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Any publication included in this abstract book/ or opinions expressed therein

remain solely those of the author(s). Such publications have been included
only for ease of reference and academic purposes.

Edited by:
Prof. Dr. Teh Lay Kek
Prof. Dr. Mohd Zaki Salleh
Prof. Dr. Jean-Frdric Weber

Published by:
Penerbit Universiti Teknologi MARA (UPENA),
Universiti Teknologi MARA,
40450 Shah Alam,
Selangor Darul Ehsan

Printed by:
Printing Unit,
Faculty of Art & Design,
Universiti Teknologi MARA (UiTM),
40450 Shah Alam, Selangor.

ISBN 978-967-363-331-9
2012 Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy,
Universiti Teknologi MARA, Malaysia.




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Welcome Note

Assalamualaikum w.b.t. and a very good day,

Distinguished guests, speakers, lecturers, MyMS members and
researchers.

It is my great pleasure to welcome you all to the 4
th
Australasian
Metabolomics Symposium and Workshops. I am pleased that the
symposium and workshops could take place here at our beloved UiTM
Puncak Alam campus and hope that you are pleased with our hospitality.

I was informed that, this is the first of its kind of event to be held here in
UiTM and in Malaysia. The 4
th
Australasian Symposium was previously
held in New Zealand and Australia. I appreciate the effort that the MyMS
committee took to realise this event and hope that this effort continues in
future.

The 4
th
Australasian Metabolomics Symposium and Workshops provides a golden platform for all to
exchange knowledge and to build networking. I would like to encourage all researchers to take advantage of
this platform to meet colleagues from your own speciality and also to discuss with experienced Professors
and researchers in Metabolomics on potential research collaboration.

Seeing the prospect of the omics field, the researchers at UiTM have come to realise the advantage of
metabolomics and are working on metabolomics based researches. To share the beauty of metabolomics, I
was told that the committee has attempted to replicate the spirit of the original Symposium written by Plato,
the great Greek philosopher for benefits of the research society as a whole. As a result, they have organised
the symposium and workshops to discuss on the selected theme of Moving Forward to Integrate Systemic
Biology into Post-Genomics Era.

Your strong support has made the 4
th
Australasian Metabolomics Symposium and Workshops a record
breaking event. The quality of technical programme is world class and the spectrum of topics is current and
broad. Two workshops were added to the programme to make this symposium strong and wholesome. The
invited speakers include Prof. Dr. David Wishart (University of Alberta, Canada), Prof. Dr. Rudolf Grimm
(Agilent Technologies, Adjunct Professor to University California Davis), Dr. Matthias Pelzing (Bruker
Daltonics, Australia), Dr. Ute Roessner (University of Melbourne), Prof. Dr. Thomas Hennessy (Adjunct
Professor to PROMISE, UiTM), Dr. Fang Fang (Bruker Biospin, Germany), Prof. Dr. Robert Trengove
(Director for the Murdoch Separation Science and Metabolomics Laboratory and the node leader for the
Murdoch Node of Metabolomics Australia) and Assoc. Prof. Dr. Markus R. Wenk (National University of
Singapore) are globally renowned for their expertise in the metabolomics field.

I hope you enjoy your participation in the 4
th
Australasian Metabolomics Symposium and Workshops and
hope that youre being here at our Puncak Alam campus would be a memorable one.

With this, I officiate the 4
th
Australasian Metabolomics Symposium and Workshops.

Thank you.




Dato Prof. Ir. Dr. Sahol Hamid Abu Bakar
FASc, PEng, DSPN, DJN, DSM, BCN
Vice Chancellor, Universiti Teknologi MARA
Malaysia


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Welcome Note

Assalamualaikum w.b.t and good day.

Distinguished speakers, researchers, lecturers and MyMS members.

I would like to welcome you to the 4
th
Australasian Metabolomics
Symposium and Workshops where this would be the first time it is held in
Malaysia, at UiTM Puncak Alam Campus. My sincere congratulation and
appreciation to the organising committee for all the hard work done in
making this event a success. Hopefully, we will continue to excel with
passion and enthusiasm in our research studies.

To all participants of the symposium and workshops, I wish you a rewarding
journey throughout the five days of learning and sharing, enriching your
minds on " The art of metabolomics: moving forward to integrate systemic
biology in post-genomics era - aptly used as the conference theme. To guests of UiTM, I wish you a
pleasant stay in our green campus here in Puncak Alam. To guests of the country, I wish you lots of
memorable experience in Malaysia, and I hope that you will come back to savour more of our Malaysian
hospitality.

The symposium and workshop mark the milestones of the Malaysian Metabolomics Society (MyMS)
gathering renowned scientists from everywhere to meet and share their knowledge and experiences. MyMS
was registered with the Registrar Office of Society and it symbolises the desire of the academics in Malaysia
to enhance metabolomics related researches and to integrate this tool into the post genomic era, to help
provide more understanding towards what is known through genomics. MyMS vowed to take the challenge
of realising the mission of gathering the scientists to share knowledge, to collaborate and to accelerate
metabolomics related researches. MyMS aims to foster and inculcate good research culture among the
young generation and provide or facilitate them with the tools they need. I cannot think of a better symbol for
the goals of this society but to seed and sustain the reformation that requires each of us who carries the
flame and serves as a torchbearer for metabolomics research in Malaysia.

