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MARINE ORGANIC CHEMISTRY

SL20103
PRACTICAL TITLE : HIGH-PERFORMANCE
LIQUID CHROMATOGRAPHY AND THINLAYER CHROMATOGRAPHY

NAME

MATRIX NO.

ENGRID PACHECO DANING TUZAN BS14161084


SHARMINA BINTI JEBIN

BS14110781

SITI SYAHIRAH BINTI MUZLI

BS14110802

MULLAISUDHARR A/L MUNIANDY

BS14110554

SHARIFAH AZIAH JASMINE BINTI


SHARIF JAKARIA

BS14110110

1.0 INTRODUCTION
Chromatography is a pretty accurate description of what happens to in on wet paper
because it literally means colour writing. Chromatography is actually a way of separating
out a mixture of chemicals which are in gas or liquid form by letting them to creep slowly
past another substance which is typically a liquid or solid. The essential thing about
chromatography is that we have some mixture in one state of matter moving over the
surface of something else in another state of matter (liquid or solid) that stays where it is.
The moving substance is called the mobile phase and the substance that stays put is the
stationary phase. As the mobile phase moves, it separates out into components on the
stationary phase.
Gas chromatography is a term used to describe the group of analytical separation
techniques used to analyse volatile substances in the gas phase. In gas chromatography, the
components of a sample are dissolved in a solvent and vaporized in order to separate the
analytes by distributing the sample between two phases which are stationary phase abd a
mobile phase. The mobile phase is a chemically inert gas that serves to carry the molecules
of the analyte through the heated column. Gas chromatography is one of the sole forms of
chromatography that does not utilize the mobile phase for interacting with the analyte. The
stationary phase is either a solid adsorbant termed gas- solid chromatography (GSC) or a
liquid on an inert support termed gas- liquid chromatography (GLC).
Liquid chromatography is a technique used to separate a sample into its individual
parts. This separation occurs based on the interactions of the sample with the mobile and
stationary phases. There are many stationary and mobile phase combinations that can be
employed when separating a mixture, there are several different types of chromatography
hat are classified based on the physical states of those phases. Liquid- solid column
chromatography is the most popular chromatography technique and the one discussed here
features a liquid mobile phase which slowly filters down through the solid stationary phase
bringing the separated components with it.

OBJECTIVE
1. To acknowledge the analytical separation techniques use to separate marine organic
compounds.
2. To study the uses and functions of each chromatography machines.

MATERIAL
1) High Performance Liquid Chromatography machine
2) Mass spectrometry machine
3) Gas chromatography machine

PROCEDURE
1. All of us are instructed to assemble at Biotechnology Research Institute (BRI),
University Malaysia Sabah (UMS) at 8:00am.
2. Dr. Shafiqquzzaman Sidiqquee then gave us a short briefing about the practical and
he led us to the BRI building into the lab.
3. Proper attire should be worn in the laboratory and a notepad with pen should be
brought.
4. A PhD student named Yong then gave us a briefing on how to use the Liquid
Chromatography machine and precautions that should be taken when handling the
machine.
5. After the briefing on the LCG, we are then brought to another lab where the Gas
Chromatography is kept and a further briefing on the machine is done by Young.
6. Pictures and notes are taken during the practical as evidence and further reference.

RESULT

Solvent Cabinet 1

Degasser 2

Capillary pump

Auto Sampler

Photodiode Detector

Figure 1 : HPLC System

Flight Tube

Glass Capillary

Double Electron System


!
Figure 2 Mass Spectrometer

Figure 3: Gas Chromatography Machine

Figure 4: Computer is used as an online system to monetize the sample

Figure 5: Computer is used as an online system to monetize the sample

Figure 6: The auto sampler and the injector.

