it final report
TRANSCRIPT
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Universiti Tunku Abdul Rahman
Faculty of Science
Bachelor of Science (HONS) Biochemistry
Year Two Semester Two
UDEE 2306 INDUSTRIAL TRAINING
REPORT
NAME : CHEW HUI SIN
ID NO : 0908196
COMPANY : BIOFACT LIFE SDN BHD
ADDRESS : LOT 5094, KAWASAN PERINDUSTRIAN
PARIT JAMIL, 84150 PARIT JAWA,
MUAR, JOHOR
SUPERVISOR (COMPANY) : Mr NG HONG SENG
SUPERVISING LECTURER : DR KHOO KONG SOO
DATE OF SUBMITION : 13 JANUARY 2010
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Table of Contents
Table of Contents ....................................................................................... 2
Acknowledgement ..................................................................................... 3
Abstract .................................................................................................. 4
Introduction ............................................................................................... 5
Goal of Industrial Traning ......................................................................... 5
Scope Of Training ................................................................................. 5
Company Background ............................................................................... 6
Mission& Vision ....................................................................................... 7
Award and Accreditation ........................................................................ 7
Product ................................................................................................... 10
Work-Based Learning Experience .......................................................... 12
Learning Outcomes ................................................................................. 19
Microbiology Lab .................................................................................... 19
Chemical Lab ...................................................................................... 22
Conclusion ............................................................................................... 23
References ............................................................................................... 24
Appendix ................................................................................................... 25
Bi-weekly reports ..................................................................................... 25
Project Assigned ................................................................................... 26
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ACKNOWLEDGEMENT
I feel so glad and appreciated with my University, Universiti Tunku Abdul
Rahman, give me an opportunity to go through this industrial training. This program
helps us to get use with the truly working environment prior our graduation to increase
our competitive force in the real world. I would like to thanks Dr Khoo Kong Soo,
lecturer supervising me in Utar that willing to guide me and giving me direction from
time to time during the industrial training.
I would like to thanks my company, Biofact Life Sdn Bhd that providing me an
excellent working environment for me learn. Not to forget the science executive of our
company, Mr. Eston Lee, which willing to hire me as a trainee and also assigned me to
different department to handle different kinds of tasks.
Last but not least, both of my supervisors which are Mr Ng Hong Seng and also
Miss Lee Chia Yen. I am so appreciate with their encouragement to share their working
experience with me. By the way, they also assisting me in complete the project offering
to me. I do learn a lot of knowledge in their department.
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ABSTRACT
Industrial training is credited course programmed that compulsory to satisfy as a
degree course work requirement for graduation. As a biochemistry student, all of us have
to undergo the 12 weeks industrial training from 4 october until 24 December. This report
will cover the essential aspects of my industrial training experience. From the goal of
which I would want to achieve from this experience to the learning and training grounds
which I have undertaken. This report will introduce the company profile first. Secondly, I
would be mentioned my learning experience within this 12 weeks of working period.
Thirdly, I will include what I had gained and applied in this report. Last but not least, the
bi-weekly report and sample of work done would be included as appendices in the end of
the report.
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INTRODUCTION
Goal of Industrial Training
The goal of this industrial training is to assist the students by exposing them to the real
working world. Within twelve weeks, students are exposed to the industrial so when it is time for
them to venture into work life it would not come as a shock.
Another goal is to gain experience in real working environment. This is so that students
are able to learn work ethics, take on and handle responsibilities and meet deadlines efficiently.
This industrial training also enhances and assists students to be aware of their strengths
and weaknesses and to grow and improve themselves from here on. With this realization, students
are able to build confidence in their capabilities and use theories learnt in classes in real life
situations.
Scope of Training
In the beginning of the internship in Biofact Life, I was assigned to the chemical
laboratory. Under the guidance of Miss Lee Chia Yen, our company chemist, I learn to handle the
main instruments in the lab. I was getting an experience on how to become a good assistant in the
lab to assist my supervisor to complete her daily routine works.
Besides, I also get a chance to learn from the microbiologist in our company. Our
company microbiologist, Mr Ng Hong Seng is also quality assurance in our company. Thus,
besides working as a trainee in the microbiology lab, he also introduces me on how a QA
department operate in the company. He also gives an opportunity for me to become a one day
microbiologist in the company in order to let me experience the real working environment more
deeply.
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My company also assigned me a project regarding to cordycepins extraction. Thus, I
planned and carried out difference type of experiment in order to complete the project. And Mr
Ng Hong Seng and Miss Lee Chia Yen will assist and guide me throughout the whole project.
