Presentation of Industrial training, for organic chemistry in MSC Department of chemistry Gujarat University Ahmedabad

1. DEPARTMENT OF CHEMISTRY
SCHOOL OF SCIENCES
A PRESENTATION ON
INDUSTRIAL TRAINING
2019

2. 25-Jul-20

3. AnSys Research Laboratories
Delivering the Science-Ensuring Pharma Solutions
2-Third Floor Ceramic Center,
Opp. HDFC Bank-Vikram Plaza,
Nr. Vishawakarma Tample,
Gota Railway Bridge
Gota, Ahmedabad-382481, Gujarat, India.
SUBMITTED TO…
Department Of Chemistry
School Of Sciences
Gujarat University,
Ahemdabad -380009
SUBMITTED BY…
Nilesh D. Parmar
M.Sc Sem – 4
Exam No. 0276
25-Jul-20

4. Contents
 Introduction Of The Company
About Research and Development
 What I have learnt ?
 Introduction to The Glassware Used in Laboratories
 Introduction to The Instruments present in Laboratories
25-Jul-20

5.  Ansys research laboratories is a contract research organization.
 AnSys provides a wide variety of pharma solution which range from api
impurities, intermediates ,bulk compound to specialty species in the
pharmaceutical, fine chemical and biotechnology industries.
 They are committed to providing our customers with the best quality
services at the most competitive prices in time.
Introduction Of The Company
25-Jul-20

6. 1. Research And Development (R&D)
2. Analytical Development Lab (ADL)
25-Jul-20
Department Of Company

7. 25-Jul-20
What is Research and Development (R&D)?
“The process of obtaining new knowledge and using it to improve
existing products and introduce new ones”
Three type of work are being done in R&D Laboratory:
(1) Discovery of a New Compounds.
(2) Synthesis of known Product which is new for Industry.
(3) Method development for own Products.

8.  Reaction
 Reaction Moniter
TLC
Stirring
Temperature
Workup of Reaction
 Solvent used
MDC & MeOH
Ethyl Acetate & Hexene, Water etc.
 Purification by Glass Manual column
 Distillation by Rotavapour
 LCMS
 Mass
25-Jul-20
R & D Work

9. 25-Jul-20
What I Have Learnt ?
R & D
1
Safety
Precaution
2
Reaction
handling
3
Reaction
monitoring
4
Separation
technique
5.
Purification

10. 25-Jul-20
1.Safety Precaution
General safety
Glassware safety
Chemical safety
Electrical & heating safety

11. 2. Reaction handling
Arrange reaction assembly
Clean glassware
Handling chemical
Temperature (heating/cooling), under Nitrogen,
stirring, observation color changes
25-Jul-20

12. 25-Jul-20
3. Reaction monitoring
TLC
TLC stain
Ex. KMno4, Ninhydrine…etc
TLCchamber

13. 25-Jul-20
4. Separation technique
Colum chromatography
Extraction
Vacuum filtration
Vacuum Filtration Assembly
Separatingfunnel
Celite filter aid

14. 25-Jul-20
5. Purification
Colum chromatography
Glass column for separating mixture

15. Round Bottom Flasks(RBF)
Single neck Double neck Triple neck
25-Jul-20
Introduction to The Glassware Used in Laboratories

16. Condenser, Joints, Pear Shape Flask & Other Equipments
25-Jul-20

17. 25-Jul-20
Introduction to The Instruments
UV-Chamber
Fume hood
Magnetic stirrer with hotplateStirr bar
Rotavap

18. 25-Jul-20

19. Objective:
‘’To separate Organic compounds with the help of Column
Chromatographic technique’’
Chromatography:
Chromatography has been developed into a new method of separation of mixture
of substances mainly when they are available in small amounts. This method is
very useful when the components of a mixture have almost the same physical and
chemical properties and hence can’t be separated by other usual methods of
separations. The term chromatography means writing in colour (in Greek:
Khromatos-colour, and graphos- written). It was discovered by Mikhail Tswett in
1906.
The methods of separation in chromatography are based on the distribution of the
components in a mixture between a fixed (stationary) and a moving (mobile)
phase. The stationary phase may be a column of adsorbent, a paper, a thin layer of
adsorbent on a glass plate, etc., through which the mobile phase moves on. The
mobile phase may be a liquid or a gas. When a solid stationary phase is taken as a
column it is known as column chromatography.
25-Jul-20

