjugated double bond in the

same ring.
Example:
Heteroannular diene It is a cyclic diene in which double bonds in conjugation are

present in different rings.
Example:
Endocyclic double bond A double bond present in a ring.

Exocyclic double bond A double bond in which one of the double bond is a part of a
ring system.
Exocyclic double bond
Example:
Endocyclic double bond

Ring A has one endocyclic
A B and one exocyclic double bond.
Ring B has only one endocyclic
double bond.
WOODWARD FIESER RULE FOR CONJUGATED DIENE, TRIENE SYSTEMS

Homoannular conjugated diene 253 nm
Heteroannular conjugated diene 214 nm
Acyclic conjugated diene 217 nm
Parent Values Acyclic triene 245 nm
Increment Each alkyl substituentor Ring residue +5 nm
Exocyclic double bond +5 nm
Double bond extending conjugation +30 nm
Auxochromes -Cl, -Br +5 nm
-OH/-OR/-SH +6 nm
-SR +30 nm
-NR2 +60 nm
-OCOCH3 +0 nm
Question: Calculate λmax Question: Calculate λmax
H 3C CH3
Parent value for Homoannular diene =
253 nm Two alkyl substituents = 2 × 5 = 10 nm
Two alkyl substituents = 2 × 5 = 10 nm Three ring residue = 3 × 5 = 15 nm
Two ring residue = 2 × 5 = 10 nm One exocyclic double bond = 5 nm
Total calculated λmax = 253 + 10 + 10 = Total calculated λ max = 214 + 10 + 15 + 5 =
273 nm 244 nm

114
WOODWARD FIESER RULE FOR α, β-UNSATURATED CARBONYL COMPOUNDS
Parent 215 nm
R = H (Aldehyde) 207 nm
Values X = OH, OR (Acid or Ester) 193 nm
X = alkyl (Ketone) or six membered ring 215 nm
Homoannular conjugated diene +39 nm
Increment Exocyclic double bond +5 nm
Double bond extending conjugation + 30 nm
Alkyl group α β γ /higher

-Cl +15 nm +12 nm +12 nm +12 nm
-OH +35 nm +35 nm +30 nm +50 nm
-SR - 85 nm - -
Auxochrome -NH2 - 95 nm -
-OCHCH3 +6 nm +6 nm +6 nm +6 nm
-Br +25 nm +30 nm +18 nm +31 nm
-OR +35 nm +35 nm +18 nm +31 nm
Question: Calculate λmax.
O
Parent value for β unsaturated 6 membered cyclic compound = 215 nm

One ring residue on α carbon = 10nm
Two ring residue on β carbon = 2 ×12 =24 nm
Double bond exocyclic to two (both) ring = 2 × 5 = 10 nm
Total calculated λ max = 215 + 10 + 24 + 10 = 259 nm
WOODWARD FIESER RULE FOR ACYL BENZENE DERIVATIVE

X = Alkyl 246 nm
Parent value X = OH/OR 230 nm
X=H 250 nm
Ortho Meta Para
Alkyl +3 nm +3 nm +10 nm
OH/OR +7 nm +7 nm +25 nm
Auxochrome Cl 0 nm 0 nm +10 nm
Br +2 nm +2 nm +15 nm
NH2 +13 nm +13 nm +58 nm
NHOCOCH3 +20 nm +20 nm +45 nm

115
Question: Calculate λmax. Question: Calculate λmax.

CH3 OH
O O
Br Br
Br OH
parent value for acyl benzene (Ketone) Parent value for aromatic carboxylic acid
derivative= 246 nm derivative = 230 nm
Br atom at para position = 15 nm Br atom at two ortho position = 2 × 2 = 4 nm
Total calculated λmax = 246 + 15 = 261 nm OH group at para position = 25 nm
calculated λmax = 230 + 04 + 25 = 259 nm
DIGESTER -79
INFRA RED SPECTROSCOPY
INFRA RED REGION

Near infrared 0.8 µm to 2 µm
(Overtone region)
Middle infrared 2 µm to 15 µm
(Fundamental region)
Functional region 2 µm to 8 µm
(1300 – 4000 cm-1)
Fingerprint region 8 µm to 15 µm
(650 – 1300 cm-1)
Far infrared (Rotational 15 µm to 1000 µm
vibration)
Vibrational frequency 1 k

2 c 
Where, = Frequency, = Wave number
C = Velocity of light, = Force constant of bond
DEGREE OF FREEDOM
Type of degree of Liner Non-liner
freedom
Fundamental 3n-5 3n-6
absorption band
Transitional degree of 3 3
freedom
Rotational degree of 2 3
freedom
Vibrational degree of 3n-5 3n-6
freedom
Total degree of freedom 3n 3n

116
TYPE OF VIBRATION
Stretching Bond length Symmetrical Two bends increase or decrease in
vibration altered. length symmetrically.
Asymmetrical Two bends increase or decrease in
length asymmetrically.
Bending Bond angle In plane bending Scissoring Bond angle decrease.
vibration altered. Rocking Bond angle maintained
but both bonds move
within the plane.
Out of plane Wagging Both atoms move to one
bending side of plane
Twisting One atom is above the
plane and the other is
below the plane.
DIGESTER -80
ABSORPTION PEAK WITH INTENSITY OF SOME SELECTED FUNCTIONAL GROUP
TYPES OF COMPOUNDS FREQUENCY RANGE CM-1

C-H stretching
Alkane 2850-2970
Alkene 3010-3095
Alkynes 3320-3310
Aromatic rings 3310-3100
Aldehydes 2900-2500
C=C & C=C bond stretching
Alkane 1680-1620
Alkynes 2300-2100
C=O stretching (Carboxyl)
Saturated aliphatic ketone 1750-1700
α, β unsaturated aliphatic ketone 1685-1660
Saturated Aliphatic Aldehyde 1740-1720
Α, β unsaturated aliphatic aldehyde 1705-1680
Aryl aldehyde 1700-1680
Saturated ester 1750-1735
Unsaturated ester 1730-1715
Saturated carboxylic acid 1725-1700
Unsaturated carboxylic acid 1715-1690
Aryl carboxylic acid 1700-1680
Amide 1680-1630
Imide 1700-1670
Lactam 1720-1660
Thiocyanate (C=S) 1200-1025
Sulphone (S=O) 1180-1140
Sulphonamide 1350-1300
O-H stretching (Hydroxyl)
Alcohol (OH) free 3650-3450
Hydrogen bonded 3570-3450
Sec Ter –OH bonding alcohol 1100-1050
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N-H stretching (Amines)

Pri, sec, ter amines 3500-3400
N-H bending (Amines)
Pri, sec 1650-1550
C-N stretching
Aliphatic 1200-1000
Aromatic 1350-1250& 860
C=N nitrile 2280-2200
Halogens Compounds
Chlorides 800-600
Bromides 650-500
Iodides 600-500
Fluorides 1400-1000
DIGESTER -81
NMR SPECTROSCOPY
Fermions Odd mass nuclei with an odd number of nucleons have

fractional spins.
I = 1/2 ( 1H, 13C, 19F, 31P )
I = 3/2 ( 11B, 33S )
I = 5/2 ( 17O )
Gives NMR spectra.
Passes angular momentum.
Bosons Even mass nuclei with odd numbers of protons and
neutrons have integral spins.
I = 1 ( 2H, 14N )
I = 0 (12C, 16O, 32S)
Not gives NMR spectra.
Not passes angular momentum.
 g 
   
Larmor  2 
equation Where, g = Gyromagnetic ratio
B = Strength of magnetic field
Chemical shift A chemical shift is defined as the difference in parts per
million (ppm) between the resonance frequency of the
observed proton and Tetramethylsilane (TMS) hydrogens.
δ is dimensionless and expressed as ppm
Tetra methyl TMS is the most common Reference compound in NMR,
silane it is set at δ=0 ppm.
TMS is used as reference in proton NMR. Because
1. TMS has 12 equivalent protons.
2. Chemically inert and very low boiling point.
3. Miscible with all organic substances.
TMS is not soluble in aqueous solution hence 2, 2
dimethyl-2-2-silapentane-5-sulphonate used.