Company Background
Biofact Life Sdn Bhd was incorporate in Malaysia under the Company Act 1965 on 1st
February 2005. Biofact Life Sdn Bhd specialises in the production of cordyceps by laboratory
cultivation. The company is one of the pioneer biotechnology companies in the world to employ
the latest biotechnology techniques in cordyceps cultivation at reduced cost and improved quality.
They culture the cordyceps under low temperature and oxygen conditions. They designed
environment that free from chemical pollution as well as all the yeast, mould and bacterial
contamination.
BioFact Life encompassed a seed-to-shelf approach as the company is fully committed to
extensive research and development in the production, manufacturing and marketing of their
cordyceps. GMP and ISO 22000:2005-certified are practise in the manufacturing division. Thus,
the quality, safety and efficacy to manufacture the product in the form if capsule, powder, tablet,
pill, liquid, and ointment are maintained.
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Vision
Quality for a quality life
Mission
y To become a key global player in the mainstream of natural healthcare markets
through vertical integration and strategic globalization.
y To develop, manufacture and promote quality natural healthcare products for
mankind.
y To keep abreast with the latest developments in natural healthcare products.
y To ensure continuous improvement in the standard of our products and services so
that our company, employees, business partners and stakeholders will benefit and
prosper together.
Award and Accreditation received by BioFact Life
Golden Bull Award 2009
BioFact Life Sdn Bhd has been awarded with the Golden Bull Award 2009, which is most
representative annual business award that accredits and honours the best efforts and achievements
if SMEs in Malaysia.
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SMEs Fast Moving Companies 2009In 2009, Biofact Life also received recognition from the SME Fast Moving Companies. The
SME100 is an Annual Recognition Award arganized by SMe & Entrepreneurship Magazine for
recognizing the achievements of SMEs in Malaysia.
ISO 22000:2005
Biofact Life Snd Bhd demonstrating their commitment to ensuring the higest standard food safety
management also received ISO 22000:2005 certified in the first quarter of 2009. Through this
certification, they ensure that the mist effective food safety systems are established operated and
updated within the framework of structured management sysyatem and incorporated into overall
management activities of BioFact Life.
The Malaysia Book of Records
BioFact Life has been accreditated by the Malaysia Book Of Records as the First Malaysian
company that successfully cultivate cordyceps in a specially designed 100K clean room where the
room conditions are regulated and controlled according to the different growth stage of
cordyceps.
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Halal Product Certified
BioFact Life also obtained a Halal Certification from the Department of The advancement of
Islam Malaysia (JAKIM).
Good Manufacturing Practice
BioFact Life is GMP- Certified. The company takes proactive steps to ensure that the products
are safe, pure and effective by enforcing a quality approach in the manufacturing, processing and
packaging processes.
Bioinno Awards 2008
BioFact Life also awarded the Gold Winner in Bioinno Award Competition organized by the
Ministry of Science, Technology and Innovation (MOSTI).
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Product Manufactured by BioFact Life
The pharmaceutical products manufactured by BioFact Life are basically added up with
cordyceps they cultivated. Those are the hot seller product in our company:-
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Work-based Learning Experience
Cordyceps Sinesis, which is known to the Chinese as Dong Cong Xia Cao and
to Japanese as Tochukuso was long been used in medicine.(John et al.,2004) The native
cordyceps are grown on insect such like worm. The insects body play role in providing
nutrient for the cordyceps to growth. The cordyceps cultured in our company are using
rice as the substrate for cordyceps. After the cordyceps being harvested, the substrate is
remained. But the substrates do absorb some unique ingredient found in cordyceps such
like cordycepins. My company would like to extract the cordycepins found in substrate
for others usage in the industrial.
So, my company assigned me the project to set the procedure for the cordycepins
extraction. Firstly, I planned the overall extraction process. The process is as showed in
the flow chart below;-
In the first step, I plan to using heat to release the cordycepins found in cordyceps
substrate. Then I will filter out the residual. The extract might full with different large
compound or molecule like polysaccharides from the substrate. So, I try to using heat to
Incubatethe
substrate
Filter Boil the
extraction
Filter
Concentratethe
extraction
Liquid Extract of
Cordycepins
Autoclavethe
Extract
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break down those molecules that consider as the impurities in the extract by using heat. The
impurities is then filter out. We will set the minimum concentration of cordycepins should
presence in the extract to 300ppm. If we could not get the concentration we desired directly
from the process, concentration method is needed to concentrate the extraction. At the end
we will autoclave the final extract to kill the bacterial thus contamination is avoided.
