MM™
TABLE OF CONTENTS
ACKNOWLEDGEMENTS 1
CHAPTER 1: INTRODUCTION 2
CHAPTER 4: ACTIVITIES 15
4.1 Chemistry Laboratory 15
4.2 Microbiology Laboratory 27
4.3 Grading Laboratory 45
4.4 Food Processing Laboratory 46
CHAPTER 6: CONCLUSION 52
EXTRAS 53
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ACKNOWLEDGEMENTS
I would like to express my gratitude to all those who gave me the possibility to complete
Zaharahtul Hidayah Mohd Yusop, for her invaluable and stimulating suggestions and
encouragement, which helped me in all the time of projects and for the writing of this
report.
namely Madam Norfazlina Abdul Wahab, Madam Noor Fadzilah Abdul Rashid, Mr.
Mohammad Safuan Abdul Rahim, and Miss Nur Hidayah Zulkhairi. Without their help, this
Downstreaming Division, Madam Linda Buang and my Academic Supervisor for their advice,
Finally yet importantly, I would like to express my gratitude for the love and
1
CHAPTER 1
INTRODUCTION
Industrial training forms part of the university requirements where students are exposed to
experiencing the real world of work in organization. The idea of having students to undergo
a short attachment programme with organization is actually to assist them for employability
once they graduated from UNiSEL. The industrial training programme is also aimed at
grooming students and ensuring that they meet the industries’ requirements for
programme for student is also intended to develop linkages with industries and this is part
The early exposure of real work life situation is deemed necessary as this prepares
students not only acquiring the theoretical aspects of the knowledge, but far more
important, is how this knowledge can be applied practically in real workplace settings.
ii. To expose students to the latest technology and management utilized by industry.
2
iii. To enable students to gain experience and exposure to the reality of working
environment in organization.
v. To develop students with the necessary technical and professional skills including
vi. To expose students to work related problems and issues and in a way develop their
vii. To enable students to apply the theories and concepts to real workplace situation.
guarantee of safety either in processed form, semi-process, or fresh form. It covers the
Quality Control Lab, activity which has existed since 1957 with the establishment of a
Grading Lab, followed by a Microbiology Lab in 1992 which is located in the MPIB old
premises (Syarikat Bandar Baru UDA or SBBU Building). In 1997, a Chemical Lab was
established in-line with the addition of activity scope which is the development and
service, the Quality Control Lab has successfully obtained MS ISO 1900:2000 certifications
for 3 years in a row (1999 to 2001). Today, the Quality Control Lab is also in the process to
obtain the MS ISO 17025:2005 accreditation certification to improve client’s trust towards
service quality.
3
Vision
Mission
• To improve focused research and development activities as well as
improving product image and quality in heading towards industry
excellence.
Motto
• “Komitmen Asas Kualiti” (Commitment is the Foundation of Quality)
Quality Policy
• Committed to provide verification inspection services for Malaysian
pineapple industry products which fulfills the requirement of international
standards
Corporate Objective
• Develop a service which fulfills the requirement of legislation,
specification and prescribed period.
General Objective
• Provide client-friendly service with a continuous improvement of service
quality in achieving zero-level complaint status.
In ensuring that quality policy goal is achieved, the Product Development and
Downstreaming Division have established a quality management system which can assist in
4
v. To improve the capability of service through continuous training for personnel and
iii. To prepare chemical, physical and microbiology service analysis to control and
determine specification, proximate, nutrition factor and food addictive detection for
iv. To improve lab analysis technology, suitable with the evolution of science and
technology.
v. Responsible to conduct study towards chemical contents in the industry product and
conduct research so that the ingredients in it can be used as an added value for
commercial usage.
vi. Responsible towards new studies and findings to ensure that pineapple is not only
sold fresh, in cans, or taboo-based industry only, but can also be result-generated
through biotechnology.
5
CHAPTER 2
eastern South America. It was introduced in Malaya in the 16th century by the Portuguese.
The canning industry has been around for more than 100 years. Commercial
pineapple canning was started by Singaporean Chinese community. Before the rise of other
commodities, the pineapple industry was the main contributor of the country's economy. In
1957, Lembaga Perusahaan Nanas Tanah Melayu (LPNTM) was established under the 1957
(LPNM)/Malaysian Pineapple Industry Board which carries the role to manage and develop
Pineapple canning industry was the second largest tropical fruits export after
watermelon. It has been and continued to contribute to country’s economy and provides
direct job opportunities in the plantation and process ing industry and indirectly in the
carry out pineapple industry research and development, Malaysia is capable of producing
6
Malaysian pineapple industry is supported by the government in aspect of land
development policy. The smallholder and farm sector is given priority by the government
A few strategies have been arranged based on long term planning that was
organized. LPNM/MPIB focus is to open new land and increase productivity and to handle
Global globalization and world economy integrations mean rivals will become the
main element in deciding the success and longevity of Malaysian pineapple industry.
marketing and trading as well as organizing strategy based on policy that has been set by
the government to ensure that the arrangement is more sustainable and effective.