Figure 7: Mass Spectrometry

DISCUSSION

Liquid Chromatography Machine

An LC-MS is an High Performance Liquid Chromatography (HPLC) system with a mass


spec detector.The HPLC separates chemicals by conventional chromatography on a column.
Components of an HPLC system are solvent cabinet, degasser, capillary pump, injector,
column, detector, fraction collector and an integrator. Usually the method will be reverse
phase chromatography, where the metabolite binds to the column by hydrophobic
interactions in the presence of a hydrophilic solvent (for instance water) and is eluted off by
a more hydrophobic solvent (methanol or acetonitrile). As the metabolites appear from the
end of the column they enter the mass detector, where the solvent is removed and the
metabolites are ionised. The metabolites must be ionised because the detector can only
work with ions, not neutral molecules. And ions only fly through a very good vacuum, so
removal of the solvent is a vital first step. The mass detector then scans the molecules it
sees by mass and produces a full high-resolution spectrum, separating all ions that have
different masses1.
LC system is responsible for the separation. The solvent cabinet is use for filtered the
sample. HPLC uses a liquid to push the sample. This liquid is called the Mobile Phase or
solvent. Before use the solvent may be filtered through micron pore size filters or the
container can have a frit filter. Based, degasser is a component of an HLPC system. The
function of degasser is degassed a solvents to eliminate formation of bubbles. Pump is to
deliver the mobile phase through the system before dilute into the auto sampler. After that,
the sample will go to the auto sampler after its break out. In the auto sampler, there is a
sample collector chamber. Some sample such as fat or organic compound was quite
impossible to ionize. Its can be ionized when add some amphoteric acid. The sample will not
recognized if the amphoteric acid is not added.
MS system is to ionize the sample again. There is double electron spray system
where there have two needles. One of the needles is for the reference of the sample mass
and the other is for the sample. The needles is a injector which is to put the sample in the
mobile phase.
Computer is used as an online system to monetize the sample. It uses an online
system to store data, temperature and to observe the ongoing process that occurs on the
sample. A chromatogram will display on the screen of the computer and it will show the
1

http://www.chemir.com/liquid-chromatography-mass-spectrometry.html

profile of the sample during the process. The chromatogram shows the good and poor
separation of the sample. Each peak shown on the chromatogram represents only one
profile of a compound. Profile of the sample depends on the volatility.

Gas Chromatography Machine


The gas chromatography machine starts with injecting the liquid sample into a small
sample bottle. It is then placed into the auto sampler. The samples are then pushed into the
injector. The injector have a temperature of about to 200C and the sample is injected into
the gas chromatography. Inside, the samples will evaporate the liquid. The separation
process is based on the temperature which usually starts from 15C and can be up to
300C. The liquid in the sample should be removed before it enters the column.
Next the sample will enter the column. The column is about 30m long with a
diameter of about 0.25m and the column has a temperature of up to 355C. It is where
the coating will be done upon the sample. The temperature has to be adjusted according to
the boiling point of sample2. If the temperature exceeds the sample then the result will be
loose.
Similar to the Liquid Chromatography Machine, it uses an online system to store
data, temperature and to observe the ongoing process that occurs on the sample. During
the process, a chromatogram will display on the screen of the computer and it will show the
profile of the sample. The chromatogram shows the good and poor separation of the
sample. Each peak shown on the chromatogram represents only one profile of a compound.
Profile of the sample depends on the volatility.

www.teaching.shu.ac.uk/hwb/chemistry/tutorials/chrom/gaschrm.htm

CONCLUSION
The analytical separation techniques used to separate marine organic compounds are
acknowledged and the functions of each chromatography machines are studied.

REFERENCES
1 . C h r o m a t o g ra p h y. ( n o d a t e ) . W h a t i s C h r o m a t o g ra p h y ? i n

http://

www.explainthatstuff.com/chromatography.html
Retrieved on 3rd October 2015.
2.UCDAVIS CHEMWiki. (no date). Liquid Chromatography in http://chemwiki.ucdavis.edu/
Analytical_Chemistry/Instrumental_Analysis/Chromatography/Liquid_Chromatography
Retrieved on 3rd October 2015.
3. Dr. Shafiquzzaman Siddiquee. (2015). Lecture 2 Separations of Marine Products.
Powerpoint slide. Malaysia, Sabah. Universiti Malaysia Sabah.

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