There are differences factors that may affect the concentration of cordycepins in the
extract. The factors I consider about are:-
y Ratio of substrate to water
y
Temperature
y Incubation time
y Concentration method using
y Pre-treatment involve
Differences types of experiment are carried out to determine whether those factors can affect
the concentration of cordycepins during the extraction.
Simple materials and methods are using throughout the project. Apparatus such
like beaker, Schott bottle, Bunsen burner, spatula, retort stand, Whatmanfilter paper, filter
bag, filter pump and thermometer are using in the few experiments. Besides, HPLC,
incubator and autoclaving machine are also needed in the project.
The first experiment I conducted is to find out which ratio of water to substrate can actually
work the best in the releasing of cordycepins in water. Firstly, a specific amount of substrate
is weighted and placed in a Schott bottle. The Schott bottle is then fill in with water before
place in a 60 C water bath to incubate for 2 hours. The residual is filtered out by using filter
bag. The extract is then sent to autoclave at 122 C for 15 minutes to break down the
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polysaccharides and also all the impurities in the extract. A thick precipitate will usually
found in the extract, hence, we need filter paper to filter the precipitate out. I will autoclave
the extract at 122C for 15 minutes to avoid contamination occurs. The final extract is then
sending to chemical for HPLC test. The variety of ratio and the result is stated in the table
below:-
As the conclusion, the ratio of substrate to water 1:7 can work the best in the
extraction process. This ratio showed us the closest figure between the actual and expected
concentration of cordycepins in the extraction. This means that the most cordycepin found in
the substrate were extracted out and only few amounts of cordycepins are remaining in the
substrate or residual.
The second experiment carried out is about the optimum temperature of
cordycepins extraction. There are data showing that the melting point of the cordycepins is
225 C. Therefore, I expect that the concentration of the cordycepins in the extract will
Substrate :
water
Substrate
weight (g)
Volume of
water (ml)
Cordycepin
being tested
(ppm)
Expected
concentration
of cordycepins
Effectivenes
(%)
1:1 250 250 762.347 762.347 100.00
1:4 100 400 132.147 190.587 69.33
1:5 60 440 113.628 152.469 74.52
1:6 70 430 99.252 127.058 78.11
1:7 80 420 94.973 108.907 87.2
1:9 50 450 68.520 84.705 80.88
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increase as the incubation time increase. The same steps in the first experimrnt are carried
out. But the temperature using are differences.
The result is as tabulate in the table above.
From the table above, we can conclude that the melting point is not as the same as
the temperature which cordycepins can resist. Its only shown that the physical change of
cordycepins change from solid to liquid. The best temperature get is 60 C. The hypothesis I
made are rejected as the concentration of cordycepins in the extract decrease when the we
set the extraction temperature into 75 C. while cordycepin also cannot be extracted well at l
room temperature.
Then, the experiment is followed by determine the best incubation time for the
extract. Same experiment are carried out the only different is the time taken for incubation.
We use 1, 2 and 3 hours respectively in this experiment. I do a similar hypothesis as the
previous experiment in this experiment. I thought that the concentration of cordycepins
might increase as the time of incubation increase at a constant temperature. I found out that
my expectation wrong once I received the chemical result. The result is as shown as below:-
Temperature (C) Concentration of cordycepin
obtained (ppm)
Percentage water
remained (%)
25 70.68 72.7
50 87.45 63.6
60 94.97 63.6
75 81.66 36.4
100 - 0
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As a conclusion, long period of incubation doesnt showing the positive result all
the time. The concentration of cordycepins shown us a slightly increase trend in the first two
hours. Unfortunately, the trend doesnt hold at the 3 hours of incubation. I belive that the
long period of incubation could not increase the concentration of cordycepins but only
degradation after long period expose to heat. For this reason, the cordycepins amount
decrease as the time increase. Consequently, the best incubation time I would suggest is
around 1-2 hours.
As I stated before, the concentration method is need to concentrate the extract in to
our minimum requirement, 300ppm. So, I had chosen 2 methods in this experiment to
concentrate our extract. I planned to use water bath and direct heating to concentrate the
extract. Before the experiment, the formula M1V1=M2V2 are using to calculate out the
expected concentration. The apparatus are showed as the picture below:-
Time ,t (hr) Concentration of cordycepin
obtained (ppm)
Percentage of water
remained,%
1 90.50 63.63
2 94.97 63.63
3 88.94 45.45
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The detail procedure is written in the report attach to the appendix. And the results are tabulated
in the table below:-
As the conclusion, water bath is a better concentration method as it can remain the quantity
as well as the quality of the extract after the concentrate process. But water bath is not consider as
an efficient method as it using a very long period to evaporate only 40 ml of water in this
experiment. Hence, it is not so practical in the field of industrial as compared with direct heating
method.