I hope events such as this will provide us with the opportunity to share our experiences, learn from one
another, and progress debates in a positive, cooperative manner in moving forward with excellence in
research.

Best of luck as we all work together in dispelling myths and increasing the credibility and reputation of
MyMS as a whole.

Thank you.






Prof. Dr. Mohd Zaki Salleh
Founding President of MyMS








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Welcome Note

A very good day to distinguished guests, speakers, MyMS members, and
researchers.

Ladies and gentlemen, welcome to the 4
th
Australasian Metabolomics
Symposium and Workshops held at UiTM Puncak Alam.

I would like to begin by thanking the people who had initiated the idea of this
symposium, Dr. Ute Roessner, Prof. Dr. Thomas Hennessy and Prof. Dr.
Mohd Zaki Salleh. Many must have wondered why the symposium is named
as the 4
th
Australasian Symposium, and what has it got to do with Malaysia.
For your information, the first three installments were conducted in New
Zealand and Australia. It is our good friend, Dr. Ute Roessner, who
suggested for the symposium to be held for the first time in Malaysia. This
happened over a coffee talk when a few of us were discussing on how to
move forward metabolomics in Malaysia. Metabolomics is truly a useful tool for developing and testing
theories and hypotheses and the number of such studies increases fast. The emergence of various omics
studies share one goal, which is to discover the cellular processes and identifying potential biomarkers and
providing answers to our research questions.

This symposium marks an important milestone for the efforts of Malaysian Metabolomics Society and
Pharmacogenomics Centre in bringing together local researchers interested in metabolomics to share and
learn from the experts we invited from overseas. We sincerely hope that this kind of platform remains
effective, robust and enduring in promoting progress in science. The keywords in the title for this gathering
The art of metabolomics: moving forward to integrate systemic biology in post-genomics era were selected
specifically to stress this very point.

I am indeed very pleased to see so many colleagues and friends together at this event. I would especially
like to welcome those attendees and speakers from abroad who have kindly come to share their wisdom
and insight on metabolomics. I hope this event marks the beginning of our friendships and more
collaboration work among us can be initiated and realised. My sincere gratitude goes to the eminent
speakers in accepting our invitation. Your presence is the climax of this event. Thank you Prof. Dr. David
Wishart (University of Alberta, Canada), Prof. Dr. Rudolf Grimm (Agilent Technologies, Adjunct Professor to
University California Davis), Dr. Matthias Pelzing (Bruker Daltonics, Australia), Dr. Ute Roessner (University
of Melbourne), Prof. Dr. Thomas Hennessy (Adjunct Professor to PROMISE, UiTM), Dr. Fang Fang (Bruker
Biospin, Germany), Assoc. Prof. Dr. Robert Trengove (Director for the Murdoch Separation Science and
Metabolomics Laboratory and the node leader for the Murdoch Node of Metabolomics Australia) and Assoc.
Prof. Dr. Markus R. Wenk (National University of Singapore). It is indeed an honour and a privilege to have
you here with us, sharing your precious time and experience with us.

Ladies and gentlemen, aside from the symposium we have also arranged workshops including the Pre-
Symposium Workshop. I do hope that all of us would take the opportunity to gain as much knowledge as we
can from the line of programs we have arranged. The event would provide an opportunity for all of us to
meet with professors and experts in metabolomics to discuss more and to integrate knowledge in order to
move forward in mastering the metabolomics field.

I would once again thank everyone for making this event a success. I would like to extend my gratitude to
the Vice Chancellor of UiTM, Assist. Vice Chancellor of Puncak Perdana and Puncak Alam of UiTM and
Dean of Faculty of Pharmacy for the endless support. I would like to extend my gratitude to our generous
sponsors ranging from companies throughout Malaysia especially our Platinum Sponsors, Agilent
Technologies and Bruker Corporation. This proves that our good effort is highly supported by the private
sector as they appreciate the benefit of the symposium as an excellent venue to discuss scientific and
technological collaboration across the Pacific.

While I am saying thanks it would be remiss of me if I did not express my gratitude for all the hard work and
commitment of the organising committee for making this event a success. In particular, I would like to thank
Prof. Dr. Mohd Zaki Salleh and Prof. Dr. Jean-Frdric Weber, who have been supportive of this event and


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given a lot of ideas to ensure smooth running of the event. My appreciation goes as well to the secretariat
steered by Fazleen, Richard, Dr. John Shia, the committee members including Assoc. Prof. Dr. Kalavathy
Ramasamy, Assoc. Prof. Dr. Tang Thean Hock, Assoc. Prof. Dr. Kamarulzaman, Dr. Siti Azma Jusoh, Dr.
Wan Iryani, Ruzianisra, Zafirah Liyana, Dr. Adzrool, Mr. Mior, Mr. Azanizam, ICT team of Faculty Pharmacy,
Puan Noridah. And last but not least, I owe my gratitude to all the students of Pharmacogenomics Centre
who had been running around and for the hard work done these past couple of months to prepare an
outstanding event for the benefit of all. Thank you Sharina, Salleh, Shafiq, Rose, Izwan Ismail, Lian Shien,
Ruhil, Erda, Hanif, Husaini, Ikhwan, Rahimi, Hafiq, Lan, Dr Zakaria, Izwan Yusof, Syafirul, Aida, Lyn,
Norleen, Leya, Lala, Irma and Azwin.

