PROCEDURE1. 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

Figure 1 : HPLC System

Solvent Cabinet 1

Degasser 2

Capillary pump

Auto Sampler

Photodiode Detector

Figure 2 Mass Spectrometer

Figure 3: Gas Chromatography Machine

Flight Tube

Glass Capillary

Double Electron System

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

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 200°C 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 15°C and can be up to 300°C. 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 0.25µm and the column has a temperature of up to 355°C. 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.

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

REFERENCE1. Teaching.shu.ac.uk. ‘Gas Chromatography’. 2015. Web. 3 Oct. 2015.2. Lecture done by PhD student, Yong.