Computers as data analysis in Preclinical

development
• Preclinical development, also named preclinical studies, is a stage of research in
drug discovery that is done before clinical trials (testing in humans)

• This helps determine the safety of the drug and the dose that can be further
used in humans
• Preclinical studies involve conduct of preliminary efficacy, toxicity,
pharmacokinetic and safety information.
• Wide doses of the drug are tested using in vitro (test tube or cell culture) and in
vivo (animal) experiments, and it is also possible to perform in silico profiling
using computer models of the drug–target interactions.
• After completing the initial study on experimental animals, an Investigational
New Drug (IND) application needs to be filed with the regulatory authorities
(FDA).
• This application along with supporting data consists of near about 50,000 pages
of supporting documentation.
There are several sources that generate large scientific data in
preclinical development . They are:

• Drug product purity, potency, and other testing

• Drug product stability testing
• Method development, validation, and transfer
• Drug product formulation development
• Drug substance/drug product manufacturing process development,
validation, and transfer
• Master production and control record keeping
• Batch production and control record keeping
• Equipment cleaning testing
Role of computers in preclinical development:
• Computers are ubiquitous and fundamental part of pharmaceutical
research and development for generating, managing and transmitting
information .
• Computers play vital role as data management and data analysis
• It acts as storehouse of drug information.
• Aids in study of toxicology and risk assessment
• Helps to develop predictive models for better decisions
• Makes pharmaceutical development more efficient
• Ensures collection and management of clinical data
• Aids in better medical decision making
• Improves quality of healthcare management.
• Successful utilization of computers leads to efficient completion of
the above mentioned steps thereby increasing the efficiency and
productivity of development.
• Considering the prevalence of computer applications scientist's daily
activities, special emphasis are put on three widely used computer
systems:
• CDS—chromatographic data systems
• LIMS—laboratory information management systems
• TIMS—text information management systems
Strict regulations are published by the FDA to check on the usage of
computers to aid pharmaceutical research and development.

• Key features of regulations:

• Computer systems must be validated to ensure consistently of intended
purpose, accuracy and reliability.
• It must provide time based audit trial to record actions for creating modifying
or deleting records
• Access to computer system used for research must be limited to authorize
personnel only
• It should have the capability to be configured specific to each user

One way or another, every single data point has to go through the acquiring,
analyzing, managing, reporting, auditing, and archiving process according to
a set of specific rules and regulations needless to say, the wide use of
computers has tremendously increased efficiency and productivity in
pharmaceutical development.
CHROMATOGRAPHIC DATA SYSTEMS (CDS)
CHROMATOGRAPHIC DATA SYSTEMS (CDS)
• The importance of CDS is directly related to the roles that
chromatography, particularly high-performance liquid
chromatography (HPLC) and gas chromatography (GC), play in
pharmaceutical analysis.
• HPLC and GC are the main workhorses in pharmaceutical analysis. In
today's pharmaceutical companies, development work cannot be
done without HPLC and GC.
• CDS are also used for several other instrumental analysis technologies
such as ion (exchange) chromatography (IC), capillary electrophoresis
(CE), and supercritical fluid chromatography (SFC).
• The chromatograms generated by the above analytical methods are displayed,
integrated and results are calculated by a software application called a
chromatographic data system.

• Definition:
• Sometimes referred to as a chromatography data management system (CDMS),
a chromatography data system (CDS) is a set of dedicated data-collection tools
that interface and/or integrate with a laboratory's chromatography equipment.
• A base CDS will set up a desired methodology to be used by the chromatography
equipment, acquire data from it, process the acquired data, store
the information in a database, and interface with other laboratory
informatics systems to import and export files and data.

• CDS helps in providing accurate and reliable data.

Development of CDS technique:
In the 1960s and early 1970s, chromatographs were relatively primitive and inefficient
and process was conducted manually.
• e.g. Chromatography used microsyringes for sample injection and stopwatches for
measurement of retention times. The chromatograms were collected with a strip
chart recorder. Data analysis was also performed manually.

However, compared with the traditional analytical methods, the adoption of

chromatographic methods such as HPLC and GC represented a significant
improvement in pharmaceutical analysis and had major advantages of method
specificity, sensitivity and reliability.

As chromatographic methods became more and more important in the pharmaceutical

industry as well as in other industries, practical needs prompted instrument vendors to
come up with more efficient ways for collecting and processing chromatographic data.
Advantages of modern CDS over traditional CDS
Chromatographic technique has many advantages over traditional method
• Method is more specific, accurate and precise
• It has wide ability of separation
• It is used to identify low level of impurities

• Mainly three advantages were observed by use of modern CDS over

traditional CDS

1. Modern CDS is most improved and efficient as compared to initial

computer data systems
• Server based computing technique
• Use of embedded data structure
• Direct instrument control
2. The earlier generations of CDS used a directory file structure,
meaning that the raw data and other files such as the instrument
method and data processing method were stored at separate locations.
There would either be no connections or only partial connections
between these files. The most significant drawback of this type of file
management was the potential for methods and raw data to be
accidentally overwritten. To prevent this from happening, the raw data
and result files must be locked.

