Post on 21-Aug-2015
Analytical Technologies
Spectroscopy (UV-Visible)
Chromatography (liquid Chromatography)
Titrimetric Analysis (Classical and Automated)
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Calibration
Wavelength Accuracy
Absorbance Accuracy
Stray Light
Resolution
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Applications
Assay
Characterization
Identification
Dissolution Assay
Dissolution Kinetic Study5/9/2015 ACI Limited, Md. Saddam Nawaz 10
Chromatography
Column Chromatography
Planer Chromatography
Super Critical FluidChromatography
Chromatography
Gas Chromatography Liquid Chromatography
High PerformanceLiquid Chromatography
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Instrumentation
Reservoir
PumpInjector
Column Detector
Data Processor
Low pressure gradient valve
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Column Selections
Luna, ODS-AQ, xTerra, Symmetry, Nucleosil, ProC18, Stable bond (low pH), Stable bond
extended (high pH)
Molecular weight < 2,000 Da
Organic Soluble
Aqueous soluble
Hexane
MethanolAcetonitrile
THF
Non-Ionic
Ionic
Peptides Proteins
Normal Phase
Reversed Phase
Reversed Phase
Reversed Phase
Reversed Phase
Reversed PhaseLuna, ODS-AQ
Luna, xTerra, Symmetry, Nucleosil, Stable bond (low pH),
Stable bond extended(high pH)
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Solvents for HPLC
0.010.20.891.33Water
3.15.80.341.34190Acetonitrile
3.05.10.541.33205Methanol
3.64.31.081.36210Ethanol
3.54.81.201.42215Dioxane
4.44.00.461.40212THF
4.23.90.821.38205Isopropanol
3.90.701.40210n-Butanol
3.10.421.42233Dichloromethane
2.70.321.50280Benzene
2.40.291.49285Toluene
2.10.251.40220n-Butyl ether
0.20.401.39200Heptane
0.10.301.37190Hexane
Solvent
Strength
for RPC
Polarity
(p)
Viscosity
(cP,25oC)
Refract.
index
UV
Cutoff
(nm)
Solvent
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Buffer for HPLC
20010.3 – 12.311.3Pyrrolidine
20010.0 – 12.011.0Triethylamine*
2057.3 – 9.38.3Tris
4.4 – 6.45.4
3.7 – 5.74.7
2302.1 – 4.13.1Citrate
2103.8 – 5.84.8Acetic acid*
2102.8 – 4.83.8Formic acid*
11.3 – 13.312.3
6.2 – 8.27.2
2001.1 – 3.12.1Phosphate
UV Cutoff(nm)
Buffer capacity
pKaBuffer
*Volatile buffers5/9/2015 ACI Limited, Md. Saddam Nawaz 18
System Suitability Test (SST)
Repeatability, %RSD (CV)
Capacity factor, K’
Selectivity/Relative Retention Time
Theoretical Plates, N (Efficiency)
Resolution
Asymmetry/Tailing Factor
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Repeatability, CV (%RSD)
X=(X1+X2……..Xn-1+Xn)/n
N=Number of Analysis
X1…..Xn: Retention time (or area or heights)
X = Average
C.V : Coefficient of Variation5/9/2015 ACI Limited, Md. Saddam Nawaz 22
Calibration
Solvent Delivery Unit (LC-20 AD) Stability Test
Flow Rate Accuracy Check
D2 Lamp Test
W Lamp Test
Wavelength Accuracy Test
Inject Volume Accuracy Test
Gradient Concentration Test (Low Pressure Gradient System)
Noise/Drift Test5/9/2015 ACI Limited, Md. Saddam Nawaz 28
Applications
Assay
Identification
Impurity Analysis
Dissolution Assay
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Advantages
There are several reasons why titration is used in laboratories worldwide:
Titration is an established analytical technique
It is fast
It is versatile
It is a very accurate and precise technique
(Reproducibility <1%, precision 0.1% achieved)
A high degree of automation can be implemented
Economical compared to more sophisticated techniques
It can be used by low-skilled and low-trained operators
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TitrantsThe titrants should be selected, The exact concentration of the titrant
is determined periodically and indicated as its titer. The titer should be stable. It is determined with the aid of primary standards.
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Different titration techniques
Indication principles Chemical reactions Applications
Potentiometry: The concentration-dependent potential (mV) of a solution is measured against a reference potential
• Aqueous acid/base titrations• Non-aqueous acid/base titrations • Redox titrations• Precipitation titrations
Content determination of pharmaceutical constituents
Acid content in fruit/vegetable juices, wine, milk, vinegar
Voltametry: The concentration dependent potential of a solution (mV) is measured at a constant electric current
• Karl Fischer Water in butter
Water in oil or gasoline
Bromine number or index of gasoline
Amperometry: The current flowing through a sample solution (μA) is measured at a constant potential
• Amperometrictitration
Iron(II) determination
Vitamin C determination
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Different titration techniques
Indication principles Chemical reactions
Applications
Photometry: The light transmission (mV or % transmission) of a coloured or turbid solution is measured with a photometric sensor
• Complexometry
• Turbidimetry
Total hardness of water
Two-phase surfactant titration
Conductivity: The change of conductivity during a titration is measured with a conductivity sensor connected to the conductivity board of an Excellence titrator
• Conductometrictitration
Sulphate content by precipitation with Barium
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Schematic representation of the different steps
in a titrator method
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Endpoint titrations (EP) &
Equivalence point titrations (EQP)
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Evaluation principles
symmetric curve asymmetric curve
minimum/maximum curvesegmented curve (1st derivative)
segmented curve (2nd derivative)5/9/2015 ACI Limited, Md. Saddam Nawaz 40
Potentiometric titrations in non-aqueous solvents
Solvotrode5/9/2015 ACI Limited, Md. Saddam Nawaz 42
Validation of Titration Methods
Accuracy,
Precision
Trueness
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Types of errors
Systematic errors
Random errors
Gross errors
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Analytical Method Validation Parameters
Precision
Repeatability
Intermediate Precision
Accuracy
Linearity and Range
Specificity
Sensitivity (LOD and LOQ)
Robustness
Ruggedness5/9/2015 ACI Limited, Md. Saddam Nawaz 47