Process Validation - ability (within given range) to obtain test results

- analysis of data gathered throughout the design which are directly proportional to the concentration
manufacturing of a product (amount) of the analyte in the sample
- ensure varied inputs lead to consistent and high
quality outputs
Range
3 Process of Process Validation - interval between upper and lower concentration of
1. Process Design the analyte in the sample
- commercial manufacturing process Robustness
- based on knowledge gained through - measure of its capacity to remain unaffected by small
development and scale-up activities but deliberate variations in method parameters
2. Process Qualification - provide indication of its reliability during normal use
- process design is evaluated to determine if the
process is capable of reproducible commercial Ruggedness
manufacturing - degree of reproducibility of test results obtained by
3. Continued Process Verification the analysis of the same sample under variety of
- on going assurance normal test conditions
- gained during routine production that the
process remain in a state of control Sensitivity
- equal to the slope of the calibration line in a linear
Assay Validation system
- process of demonstrating that analytical procedures
are suitable for their intended use Qualification
- ensure analytical methodology is accurate, specific, - normally used for equipment, utilities, and systems
reproducible and rugged over the specified range that - qualification is part of validation
a target will be analyzed - proving and documenting that any premises, systems
and equipment are properly installed, and or work
Accuracy correctly and lead to the expected results
- closeness of agreement between value which either - pre-requisite of validation
as a conventional true value or an accepted reference
value and the value found Calibration
- set of operation that establish, under specified
Precision conditions the relationship between values indicated by
- closeness of agreement between a series of an instrument or system for measuring , recording, and
measurements from multiple sampling of the same controlling by a material measure, and the
homogeneous sample under prescribed conditions corresponding is known values of a reference standard
- Always remember any reference standard is always
Repeatability used in calibration
- precision under the same operating condition over a
short interval of time Validation
- action of providing and documenting that any
Reproducibility process, procedure or method actually and consistently
- precision between laboratories leads to the expected results
- a documented evidence to and done to prove the
Specificity consistency of the expected results of any process,
- assess unequivocally the analyte in the presence of procedure or method
components which may be expected to be present
Equipment
Detection Limit - designed to minimize risk of error and permit
- the lowest amount of analyte in a sample; which can effective cleaning and maintenance
be detected but not necessarily quantitated as exact - installed to minimize risks of error and minimize risks
value of contamination

Quantitation Limit Design Qualification (DQ)

- lowest amount of analyte in a sample which can be - compliance with GMP should be demonstrated
quantitatively determined with suitable precision and - principle: achieve the objectives of GMP with regard
accuracy to equipment
Linearity - mechanical drawings & design features provided buy
the manuf should be examined
Installation Qualification (IQ) - number of wave per cm and is = to 1 wavelength
- carried out on a new or modified facilities, systems & (CM)
equipment
Frequency
Operational Qualification (OQ) - number of complete cycles which pass a given point
- should follow IQ per second

Performance Qualification (PQ) Chromophore

- Tests to include conditions with upper and lower - functional group absorbs radiant energy in the UV or
limits visible regions of the spectrum