Since the releasing rate of the cordycepins from the substrate has not achieve the
maximum concentration we predict. Thus, I try to find a way to increase the releasing of
cordycepins in the extract. I try to change the form or state of the substrate through pre-treatment.
Method Time taken Final volume
(ml)
Expected concentration of
cordycepins (ppm)
Actual concentration of
cordycepin (ppm)
Water Bath 2hrs40mins 7 646.43 655.00Direct Heating 38minutes 10 452.50 365.12
Direct Heating Water Bath
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I compared the concentration of autoclaved substrate and powder form of the substrate with
normal substrate. The chemical results are as shown below:-
Type of pre-treatment
Cordycepin being tested,
(ppm)
autoclaving 576.67
powdery 762.37
Without pre-treatment 657.63
It is not surprise that the powder form substrate give us the highest concentration of
cordycepins, as the size of the substrate decrease, the surface area increase, hence, releasing rate
of cordycepins increase. But the result of the autoclaved substrate showing us that the high
temperature during the autoclaving may broke down the structure of cordycepins in the substrate
as well as in the extract. So, the use of autoclaved machine should lesser in order to maintain the
quantity of the cordycepins.
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LEARNING OUTCOME
Different type of skill and knowledge I had applied and gained during the internship. The
basic knowledge that taught in our degree and also foundation helps me to perform well in the
working environment. Besides, those basic also help me to understand all the theories that are
new to me such like the HPLC, AAS and also microbiology theory and techniques that still not
yet learn in our syllabus.
MICROBIOLOGY LAB
I learn quite a lot of microbiology techniques in the microbiology. The first technique I
learn in microbiologist lab is how to serial dilution the sample product. Usually we will add one
ml of original sample are taken and add it to 90 ml of Tryptic Soy Broth (TSB), to give 10-1
dilution of original sample are taken and add it to 9ml of TSB. Similarly we can prepare 1:100,
1:1000, 1:10000 and so on to dilution of the original. Finally, one ml of any dilution is added to
sterile petri dish to which are added with sterile, cool, molten agar medium. The dishes are then
incubating at a suitable temperature within some period. After the incubation, colonies of each
microbe grow in the dish. The number of colonies of each kind is counted. The number is then
multiplied by the dilution factor to find the total number of cells per ml of the original sample.
Usually we will use the formula below to count:-
CFU (colony formed per unit)/ ml or g =
Besides, I also learn about different method of pure culture that usually use in routine
laboratory work. Before learning about the pure culture method, I learned about how to prepare
and store different type of medium and agar such like TSB, Sabouraud Dextrose Agar (SDA),
Baird Parker (BP) Agar and Levine Eosin Methylene Blue (LEMB) agar, and also the uses of
those medium and agar. Pour plate method and streak plate are usually use in our company. In
pour plate method, the sample is diluted in to a broth. The agar medium is maintained in molten
Colonies counted (average) Dilution Factor
actual volume of sample in dish, ml
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state at 45 C. One ml of each dilution is added to each sterile Petri dish to which is then poured
9ml of sterile, cool and molten agar medium. The contents are thoroughly mixed and allowed to
solidify. The dish is then incubating in a suitable temperature. In streak plate method, a small
amount of a sample is transferred into the surface of a suitable, solid agar medium either by loop
or transfer needle. This is then streaked in such a way as to provide successive dilution and
ultimately to have well isolated colonies. Streaking may be done in any do the ways. The sample
in each case will becomes progressively diluted and at the end of streak one would expect the
well isolated colonies. The result is of streak plate I did is shown below:-
Staphylococcus on Baird- Parker agar
Black shinny colony formed.
(Merck, 2005)
E.Coli on LEMB
Metallic shinny sheen in reflected light
(Merck, 2005)
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I also learn about carry out conformational test. We can confirm the presences of
staphylococcus aureus from coagulase-negative staphylococci by using Coagulase. By the way, I
also learn how to differentiate Grams Positive bacterial and Grams Negative Bacterial. Before
carry out the Gram stain, I try to prepare Bacterial Smears (Cappuccino&Sherman, 2004). The
procedure is basically as shown in the flow chart below:-
Then, the smear is flood with crystal violet. Wash it off after I minute by using tap water.