Lastly, I hope that this two-day symposium and the Pre-Symposium Workshop and Workshop will be a
fruitful one. In short, please enjoy this gathering and to the young scientists, do learn from the eminent
researchers. Many of you must have heard of the eminent speakers through their publication and I believe
for some of you it is really an ecstasy to be able to meet them in person.

I sincerely apologise if there is any shortcoming of this event. Lets make this symposium a platform to share
and exchange information for better research.

Thank you.



Prof. Dr. Teh Lay Kek
Chairperson Organising Committee















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Welcome Note

With the greatest pleasure I welcome you to the symposium in conjunction
with the 4
th
Australasian Metabolomics Symposium hosted by Prof. Dr.
Mohd. Zaki and Prof. Teh from the Pharmacogenomics Centre
(PROMISE), Faculty of Pharmacy at the Universiti Teknologi MARA. We
are especially excited to bring the newly founded Malaysian Metabolomics
Society (http://www.mymetabolomics.org/) alive by bringing metabolomics
experts together with researchers interested in using metabolomics as a
new tool for their research programs.

The Australian and New Zealand Metabolomics Network
(http://www.anzmn.com.au) is proud to be part of the exciting
developments in the metabolomics field in Malaysia and are looking
forward to the event allowing exchange of knowledge and building new
bridges and friendships!

We hope you have a great time and enjoy the Symposium!

Thank you.




Dr. Ute Roessner
Co-Chairperson Organising Committee
Australian and New Zealand Metabolomics Network


























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Acknowledgements

The MyMS Organising Committee expresses its sincere appreciation to everyone who has given invaluable
support, time and cooperation:

Y. Bhg. Dato Prof. Ir. Dr. Sahol Hamid Abu Bakar
Vice-Chancellor, UiTM Malaysia

Y. Bhg. Prof. Dr. Azni Zain Ahmed
Deputy Vice-Chancellor (Academic and International), UiTM Malaysia

Y. Bhg. Prof. Ir. Dr. Hj. Abdul Rahman Omar
Deputy Vice-Chancellor (Research and Innovation), UiTM Malaysia

Y. Bhg. Prof. Dr. Azizul Halim Yahya
Assistant Vice Chancellor (Puncak Alam and Puncak Perdana Campus), UiTM Malaysia

Y. Bhg. Prof. Dr. Abu Bakar Abdul Majeed
Assistant Vice Chancellor (Research), UiTM Malaysia

Y. Bhg. Prof. Dr. Aishah Adam
Dean of Faculty of Pharmacy, UiTM Malaysia

Y. Bhg. Prof. Dr. Mohd Zaki Salleh
Head of PROMISE, (Puncak Alam Campus), UiTM Malaysia

Plenary Speakers:-

Prof. Dr. David Wishart
Dr. Fang Fang
Assoc. Prof. Dr. Markus R. Wenk
Dr. Matthias Pelzing
Assoc. Prof. Dr. Robert Trengove
Prof. Dr. Rudolf Grimm
Prof. Dr. Teh Lay Kek
Prof. Dr. Thomas Hennessy
Dr. Ute Roessner

Pre- workshop speaker:-

Prof. Dr. Jean-Frdric Weber

Workshop Speakers:-

Christopher Bowen
Robin Philp
Assoc. Prof. Dr. Robert Trengove
Prof. Dr. Thomas Hennessy
Dr. Ute Roessner

Platinum sponsors

Agilent Technologies (M) Sdn. Bhd.
Bruker (M) Sdn. Bhd.

Sponsors

Next Gene Scientific Sdn. Bhd.
Analisa Resources (M) Sdn. Bhd.
Medigene Sdn. Bhd.

Leco
Saintifik Maju Sdn. Bhd.