20. Column Chromatography:
Column chromatography is one of the most useful methods for the separation and
purification of both solids and liquids. This is a solid - liquid technique in which the
stationary phase is a solid & mobile phase is a liquid. The principle of column
chromatography is based on differential adsorption of substance by the adsorbent.
The usual adsorbents employed in column chromatography are silica, alumina,
calcium carbonate, calcium phosphate, magnesia, starch, etc., selection of solvent is
based on the nature of both the solvent and the adsorbent. The rate at which the
components of a mixture are separated depends on the activity of the adsorbent
and polarity of the solvent. If the activity of the adsorbent is very high and polarity
of the solvent is very low, then the separation is very slow but gives a good
separation. On the other hand, if the activity of adsorbent is low and polarity of the
solvent is high the separation is rapid but gives only a poor separation, i.e., the
components separated are not 100% pure.
The adsorbent is made into slurry with a suitable liquid and placed in a cylindrical
tube that is plugged at the bottom by a piece of glass wool or porous disc. The
mixture to be separated is dissolved in a suitable solvent and introduced at the top
of the column and is allowed to pass through the column.
25-Jul-20

21. As the mixture moves down through the column, the components are adsorbed at
different regions depending on their ability for adsorption. The component with
greater adsorption power will be adsorbed at the top and the other will be adsorbed
at the bottom. The different components can be desorbed and collected separately
by adding more solvent at the top and this process is known as elution. That is, the
process of dissolving out of the components from the adsorbent is called elution and
the solvent is called is called eluent. The weakly adsorbed component will be eluted
more rapidly than the other. The different fractions are collected separately.
Distillation or evaporation of the solvent from the different fractions gives the pure
components.
Intermolecular forces, which vary in strength according to their type, make organic
molecules to bind to the stationary phase. The stronger the intermolecular force, the
stronger the binding to the stationary phase, therefore the longer the compound
takes to go through the column.
Intra-molecular hydrogen bonding is present in ortho- nitro phenol. This is due to
the polar nature of the O-H bonds which can result in the formation of hydrogen
bonds within the same molecule. But in para-nitro phenol, inter molecular
hydrogen bonding (between H and O atoms of two different para-nitro phenol
molecules) is possible. As result of inter molecular hydrogen bonding para-nitro
phenol undergo association that increases the molecular weight, whereby
decreasing volatility.
25-Jul-20

22. Introduction
Thin-layer chromatography (TLC) is a very commonly
used technique in synthetic chemistry for identifying compounds,
determining their purity and following the progress of a reaction. It also
permits the optimization of the solvent system for a given separation
problem. In comparison with column chromatography, it only requires
small quantities of the compound (~ng) and is much faster as well.
Stationary Phase
As stationary phase, a special finely ground matrix (silica gel, alumina,
or similar material) is coated on a glass plate, a metal or a plastic film as
a thin layer (~0.25 mm). In addition a binder like gypsum is mixed into
the stationary phase to make it stick better to the slide. In many cases, a
fluorescent powder is mixed into the stationary phase to simplify the
visualization later on (e.g. bright green when you expose it to 254 nm UV
light).
25-Jul-20

23. Preparing the Plate
Do not touch the TLC plate on the side with the white
surface. In order to obtain an imaginary start line, make two notches
on each side of the TLC plate. You can also draw a thin line with pencil.
‘’Do not use pen. Why? ’’
The start line should be 0.5-1 cm from the bottom of the plate.
Capillary spotters
Place a melting point capillary and in the dark blue part of the
Bunsen burner flame. Hold it there until it softens and starts to sag.
Quickly remove the capillary from the flame and pull on both ends to
about 2-3 times its original length. If you pull the capillary inside the
flame, you will have a "piece of art", but not a good spotter. Allow the
capillary to cool down, and then break it in the middle. Make sure that
you break off the closed end on one of them. Do not use gloves when
you pull capillaries. You will have much better control without them!
25-Jul-20