The smear is now purple-blue in color. Then, stain it with Grams Iodine for another 1 minutes.
The Grams Iodine is then washing off after 1 minute. The smear is then added with Decolorize
agent (95% ethyl alcohol) until showing only blue tinge. Gently wash the decolorized smear with
tap water and counterstain with safranin for 45 seconds. Lastly wash the smear with tap water and
blot dry with bibulous paper before examine under a microscope. Grams positive cell such likestaphylococcus have thick peptidoglycan layer, whereas the peptidoglycan layer in Grams
negative organism is much thinner and surrounded by outer lipid-containing layers. Grams
positive bacterial will shown blue color when we exam it under microscope. Grams negative
bacterial will counterstained by red safarnin, thus, pink color showed under a microscope.
Prepare a clean
glass slide
Drop 2 loopfuls of water on the
center of the glass slide and dilute
the culture placed.
Transfer the
culture to the
slide
Air dries for few minutesHeat Fixation
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CHEMICAL LAB
I applied the thing I learned in school life in chemical lab. I know how to prepare solution
from their stock solution. I know how to dilute the solution in to the concentration we need by
using volumetric flask.
Besides that, I also learned about the new equipment such like AAS, HPLC and also
Microwave digester in the lab. These 3 equipments play an important role in our chemical lab.
The learn how to operate the microwave digester and also how to clean it after use to maintain its
performances. The Microwave digested always deal with concentrate acid, must be very careful
when handling it.
Our company uses AAS (Atomic absorbance spectrometer) determine the concentration
of heavy meter found in the final product manufactured by our company to ensure that all of the
products are safe to be consume. AAS is an equipment special designed for determine the
concentration of different heavy metal in a sample. The presence of Cadmium, mercury,lead and
arsenic are determined in the product. Usually I will prepare the different concentration of
standard solutions which will use to calibrate the standard curve. The sample is only allowed to
enter the system for AAS test after the standard graph is plotted.
High- Performance Liquid Chromatography (HPLC), is widely use to separate and
determine species in a variety of organic, Inorganic and also biological material. (Leong, n.d) We
usually use it o determine the concentration of cordycepins and adenosine in our product as well
as the cordyceps we cultured. Varieties of detectors are found in the market. But BioFact Life
only uses UV detector and refraction detector in their daily routine works. I am so glad that I have
an experience on how to operate HPLC.
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CONCLUSION
As a conclusion, industrial training has introduced me the real working environment I
will face in the future. I have gained a lot of knowledge which I might not have an opportunity to
learn in book and class. For example, I learned how to communicate with people and how toadapt with the company environment and culture once I step out the University. I learned
different types of new skill which are very useful not only in my studies but also in the future
when I work. For example, have a chance to operate the equipment in chemical lab such like
AAS and HPLC give me advance knowledge. Through the industrial training, I have developed
new skill and also polished the skills that I am weak at.
In the twelve week of industrial training, I made new friends. I learn how to interact with
new people and how to cooperate with my colleague in the working area. From time to time, I
saw my weaknesses in communication with other and also how to express myself. My supervisor
and all the colleagues in the company have given me pieces of useful advices for me to improve
myself. Such like my communicating skill and also presentation skill as I am very shy when
needed to talk in front of new people. I will try my best to improve all the weaknesses.
I believe that the unforgettable experience I gained in BioFact Life has prepared me well
for my employment. Industrial training also helps me to identify my future career. I become more
confidence and skillful after the industrial training. For me, working in laboratory is so
challenging and interesting. In my opinion, gain knowledge through the industrial training is
more interesting compare to University learning environment. I hope that I will have an
opportunity to having this kind of experience in the time remaining at UTAR.
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REFERENCES
1. Leong,L.K.(n.d).Chapter 3: HPLC[powerpoint slides].Unpublished manuscript, UESC
3254, University Of Tunku Abdul Rahman,Malaysia
2. John,C.H.,Phillip,C.,Megan,L.P.,Dinesh,P.(2004).Analysis of Quality And Techniques
For Hybridization Of Medicinal Fungus Cordyceps Sinesis (Berk.)Sacc.(Ascomycetes),1-
2.
3. J.G.Cappccino.,N.Sherman.,(2004)Microbiology A Laboratory Mannual(7th ed.) . San
Francisco: Benjamin Cummings.
4. Microbiology Manual (12
th
ed.).Germany: Merck.
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