Malaysia is one of the world major producers of pineapple other than Thailand,
Philippines, Indonesia, Hawaii (USA), Ivory Coast, Kenya, Brazil, Taiwan, Australia, India and
South Africa. Canned pineapple fruit have high market demand in countries like Japan,
7
2.2: History of MPIB
Ministry of Primary Industries, formed under the Pineapple Industry Act 1957 which was
previously known as Lembaga Perusahaan Nanas Tanah Melayu. In keeping with the
development of the pineapple industry, the agency name was altered to Lembaga
Perindustrian Nanas Malaysia/Malaysian Pineapple Industry Board in 1992 (Act 105- Revised
On May 29th 1996, the Cabinet made a decision to disband the Pineapple Board, and
its functions were transferred to the Department of Agriculture, FAMA and MARDI. The
The Pineapple Board was placed under the Ministry of Agriculture on 1st June 1999
to complete the disbanding implementation. The Ministry of Agriculture was given the role
8
distribution, staff, property, liability, authority and Pineapple Board legislation among
involved agencies. Fifty six staff from the Pineapple Board Development Section was
transferred to Johor Pineapple Industry Development Section. Eight staff from the Pineapple
Board Administrative Division was transferred to Johor Civil Service and 14 staff from the
Inspector Section, Planning and Marketing Development Section was transferred to FAMA
office, Johor branch. As many as 22 staff remained in the Pineapple Board to carry out
However, on 14th June 2000, the Cabinet has agreed to revise the disbanding order.
Therefore, on 15th September 2004, the ministers has come to a conclusion that the
Pineapple Board were to remain as the leader in pineapple industry development and the
disbanding decision on the Pineapple Board was called off on 29th May 1996. Apart from
that, movable and unmovable property that belonged to the Pineapple Board and all the
managing Malaysian pineapple industry. In a long term, MPIB strives to be the global leader
globalize the market by offering investment opportunities further increasing the standard of
9
2.4: Organization Chart
MANAGING
DIRECTOR
INFORMATION
TECHNOLOGY UNIT
CORPORATE EXPANSION
MPIB BRANCH
QUALITY CONTROL/ PLANNING
OFFICES
FACTORY
PINEAPPLE
FINANCE DEVELOPMENT
FOOD STATIONS
TECHNOLOGY
PUBLIC RELATION UNIT FERTILIZERS
LICENCING & SUBSIDISATION
FRUIT
SEPARATION PINEAPPLE
ADMINISTRATION
/SECRETARIAT PLANTING
ADVANCEMENT
10
CHAPTER 3
GENERAL OPERATION
21week 1 22 23 24 25
report to MPIB; microbial analysis of field trip to Josaphine preparing media: PDA,
introduction to the samples from Salija & model farm at Nilai, BCA, TPC; preparing
department & labs; Lee; clean-up lab Negeri Sembilan diluent: Ringer
review instrument’s apparatus
manual, analysis
methods & previous
internship reports
28 week 2 29 30 31 01january
microbial analysis of microbial analysis of microbial analysis of moisture & ash content grading analysis of
samples from Al samples from Al samples from Lee determination of various pineapple
Barkah; preparing Barkah; preparing samples from Al canned products
lauryl sulphate broth & lauryl sulphate broth & Barkah, Mastuti &
dextrose tryptone dextrose tryptone Salija; preparing media:
broth broth; preparing 70% PDA,BCA, TPC
alcohol solution
11
04 week 3 05 06 07 08
microbial analysis of preparing reports microbial analysis of preparing media: PDA, preparing diluent:
samples from Lee; samples made by MPIB; BCA, TPC; preparing Ringer; recording
preparing diluent: preparing reports reports microbial analysis
Ringer results
11 week 4 12 13 14 15
microbial analysis of microbial analysis of microbial analysis of preparing reports microbial analysis of
samples made by MPIB samples made by MPIB; samples made by MPIB; samples made by MPIB;
preparing media: PDA, preparing media: PDA, recording results;
BCA, TPC BCA; recording results clean-up lab apparatus
18 week 5 19 20 21 22
microbial analysis of microbial analysis of microbial analysis of preparing media: PDA, microbial analysis of
samples from Lee; samples from Lee; samples from Lee; BCA, TPC; preparing samples from Lee;
preparing media: PDA, diluent: Ringer; recording results;
BCA, TPC recording results clean-up lab
25 week 6 26 27 28 29
recording results; preparing 0.5M NaOH; preparing reports; preparing reports; preparing reports
preparing media: TPC, learn how to use clean-up lab; making making pineapple jams
PDA; preparing reports Petrifilm™; recording pineapple cookies
results; preparing
reports
12
01 week 7 02 03 04 05
field trip to Lee grading analysis of microbial analysis of grading analysis of microbial analysis of
Pineapple Co Pte Ltd various pineapple samples from Shamu & various pineapple samples from Shamu;
factory; acidity analysis canned products; QC; preparing media: canned products; preparing reports;
of various pineapple preparing reports PDA, BCA, TPC preparing reports clean-up lab
canned products
08 week 8 09 10 11 12
attending seminar microbial analysis of preparing lauryl participate in Method recording results;
Method Validation in samples from Ayam sulphate broth; clean- Verification of preparing reports
Analytical Chemistry; Brand & Cheung; up lab apparatus Microbiological
recording results; preparing media: PDA, Methods
clean-up lab BCA; clean-up lab
15 week 9 16 17 18 19
preparing reports preparing reports; microbial analysis of
preparing media: PDA, samples from Ctron;
BCA, TPC; clean-up lab clean-up lab
apparatus
22 week 10 23 24 25 26
microbial analysis of microbial analysis of preparing media: BCA, recording results
samples from Josa; samples from Salija; PDA, TPC; preparing
preparing media: PDA, recording results; diluent: Ringer;
BCA, TPC; preparing clean-up lab recording results
diluent: Ringer
13
01 week 11 02 03 04 05
preparing media; microbial analysis of microbial analysis of preparing reports
preparing lauryl samples from Al samples from Aza Pine;
sulphate broth; Rauzah; clean-up lab preparing media; clean-
recording results up lab
08 week 12 09 10 11 12
preparing reports microbial analysis of microbial analysis of preparing media; clean- microbial analysis of
samples from Al Rauzah samples from Al Rauzah up lab samples from Maqh
Halwa & SHM Global
15 week 13 16 17 18 19
microbial analysis of microbial analysis of preparing media; preparing media; preparing reports;
samples from Maqh; samples from Maqh; preparing reports preparing reports; recording results
clean up lab recording results
22 week 14 23 24 25 26
microbial analysis of microbial analysis of microbial analysis of preparing media preparing reports;
samples from Maqh samples from Aza Pine samples from Aza Pine clean-up lab
29 week 15 30 31 01&02april
preparing reports preparing reports preparing reports preparing and submitting the reports
14
CHAPTER 4
ACTIVITIES
Introduction
Moisture content is one of the most commonly measured properties of food materials. It is
important to food scientists for a number of different reasons:
Legal and Labeling Requirements. There are legal limits to the maximum or minimum
amount of water that must be present in certain types of food.