118
Shielding of High electron density around a nucleus shields the

protons nucleus from the external magnetic field and the signals
are up field in the NMR spectrum
DE shielding of Lower electron density around a nucleus Deshields the
protons nucleus from the external magnetic field and the signals
are downfield in the NMR spectrum
𝜏 = 10 – δ
δ = Chemical shift
DIGESTER -82
SOME COMPOUNDS WITH THEIR NMR SIGNAL
NAME OF COMPOUNDS NUMBER OF NMR SIGNALS

Acetone 01
1,2-dibromoethane 01
Cyclobutane 01
Ethyl chloride 02
Methanol 02
Isopropyl chloride 02
Tertiary butyl amine 02
1,1-dibromoethane 02
Methylene 02
Diethyl ether 02
Ethyl benzene 03
Diethyl succinate 03
Ethyl amine 03
2-propanol 03
n-propyl chloride 03
2-bromopropene 03
Vinyl chloride 03
1,2-dichloropropane 04
Methyl cyclopropane 04
Ethoxy acetic acid 04
Propyl formate 04
DIGESTER -83
ABSORPTION POSITION OF SOME PROTON
TYPE OF PROTON CHEMICAL SHIFT

HC CH 2 to 3
3 to 4
CH Cl
3.4 to 4
CH OH
3.3 to 4
CH O R

119
O 2 to 2.7
CH C OH
O 9 to 10
R C H
R NH2 1 to 5
DIGESTER -84
MASS SPECTROSCOPY
Molecular ion Ion formed from a molecule by removal of one electron of lowest
peak (parent ionization potential.
peak) Height of parent peak = Aromatic˃ Acylic ˃ Ketone ˃ Amine ˃ Ether ˃
Carboxylic acid
Base peak Most intense peak.
It is considered as 100%
Most abundant peak
Fragment ion The ions produced from the molecular ion by cleavage of bonds are
called fragment ions.
Metastable Mass spectrum of molecule shows sharp peaks at m/z integrals.
Ions
M+1 peak It occurs due to presence of C13 , H2 , N13
M+2 peak It occurs due to presence of O18 , S34 , l37 ,Br31
DIGESTER -85
TYPE OF MASS ANALYZER AND THEIR WORKING
MASS ANALYZER WORKING

Quadrupole Scan radio frequency field
Time Of Flight (Tof) Time Of Flight correlate directly to ion’s m/z
Magnetic Sector molecular ions are separated according to their masses
and collected
Double Focusing Magnetic differentiate the small mass differences of the fragment
Quadruple Ion Trap Scan radio frequency field
Ion Cyclotron Resonances Translate ion cyclotron motion to m/z
DIGESTER -86
TYPE OF IONIZATION SOURCE AND THEIR PRINCIPLE
IONIZATION SOURCE PRINCIPLE

Electron Impact (El) Electron transfer
Chemical Ionization (CI) Proton transfer
HPLC Matrix-Assisted Laser Desorption (MALDI) Photon transfer
Fast Atom Bombardment (FAB) Ion desorption
Atmospheric Pressure Chemical Ionization(APCI) Corona discharge
Electrospray Ionization (ESI) Evaporation of charged droplet

120
DIGESTER -87
MC-LAFFERTY REARRANGEMENT REACTION
Mc-Lafferty rearrangement reaction - It involves the migration of γ-hydrogen atom

followed by cleavage of β-bond then rearrangement leads to the elimination of neutral
molecule. Elimination of neutral molecules from Aldehydes, Ketone, Amine,
Unsaturated compounds, Substituted aromatic compound etc takes place. The
rearrangement proceeds through a sterically hindered 6 membered transition state.
H H + +
: O: .O: O.
+
.
Molecular ion
DIGESTER -88
COUPLING CONSTANT
Coupling constant • Distance between two split lines.

• Expressed in Hertz.
• Ortho, meta, para-isomers are differentiated on the
basis of Coupling constant.
Compounds Coupling constant values
Anti 5 - 12
Gauche 2-4
Cis 6 - 14
Trans 11-18
Geminal protons 0 - 20
Vicinal protons 2 - 18
DIGESTER -89
INFORMATION FROM PMR
Area of Peak Number of absorbing protons giving rise to a signal

Intensity of Signal Relative number of protons of different kinds
Number of Signals Different sets of equivalent protons in molecule
Splitting or Environment of proton with respect to neighboring proton
Multiplicity of Signal

121
DIGESTER -90
X RAY
X ray was discovered by William K. Roentgen.

Nλ = 2d sinθ
Bragg’s equation Where,
N = integer
d = distance between crystal
θ = Angle of diffraction
X-ray technique Principle
X-ray absorption method Based on absorption of x-ray
X-ray diffraction method Based on scattering of x-ray
X-ray fluorescence method Based on emit x-ray of characteristic
wavelength
DIGESTER -91
COMPARISON OF ATOMIC ABSORPTION AND FLAME EMISSION SPECTROSCOPY
ATOM ABSORPTION SPECTROSCOPY FLAME EMISSION SPECTROSCOPY

Amount of light absorbed by ground state Amount of light emitted by excited atom is
atom is measured. measured.
Absorption intensity a. signal response Does Absorption intensity and signal response
not depend upon temperature. greatly influenced by temperature
variation.
Beers law is obeyed. Beer law is not obeyed.
Intensity of emitted radiation is directly Intensity of absorbed radiation is directly
Proportional to the number of atoms in proportional to the number of atoms in
ground state. excited state.
DIGESTER -92
COMPARISON OF NEPHELOMETRY AND TURBIDIMETRY
NEPHELOMETRY TURBIDIMETRY
Nephelometry deals with measurement of Turbidimetry deals with measurement of
Intensity of scattered light Intensity of transmitted light
In Nephelometry, the intensity of the Turbidimetric measurements are made at
scattered light is measured, usually at right 180o from the incident light beam.
angles to the incident light beam.
Similar to Fluorimetry because both Similar to Colorimetry because both
measure scattered radiations but elastic measure transmitted radiations but light
scattering in Fluorimetry while non-elastic intensity decreased by scattering in
scatter in Nephelometry. Turbidimetry while by absorption in
Colorimetry.
Low concentrated suspension High concentrated suspension