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Organising Committee

Advisor : Prof. Dr. Mohd Zaki Salleh
: Prof. Dr. Aishah Adam
Chairperson : Prof. Dr. Teh Lay Kek
Co-chairperson : Dr. Ute Roessner
Secretary : Ms. Fazleen Haslinda Mohd Hatta
: Mr. Richard Muhammad Johari James
Treasurer : Assoc. Prof. Dr. Kalavathy Ramasamy
: Assoc. Prof. Dr. Tang Thean Hock
Scientific committee : Prof. Dr. Mohd Zaki Salleh
: Prof. Dr. Jean-Frdric Weber
: Dr. Wan Iryani Wan Ismail
: Dr. John Shia Kwong Siew
: Dr. Zakaria Bannur
: Mrs. Ruzianisra Mohamed
Program and abstract book : Ms. Erda Syerena Rosli
: Ms. Nur Jalinna Abdul Jalil
: Ms. Asbiyatulaida Derahman
Registration : Mrs. Zafirah Liyana Abdullah
: Mrs. Siti Nooraishah
: Ms. Nur Azwin Ismail
: Ms. Nornazliya Mohamad
Protocol : Mr. Azanizam Ismail
: Ms. Irma Syakina Mohd Zaki
: Mr. Mohd Shafiq Aazmi
Hospitality : Mrs. Nurul Aqmar Mohd Nor Hazalin
: Ms. Sharina Hamzah
: Mr. Mohd Syafirul Shamsuddin
: Ms. Rose Iszati Ismet Nayan
Exhibition (sponsors) : Mrs. Lee Lian Shien
: Mrs. Norleen Mohamed Ali
: Mr. Muhammad Husaini Ismail




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ICT, Art & Design : Mr. Mohd Izwan Mohamad Yusof
: Mr. Mohd Ikmal Hanif Abdul Khalid
: Mr. Mohamad Izwan Ismail
: Dr. Adzrool Idzwan Ismail (Photography and video)
: Assoc. Prof. Dr. Kamarulzaman Md. Isa
(Photography & video)
Publicity : Dr. Siti Azma Jusoh
: Mr. Mohd Salleh Rofiee
: Mr. Mohamad Izwan Ismail
: Mrs. Noridah Md Lasam
Social and tour : Ms. Ruhil Nadirah Che Omar
: Ms. Rose Iszati Ismet Nayan
Conference workshop : Mr. Mohd Ikhwan Ismail
: Mr. Mohd Syafirul Shamsuddin
: Mr. Mohamad Izwan Ismail
: Mr. Mohd Salleh Rofiee
: Mr. Hasbullani Zakaria
: Mr. Muhamad Hafiq Zulkifli
Food and beverages : Mr. Mohd Rahimi Muda
: Ms. Sharina Hamzah
: Mr. Mohd Syafirul Shamsuddin






















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Malaysian Metabolomics Society (MyMS)

About

Metabolomics related research is dated back in 1970 by the first paper published by Pauling. In 2001, the
term "metabolomics" was introduced by Oliver Fiehn and defined as "a comprehensive and quantitative
analysis of all metabolites" in a system. Later in 2004, Metabolomics Society was founded by Rima
Kaddurah-Daouk, Associate Professor, Duke University Medical Center. And in 2007, Professor Dr David
Wishart, project leader for The Human Metabolome Project released the first draft of the human
metabolome comprising of 500 metabolites, 1,200 drugs and 3,500 food components. Now the database
has grown to cover more than 7900 metabolites (http://www.hmdb.ca/).

Watching the impressive movement in metabolomics by scientists all over the world and with the potential of
metabolomics in closing the gaps of information currently faced with other omics, a group of researchers in
Malaysia came together to establish the Malaysian Metabolomics Society (MyMS). The founding President,
Prof. Dr. Mohd Zaki Salleh together with Prof. Dr. Teh Lay Kek, advised by Dr. Ute Roessner and Prof. Dr.
Thomas Hennessy took the challenge to steer the protemp committee comprised of 20 academics with 30
members from all background. The society is registered later with the Registry of Societies of Malaysia in
2011. The members are professionals working in various fields including Life Sciences, Chemistry,
Pharmacy and Medicine. It is an independent, Non-Governmental Organisation and a Non-Profit
Organisation governed by Registry of Societies of Malaysia. One of the important function of this society is
to gather scientists in Malaysia interested in metabolomics to work together and establish or contribute to
the metabolome database, to help the scientific committee move further in unravelling disease mechanisms,
discover new and more sensitive and specific biomarker to help diagnose disease quicker and monitor drug
responses and disease progresses in patients.

Vision
To become a platform for gathering scientists and enhance metabolomics-based researchers

Missions
To enhance metabolomics-related researchers in Malaysia.
To become the platform for knowledge sharing among the researchers in the field of metabolomics.
To organise seminars, symposiums, workshops.
To facilitate networking between scientists and researchers in different organisations locally and
internationally.
Committee
President
Prof. Dr. Mohd Zaki Salleh

Deputy President
Assoc. Prof. Dr. Zainul Amiruddin Bin
Zakaria

Vice President
Prof. Dr. Teh Lay Kek

Secretary
Dr. John Shia Kwong Siew

Deputy Secretary
Richard Muhammad Johari James


Treasurer
Assoc. Prof. Dr. Kalavathy Ramasamy

Deputy Treasurer
Selene Tan Shiau Leng

International Advisors
Prof. Dr. Thomas Hennessy
Dr. Ute Roessner

Committee members
Dr. Wan Iryani Wan Ismail
Dr. Siti Azma Jusoh
Dr. Mohd Shihabuddin Mohamed
Noorden
Fazleen Haslinda Mohd Hatta
Seetha Ramasamy
Tee Ting Yee



