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

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 sample 2. 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.

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

REFERENCE
1. Teaching.shu.ac.uk. Gas Chromatography. 2015. Web. 3 Oct. 2015.
2. Lecture done by PhD student, Yong.