Goal of Cleaning Validation Change of Spin

- verify that no gross contamination remains after - NMR, ESR: Low energy
cleaning and any process residues do not jeopardize
the safety of the patient and the quality of the next Change of orientation
product - Microwave: Low energy
Change of configuration
4 Principles with relevance to cleaning - Infrared: Low Energy
1. Encourage the early adoption of new
technological advances Change of electron distribution
2. Facilitate industry application of modern quality - Visible light, UV, X-ray
management techniques
3. Encourage implementation of risk-based Change of nuclear configuration
approaches - Gamma ray: High energy
4. Ensure that regulatory review, compliance, and
inspection policies are based on state-of-the-art Electric field
pharmaceutical science - a vector whose length is proportional to the field
strength
Spectrometry
- deal with the measurement of spectra, power, Emission Spectroscopy
transmitted, fluorescent, reflected, or emitted energy - methods in which the stimulus is heat or electrical
energy
Spectrophotometry
- branch of spectrometry Chemiluminescence Spectroscopy
- measurement of absorption by chemical species of - excitation of an analyte by chemical reaction
radiant energy of definite and narrow wavelength
Absorption Spectroscopy
Electromagnetic Spectrum - measure the amount of light absorbs as a function of
- the complete system of energy propagated in the wavelength
wave form
- form of energy transmitted through space at Photoluminescence Spectroscopy
enormous velocities - emission of photons is measured following absorption
- important forms: fluorescence and phosphorescence
Visible Radiation spectrum
- UV, Infrared, Visible light Absorption Process
- as light traverses a medium containing an absorbing
Electromagnetic radiation analyte, the intensity decreases and the analyte
- wave with properties of wavelength, frequency, becomes more excited
velocity and amplitude
Absrobance
Colorimetric - related to the transmittance in a logarithmic manner
- visible region of the spectrum - as absorbance increases the transmittance decreases