4
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Table of Contents

Page
Welcome Note
Vice Chancellor Universiti Teknologi MARA i
President of MyMS ii
Chairperson Organising Committee iii
Co-chairperson Organising Committee v

Acknowledgements vi

Organising Committee vii

Malaysian Metabolomics Society (MyMS) ix

Symposium Programme
Tuesday (4
th
September 2012) 1
Wednesday (5
th
September 2012) 2

Plenary Speakers 3

Abstract
Plenary Lectures 7
Oral Communications
Day 1 18
Day 2 26
Poster Communications 36

Appendices
List of Participants xi
Floor Plan xvii
Contacts Number (Secretariat of Symposium) xviii
Conference Evaluation Form -
MyMS Membership Form -







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Symposium Programme

Tuesday (4
th
September 2012)


Time Programme
0830 - 0930 Registration
0930 - 1015
Plenary Lecture 1: Prof. Dr. David Wishart
Comprehensive Characterisation of the Human Biofluid Metabolomes
1015 - 1100
Plenary Lecture 2: Prof. Dr. Rudolf Grimm
Multi-omics Studies of the Oldest Human Mummy the Tyrolean Iceman or Oetzi
1100 - 1130 Tea Break/Poster Session
1130 - 1215
Plenary Lecture 3: Dr. Matthias Pelzing
Ultrafast Statistical Profiling: FT-ICR Based Profiling of Myxobacteria Natural
Products for Rapid Detection and Identification of Marker Compounds
1215 - 1300
Plenary Lecture 4: Dr. Ute Roessner
The Potential of Metabolomics in Systems Biology
1300 - 1430 Lunch Break/Poster Session
1430 - 1515
Plenary Lecture 5: Dr. Fang Fang
Innovative NMR Metabolomics for Food Safety and Quality Control and Newborn
Urine Screening
1515 - 1530 Tea Break/Poster Session
1530 - 1600 Talk by Representative of Sponsor (platinum)
1615 - 1730 Oral Communication 1
1800 - 2230
Symposium and Workshops Officiated by Vice Chancellor UiTM/Cultural
Dance/Dinner (BBQ)/Social Events












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Symposium Programme

Wednesday (5
th
September 2012)


Time Programme
0830 - 0930 Registration
0930 - 1015
Plenary Lecture 6: Assoc. Prof. Dr. Robert Trengove
Metabolomics and the Nexus with Residue Analysis: The Study of Health Impacts of
Residue Exposure
1015 - 1100
Plenary Lecture 7: Assoc. Prof. Dr. Markus R. Wenk
Lipidomics, New Tools and Applications
1100 - 1130 Tea Break/Poster Session
1130 - 1215
Plenary Lecture 8: Prof. Dr. Thomas Hennessy
The Application of Mass Profiler Professional in Biomarker Discovery: A Statistical
Analysis & Visualisation Software Tool in PTSD Metabolomics
1215 - 1300
Plenary Lecture 9: Prof. Dr. Teh Lay Kek (PROMISE, UiTM)
Moving Forward with Metabolomics: Scenario in Malaysia
1300 - 1430 Lunch Break/Poster Session
1430 - 1500 Talk by Representative of Sponsor (Platinum)
1515 - 1630 Oral Communication 2
1630 - 1730 Round up Discussion
Tea Break/Adjourn















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Plenary Speakers


Prof. Dr. David Wishart
A professor in the Department of Computing Science and Biological Sciences
at the University of Alberta, Edmonton.

Wishart's research interests are very broad, touching on areas relating to
nanobiology, genomics, proteomics, metabolomics, bioinformatics and
systems biology. His particular interests lie in developing novel methods or
improved experimental techniques to facilitate the understanding of biological
systems at the molecular (or nano) scale. His research includes the Human
Metabolome Project and the Human Metabolite Database Project, among
others.

Dr. Fang Fang
An application Chemist for Bruker BioSpin since 2008.

Her research interests include Metabonomics and Mixture Analysis. She is the
expert in the areas related to High Resolution Magic Angle Spinning (HR-MAS-
NMR). She is involved in studies of food safety and quality control (juice,
honey, beer, wine, milk powder, cheese, gutter oil), as well as in clinical
biofluid screening (inborn errors, epidemiology, infections and cancer).

Assoc. Prof. Dr. Markus R. Wenk
Associate Professor of Biochemistry at National University of Singapore,
Director of SLING and Privatdozent at the University of Basel.