122
DIGESTER -93
COMPARISON OF FLUORESCENCE AND PHOSPHORESCENCE
FLUORESCENCE PHOSPHORESCENCE
Life time Short, < 10-5 Sec Long, Several Seconds
Electron spin No Yes
Excited states Singlet Triplet
Quantum yield High Low
Temperature Most Temperature Proffered in Low Temperature
DIGESTER -94
APPLICATION OF ANALYTICAL TECHNIQUE
TECHNIQUE APPLICATION
 Detection of impurities
UV VISIBLE  Structure elucidation of organic compounds
 Distinction between conjugated and non-conjugated system
 Identification of functional group
 Study of polymer
IR  Structure determination
 Distinction between inter and intramolecular H-bonding
 Study of keto-enol Tautomerism
 Identification of Geometrical isomer
 Detection of H-bonding
NMR  Detection of aromaticity
 Distinction between cis-trans isomer
 Detection of electronegative atom
 Molecular mass determination
 Identification of unknown compound
MASS  Drug metabolism study
 Clinical, toxicological and forensic application
 Presence of isotopes
AAS  It can be used to determine Al, Mg, Cu, Zn, Pub, and Ni.
 Estimation of trace elements in biological fluids (blood).
ESR  Study of free radical.
 Determination of reaction rate and mechanism.
 Study of inorganic compound.
X-ray  Determination of crystalline structure
 Determination of particle size
 Property of metal
Raman  Structure elucidation of molecules
spectroscopy  Presence of Tautomerism
 Study of ionic equilibrium and chemical bond

123
Nephelometry  Turbidimetric titration

Turbidimetry  Clinical immunology (Ag- Ab. detection, Determination of CSF,
Rheumatoid factor detection)
 Determination of hardness of water
 Analysis of sugar product
 Determining of concentration of smog, fog and aerosol.
Fluorimetry  Determination of Uranium in salt
 Determination of vitamin
 Fluorescence indicator
Eosin pH 3 to 4 (Colourless to green)
Fluorescein pH 4 to 6 (Colourless to green)
DIGESTER -95
ELECTRO ANALYTICAL TECHNIQUE
BASIC PRINCIPLE CONSTANT ELECTRODE

Conductometry Conductance V/S Conductance Platinum
volume of Titrant electrode
added
Potentiometry Potential V/S volume of Potential
Titrant added (No current
flow i.e., I = 0)
Polarography Current V/S Applied Half way potential Dropping
Potential mercury
Amperometry Current V/S volume of Conductance Rotating
Titrant added platinum
(V=Constant) electrode
DIGESTER -96
TYPES OF ELECTRODE
Reference electrode Standard Hydrogen Electrode (SHE)

Saturated Calomel Electrode (SCE)
Silver-Silver Chloride Electrode
Indicator electrode Metal indicator
Membrane Idicator Electrode
ELECTRODE SYSTEM AND THEIR CONSTRUCTION
Electrode System Construction
Ag/AgCl electrode Silver wire is coated with thin film of silver chloride and kept in
saturated solution of KCl.
Calomel ( (Hg/Hg2 Cl2 ) Solid mercury surrounded with Hg, Hg2 Cl2 Paste and kept in
electrode saturated solution of KCl.
Glass electrode Most widely used H+ ion sensitive electrode used in pH meter.
Standard Hydrogen It is a primary reference electrode. It consists of Pt electrode
electrode (SHE) immersed in a solution whose hydrogen ion activity is 1.0 and
in which H2 gas is Bubbled at 1 atm Pressure.

124
DIGESTER -97
THERMO ANALYTICAL TECHNIQUE
METHODS PARAMETER MEASURED GRAPH

Differential thermal analysis (DTA) Temperature difference ∆T v/s temp.
Differential scanning Colorimetry Enthalpy dH/dt v/s temp.

(DSC)
Thermo gravimetric analysis (TGA) Mass Mass v/s temp.
Dynamic mechanical analysis (DMA) Deformation -
Dielectric thermal analysis (DETA) Deformation -
Evolved gas analysis (EGA) Gaseous decomposition Thermal
conductivity v/s
temp.
Thermo-optical analysis (TOA) Optical properties -
DIGESTER -98
TYPE OF TITRATION
TITRATION DESCRIPTION APPLICATION

Neutralization in which  To analyses mixture of acids
Acid base titration titrant is delivered into an  To control acidity or alkalinity
analyte until the unknown
solution is completely
neutralized.
Titration of oxidizing  Determine oxidation state of element
agent by reducing agent  Quantitative determination of metal
Redox titration or titration of reducing Ca, Mg, Zn, Co, Ni
agent to oxidizing agent.
Complexometric Titration of metal ions Determination of hardness of water
titration with a complexing agent
or chelating agent.
Appearance or  Measurement of concentration of
disappearance of chlorine in water
elementary iodine  Analysis of hydrogen peroxide
Iodimetry titration indicates end point.
Iodimetry – Titration with standard solution of iodine.
I2 + 2S2O3-2 → S4O62-
Iodometry – Titration in which iodine is liberated in with chemical
reaction. Br2 + 2I- → 2Br - + I2

125
One product is a  To determine electrode potential

precipitated.  For determination of chloride,
(Known as Argentometric cyanide and thiosulphite
titration)
Mohr titration Titration of sodium chloride with
standard silver nitrate in neutral solution
giving a precipitate of red silver chromate
at the end point.
Ag+ + Cl- → AgCl ↓
Precipitation titration 2Ag+2 + CrO-4 (Red colour precipitate)
Fajan titration Involve titration of NaCl with standard of
silver nitrate using adsorption indicator.
Adsorption indicator are: Fluorescein,
Eosin
Volhard’s titration AgNo3 + NH4SCN → AgSCN + NH4NO3
NH4SCN + Fe2(SO4)3(NH4)S04 → Fe [
Fe(SCN)6] ↓ (Red colour complex
indicate end point)
DIGESTER -99
INDICATORS FOR ACID BASE TITRATION
COLOUR ON COLOUR ON
INDICATOR PH RANGE OF
ACIDIC SIDE BASIC SIDE
Methyl Violet 0.0 to 1.6 Yellow Violet
Bromophenol Blue 3.0 to 4.6 Yellow Blue
Methyl orange 3.1 to 4.4 Red Yellow
Methyl red 4.4 to 6.2 Red Yellow
Phenol red 6.8 t0 8.4 Yellow Red
Cresol red 7.2 to 8.8 Yellow Red
Naphtholphthalein 7.3 to 8.7 Yellow Blue
phenolphthalein 8.3 to 10.0 Colourless Pink
Alizarin yellow 10.1 to 12.0 Yellow Red
DIGESTER -100
INDICATORS FOR COMPLEXOMETRIC TITRATION
INDICATOR PH RANGE COLOUR CHANGE

Mordant Black II 6 to 7 Red to Blue
Enoehrome Black T 6 to 7 Red to Blue
Solochrome black T 6 to 7 Red to Blue
Murexide 12 Violet to blue
Catechol Violet 8 to 10 Violet to red
Methyl blue 4 to 5 Blue to yellow

126
Thymol blue 10 to 12 Blue to gray

Alizarine 4.3 Red to yellow
Sodium alizarie sulphate 4 Blue to red
Xylenol 1 to 6 Lemon to yellow
DIGESTER -101
PRIMARY STANDARD
TITRATION METHOD PRIMARY STANDARDS

Acid-base reactions Na2CO3, Na2B4O7, KH (C8H4O4), HCl
Complex formation reactions AgNO3, NaCl
Precipitation reactions AgNO3, KCl
Redox reactions K2Cr2O7, Na2C2O4, I2
DIGESTER -102
POINTS IN COMPLEXOMETRIC TITRATION
EDTA EDTA is used as chelator (Hexadentate).