Economic. The cost of many foods depends on the amount of water they contain -
water is an inexpensive ingredient, and manufacturers often try to incorporate as much
as possible in a food, without exceeding some maximum legal requirement.
Microbial Stability. The propensity of microorganisms to grow in foods depends on their
water content. For this reason many foods are dried below some critical moisture
content.
Food Quality. The texture, taste, appearance and stability of foods depend on the
amount of water they contain.
Food Processing Operations. Knowledge of the moisture content is often necessary to
predict the behaviour of foods during processing, e.g. mixing, drying, flow through a
pipe or packaging.
MC in this experiment was expressed as a percentage of the wet weight of the material.
Objective
To determine MC in processed pineapple products
Materials
Processed pineapple products: Sos Nanas Salija, Sos Nanas Pedas Al Barkah Enterprise, and
Kordial Nanas Mastuti; porcelain dish; desiccator; digital weighting machine; oven.
Methodology
Three porcelain dishes were dried in oven at 105°C overnight. The next day, before weight
was measured, the dishes were put in desiccator (Figure 4.1) for several minutes to ensure it
attains room temperature. The dishes were weight several times for every 3 hours until
consistent readings were obtained. The following amount of samples was inserted into
three porcelain dished respectively: 10g of Sos Nanas Salija, 10g of Sos Nanas Pedas Al
Barkah Enterprise, and 5g Kordial Nanas Mastuti (Figure 4.2). Then the dishes were put into
the oven at 105°C overnight. The next day, the dishes filled with samples were weight
several times for every 3 hours until consistent readings were obtained. Results recorded.
16
Figure 4.1: Dishes were dried in the desiccator
17
Results and Discussion
Kordial Nanas
22.58 10.14 28.43 5.85 57.69
Mastuti
Level of moisture content in Kordial Nanas Mastuti is high due to its nature that is liquid and
intrinsically a concentrated pineapple juice. Sauce (Sos Nanas Salija and Sos Nanas Pedas Al
Barkah) in the other hand, is semi-solid - partly due to boiling or high-pressure treatment
during production, where some of water content was removed.
Food Act 1983 (amended 1985) and Food Regulation 1985 does not specify acceptable level
of moisture content in pineapple sauces and cordials. Oven drying process was considered
standard in industry and has the accuracy of a few tenth of one percent (Kraszewski, 1991).
Conclusion
Objective of this experiment was reached.
Reference
Kraszewski, A.W. 1991. Microwave Aquametry – Needs and Perspectives. IEEE Transactions
of Microwave Theory and Techniques. 39(5): 828-835.
18
4.1.2 Determination of Ash in Processed Pineapple Products
Introduction
The ash of foodstuff is the organic residue (such as Ca, Na, Cl and K) remaining after the
organic matter has been burnt away. Hence, ash content can be determined by incinerating
a known quantity of foodstuff, previously dried until constant weight is obtained. Ashing
should be done at temperature exceeding 550°C, at which inorganic salt like alkali chlorides
will volatilize. Moreover, a portion of the ash will fuse and enclose some carbon, preventing
them from being ignited. Continuous heating of sample ensures final and constant weight
was obtained.
Analytical techniques for providing information about the total mineral content are based
on the fact that the minerals (the “analyte”) can be distinguished from all the other
components (the “matrix”) within a food in some measurable way. The most widely used
methods are based on the fact that minerals are not destroyed by heating, and that they
have a low volatility compared to other food components. The three main types of
analytical procedure used to determine the ash content of foods are based on this principle:
dry ashing, wet ashing and low temperature plasma dry ashing. The method chosen for a
particular analysis depends on the reason for carrying out the analys is, the type of food
analyzed and the equipment available. Ashing may also be used as the first step in preparing
samples for analysis of specific minerals, by atomic spectroscopy or the various traditional
methods described below. Ash contents of fresh foods rarely exceed 5%, although some
processed foods can have ash contents as high as 12%, e.g., dried beef.
Objective
To obtain ash content in processed pineapple products
Materials
Processed pineapple products: Sos Nanas Salija, Sos Nanas Pedas Al Barkah Enterprise and
Kordial Nanas Mastuti; porcelain dish; desiccator; digital weighting machine; oven, Bunsen
burner.
19
Methodology
Three porcelain dishes were dried in oven at 105°C overnight. The next day, before weight
was measured, the dishes were put in desiccator for several minutes to ensure it attains
room temperature. The dishes were weight several times for every 3 hours until consistent
readings were obtained. The following amount of samples was inserted into three porcelain
dished respectively: 10g of Sos Nanas Salija, 10g of Sos Nanas Pedas Al Barkah Enterprise,
and 5g Kordial Nanas Mastuti. Samples were gently heated over the Bunsen burner until it
stops fuming. The samples were then transferred into muffle furnace at 550°C until greyish
or whitish ashes were obtained. The dishes were cooled in the desiccator and weighted
soon after it had attained room temperature. The samples were weighted several times
until constant reading was obtained. Results recorded.