Radiant energy Beer’s Law

- energy transmitted through EM Waves - absorbance is directly proportional to the
concentration of the absorbing species
Wavelength
- length of a complete wave or cycle Waveleng Color Light Color Transmitted
th Absorbed
Wavenumber 400-435 Violet Yellow-green
 435-480 Blue Yellow Infrared Spectroscopy
480-490 Blue-green Orange - aka Vibrational Spectroscopy
490-500 Green-blue Red - interaction of infrared radiation with matter
500-560 Green Purple - conducted w/ infrared spectrometer or
560-580 Yellow-green Violet spectrophotometer
580-595 Yellow Blue - identity of raw materials in manuf of drugs
595-650 Orange Blue-green - preliminary check for compound identity in synthetic
650-750 Red Green-blue chemistry
- fingerprint test
UV/Visible Spectroscopy - detects polymorphs of drugs
-absorption spectroscopy/reflectance spectroscopy in
UV-VIS spectral region Atomic Mass Spectroscopy
- EM radiation absorbed and emitted by atoms it is for
Components of UV-Vis Spectroscopy: determination of elemental components
1. Spectroscopic source - only performed on gaseous medium
- generate a beam of radiation that is sufficiently - determine nearly all elements in the periodic table
powerful enough to be detected
Classification of Atomic Spectroscopic Methods
Atomization Temp Type of
2. Wavelength detector method conditions Spectroscopy
- uses monochromators or polychromators to isolate Inductive
6000-8000 C Emission, Mass
specific wavelength in the instrument coupled plasma
Absorption,
3. Sample Container Flame 1700-3150 Emission,
- Cuvette: container in which the sample or reference Fluorescence
sol’n is place Absorption,
Electrothermal 1200-3000
- must be transparent to the radiation Fluorescence
- cuvette is made up of quartz Direct current-
5000-10000 Emission
4. Detector plasma
- indetifies, records, or indicates a change in one of the Electric arc 3000-8000 Emission
Electric spark Varied Emission, Mass
variables in its environment such as pressure,
Nuclear Magnetic resonance (NMR)
temperature, or EM radiation
- determine the content and purity of a sample as well
- Human eye: converts visible radiation into an
as its molecular structure
electrical signal
- uses: exploits magnetic properties of atomic nuclei
- Transducer: converts nonelectrical quantities such
- determine physical and chemical properties of atoms
as light intensity, pH, mass, and temperature into
of the molecules
electric signals
- applied in MRI
5. Signal processor and readout
Mass Spectroscopy
- signal process: e-device that may amplify the electric
- determine identity of an unknown compound
signal from the detector
- analyte are converted to ions by applying energy to
- eg. of readout materials: computers and digital
them
meters
- Pharmaceutical, Environmental, and Forensic analysis
Spectroscopic Waveleng
Spectroscopy Molecular Mass Spectrometry
Source th (nm)
Xenon arc - capable of providing information about structure of
250-600 Molecular fluorescence inorganic, organic, and biological molecules
lamp
H2 and D2 UV molecular
160-380 Mass Spectrum
lamps absorption
Tungsten/ UV/VIS/IR molecular - ion abundance plotted against mass-to-charge ratio
240-2500
Halogen absorption
Vis/IR molecular AES: Flame Photometer
Tungsten lamp 350-2200
absorption - determine alkali and alkaline earth metals
400- - determine led in petrol
Nernst glower
20000 - determine Ca and Mg in cement
750- IR molecular
Nichrome wire
20000 absorption AES: Inductively Coupled Plasma
1200- - detect and quantify elements in a sample
Globar
40000 - determines all elements except gases and C,N,F,O,H
Atomic Absorption Spectroscopy (AAS) - aka Rf value
- food, water and clinical analysis - ratio of the distance travelled by the solute to the
- additive analysis in lubricating oils and greases distance travelled by the solvent
- unitless quantity
UV and Vis Light
- detect geometrical isomers, functional group, and Factors affecting Rf Value
impurities - thickness of stationary phase
- quali and quanti analysis - moisture on TLC plate and its nature
- saturation and volume of mobile phase
Light-scattering spectrometry - temp of the environment
- analysis on sol’ns of marcomolecules - size of the sample taken
- molecular wt. determination
- air pollution analysis Reverse Phase Chromatography (RPC)
- diffusion phenomena - partition phenomenon
- Non-polar solvent is fixed to the paper or solid column
Chromatography material as SP
- process in which a solution of a mixture containing - Polar solvent as MP
inert materials, drug principle, and impurities is
separated into its component while moving through a Gas Chromatography (GC)
bed of fixed porous solid - carrier gas an inert gas used as MP
- separating a mixture samples is separated into - SP is referred as liquid substrate has high-boiling
components liquid used to coat granular particles made of siliceous
earth or firebrick housed in a column a constant temp
Principles of Chromatography: chamber
1. Resolution of mixtures
2. Determination of homogeneity Factors Affecting GC Separation
3. Comparison of substances suspected of being - Volatility & Polarity
identical - Column length, temperature & packing polarity
4. Purification - Flow rate of the gas
5. Concentration of substance from dilute solutions
6. Identification and control of technical products Application of GC
7. Quantitative separation from complex mixtures - Separation and analysis of organic compounds
8. Indication of molecular structure - Testing purity of compounds
Stationary phase/Adsorbent - Determine relative amounts of components in mixture
- phase that doesn’t move - Compound identification
- Isolation of pure compounds
Mobile phase
- phase that moves through the stationary phase Component of GC
1. Carrier Gas
Column Chromatography (CC) - carries the sample vapor through the column
- simplest to the detector
- aka elution chromatography - Helium commonly used, N2 and H2 pressured
- SP/Adsorbent: purified siliceous earth, activated tank
ammonia, silica gel, calcium carbonate
- product: Eluate 2. Flow Rate controller
- must be precisely controlled
Thin Layer Chromatography (TLC) - depends on the type of column
- spotting of a sample of a mixture of components at - decreases as column temp increases
one end of an adsorbent-coated glass - viscosity of carrier gas increase w/ temp
- for organic substances
- advantages over PPC is that it requires short time, 3. Sample injector
small amount of material, provides complete - 2 mode: split and splitless modes
separation of components in complex mixtures,
sensitive 4. Column oven
- SP/Adsorbent: thin plate of silica gel G - where the column sits in
(G=gypsum, binder of silica) - isothermal if temp is constant within duration
- best solvent of exploratory run: benzene, or of analysis
chloroform w/ 10% ethanol
5. GC columns
Retention Factor - where separation takes place
 - packed or capillary columns - separation is based on molecular size
- has network-like structure of polymer materials
6. GC detectors having pores of various sizes
- highly sensitive
- rapidly responds to conc changes pH meter
- stable w/ respect to noise and drift - measures electro-chemical potential between a
- low sensitivity to variations in flow, pressure, known liquid inside the glass electrode and an
and temp unknown liquid outside
Type of detector Definition - measures the potential of hydrogen ions or potential
Thermal Conductivity Non-destructive, moderate of hydrogen
Detector (TCD) sensitivity - only measure electric voltage
Flame Ionization Destructive, for detecting - not used in moving liquids of low conductivity
Detector (FID) hydrocarbons, weakly
sensitive to carbonyl, amine, Viscosity
alcohol and amine but not to - a liquid’s resistance to flow or shear stress
non-combustibles - Quantitative measure of a fluid’s resistance to flow
Nitrogen Phosphorous Highly sensitive, specific
Detector (NPD) detector, has strong
Dynamic or Absolute Viscosity
response to compound that
contain N or P, destructive - measure of the resistance it offers to relative shearing
Electron Capture Detects electron-absorbing motion
Detector (ECD) components (high-electron-
negativity) such as Kinematic Viscosity
halogenal compounds in the - ratio of absolute viscosity to the density of fluid
output stream of a gas
chromatograph, non- Viscometer
destructive - measures viscosity and flow properties of fluids
Photo-ionization Volatile organic compounds
Detector (PID) and gases, uses UV light Capillary Viscometer
- gives the kinematic viscosity of the fluid
High-Pressure Liquid Chromatography (HPLC) - based on Poseuille’s law
- depends on adsorption, partition, ion-exchange, and
molecular exclusion Rotational Viscometer
- has greater speed, precision and accuracy - gives the dynamic viscosity
- MP varies from aq to non-aq solvents (must be - principle: the fluid whose viscosity is being measure is
degassed) to remove bubbles) sheared between 2 surfaces
- uses detectors (UV absorbance and refractive index) - one surface is stationary the other is rotated