Markus Wenk obtained his PhD in Biophysics from the Biozentrum at the
University of Basel, Switzerland in 1997. He then moved to Yale University
where he initially was a postdoctoral fellow and subsequently a research
faculty scientist in the Department of Cell Biology. In 2004, he took up a
position as an Assistant Professor at the National University of Singapore
(Yong Loo Lin School of Medicine, Department of Biochemistry and Faculty of
Science, Department of Biological Sciences). He is currently an Associate
Professor, Assistant Head of Department of Biochemistry and member of the
Executive Committee of NGS, the NUS Graduate School for Integrative
Sciences and Engineering. Since 2005, he is Privatdozent in the Faculty of
Science at the University of Basel. In this capacity, he has initiated and is co-
directing an international joint graduate program between Singapore and
Switzerland. He is also founder and organiser of the biennial International
Singapore Lipid Symposium and Executive Editor of Progress in Lipid
Research. Dr. Markus had introduced and established novel techniques for
analysis of phospholipids metabolism pertinent to membrane dynamics at the
neurological nerve terminal. His work resulted in scientific publications which
have major impact on conceptual advancements in the field of neurobiology
and lipid metabolism. He is now spearheading novel approaches in global
analysis of lipids (lipidomics) for applications in drug and biomarker
development with relevance to various disease areas.


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Dr. Matthias Pelzing
An application Manager for Bruker Daltonics Asia Pacific.

Matthias Pelzing received his diploma degree in chemistry and his Ph.D.
degree in mass spectrometry at the University of Leipzig, Germany in 1991
and 1994. In his Ph.D. thesis he studied waste products in semiconductor
industry with mass spectrometric methods in the group of Prof. R. Herzschuh.
From 1995 to 1998 he worked as a Postdoctoral Fellow at Institute of
Tropospheric Research on characterisation of atmospheric aerosol particles
with different mass spectrometric techniques. In 1999 he joint Bruker Daltonik
where he worked as applications scientist with focus on coupling techniques of
separation systems to API-MS systems. Since January 2006 he has been
working as Application Manager for Bruker Daltonics Asia Pacific/ Japan
based in Melbourne/Australia. Over the last 10 years, his main research
interests are in method development of hyphenated separation techniques
coupled to mass spectrometry.

Assoc. Prof. Dr. Robert Trengove
Director for the Murdoch Separation Science and Metabolomics Laboratory
and the node leader for the Murdoch Node of Metabolomics Australia.

Robert Trengove has pioneered the development of MS-based omics
techniques for more than 20 years, collaborating with Australian and
International researchers and industry. He currently leads a team of more than
15 researchers working on a diverse range of topics including HIV/AIDs, iron
disorders, desalination, microalgae lipidomics, fungal and bacterial
metabolomics. Professor Trengove has published many high-impact journal
articles and has ongoing commercial contract research arrangements with
major industry players in the petrochemical, clinical and animal health
pharmaceutical sector.

Prof. Dr. Rudolf Grimm
Worldwide Proteomics and Metabolomics Market Development Manager and
Director of Science and Tech at Agilent Technologies. Adjunct Professor at the
University of California in Davis.

Rudolf Grimm received his Ph.D. in Biology from the University of
Munich/Germany. After completing a post-doc at the University of
Freiburg/Germany and Riken Institute in Tokyo/Japan he joined Hewlett-
Packard as a senior life science application chemist in 1991. In 1998, he
became the head of protein chemistry at the Munich based proteomics
company Toplab. In 1999, he joined Hexal Pharma to establish the Biotech
Laboratories for the successful development of generic recombinant protein
drugs (Biosimilars). In 2002, he re-joined Agilent Technologies as the
worldwide proteomics and metabolomics market development manager. In
2009 he became a director of science and technology at Agilent Technologies.
In 2010, Rudi became an Adjunct Professor at the University of California in
Davis and in 2011 an Adjunct Professor at Chungnam National University in
Daejeon/South-Korea. Since 2010, he managed the strategic academic
collaborations in Asia-Pacific.






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Prof. Dr. Teh Lay Kek
A Professor of the Faculty of Pharmacy at University Teknologi MARA (UiTM),
Puncak Alam Campus, Selangor, Malaysia.

She graduated with a BPharm (Hons.) from the University of Nottingham, UK
in 1995 and completed her Master of Clinical Pharmacy in 1997 from Universiti
Sains Malaysia (USM). In 2002, she obtained her Doctor of Philosophy
certificate in Pharmacology from USM. She started her career as a lecturer in
University of Malaya in 1998. She moved to UiTM as a senior lecturer in 2003
and was promoted to position as an associate professor in 2008 and to
professor in 2012. She is one of the active founding members of the
Pharmacogenomics Centre (PROMISE) established in 2008. She is also one
of the founding members of the Malaysian Society for the Advancement of
Pharmacogenomics and Malaysian Metabolomics Society (MyMS). Her
research interests are in experimental and clinical pharmacogenomics as well
as metabolomics.

Prof. Dr. Thomas Hennessy
Application scientist for life sciences high-end mass spectrometry
applications. Adjunct Professor at Universiti Teknologi MARA, Malaysia.

Thomas Patrick Hennessy graduated with a M.Sc in Chemistry (Diplom) in
1999 and received his PhD in Chemistry with the dissertation title Peptide
mapping employing analytical & micro-separation techniques in 2002. Key
aspects of his research comprise translational research and clinical
applications using -omics technologies. His tutoring and teaching experience
features advanced vocational training at international symposia and
conferences as well as international workshops in separation science in the
post-genome era. The requirements of increasingly complex separation
systems and platforms frequently result in organising advanced vocational
short courses and workshops co-lecturing alongside renowned experts from
industrial co-operation partners and academic experts on multi-dimensional
separation technologies hyphenated to mass spectrometry. Moreover, his
lectures cover biochemistry, inorganic & analytical chemistry as well as basic
quantum mechanics.