EDTA has six potential sites for bonding a metal ion; the four
carbonyl groups and two amino groups.
INDICATOR - Erichrome Black T
Masking Agent Masking agent is a reagent that protects some component of the
analyte from reaction with EDTA.
Demasking Demasking Agent is a reagent that release of a metal ion from a
Agent masking agent.
DIGESTER -103
NON AQUEOUS TITRATION TYPES OF SOLVENT
SOLVENT DESCRIPTION EXAMPLE

Aprotic Solvent chemically inert and are not Chloroform, Benzene
solvent involved in any chemical reaction.
Photogenic These are acidic substance and yield Sulphuric acid,
solvents protons. Hydrochloric acid, Nitric
acid
Photophilic These are basic in nature and can abstract Pyridine, n-Butyl amine
solvents proton from acids to give solvated protons.
Amphiprotic Both photogenic and photophilic. Water, Acetic acid, Alcohol
solvents

127
DIGESTER -104
CHRMATOGRAPHIC TECHNIQUES
PRINCIPLES TECHNIQUES
 Thin Layer Chromatography (TLC)
 High Performance Liquid Chromatography (HPLC)
 Adsorption Column Chromatography
Adsorption
 Gas Solid Chromatography
Partition  Paper Chromatography
 Gas Liquid Chromatography
Ion-Exchange  Ion Exchange Chromatography
Based on molecular size  Size-Exclusion Chromatography
(Exclusion)  Gel Filtration and Gel-Permeation
 Chromatography (GPC)
Analyte and ligand interaction  Affinity Chromatography
is utilized for the separation.
DIGESTER -105
CLASSIFICATION OF CHROMATOGRPHIC SEPARATIONS
TYPE OF MOBILE TYPE OF METHOD OF FIXING THE

NAME
PHASE STATIONARY PHASE STATIONARY PHASE
Adsorption Liquid Solid In a tubular column
Gas-solid Gas Solid In tubular column
Adsorbed on a porous solid
Gas-liquid Gas Liquid held in a tube or on the inner
surface of a capillary tube
Gel Liquid Liquid In the interstices of a
polymeric solid
Ion Liquid Solid Finely divided ion-exchange
exchange resin in a tubular column
Paper Liquid Liquid In the pores of a thick paper
Partition Liquid Liquid Adsorbed on a porous Solid in
a tubular column
Thin layer Liquid Liquid or solid Finally divided solid on a glass
plate
DIGESTER -106
NORMAL PHASE V/S REVERSE PHASE
CHARACTERISTIC NORMAL PHASE REVERSE PHASE

Stationary phase Polar (silica gel) Non polar (octa decyl silane C18)
Mobile phase Non polar Polar
Mechanism Adsorption Partition
Compound eluted first Non polar Polar

128
DIGESTER -107
TYPE AND THEIR STATIONARY AND MOBILE PHASE
TECHNIQUE STATIONARY PHASE MOBILE PHASE

Adsorption Silica gel, alumina, Nonpolar/Polar organic
charcoal, solvents
polyamide
Partition Cellulose, silica gel Mixed aqueous, organic
solvents
Reversed Phase ODS silica gel, coated silica, Mixed aqueous, polar solvents
acetylated cellulose
Ion exchange Ion exchange resins, Buffered aqueous solutions
cellulose,
Diethyl amino ethyl
Size exclusion Dextran gels, Agarose, Buffered aqueous solutions
Polyacrylamide
DIGESTER -108
VISUALIZING AGENT
COMPOUND REAGENT COLOUR

General Iodine vapour Brown
Acids Bromo cresol green Yellow
Aldehyde and Ketone 2, 4-Di nitro phenyl hydrazine Yellowish red
Amino acid Ninhydrin Purple
Alkaloids Mercuric nitrate Yellowish brown
Carbohydrate Aniline phthalate Gray black
Lipids Bromothymol blue Light green
Steroids Antimony trichloride Purple
Phenolic compound FeCl3 Purple
REAGENTS USED FOR

10% Sulphuric acid in ethanol All type of compounds.
Cobalt Chloride (CoCl2) Universal Stain
4-amino Antipyrine Phenols and aryl amines
Ammonium molybdate Phosphoric acid derivative
Aniline phthalate Reducing sugar
Anisidine phthalate Carbohydrates
Chloranil reagent Phenols
Dithiazone Metal ions
Ehrlich’s reagent Amine, indole derivatives
Dichlorofluorescin / fluorescin sod. Salt N-substituted barbiturates
Formaldehyde / sulphuric acid (1:10) Antihypertensive drugs
Gibb's reagent Phenols
Methyl yellow Chlorinated insecticide

129
DIGESTER -109
TYPE OF STATIONARY PHASE
Silica gel Most widely used adsorbent

(SiO2) Surface of the silica gel is acidic in nature.
Alumina (Al2O3) The active groups on the surface of chromatography alumina are
hydroxyl groups, oxide (O2–) ions and an aluminium cation.
Diatomaceous earth Chemical composition of oven-dried diatomaceous earth is 80-
(Kieselguhr) 90% silica, with 2-4% alumina and 0.5-2% iron oxide.
Magnesium Silicate Used for separation of sugars.
(MgSiO3)
Cellulose Commonly used for separating hydrophilic substances like amino
acids, sugars.
Charcoal Specific property of adsorbing strongly aromatic substances.
DIGESTER -110
PURE CELLULOSE PAPERS (WHATMAN PAPERS)
These types of papers are prepared from cotton linters selected to be especially low in
organic and inorganic impurities and uniform in physical characteristics.
Components Percentage
α Cellulose 98-99
β Cellulose 0.3-1.0
Pentosans 0.4-0.8
Ether soluble matter 0.015-0.03
Ammonia 0.001-0.06
Organic nitrogen <0.01
Inorganic material 0.008-0.06
Mostly used whatmann chromatographic filter papers are
Whatmann 31ET Used for separation of substances having
sufficiently wide apart Rf
Whatmann 3MM Generally used for preparative purposes
Whatmann 20 Slow paper
Whatmann 540-42 Acid washed paper
DIGESTER -111
COMPARISON ON COLUMN, HPLC AND GAS CHROMATOGRAPHY
Column chromatography HPLC

Stationary phase (particle size) 70 – 150 µm 3 – 20 µm
Column size Large Small
Pressure 50 – 150 psi 500 – 3000 psi