Falling ball viscometer

Detectors of HPLC - measure resistance to the piston falling through the
- Spectrophotometric, Fluorescence, Refractive index, material
Electrochemical, Radiochemical, Mass spectroscopy,
NMR
Amperometry
Adsorption Chromatography - detect ion presence on a solution on the basis of
- aka Liquid-Solid Chromatography (SLC) electric current or change in electric current
- mobile phase and adsorbent interaction is strong, - Uses; redox titration, study complex formation,
only a small amount of analyte will be adsorbed quantification of ions or mixture of ions, determine
- mobile phase and the adsorbent interaction is weak, water using KF reagent (biamperometry)
greater amounts of analyte will be adsorbed
- require polarity scale known as eluotropic scale Amperometric Titration
- a class of titrations in which the equivalence point is
Ion-exchange Chromatography (IEC) determined through measurement of the electric
- uses cation or anion resins as columns current produce by the titration reaction
- Cation exchanger: capable of quantitatively - quantitative analysis
removing alkaloid bases from sol’n
- Anion exchanger: used to liberate bases from Coulometry
alkaloidal salts - determine the amount of matter transformed during
electrolysis reaction
Molecular Exclusion Chromatography (MEC) - measure the amount of electricity consumed or
- aka Gel filtration or gel permeation produced
chromatography
- uses coulombs

Controlled-potential coulometry
- apply a constant potential to electrochemical cell

Controlled-current coulometry
- pass a constant current through the electrochemical
cell

Faraday’s Law
- amount of substance deposited during electrolysis is
proportional to the quantity of electrical charge passed
and to the equivalent wt. of the substance
- total charge passing through the electrochemical cell
is proportional to the absolute amount of analyte

Voltammetry
- application of time-dependent potential excitation
signal to the working electrode
- auxiliary electrode: platinum wire
- reference electrode: Ag/AgCl electrode
- working electrode: mercury, platinum, gold, silver,
carbon

Cyclic Voltammetry
- measures the current that develops in an
electrochemical cell under conditions where voltage in
excess of that predicted by the Nernst equation
- performed by cycling the potential of a working
electrode, and measuring the resulting current

Potentionmetry
- deals with the study and measurement of electrode
potential

Electrode Potentials
- determined by the relative tendencies which different
metals, ions, and molecules have to gain or lose
electrons under controlled conditions