Dr. Ute Roessner
Head of the Metabolomics Facility at the Australian Centre for Plant Functional
Genomics. One of the pioneers of metabolomics technology developments
especially in the plant research / agricultural fields.

Dr. Roessner obtained her PhD in Biochemistry at the MPI for Molecular Plant
Physiology in Germany, where she developed novel GC-MS method to
analyse metabolites in plants. The field of metabolomics emerged together
with the application of sophisticated data mining, and it is today an important
tool in biological sciences, systems biology and biomarker discovery. In 2003,
Dr. Roessner moved to Australia where she established a GC-MS and LC-MS
based metabolomics platform at the University of Melbourne, node of the
Australian Centre for Plant Functional Genomics. Since 2007, Dr. Roessner
has also been involved in the setup of Metabolomics Australia (MA), an
Australian federal government investment, through Bioplatforms Australia Ltd.
and currently she leads the MA node at the University of Melbourne.




4
th
Australasian Metabolomics
Symposium 2012



















ABSTRACTS
Plenary Lectures















7

4
th
Australasian Metabolomics
Symposium 2012



Plenary Lectures


No. Name Title
1. David Wishart Comprehensive Characterisation of the Human Biofluid Metabolomes
2. Rudolf Grimm
Multi-omics Studies of the Oldest Human Mummy the Tyrolean Iceman or
Oetzi
3. Matthias Pelzing
Ultrafast Statistical Profiling: FT-ICR Based Profiling of Myxobacteria
Natural Products for Rapid Detection and Identification of Marker
Compounds
4. Ute Roessner The Potential of Metabolomics in Systems Biology
5. Fang Fang
Innovative NMR Metabolomics for Food Safety and Quality Control and
Newborn Urine Screening
6. Robert Trengove
Metabolomics and the Nexus with Residue Analysis: The Study of Health
Impacts of Residue Exposure
7. Markus R. Wenk Lipidomics, New Tools and Applications
8. Thomas Hennessy
The Application of Mass Profiler Professional in Biomarker Discovery: A
Statistical Analysis & Visualisation Software Tool in PTSD Metabolomics
9. Teh Lay Kek Moving Forward with Metabolomics: Scenario in Malaysia






















8

4
th
Australasian Metabolomics
Symposium 2012



Comprehensive Characterisation of the Human Biofluid Metabolomes

David Wishart

Depts. of Computing Science and Biological Sciences, University of Alberta, Edmonton, AB,
Canada.

Corresponding author: dwishart@ualberta.ca

Abstract

The most accessible material for metabolomic studies in humans are biofluids. In particular,
blood, urine, saliva and cerebrospinal fluid (CSF) are relatively easy to obtain and are routinely
used in many clinical and metabolomic studies. However, despite their ubiquity and clinical utility,
surprisingly little is (or has been) known about their full chemical composition. As part of an
ongoing effort begun in 2005, we have been comprehensively characterising the metabolomes of
human CSF, blood, urine and saliva using a broad array of experimental techniques. This
experimental work has been closely coupled with an extensive review of the literature. In this
presentation I will describe our progress towards this goal and our specific methodologies. In
particular I will focus on our recent advances in characterising the human CSF and human urine
metabolomes. Comprehensive knowledge of human biofluid metabolomes, along with their
normal concentration ranges opens the door to the discovery of novel disease biomarkers and
biomarker profiles. In this presentation I will also describe the potential impact of knowing what is
detectable at certain concentration thresholds in terms of the development of automated
metabolomic detection and quantification for different platforms.


Keywords: biomarker; human biofluids; metabolomic

























9

4
th
Australasian Metabolomics
Symposium 2012



Multi-omics Studies of the Oldest Human Mummy the Tyrolean Iceman
or Oetzi

Rudolf Grimm
1,3,6
*,

Rick Reisdorph
2
, Roger Powell
2
, Spencer Mahaffey
2
, Hyun Joo An
3,6
,
Grace Ro
3
, Sureyya Ozcan
3
, Carlito Lebrilla
3
, Christian Reiter
4
and Thomas Bereuter
4,5
,
Rob Moritz
7
, Michael Hoopman
7
, Holly Ganz
3
, Bart Weimer
3
, David Bishop
8
, Philip Doble
8
,
Amaury Gassiot
9
, Markus Wenk
9
, Theo Sana
1
, Frank Maixner
10
, Albert Zink
10


1
Agilent Technologies, Santa Clara, USA;
2
National Jewish Health Institute, Denver, USA;
3
UC
Davis, Davis, USA;
4
Medical University of Vienna, Austria;
5
Technical University Graz, Austria;
6
Chungnam National University, Daejeon, South-Korea;
7
ISB, Seattle, USA;
8
University of
Technology Sydney, Australia;
9
National University of Singapore, Singapore,
10
Institute for
Mummies and the Iceman, EURAC, Bolzano, Italy.