130
Column (internal diameter) HPLC Gas chromatography

Preparative 100 mm -
Normal bore 3.9 to 5.0 mm 0.53 mm
Mini bore 2.1 to 3.9 mm 0.18 mm
Micro bore 1.0 to 2.1 mm 0.1 mm
Capillary 50 µm to 1 mm 0.2, 0.25
DIGESTER -112
COMPARISON BETWEEN TLC AND HPTLC
PARAMETER TLC HPTLC

Technique Manual Instrumental
Efficiency Less High
Layer Lab made/ pre-coated Pre-coated
Mean particle size 10-12µm 5-6 µm
Layer thickness 250 µm 100 µm
Plate height 30 µm 12 µm
Solid support Silica gel, Alumina, Silica gel – normal phase
Kieselguhr
C8 and C18 – reverse phase
Sample spotting Manual spotting (capillary) Auto sampler (syringe)
DIGESTER -113
DETECTORS IN HPLC
DETECTORS DESCREPTION
Uv-visible spectroscopic Measures the ability of solutes to absorb light at a
detectors particular wavelength(s) in the ultraviolet (uv) or visible
(vis) wavelength range.
Refractive index detector Measures the molecule’s ability to deflect light in a flowing
mobile phase in a flow cell relative to a static mobile phase
contained in a reference cell.
Photo diode array detector Monitoring absorbance of solutes at several different
wavelengths
Fluorescence detectors Sensitivity depends on the fluorescence properties of the
components in the elute
Electrical conductivity Conductivity detectors measures electronic resistance and
detector measured value is directly proportional to the
concentration of ions present in the solution.

131
DIGESTER -114
DETECTORS IN GAS CHROMATOGRAPHY
SUPPORT
DETECTORS TYPE SELECTIVITY DETECTABILITY
GASES
Flame ionization Mass flow Hydrogen Most organic 100pg
(FID) and air compounds
Thermal Concentration Reference Universal 1ng

conductivity
(TCD)
Electron capture Concentration Make up Halides, nitrates, 50fg
nitriles,
peroxides,
anhydrides,
organometallics
Nitrogen Mass flow Hydrogen Nitrogen, 10pg
phosphorus and air phosphorus
Flame Mass flow Hydrogen Sulphur, 100pg
photometric and air phosphorus, tin,
possibly boron, arsenic,
oxygen germanium,
selenium,
chromium
Photoionization Concentration Make up Aliphatic, 2pg
aromatics,
ketones,
esters,
aldehydes,
amines,
heterocyclic
DIGESTER -115
IMPORTANT POINT IN GAS CHROMATOGRAPHY
Carrier gas used in gas Helium, Argon, Nitrogen,

chromatography
Flow regulator used in gas Rotameter, Soap bubble flow meter
chromatography
Column type in in gas (a) Capillary column – Diameter 0.2 to 0.7 mm
chromatography length – 150 meter
(b) Packed column – Diameter 2 to 5 mm length 1
to 3 meter

132
Parameter
Retention time Difference between point of injection and
appearance of peak maxima.
Retention volume Volume of carrier gas required to elute
50 % of component from the column.
HETP (Height equivalent to If HETP is less – column is more efficient
theoretical plate) If HETP is more – Column is less efficient
Number of theoretical plate If number of theoretical plate is high than column is
highly efficient
If number of theoretical plate is less than column is
less efficient
B
Van Deemter Equation H = A + + Cμ
μ
Where, H = Height of theoretical plate,
= Average liner velocity of mobile phase
A = Eddy diffusion
B = Longitudinal or ordinary diffusion term
C = Non equilibrium or resistance to mass transfer
Van Deemeter plots • The term ‘A’ is independent of flow of
the mobile phase
12
• The term B/u decrease drastically in
10
the beginning with increase in the
HETP-AB/v+Cv
08 flow rate of mobile phase. Increase in
Cv the flow rate beyond particulars
HETP (mm)
06
value , leads to slow decrease value
04 in the value of B/u
B/v
02
• The term Cu increase with increase
A in the rate
00
0 10 20 30 40 50 60 70 80
DIGESTER -116
CHIRAL CHROMATOGRAPHY
Chiral chromatography - Separation of particular isomer from enantionmeric mixture

involves formation of Diastereomers
Stationary Naphthyl Alanine, Naphthyl Leucine, Dinitro benzoyl phenyl glycine, β-
Phase Cyclodextrin
Mobile phase Non polar solvents

133
DIGESTER -117
SIZE EXCLUSION CHROMATOGRAPHY
Size Exclusion Chromatography - Larger molecule unable to fit into pores are eluted
first while small molecules enters into pores and are eluted later.
Gel Permeation Stationary phase used are semi-rigid or rigid gel.
Example: Cross-linked polystyrene, Alkylated Dextran,Controlled
porosity Glass beads
Gel Filtration- Stationary phase used are cross-linked carbohydrates.
Example: Sephadex (Cross linked dextran), Agarose (Sepharose),
Polyacrylamide (Bio-gel)
DIGESTER -118
ION EXCHANGE RESINS
TYPE STATIONARY PHASE MOBILE PHASE

Strongly acidic cation Sulphonated polystyrene Cations, inorganic separations,
exchange resin resins vitamins, amino acids
Weakly acidic cation Carboxylic polymethacrylate Cations, transition elements,
exchange resin resins amino acid, antibiotic
Strongly basic anion Quarternary ammonium Anion, halogen, alkaloids,
exchange resin polystyrene resin vitamin B complex
Weakly basic anion Phenol formaldehyde and Anionic complexes of metals,
exchange resin polyamide polystyrene vitamins and amino acid
DIGESTER -119
IMPORTANT POINTS TO REMEMBER
Edge effect The solvent front in the middle of TLC plates moves faster than that
edge. Therefore the spot are distorted and not regular.
Preparation of (a) Pouring
thin layer plate (b) Spraying
(c) Dipping
(d) Spreding (Best technique) Spreder is 0.25 mm
Retention factor Distance travelled by solute/ Distance travelled by solvent.
(Rf ) Rf Value cannot be greater than 1.
Mobile phase in CCl4 , Cyclohexane, Ether, Acetone, Benzene, Toluene, Chloroform,
TLC Alcohol
Hydrophilic Isopropanol : Ammonia : Water (9:1:2)
Mobile phase in n-Butanol : glacial acetic acid : Water (4:1:5)
paper Methanol : Water (3:1)
chromatography Hydrophobic Kerosene : 70% Isopropanol
Dimethyl ether : Cyclohexane

134
Elution technique (a) Isocratic elution – Single solvent run through column.
(b) Gradient elution – Solvents with increasing polarity
Detection of (a) Uv-visible detector
compound in (b) Fluorescence detector
column (c) Flame ionization detector
chromatography (d) Refractive index detector
Pump used in (a) Pneumatic pump
HPLC (b) Reciprocating pump
(c) Displacement pump
DIGESTER -120
NON AQUEOUS TITRATIONS
S. NO. NAME OF THE DRUG TITRANT