Kordial Nanas
20.95 5.02 20.96 0.01 0.20
Mastuti
Dry ashing method was used in this experiment. Ash content varies on different products;
mainly influenced by the mineral contents of the fresh fruits used and the manufacturing
processes. Food Act 1983 (amended 1985) and Food Regulation 1985 does not specify
acceptable level of ash content in pineapple sauces and cordials. Due to its nature that are
semi solid, ash content of the sauce will be higher than cordials and juices.
20
Research by Camara et al. (1994) indicated ash content of fresh pineapple juices is between
0.163% and 0.414%. Fresh pineapple juices are high with K (average 96.68mg/100mL), Ca
(14.59mg/100mL) and Mg (13.36mg/100mL). Other minerals traced in pineapple juices are
Na, P, Fe, Mn and Zn. However, amount of these minerals as well as the ash content are
slightly lower in processed pineapple products including sauces and cordials.
Conclusion
Objective of this experiment was reached.
Reference
Camara, M., Diez, C., and Torija, E. 1994. Chemical Characterization of Pineapple Juices and
Nectars: Principal Component Analysis. Food Chemistry. 54(1): 93-100.
21
4.1.3 Determining pH Value using Universal pH Indicator
Introduction
Universal pH indicator (Figure 4.3) is a pH indicator that ranges from the number 1-14 (on
the pH chart); to indicate the acidity or basicity of solutions. A universal indicator typically
composed of water, methanol, propanol, phenolphthalein sodium salt, methyl red,
bromothymol blue monosodium salt and thymol blue monosodium salt.
The colours that indicate the pH of a solution, after adding a universal indicator are:
Wide range of pH test papers with distinct colours for each pH from 1 to 14 is available.
Colour matching charts are supplied with the specific test strips. In this experiment pH
indicator strips in Figure A are used.
22
Materials
Pineapple processed products (sauces, jams, cordial and canned products) and/or fresh
pineapple; pH indicator strips.
Methodology
Approximately 5g of processed pineapple products were taken. For fresh pineapple, it was
first made into juices and approximately 10mL of juice will be used. Products were stirred
well and the pH indicator strips are then dipped and left for a minute. The colour changes in
the pH indicator strips are compared with the colour matching indicator provided to
determine the pH value of the sample. The readings are repeated three times and average
are calculated and recorded down. Aseptic measures applied throughout the process.
Reference
http://en.wikipedia.org/wiki/Universal_indicator (170210)
23
4.1.4 Determination of Acidity Level Using Titration Method
Introduction
Titration is a common laboratory method of quantitative chemical analysis that is used to
determine the unknown concentration of a known reactant. Becaus e volume measurements
play a key role in titration, it is also known as volumetric analysis. A reagent, called the
titrant or titrator, of a known concentration (a standard solution) and volume is used to
react with a solution of the analyte or titrant, whose concentration is not known. Using a
calibrated burette to add the titrant, it is possible to determine the exact amount that has
been consumed when the endpoint is reached. The endpoint is the point at which the
titration is complete, as determined by an indicator This is ideally the same volume as the
equivalence point the volume of added titrant at which the number of moles of titrant is
equal to the number of moles of analyte, or some multiple thereof (as in polyprotic acids). In
the classic strong acid-strong base titration, the endpoint of a titration is the point at which
the pH of the reactant is just about equal to 7, and often when the solution permanently
changes colour due to an indicator. There are however many different types of titrations
Many methods can be used to indicate the endpoint of a reaction; titrations often use visual
indicators (the reactant mixture changes colour). In this experiment, the simple acid-base
titrations a pH indicator are used, such as phenolphthalein, which becomes pink when a
certain pH (about 8.2) is reached or exceeded. Another example of indicator that can be
used is methyl orange, which is red in acids and yellow in alkali solutions.
Objective
To determine acidity level of processed pineapple products or fresh pineapple using
titration method
Materials
Burette; retort stand; conical flask and measuring cylinder; sodium hydroxide (NaOH) 0.1M
(prepared prior of this experiment); distilled water and processed pineapple products
(sauces, jams, cordials, marmalade and canned products) or fresh pineapple.
24
Methodology
0.1M of sodium hydroxide are prepared prior of this experiment. For cordials and canned
pineapple products (only the syrup are taken), 10mL of samples were taken while for others,
10g. Taken samples were transferred into 250ml conical flask. Two or three drops of
phenolphthalein were added into the conical flask. With constant agitation, conical flask is
titrated with 0.1M sodium hydroxide till the colour of the solution turns into pink. Results
recorded.
25
NaOH NaOH NaOH Calculation
Sample/Code Average
Initial (mL) Final (mL) Usage (mL) (%)
0.0 16.0 16.0 0.17
Sauce A
16.0 33.2 17.2 0.18 0.17%
QC/IKS20/10
33.2 47.0 14.0 0.15
47.0 59.4 12.4 0.13
Sauce B
59.4 72.6 13.4 0.14 0.13%
QC/IKS21/10
0.0 12.6 12.6 0.13
12.6 22.4 9.8 0.10
Jam A
22.4 33.6 11.2 0.12 0.11%
QC/IKS124/10
33.6 44.2 10.6 0.11
44.2 54.6 10.4 0.11
Jam B
54.6 65.8 11.2 0.12 0.11%
QC/IKS125/10
65.8 76.4 10.6 0.11
Pineapple sauces are more acidic compared to pineapple jams. The acidity of pineapple
sauces is not due to the presence of microbes or free organic acids but to vinegar or other
weak acids (as preservatives) typically added as the ingredients of sauces. High
sugar/sorbitol content lowers the acidity of the jams.