24. Spotting the plate
The thin end of the spotter is placed in the dilute solution; the solution
will rise up in the capillary (capillary forces). Touch the plate briefly at
the start line. Allow the solvent to evaporate and spot at the same place
again. This way you will get a concentrated and small spot. Try to avoid
spotting too much material, because this will deteriorate the quality of
the separation considerably (‘tailing’). The spots should be far enough
away from the edges and from each other as well. If possible, you should
spot the compound or mixture together with the starting materials and
possible intermediates on the plate. They will serve as internal reference
since every TLC plate is slightly different.
25-Jul-20

25. Developing a Plate
A TLC plate can be developed in a beaker or closed jar (see picture
below). Place a small amount of solvent (= mobile phase) in the
container. The solvent level has to be below the starting line of the TLC,
otherwise the spots will dissolve away. The lower edge of the plate is then
dipped in a solvent. The solvent (eluent) travels up the matrix by
capillarity, moving the components of the samples at various rates
because of their different degrees of interaction with the matrix
(=stationary phase) and solubility in the developing solvent. Non-polar
solvents will force non-polar compounds to the top of the plate, because
the compounds dissolve well and do not interact with the polar
stationary phase. Allow the solvent to travel up the plate until ~1 cm
from the top. Take the plate out and mark the solvent
front immediately. Do not allow the solvent to run over the edge of the
plate. Next, let the solvent evaporate completely.
25-Jul-20

26. TLC chamber for development
e.g. beacher
with a lid or a closed jar
after ~5 min after ~10 min after drying
25-Jul-20

27. Reagent Works well for Colors
Iodine Unsaturated and aromatic compounds Brown spots
Sulfuric acid General stain Brown or black spots
Chromic acid For difficult to stain compounds Black spots
UV light Compounds with extended conjugation
like aromatic compounds
Pink on light green background
Cerium sulfate Good general stain, very well for
alkaloids
Ferric chloride Phenols Purple
Ninhydrin Amino acids, amines Purple
2,4-Dinitrophenylhydrazine Aldehydes, ketones Yellow/orange
Vanillin Good general stain, very well for
hydroxyl or carbonyl compounds
Colors vary
Potassium permanganate Works well for all compounds that can
be oxidized
Yellow on purple
Yellow or light brown on purple
Bromocresol Green Carboxylic acids (pKa<5) Yellow spot on blue background
p-Anisaldehyde Good general stain, particularly sensitive
towards nucleophiles
Varying colors on light pink plate
upon heating
Phosphomolybdic acid (PMA) Very sensitive Dark green spot on light green plate
Ehrlich’s Reagent
(Dimethylaminobenzaldehyde)
Indoles, amines Pink or red-violet
Dragendorff-Munier Stain Amines even the ones that are low in
reactivity
Various colors
25-Jul-20

28. 25-Jul-20
In either way, the spots on the TLC plate should be circled (marked) to
have a permanent record how far the compound traveled on the plate.
Asketch of the developed plate should be placed in your lab notebook. A
picture (cell phone) could not hurt either.
Analysis
The components, visible as separated spots, are identified by comparing
the distances they have traveled with those of the known reference
materials. Measure the distance of the start line to the solvent front (=d).
Then measure the distance of center of the spot to the start line (=a).
Divide the distance the solvent moved by the distance the individual spot
moved. The resulting ratio is called Rf-value. The value should be
between 0.0 (spot did not moved from starting line) and 1.0 (spot moved
with solvent front) and is unitless.

29. 25-Jul-20
The Rf (=retardation factor) depends on the following parameters:
 Solvent System
 Absorbent (Grain Size, Water Content, Thickness)
 Amount Of Material Spotted
 Temperature
Due to the fact that all those variables are difficult to keep constant, a reference
compound is usually applied to the plate as well.

30. 25-Jul-20
A fume hood (sometimes called a fume cupboard or fume closet) is a type
of local ventilation device that is designed to limit exposure to hazardous or toxic
fumes, vapors or dusts.
This is used to:
protect the user from inhaling toxic gases (fume hoods, biosafety cabinets, glove
boxes)
protect the product or experiment (biosafety cabinets, glove boxes)
protect the environment (recirculating fume hoods, certain biosafety cabinets, and
any other type when fitted with appropriate filters in the exhaust airstream)