1 Acetazolamide 0.1 MN (C4H9)4 OH [tetrabutyl
ammonium hydroxide]
2 Adrenaline 0.1 MHClO4
3 Albendazole 0.1 MHClO4
4 Allopurinol 0.1 MN(C4H9)4 OH
5 Amiloride HCl 0.1 MHClO4
6 Aminocaproic acid 0.1 MHClO4
7 Amitripty line HCl 0.1 MHClO4
8 Astemizole 0.1 MHClO4
9 Atenolol 0.1 MHClO4
10 Atropine metuonitrate 0.1 MHClO4
11 Benzhexol HCl 0.1 MHClO4
12 Bisacodyl 0.1 MHClO4
13 Bromocriptinemesylate 0.1 MHClO4
14 Bupremorphine HCl 0.1 MHClO4
15 Caffeine 0.1 MHClO4
16 Calcium Pantothenate 0.1 MHClO4
17 Carbenoxolone sodium 0.1 MN(C4H9)4 OH
18 Carbidopa 0.1 MHClO4
19 Chlordiazepoxide 0.1 MHClO4
20 Chlorhexidine gluconate 0.1 MHClO4
21 Chloroquine PO43- 0.1 MHClO4
22 Chlorpheniramine maleate 0.1 MHClO4
23 Chlopromazine HCl 0.1 MHClO4
24 Chlorthalidone 0.1 MN(C4H9)4 OH
25 Cimetidine 0.1 MHClO4
26 Clofazimine 0.1 MHClO4
27 Clotrimazole 0.1 MHClO4
28 Codeine PO43- 0.1 MHClO4
29 Colchicine 0.1 MHClO4
30 Cyclizine HCl 0.1 MHClO4
31 Cyproheptadine HCl 0.1 MHClO4

135
32 Cytarabine 0.1 MHClO4

33 Dehydroemetine HCl 0.1 MHClO4
34 Dequalinium Cl 0.1 MHClO4
35 Diazepam 0.1 MHClO4
36 Diclofenaec sodium 0.1 MHClO4
37 Dicyclomine HCl 0.1 MHClO4
38 Diethyl carbamazine 0.1 MHClO4
39 Di- iodohydroxyquinoline 0.1 MN(C4H9)4 OH
40 Diloxanide furoate 0.1 MN(C4H9)4 OH
41 DiphenhydramineHCl 0.1 MHClO4
42 Dothiepin HCl 0.1 MHClO4
43 Doxepin HCl 0.1 MHClO4
44 Econazole NO3 0.1 MHClO4
45 Emetine HCl 0.1 MHClO4
46 Ergometrine maleate 0.1 MHClO4
47 Ergotaminetartarate 0.1 MHClO4
48 Ethambutol HCl 0.1 MHClO4
49 Ethionamide 0.1 MHClO4
50 Ethopropazine HCl 0.1 MHClO4
51 Ethosuximide 0.1 MN(C4H9)4 OH
52 Fenfluramine 0.1 MHClO4
53 Fluorouracil 0.1 MN(C4H9)4 OH
54 Fluphenazine decanoate 0.1 MHClO4
55 Gallamine triethiodide 0.1 MHClO4
56 Glycine 0.1 MHClO4
57 Haloperidol 0.1 MHClO4
58 Histamine PO43- 0.1 MHClO4
59 Homatropine HBr 0.1 MHClO4
60 Hydro chlorothiazide 0.1 MN(C4H9)4 OH
61 Hyoscine butyl bromide 0.1 MHClO4
62 Idoxuridene 0.1 MN(C4H9)4 OH
63 Imipramine HCl 0.1 MHClO4
64 Isoprenaline 0.1 MHClO4
65 Isoxsuprine 0.1 MHClO4
66 Ketamine HCl 0.1 MHClO4
67 Ketaconazole 0.1 MHClO4
68 Labetalol HCl 0.1 MHClO4
69 Levodopa 0.1 MHClO4
70 Lignocaine HCl 0.1 MHClO4
71 Mebendazole 0.1 MHClO4
72 Mebeverine HCl 0.1 MHClO4
73 Meclizine HCl 0.1 MHClO4
74 Mepyraminemaleate 0.1 MHClO4
75 Mercaptopurine 0.1 MN(C4H9)4 OH
76 Metformin HCl 0.1 MHClO4
77 Methadone HCl 0.1 MHClO4

136
78 Methoxamine HCl 0.1 MHClO4

79 Methyldopa 0.1 MHClO4
80 Metoprololtartarate 0.1 MHClO4
81 Metronidazole
0.1 MHClO4
82 Miconazole NO3-
83 Morphine SO42- 0.1 MHClO4
84 Neostigmine Br- - do-
85 Niclosamide 0.1 MN(C4H9)4 OH
86 Nicolinamide 0.1 MHClO4
87 Nikethamide 0.1 MHClO4
88 Nitrazepam - do-
89 Noradrenalinebirartarate - do-
90 Norfloxacin - do-
91 Nortriptyline HCl - do-
92 Noscapine 0.1 MHClO4
93 Oxprenolol HCl - do-
94 Penicillamine - do-
95 Pentamidineissothionate 0.1 MN(C4H9)4 OH
96 Pentazocine 0.1 MHClO4
97 Pethidine HCl - do-
98 Phenformin HCl - do-
99 Phenindamine maleate 0.1 MHClO4
100 Pheniramine maleate 0.1 MHClO4
101 Phentolamine mesylate 0.1 MN(C4H9)4 OH
102 Pholcodine 0.1 MHClO4
103 Physostigminesalicylate 0.1 MHClO4
104 Pilocarpine NO3 0.1 MHClO4
105 Piperazine adipate 0.1 MHClO4
106 Potassium citrate - do-
107 Prazosin 0.1 MHClO4
108 Primaquin PO43- - do-
109 ProchlorperazineMaleate - do-
110 Proguanine HCl - do-
111 Propantheline Br- - do-
112 Propylphenazone - do-
113 Psuedo ephedrine HCl - do-
114 PYridenine HCl - do-
115 Pyrimethamine - do-
116 Quinidene SO42- - do-
117 Quinine bisulphate - do-
118 Quiniodochlor 0.1 MN(C4H9)4 OH
119 Salbutamol 0.1 MHClO4
120 CH3COONa - do-
121 Sodium benzoate - do-
122 Sodium CMC - do-
123 Sodium citrate - do-

137
124 Sodium cromoglycate - do-

125 NaF - do-
126 Sodium salicylate - do-
127 Sodium valproate - do-
128 Succiyl choline chloride - do-
129 Sulfafurazole 0.1 MN(C4H9)4 OH
130 Tamoxifen citrate 0.1 MHClO4
131 Terbutaline SO 2-4 - do-
132 Thiobendazole - do-
133 Thiamine HCl - do-
134 Thiopentone NA 0.1 M Limethoxide
135 Timolol maleate 0.1 MHClO4
136 Tinidazole - do-
137 Trimterene - do-
138 Trifluoperazine HCl - do-
139 Triflupromazine HCl - do-
140 Trimethoprim 0.1 MHClO4
141 Tripolidine HCl - do-
142 Tropicamide - do-
143 Verapamil HCl - do-
144 Xylometazole - do-
DIGESTER -121
COMPLEXOMETRIC TIRATIONS
S.NO CHEMICAL TITRANT