Conclusion
Objective of this experiment was reached.
Reference
Ranken, M.D. and Kill, R.C. 1997. Food Industries Manual. Blackie Academic and
Professional, London, UK.
26
4.2 MICROBIOLOGY LABORATORY
Food safety involves safeguarding food from anything that could harm the health of
consumers. While high standards enables everyone to enjoy their food without illness,
injury or other problems, poor standards can led to all kinds of harms and even death. As
food safety is so important to everyone, food handler has legal obligations for keeping food
safe to eat.
Most cases of food poisoning resulted from presence of large number of pathogenic
bacteria that are living on the food. Other causes of food poisoning including viruses, mould
and yeast, poisonous plant and chemicals or metals.
There are two types of illness linked to food: food poisoning and food-borne disease. Food
poisoning is caused by consuming food contaminated by harmful substances or by harmful
bacteria that are living on the food, while a food-borne disease is caused by consuming food
or water that carries harmful microorganism.
The incidence of spoilage in canned foods is very low, but when it does occur, it is important
to proceed with the investigation. Abnormalities on can/container usually indicate an
abnormal product. During progressive spoilage, shape of the container may progress from
normal to flipper, to springer, to soft or to hard swell. But it is noteworthy that buckling or
denting, closing while cool, overfilling, or prolonged storage may also cause flipper or
springer.
Microbial spoilage or hydrogen from reaction of acids with the metals of the cans may
produce flipper, springer or swell. Summer temperature and high altitudes accentuate the
27
degree of swelling. Not all microorganisms that grow in canned foods causing can
abnormality.
Spoilage within the can may also result from leakage or under-processing. Leakage occurs
from cans defects, punctures or rough handling. Contaminated cooling water sometimes
enters the interior through pinholes or poor seams. A viable mixed flora or rods and cocci
are indicative of leakage and can be determined by can examination. Under-processing may
occur because of deliberate undercooking to preserve a “fresher product”, fatly retort
operations, excessive contamination for which normally adequate process are insufficient or
accidental by passing retort operations. When the can contains a spoiled product and no vial
microorganism, spoilage may have occurred before processing or the microorganism may
have died during storage.
The parameters for the analysis and simple method to run the microbiological analysis are
as follows:
Microbiological
Purpose Media Organism
analysis
Total bacteria General viable and Standard plate All bacteria and
count/total plate cultivation of count agar (TPC) microorganism
count microorganisms
Moulds and yeasts Cultivation and Potato dextrose Moulds and yeasts
counting enumeration of agar (PDA)
moulds and yeasts
Flat sour bacteria For the detection of Dextrose tryptone Flat sour bacteria
flat-sour broth
thermophiles and
mesophiles in food
products
28
The following specification for microbiological analysis is based on Malaysian Food
Regulation 1985 and Official Method of Analysis AOAC International 17 th edition:
Minimum limit
Analysis parameter Canned Pineapple/ Jam/Marmalade/
Pineapple Juice Pineapple Sauce
Enumeration of
coliform (MPN method, <10 cfu/100 ml sample <10 cfu/100 ml sample
LST: 37°C/48H)
Detection of E.coli
(MPN method, BGLB: nil nil
44°C/48H)
Enumeration of
Bacillus cereus < 1.0 x 10-1 cfu/g sample < 1.0 x 10-1 cfu/g sample
(BCA: 37°C/48H)
29
4.2.1 Preparation of Microbial Culture Media
Introduction
Bacillus cereus Selective Agar developed by R. Holbrook and J.M. Anderson for the isolation
and enumeration of Bacillus cereus in foods. It meets the requirements for a medium that is
sufficiently selective to be able to detect small numbers of Bacillus cereus cells and spores in
the presence of large numbers of other food contaminants. The medium is also sufficiently
diagnostic that colonies of Bacillus cereus are readily identified and confirmed by
microscopic examination.
Potato Dextrose Agar (PDA) is recommended by the APHA for plate counts of yeasts and
moulds in the examination of foods and dairy products. It is also used for the stimula tion of
sporulation (slide preparations), maintenance of stock cultures of certain dermatophytes
and for differentiation of atypical varieties of dermatophytes by pigment production. PDA is
a fungal media (common organisms that can be cultured on PDA are yeasts such as Candida
albicans and Saccharomyces cerevisiae and moulds such as Aspergillus niger) and does not
contain agar.
Plate Count Agar (PCA) also known as Total Plate Count (TPC); is made according to the
American Public Health Association (APHA) formulation. The composition of plate count
agar may vary, but typically it contains 0.5% peptone, 0.25% yeast extract, 0.1% glucose,
1.5% agar; pH adjusted to neutral at 25°C. They are recommended to assess or to monitor
"total" or viable bacterial growth of a sample and may also be used to determine the
sanitary quality of foods, water and other materials. PCA is not a selective medium. Plate
count broth does not contain agar.
Lauryl Sulphate Broth, which is also known as Lauryl Sulfate Tryptose Broth, is used for the
detection of coliform organisms in materials of sanitary importance. Peptone (the main
component) provides essential growth substances, such as nitrogen and carbon compounds,
sulfur and trace ingredients. The potassium phosphates provide buffering capacity. Sodium
chloride maintains osmotic equilibrium. Lactose provides a source of fermentable
30
carbohydrate for coliform organisms. The fermentation of lactose with gas formation is a
presumptive test for coliforms.