Modern day CDS applied lock or encryption system to raw data and
result files which provided system security and user privileges.
3. Older CDS lacked direct instrument control. The modern CDS
detector channels were introduced to connect all CDS workers in
analytical laboratories across the entire pharmaceutical industry. This
helped to collect all data into CDS from several instruments including
GC, HPLC etc.
INTRODUCTION TO CHROMATOGRAPHY
• Chromatography is a laboratory technique for the separation of a mixture.
Principle:
The sample mixture to be analyzed is dissolved in a fluid called the mobile
phase.
Mobile phase carries the sample through a structure called the stationary
phase.
The various constituents of the mixture travel at different speeds, causing
them to separate.
The separation is based on differential partitioning between the mobile and
stationary phases.
Subtle differences in a compound's partition coefficient result in differential
retention on the stationary phase and thus affect the separation.
• Retention Factor, Rf ?
• The retention factor, Rf, is a quantitative indication of how far a
particular compound travels in a particular solvent.
• The Rf value is a good indicator of whether an unknown compound
and a known compound are similar, if not identical.
• If the Rf value for the unknown compound is close or the same as the
Rf value for the known compound then the two compounds are most
likely similar or identical.
• The retention factor, Rf, is defined as Rf = distance the solute (D1)
moves divided by the distance traveled by the solvent front (D2) Rf =
D1 / D2
• Stationary phase in chromatography, is a solid phase or a liquid phase
coated on the surface of a solid phase. Mobile phase flowing over the
stationary phase is a gaseous or liquid phase. If mobile phase is liquid it is
termed as liquid chromatography (LC), and if it is gas then it is called gas
chromatography (GC). Gas chromatography is applied for gases, and
mixtures of volatile liquids, and solid material. Liquid chromatography is
used especially for thermal unstable, and non-volatile samples.

• The purpose of applying chromatography which is used as a method of

quantitative analysis apart from its separation, is to achieve a satisfactory
separation within a suitable time interval. Various chromatography
methods have been developed to that end. Some of them include column
chromatography, thin-layer chromatography (TLC), paper chromatography,
gas chromatography, ion exchange chromatography, gel permeation
chromatography, high-pressure liquid chromatography, and affinity
chromatography.
Different types of chromatographyu based on separation principle,
geometry of method, mode of chromatography
• Column chromatography
• Ion-exchange chromatography
• Gel-permeation (molecular sieve) chromatography
• Affinity chromatography
• Paper chromatography
• Thin-layer chromatography
• Gas chromatography
• Dye-ligand chromatography
• Hydrophobic interaction chromatography
• Pseudoaffinity chromatography
• High-pressure liquid chromatography (HPLC)
• Chromatography methods based on partition are very effective on
separation, and identification of small molecules as amino acids,
carbohydrates, and fatty acids.

• However, affinity chromatography (ie. ion-exchange chromatography)

are more effective in the separation of macromolecules as nucleic
acids, and proteins.
• Paper chromatography is used in the separation of proteins, and in
studies related to protein synthesis;
• gas-liquid chromatography is utilized in the separation of alcohol,
esther, lipid, and amino groups, and observation of enzymatic
interactions, while
• molecular-sieve chromatography is employed especially for the
determination of molecular weights of proteins.
• Agarose-gel chromatography is used for the purification of RNA, DNA
particles, and viruses
Paper chromatography
• In paper chromatography support material consists of a layer of cellulose highly
saturated with water. In this method a thick filter paper comprised the support,
and water drops settled in its pores made up the stationary “liquid phase.”
Mobile phase consists of an appropriate fluid placed in a developing tank. Paper
chromatography is a “liquid-liquid” chromatography.

Thin-layer chromatography
• Thin-layer chromatography is a “solid-liquid adsorption” chromatography. In this
method stationary phase is a solid adsorbent substance (alumina or silica) are
coated on glass plates, aluminium foil or silica.
• In cases where molecules of the sample are colorless, florescence, radioactivity
or a specific chemical substance can be used to produce a visible coloured
reactive product so as to identify their positions on the chromatogram.
• Formation of a visible colour can be observed under room light or UV light. The
position of each molecule in the mixture can be measured by calculating the ratio
between the distances travelled by the molecule and the solvent. and expressed
with a symbol Rf. Rf. value is used for qualitative description of the molecules.
• Gas chromatography
is a “gas-liquid” chromatography. Its carrier phase consists of gases as
He or N2. Mobile phase which is an inert gas is passed through a
column under high pressure. The sample to be analyzed is vaporized,
and enters into a gaseous mobile phase. The components contained in
the sample are dispersed between mobile phase, and stationary phase
on the solid support. Gas chromatography is a simple, multifaceted,
highly sensitive, and rapidly applied technique for the extremely
excellent separation of very minute molecules. It is used in the
separation of very little amounts of analytes
HPLC: (High performance liquid chromatography)
Using this chromatography technique it is possible to perform
structural, and functional analysis, and purification of many molecules
within a short time,
This technique yields perfect results in the separation, and
identification of amino acids, carbohydrates, lipids, nucleic acids,
proteins, steroids, and other biologically active molecules.
In HPLC, mobile phase passes through columns under 10–400
atmospheric pressure, and with a high (0.1–5 cm/sec) flow rate. In this
technique, use of small particles, and application of high pressure on
the rate of solvent flow increases separation power, of HPLC and the
analysis is completed within a short time.
• High performace thin layer chromatography: (HPTLC)
• Is an advanced form of TLC. It provides superior separation power
using optimized coating material, automated procedures for mobile
phase feeding, layer preconditioning, precise sample application,
chromatogram development scanning and photo documentation.

• It has several advantages like it promotes better separation

efficiencies, shorter analysis time, lower amount of mobile phase,
efficient data acquisition and processing.
Applications: Validation of two or more drug combinations in a dosage
form, identification of constituents, impurities and active substances.
• Online link on chromatography for reference if required
• https://youtu.be/XMtmSz_9umk