1 Al2(SO4)3 0.5 M disod. edetate
2 Bismuth subcarbonete 0.1 M disod. edetate
3 CaCO3 0.05 M disod. edetate
4 CaCl2 0.05 M disod. edetate
5 Calcium gluconate 0.05 M disod. edetate
6 Calcium Lactate 0.05 M disod. edetate
7 Calcium levulinate 0.05 M disod. edetate
8 Dibasic Ca phosphate 0.1 M disod. edetate
9 Heavy MgCO3 0.05 M disod. edetate
10 MgCl2 0.05 M disod. edetate
11 Mg(OH)2 0.05 M disod. edetate
12 Heavy MgO 0.05 M disod. edetate
13 Magnesium stearate 0.1 M disod. edetate
14 MgSO4 0.05 M disod. edetate
15 Magnesium trisilicate 0.05 M disod. edetate
16 ZnCl2 0.1 M disod. edetate
17 ZnO 0.1 M disod. edetate
18 Zinc stearate 0.1 M disod. edetate
19 ZnSO4 7H2O 0.1 M disod. edetate

138
DIGESTER -122
ACID BASE TITRATIONS
S.NO NAME OF DRUGS TITRANT

1. Amantadine 0.1 M NaOH
2. NH4Cl 0.1 M NAOH
3. Aspirin 0.5 M NaOH (back titration)
4. Benzoic acid 0.5 M NaOH
5. Boric acid 1 M NaOH
6. Bromohexine HCl 0.1 M NaOH
7. Bupivocaine 0.01 M ethanolic NaOH
8. Busulphan 0.1 M NaOH
9. Calamine 0.5 M H2SO4
10. Chlorambucil 0.1 M NaOH
11. Chlorpropamide 0.1 M NaOH
12. Citric acid 1 M NaOH
13. Clonidine 0.1 M ethanolic NaOH
14. Cycloserine 0.1 M NaOH
15. Dextromethorphan 0.1 M NaOH
16. Ephedrine 0.1 M NaOH
17. Ethacrynic acid 0.1 M HCl
18. Ethinyl oestradiol 0.1 M NaOH
19. Flurbiprofen 0.1 M NaOH
20. Frusemide 0.1 M NaOH
21. Fusidic acid 0.1 M NaOH
22. Glibenclamide 0.1 M NaOH
23. Glycerime 0.1 M NaOH
24. HCl 0.1 M NaOH
25. Ibuprofen 0.1 M NaOH
26. Indomethacin 0.1 M NaOH
27. Ketoprofen 0.1 M NaOH
28. Lactic acid 1 M NaOH
29. Levamisole HCl 1 M NaOH
30. Lithium Carbonate 1 M HCl
31. Mefenamic acid 0.1 M NaOH
32. MephentermineSO42- 0.05 M H2SO4
33. Methylsalicylate 0.1 M NaOH (back titration)
34. MetoclopromideHCl 0.1 M NaOH
35. Mianserin HCl 0.1 M NaOH
36. Nalidixic acid 0.1 M ethanolic NaOH
37. Nicolinic acid 0.1 M NaOH
38. Norethisterone 0.1 M NaOH
39. Oxypnenbutazone 0.1 M NaOH
40. Pentobarbitontesodium 0.1 M ethanolic NaOH
41. Phenobarbitone 0.1 M ethanolic NaOH
42. Phosphoric acid 1 M NaOH
43. Phthalyl sulphathiazole 0.1 M NaOH
44. Probenecid 0.1 M NaOH
45. Promethazine 0.1 M NaOH

139
46. Propranolol HCl 0.1 M NaOH

47. Propyl thiouracil 0.1 M NaOH
48. Pyrazinamide 0.05 M H2SO4 (back titration)
49. Quinal barbitone sodium 0.1 M ethanolic NaOH
50. Saccharin 0.1 M NaOH
51. Salicylic acid 0.1 M NaOH
52. NaHCO3 1 M HCl
53. NaH2PO4.2H2O 1 M NaOH
54. Sodium fusidate 0.1 M HCl
55. NaOH 1 M HCl
56. Sod. Lactate injection 0.05 M H2SO4 (back titration)
57. Urea 0.1 M HCl
58. Sorbic acid 0.1 M NaOH
59. Tartaric acid 1 M NaOH
60. Theophylline 0.1 M NaOH
61. Thiotepa 0.1 M HCl
62. Tolbutamide 0.1 M NaCOH
63. Undeconic acid 0.5 M NaCOH
64. Warfarin sodium 0.01 M NaOH
65. Vanillin 0.1 M NaOH
DIGESTER -123
REDOX TITRATIONS
1 Ascorbic acid Tablets 0.1 M cerric ammonium sulphate

2 Ferrous fumarate -do-
3 Ferrous gluconate 0.1 M cerric ammonium nitrate
4 Ferrous sulphate - do –
5 H2O2 soln. (100vol) 0.02 M KMnO4
6 Menadione 0.1 M cerric ammonium sulphate
7 Nifedipine - do –
8 Paracetamol - do –
9 Tocopheryl acetate - do –
DIGESTER -124
NITRITE TITRATIONS
1 Benzocaine 0.1 M NaNO2

2 Dapsone 0.1 M NaNO2
3 Procainamide HCl 0.1 M NaNO2
4 Procaine HCl 0.1 M NaNO2
5 Sod PAS 0.1 M NaNO2
6 Succinylsulphathiazole 0.1 M NaNO2
7 Sulphacetamidesodium 0.1 M NaNO2
8 Sulphadiazine 0.1 M NaNO2
9 Sulpha dimethoxine 0.1 M NaNO2

140
10 Sulphadimidine 0.1 M NaNO2

11 Sulphadoxine 0.1 M NaNO2
12 Sulphamethizole 0.1 M NaNO2
13 Sulphamethoxazole 0.1 M NaNO2
14 Sulfaphenazole 0.1 M NaNO2
DIGESTER -125
POTENTIOMETRIC TITRATIONS
1 Acebutolol HCl 0.1M NaOH

2 Dipenoxylate HCl 0.1M rhanolic NaOH
3 Hydralazine HCl 0.05M HIO3
4 Phenytoin Sodium 0.1M NaOH
DIGESTER -126
ARGENTOMETRIC TITRATIONS
1 NaCl 0.1M AgNO3

2 AgNO3 0.1M Ammonium thiocyanate
3 KCl 0.1M AgNO3
4 Melphalan 0.1M AgNO3
5 Disulfiram 0.1M AgNO3
6 Lomustine 0.05M AgNO3
DIGESTER -127
IODOMETRY & IODIMETY TITRATIONS
1 Analgin Iodimetry 0.05M I2