Brilliant Green Bile Broth (or Brilliant Green Lactose Bile Broth) is used for the detection of
coliform organisms in foods, dairy products, water and wastewater, as well as in other
materials of sanitary importance. Brilliant Green Bile Broth contains two inhibitors of both
gram-positive and selected gram-negative organisms: i.e. oxgall and brilliant green dye.
Organisms, primarily coliforms, which are resistant to the action of the inhibitors and which
ferment the lactose, are able to replicate in this medium. Fermentation is detected by gas
production. Gas production within 48 ± 3 hours is considered positive evidence of
fermentation by coliform bacilli.
Materials
Plate count agar powder, potato dextrose agar powder, Bacillus cereus selective agar
powder, lauryl sulphate broth powder, Brilliant Green bile broth powder, 1000mL and
600mL beakers, screw cap test tube, Durham’s tube, spatulas, hot plate/magnetic stirrer,
digital weighting machine, sterile distilled water, pH meter, 0.5M NaOH, 0.5M HCL, 1000mL
Schott bottles.
Methodology
Bacillus cereus Selective Agar
47.8g of Bacillus cereus selective agar powder (Figure A) was dissolved in a litre of sterile
distilled water on the hot plate. pH of the solution was adjusted to 7.2 ± 0.2 by adding NaOH
or HCL and was immediately transferred into the Schott bottle to be autoclaved at 121°C for
15 minutes. Prepared medium was stored in 4°C chiller.
31
Plate Count Agar or Total Plate Count
22.5g of plant count agar powder was dissolved in a litre of sterile distilled water on the hot
plate. pH of the solution was adjusted to 7.0 ± 0.2 by adding NaOH or HCL and was
immediately transferred into the Schott bottle to be autoclaved at 121°C for 15 minutes
(Figure C). Prepared medium was stored in 4°C chiller.
32
Figure 4.5 Figure 4.6 Figure 4.7
Powders of microbial culture media were first weighted (BCA shown in Figure 4.5) then
dissolved in sterile distilled water on the hot plate (Figure 4.6 shows PDA). After pH was
adjusted, medium were ready to be transferred (TPC shown in Figure 4.7) into Schott bottles
for autoclaving followed by storage in the chiller.
References
http://www.bd.com/ds/technicalCenter/inserts/Plate_Count_Agar_Standard_Methods_Aga
r.pdf (080110)
http://www.bd.com/ds/technicalCenter/inserts/Potato_Dextrose_Agar.pdf (080110)
http://www.oxoid.com/UK/blue/prod_detail/prod_detail.asp?pr=CM0617&org=9&c=UK&la
ng=EN (080110)
http://www.bd.com/ds/technicalCenter/inserts/Lauryl_Tryptose_Broth.pdf (080110)
33
4.2.2 Preparation of Ringer Solution, Food Homogenate, and Dilution Series
Introduction
Ringer solution is an aqueous solution containing the chlorides of sodium and potassium
and calcium that is isotonic to animal tissues; used to correct dehydration and (in
physiological experiments) as a medium for in vitro preparations. This solution was invented
by Sydney Ringer - she discovered this solution keeps an isolated frog heart beating for a
longer period of time (Beck, 2000). However, according to Frank (1992), it may not optimal
for microbiological purposes. He suggested 0.1% casein peptone (tryptically digested),
0.85% NaCl and pH 0.7 after sterilization.
A serial dilution is simply a series of simple dilutions which amplifies the dilution factor
quickly beginning with a small initial quantity of material (i.e. bacterial culture, a chemical,
orange juice, etc.). The source of dilution material for each step comes from the diluted
material of the previous. In a serial dilution, the total dilution factor at any point is the
product of the individual dilution factors in each step up to it.
Materials
Stomacher, Stomacher bags, digital weighting machine, shaker, magnetic stirrer, pH meter,
pipette (1-10mL), test tube, universal bottles, 250mL Schott bottles, Ringer solution tablets,
alcohol (for swabbing), 0.5M NaOH, 0.5M HCL, and processed pineapple products such as
sauces, cordial, or canned products. Optional: can opener, knife, spatula, and forceps.
34
Methodology
Preparing Diluent Solution - Ringer Solution
Two Ringer tablets were diluted in a litre of sterile distilled water by using magnetic stirrer.
pH was then adjusted to 7.0 ± 0.2 by adding NaOH or HCL. The solution was then distributed
into Schott bottles (225mL per bottle) and universal bottles (9mL per bottle) (Figure 4.8),
before autoclaved at 121°C for 15 minutes. Ringer solution was then stored in 4°C chiller.
All processes were done in laminar airflow and aseptic methods applied.
References
Andrews, W.H., Wilson, C.R., Poelma, P.L., Romero, A., Rude, R.A., Duran, A.P., McClure, F.D.
and Gentile, D.E. 1978. Usefulness of the Stomacher in a Microbiological Regulatory
Laboratory. Applied Environmental Microbiology. 35(1): 89–93.
Frank, H.K. 1992. Dictionary of Food Microbiology. Behr’s Verlag, Hamburg, Germany.
Ito, K. and Downes, F.P. 2001. Compendium of Methods for the Microbiological Examination
of Foods. American Public Health Association, Washington, USA.
http://abacus.bates.edu/~ganderso/biology/resources/dilutions.html (080110)
37
4.2.3 Microbial Identification and Enumeration using Pour Plate Method
Objective
To identify availability and quantity of various microbes in different types of microbial
culture media.