2 Ascorbic acid Iodimetry 0.05M I2
3 Cephalexin Iodimetry 0.02M Na2S2O3
4 Chlorocresol Iodimetry 0.1M Na2S2O3
5 Chloroxylenol Iodimetry 0.1M Na2S2O3
6 Dimercaprol Iodimetry 0.1M Na2S2O3
7 Guaiphenesin Iodimetry 0.05M I2
8 I2 Iodimetry 0.1M Na2S2O3
9 Mannitol Iodimetry 0.05M I2
10 Methyl paraben Iodimetry 0.1M Na2S2O3
11 Monothioglycerol Iodimetry 0.05M I2
12 Phenol Iodimetry 0.1M Na2S2O3
13 KI Iodimetry 0.05M KIO3
14 KMnO4 Iodimetry 0.1M Na2S2O3
15 Sod. ascorbate Iodimetry 0.05M I2

141
16 Sod. diatrizoate Iodimetry 0.05M KIO3

17 Sorbitol Iodimetry 0.05M I2
18 Na2S2O35H2O Iodimetry 0.05M I2
19 Sodium metabisulphite Iodimetry 0.1M I2
20 Thyroxine sodium Iodimetry 0.1M Na2S2O3
DIGESTER -128
DRUGS ANALYZED BY GRAVIMETRIC ANALYSIS
1. Thioacetazone sodium. 2. Fluorescein 3. BaSO4
DIGESTER -129
DRUGS ANALYZED BY HPLC
1. Alprazolam 2. Cefadroxil 3. Cefazolin sodium 4. Cefotaxime sodium

5. Ceftazidime 6. Cefuroxim 7. Ca Folinate 8. Cholecalciferol
9. Ciprofloxacin 10. Cisplatin 11. Diltiazem 12. Doxorubicin HCl
13. Enalapril 14. Ergocalciferol 15. Guaggul resin 16. Insulin
maleate
17. Methotrexate 18. Opium 19. Omeprazole 20. Ormeloxifene HCl
21. Piroxica 22. Ranitidine 23. Vinblastin SO 2- 4 24. Vincristine SO 2- 4
DIGESTER -130
MICROBIOLOGICAL ASSAYS
NAME OF THE DRUG MICRORGANISM USED

Amikacin Staphylococcus aureus
Amphotericin B Saccharomyces cerevisiae
Erythromycin Micrococcus luteus
Doxycycline HCl Staphylococcus aureus
Gentamicin SO42- eye drops Staphylococcus epidermidis
Kanamycin SO42- B. pumilus, S. aureus
Kanamycin B B. subtilis
Novobiocin Na S. epidermidis
Nystatin Saccharomyces cervisiae
Neomycin Sraphylococus epidermides
Rifampicin B. subtilis
Streptomycin SO42- Klebsiella pneumonia & B. subtilis
Tetracycline Staphylococcus aureus, B. cereus

142
DIGESTER -131
SPECTROMETRY
NAME OF THE DRUG ABSORBANCE

1. Amodiaquine HCl 343nm
2. Calciferol Capsules 500 & 550nm
3. Carbamezapine 285nm
4. Carbimazole 291nm
5. Chloramphenicol 278nm
6. Cyanocobalamn 361nm
7. Danazol 285nm
8. Deslanoside 420nm
9. Dienoestrol 245nm
10. Digitoxin 495nm
11. Dydrogesterone 286nm
12. Folic acid 550nm
13. Furazolidine 367nm
14. Gresiofulvin 291nm
15. Hydroxocobalamin 351nm
16. Hydroxyprogesterone hexanoate 240nm
17. Lanatoside C 484nm
18. Levonorgestrol 241nm
19. Megesterol acetate 287nm
20. Methylergometrine maleate 550nm
21. Nalorphine HCl 285nm
22. Nandrolone decanoate 239nm
23. Nitrofurantoin 367nm
24. Nitrofurazone 375nm
25. Norgesterol 241nm
26. Oestradiol benzoate 231nm
27. Phenolphthalein 555nm
28. Reserpine 388nm
29. Riboflavin 444nm
30. Rifampicin 475nm
31. Spironolactone 238nm
DIGESTER -132
MISCELLANEOUS
NAME OF THE DRUG TITRANT
Thiomersal 0.1 M ammonium thiocyanate
Sodium stilbogluconate 0.05 M ferric ammonium SO42-
Sodium lauryl sulphate 0.04 Benzethomium Cl-
Phenyl mercuric acetale 0.1 M amm thiocyanate
Isoniazid 0.0167 M KBrO3
Desferrioxamine mesylate 0.1 M ferric ammonium sulphate
Cyclophosphamide 1.1 M AgNO3
Cloxacillin sodium 1.2 M mercuric NO -3
Cetrimide 0.05 M KIO3
Captopril 0.025 M KIO3
143

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DIGESTER : PHARMACEUTICAL ANALYSIS DIGESTER

Accuracy Accuracy is the closeness of experimental value to the true value.

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Mean or Mean is obtained by dividing the sum of a set of measurements by the

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REGION λ WAVELENGTH ν FREQUENCY (HERTZ)

PRINCIPLE SPECTROSCOPIC METHOD

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Refraction of  Refractometry and Interferometry

ANALYTICAL TECHNIQUE MOLECULAR CHANGES

SPECTROSCOPIC METHODS PRINCIPLE

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Fluorimetry Measures the intensity of fluorescence caused by

SPECTROSCOPY SOURCE OF RADIATION

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SPECTROSCOPY DETECTOR MADE UP OF WITH

ANALYTICAL TECHNIQUE REFERENCE STANDARD GRAPH

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Beer’s law Find relationship between transmittance and concentration of

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Homoannular diene: It is a cyclic diene having conjugated double bond in the

Heteroannular diene It is a cyclic diene in which double bonds in conjugation are

Endocyclic double bond A double bond present in a ring.

Endocyclic double bond

WOODWARD FIESER RULE FOR CONJUGATED DIENE, TRIENE SYSTEMS

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WOODWARD FIESER RULE FOR α, β-UNSATURATED CARBONYL COMPOUNDS

Alkyl group α β γ /higher

Parent value for β unsaturated 6 membered cyclic compound = 215 nm

WOODWARD FIESER RULE FOR ACYL BENZENE DERIVATIVE

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Question: Calculate λmax. Question: Calculate λmax.

INFRA RED REGION

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TYPES OF COMPOUNDS FREQUENCY RANGE CM-1

N-H stretching (Amines)

Fermions Odd mass nuclei with an odd number of nucleons have

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Shielding of High electron density around a nucleus shields the

NAME OF COMPOUNDS NUMBER OF NMR SIGNALS

TYPE OF PROTON CHEMICAL SHIFT

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MASS ANALYZER WORKING

IONIZATION SOURCE PRINCIPLE

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Mc-Lafferty rearrangement reaction - It involves the migration of γ-hydrogen atom

Coupling constant • Distance between two split lines.

Area of Peak Number of absorbing protons giving rise to a signal

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X ray was discovered by William K. Roentgen.

ATOM ABSORPTION SPECTROSCOPY FLAME EMISSION SPECTROSCOPY

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Nephelometry  Turbidimetric titration

BASIC PRINCIPLE CONSTANT ELECTRODE

Reference electrode Standard Hydrogen Electrode (SHE)

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METHODS PARAMETER MEASURED GRAPH