Materials
Micropipette (100-1000µL), consumable petri dishes, Bunsen burner, shaker, incubator 25°C
and incubator 37°C. The following materials were prepared prior of this experiment:
universal bottles filled with Ringer solution; food homogenate of canned pineapple slices
Lee Pineapple Co. Pte. Ltd. - reference number L34/34, L60/60, L96/96, and L101/102;
microbial culture media: BCA, PDA, TPC and lauryl sulphate broth.
Methodology
Consumable petri dishes were first labelled as follows:
L34 P-1 3.
…where L34 represent sample reference number; P-1 referring to medium-dilution series:
P-1 is PDA and Dilution 1 (10-1), P-2 is PDA and Dilution 2 (10-2), and so on (BCA = B; TPC = T);
while 3. loosely refers to experiment date (January 3rd). Similar labelling methods applied on
screw cap test tubes (filled with lauryl sulphate broth) as well. Three dilution series are
made for all four samples tested.
As the serial dilutions of the materials tested are prepared, 1mL volume are transferred into
the petri dishes and mixed with liquefied medium as labelled. Petri dishes filled with TPC
and BCA medium were then incubated in incubator 37°C for 3-4 days while PDA were
incubated for 4-5 days in incubator 25°C.
Colonies are formed both on surface and in the depth of the medium. The count is
calculated from the mean count of a number of plates. For lauryl sulphate broth in screw
38
cap test tubes, medium turbidity and presence of bubbles were observed. All processes
were done in laminar airflow and aseptic methods applied.
Figure 4.10: Petri dishes after several days of incubation. Plate with numerous microbial
growth showed changes in colour
39
Figure 4.11: Colonies formed several days after incubation. Presence of microbes in Dilution
2 (10-2) and Dilution 3 (10-3) was considered insignificant if: a) Dilution 1 (10-1) does not
exhibit any microbial growth, b) number of colonies is larger than the previous dilution series
BCA
PDA
40
TPC
Conclusion
Objective of this experiment was reached.
41
4.2.4 The Use of 3M™ Petrifilm™ in Identification of E. coli/Coliform, Aerobic Microbes
and Yeast and Mould. .
Introduction
The 3M™ Petrifilm™ E. coli/Coliform Count (EC) Plate is a sample-ready-culture-medium
system which contains Violet Red Bile (VRB) nutrients, a cold-water-soluble gelling agent, an
indicator of glucuronidase activity, 5-bromo-4-chloro-3-indolyl-b-D-glucuronide (BCIG) and a
tetrazolium indicator that facilitates colony enumeration. Petrifilm EC plates are used for
the enumeration of E. coli or coliform in the food and beverage industries. Petrifilm EC plate
components are decontaminated though not sterilized.
The 3M™ Petrifilm™ Yeast and Mould (YM) Count Plate is a sample-ready culture medium
system which contains nutrients supplemented with antibiotics, a cold-water-soluble gelling
agent, and an indicator that facilitates yeast and mould enumeration. Petrifilm YM plates
are used for the enumeration of yeast and mould in the food and beverage industries.
Petrifilm YM plate components are decontaminated though not sterilized.
The use of 3M™ Petrifilm™ is convenient especially when fast results are needed and there
are limited spaces in incubators, since the petrifilm is a small and thin layer of paper.
Petrifilm plates are designed to be as accurate as conventional plating methods. However, it
is very expensive.
Materials
3M™ Petrifilm™ E. coli/Coliform Count (EC) Plate, 3M™ Petrifilm™ Aerobic Count (AC) Plate,
The 3M™ Petrifilm™ Yeast and Mould (YM) Count Plate, food homogenate (prepared prior
of this experiment)
42
Methodology
3M™ Petrifilm™ E. coli/Coliform Count (EC) Plate
Petrifilm EC plate was put on a flat, level surface. Top film was lifted and at the same time,
1mL of sample suspension was dispensed onto the centre of bottom film (Figure 4.12). The
top film was then rolled onto the sample to prevent air bubbles. Plastic s preader with the
flat side was placed down on the centre of the plate. It was then pressed gently on the
centre to distribute the sample over the entire plate growth area before the gel is formed.
Several minutes after the gel was formed, plate was incubated in a horizontal position.
Figure 4.12: 1mL of food homogenate was dispensed onto the centre of bottom film.
43
Figure 4.13: Plastic spreader was used to evenly distributing the sample throughout
the petrifilm.
All processes were done in laminar airflow and aseptic methods applied.
Reference
Manuals of 3M™ Petrifilm™ Aerobic Count (AC) Plate, 3M™ Petrifilm™ Yeast and Mould
(YM) Count Plate and 3M™ Petrifilm™ E. coli/Coliform Count (EC) Plate printed by 3M (2008)
http://en.wikipedia.org/wiki/Petrifilm (260110)
44
4.3 GRADING LABORATORY
MPIB accepts 30 canned pineapple products every week from the registered factory. The
sample taken from an inspector from a cannery shall be grouped, examined and tested.
Every canned pineapple is graded continuously start from “Fixed Standard” till “Quality
Standard” until finish based on grade whether it is “Fancy”, “Canadian” and “Choice”.
Grading analysis is done to ensure the quality of canned pineapple product based on “Fixed
Standard” and “Quality Standard” parameter.
45
4.4 FOOD PROCESSING LABORATORY
Introduction
Seven grams of less-sugar jams or fruit spread will have about 8 calories per teaspoon,
compared with the 16 calories of full-sugar jams/spreads (Schmidt, 2003). As an energy
source, sorbitol (sugar replacer in this experiment) was absorbed slowly and incompletely,
and requires little or no insulin for metabolism. That’s why it has special importance for
people suffering from adiposity and pancreatic diabetes.
Sweetness of sorbitol varies from 25 percent to 100 percent as sweet as sugar. They also
aren’t cavity-promoting either. However it costs almost twice the price of sugar (Duyff,
2006)
Pineapple contains little or no pectin so pectin must be added when making jam. Pineapples
contain the enzyme bromelain, which is a proteolytic enzyme that breaks down proteins.
This can cause problems for operators’ hands which are in contact with the juice for long
periods. During cutting operations, gloves should therefore be worn and washed each day.
According to Food Act 1983 and Food Regulation 1985, jams may contain permitted
preservative, colouring, flavouring and food conditioner. Jams must also contain not less
than 35% fruit. Citric acid is not a preservative; it is added to adjust the pH. Jams give a gel
when there is the correct ratio of pectin to water and the pH is between 2.5-3.45. The
optimum pH to give a good gel is pH 3.0.
Objective
To develop less-sugar pineapple jams that maintains natural taste of pineapple and has
favourable attributes as regular jams.
46
Materials
Pineapple cultivar Josaphine, sorbitol, pectin, citric acid, potassium sorbate, purified water.
Other apparatus: jam bottles, pH paper, stove and pot (to sterilize jam bottles), blender.
Methodology
Two formulations of less- sugar pineapple jams were developed:
Formulation A
Pineapple 350.0g (35%)
Sorbitol 530.0mL
Pectin 5.0g
Citric acid 3.5g
Potassium sorbate 2.5g
Water 101.5mL
Formulation B
Pineapple 430.0g (45%)
Sorbitol 530.0mL
Pectin 36.65g
Citric acid 3.5g
Potassium sorbate 0.25g
Water -
Pineapple pieces (in Formulation A), already made into fruit pulp, was slowly boiled with
water for approximately 10 minutes. For Formulation B, pineapple fruit pulp was warmed
for several minutes. With constant agitation, sugar/sorbitol were gradually added and
heating temperature was closely monitored to be around 40-50°C. After all sugar/sorbitol
were added, citric acid (pH was adjusted to 3), pectin and potassium sorbate were added.
The “paste” was continuously stirred for several minutes, and then was let cool for several
minutes before it was transferred into jam bottles.
47
Figure 4.15: Jam of Formulation B
Jams of Formulation B (final weight: 500g/2 jars) are fully solidified (due to higher amount of
pectin used) and light yellow in colour. It is sweeter than Formulation A as higher amount of
pineapple were used and no water added. This also resulted in relatively strong pineapple
flavour and odour which may not good for the shelf life and consumer preferences to the
product.
Conclusion
Objective partially reached: Formulation A produced undesirable product characteristics
while relatively strong pineapple flavour and odour produced by Formulation B may have
impact in its marketability and shelf life. Further research needed.
References
Schmidt, A. 2003. Chef’s Book of Formulas, Yields and Sizes. John Wiley & Sons, New Jersey,
USA.
Duyff, R.L. 2006. American Dietetic Association Complete Food and Nutrition Guide. John
Wiley & Sons, New Jersey, USA.
48
4.4.2 Development of High-Fibre Pineapple Cookies
Introduction
Malaysian eats a lot of refined foods such as white rice and flour. These foods do not
contain much fibre, and so many people suffer from health problems such as constipation.
Pineapple contains fibre. Fibre is needed to help the intestines and bowels work properly.
Eating foods high in fibre, such as vegetables and fruits, gives the body the fibre it needs.
Objective
To develop high-fibre pineapple cookies that maintains natural taste of pineapple, crunchy,
and has favourable attributes as regular pineapple cookies.
Materials
Butter, sugar, salt, egg, pineapple flavouring, flour, baking soda, blended and dried
pineapple pulp (prepared prior of this experiment). Other apparatus: oven, bowl,
mixer/blender.
Methodology
Ingredients were weighted:
Butter, sugar and salt were mixed in the bowl using mixer/blender, followed by egg and
pineapple flavouring. With constant stirring, flour, baking soda and pineapple pulp were
added. Dough produced then shaped into cookie form and were baked in oven at 180-200°C
for 15-20 minutes.
49
Result and Discussion
Figure 4.16: Cookies are golden yellow in colour, sweet and crunchy.
Conclusion
Objective of this experiment was reached.
Reference
http://www.fao.org/wairdocs/x5425e/x5425e0a.htm (040210)
50
CHAPTER 5
51
CHAPTER 6
CONCLUSION
During 15 weeks of industrial training at MPIB, I have learned a lot of things that
prepared me with working experiences such as how microbiology laboratory operates, how
to do chemical analysis and how to grade canned pineapple products based on rules and
regulation in Food Act (1983) and Food Regulation (1985). All analysis were run to ensure
locally-distributed or exported pineapple products are safe and in good condition.
Moreover, I also had given an opportunity to visit the pineapple farm at Nilai and
Malaysia’s biggest canned pineapple manufacturer, Lee Pineapple Co Pte. Ltd. - both
receives advice, consultation and distribution assistance from the MPIB. I’ve also taken part
in mini seminar of method validation in chemistry and microbiology analysis .
I also learned how to work as a team and other practical and real-world situation of
which are not taught in the classroom.
52
Josapine model farm at Nilai, Negeri Sembilan.
53
Lorries were weighed Grader machine Pineapple separated
Lee Pineapple office Grader machine
at the cannery gate viewed from left according to sizes.
54
Fruits are peeled and cored Fruit cutting done by ...filled into ...filled ...pass through steam
using peeling machine machines cans with syrup exhauster
55
Cans are sealed by machine with Sealed cans are pasteurized, Final products should be stored at
the capacity of 60 to 65 cans per cooled and dried in the cooker- 28-30°C for 2 weeks to ensure the
minute cooler dryer unit quality before delivery
56