*Corresponding author: rudolf_grimm@agilent.com

Abstract

tzi the Iceman is a well-preserved natural mummy of a man who lived about 5300 years ago.
The mummy was found in September 1991 in the tztal Alps, hence tzi, near the Similaun
mountain and Hauslabjoch on the border between Austria and Italy. He is the oldest known natural
human mummy. We performed the first multi-omics study of this famous mummy including
genomics, proteomics, glycomics, metabolomics, lipidomics and metallomics approaches.
Complex multi-omics data will be presented from samples taking out of his stomach. Interesting
insights will be given in his last meal(s) and health conditions before he got killed. Glycomics data
will be also presented from a tissue sample taken out of his hip region and compared to data
generated from two 2400 years old Siberian ice mummies and a contemporary mummy.


Keywords: multi-omics; tzi the Iceman






















ii


10

4
th
Australasian Metabolomics
Symposium 2012



Ultrafast Statistical Profiling: FT-ICR Based Profiling of Myxobacteria
Natural Products for Rapid Detection and Identification of Marker
Compounds

Matthias Pelzing
1
*, Daniel Krug
2,3
, Thomas Hoffmann
2,3
, Aiko Barsch
4
, Matthias Witt
4
, Rolf
Mller
2,3

1
Bruker Biosciences Pty Ltd, Australia;
2
Helmholtz Institute for Pharmaceutical Research Saarland
(HIPS), Saarbrcken, Germany;
3
Saarland University, Saarbrcken, Germany;
4
Bruker Daltonik
GmbH, Bremen, Germany.

*Corresponding author: matthias.pelzing@bdal.de

Abstract

Myxobacteria represent an important source of novel natural products exhibiting a wide range of
biological activities. Some of these so-called secondary metabolites are investigated as potential
leads for novel drugs. Traditional approaches to discovering natural products mainly employ
bioassays and activity-guided isolation from different myxobacterial isolates, but genomics-based
strategies become increasingly successful to reveal additional compounds. These "metabolome-
mining" approaches hold great promise for uncovering novel secondary metabolites from
myxobacterial strains, as the number of known compounds identified to date is often significantly
lower than expected from genome sequence information. Currently, successfully established
methods are based on LC-QTOF-MS analysis requiring about 20 min analysis times per sample.
Facing several thousand myxobacterial strain isolates as well as numerous genetic knock out
mutant strains available to analyse for the presence of novel secondary metabolites, analysis time
becomes a bottleneck. In this proof of concept study we analysed several metabolite extracts from
genetic knock-out mutants by ultra-high resolution FT-ICR direct infusion measurements, requiring
only about 1 min / sample to measure. Principal Component Analysis (PCA) of the acquired MS
spectra showed a clustering of the samples according to the bacterial genetic background, i.e. wild
type and mutants could be differentiated. PCA loadings pointed to compounds responsible for this
differentiation. By measuring these regions of interest in continuous accumulation of selected ions
(CASI) mode, the sensitivity and resolution of these mass ranges could be enhanced significantly.
The instrument resolving power of R>750.000 provided isotopic fine structure information. This
enabled to literally read out the correct elemental composition for the target compounds. We
have demonstrated for myxoprincomide, a recently discovered myxobacterial secondary
metabolite, that this approach enables an unequivocal molecular formula generation for a
compound with MW >1000. The ultra high resolution of an FT-ICR instrument enables for the first
time an elemental composition determination for a molecular weight range where mass accuracy
of existing mass spectrometers would be not enough for an unequivocal decision. The presented
Ultrafast Statistical Profiling workflow enables a rapid profiling of complex metabolite extracts
and identification of relevant marker compounds by making use of the ultrahigh resolution and the
wide dynamic range provided by the FT-ICR technology addressing two of the major bottleneck
in metabolomics, sample throughput and compound identification.


Keywords: FT-ICR technology; myxobacteria; Ultrafast Statistical Profiling







11

4
th
Australasian Metabolomics
Symposium 2012



The Potential of Metabolomics in Systems Biology

Ute Roessner

Australian Centre for Plant Functional Genomics and Metabolomics Australia, School of Botany,
the University of Melbourne, 3010 Victoria, Australia.

Corresponding author: u.roessner@unimelb.edu.au

Abstract

In the last two decades, significant progress has been made in the sequencing of the genomes of
a number of different organisms. Simultaneously, large investments have occurred in the
development of high throughput analytical approaches to analyse different cell products, such as
those from gene expression (transcriptomics) as well as proteins (proteomics) and metabolites
(metabolomics). These omics approaches, when used in combination with sophisticated
bioinformatics tools for data mining and interpretation, are considered important tools to be applied
and utilised to understand the biology of an organism and its response to environmental stimuli or
genetic perturbation. The ability to measure all elements of a biological system, such as DNA,
mRNA, proteins, metabolites and structural elements such as the cytoskeleton, cell walls and
membranes, and also to determine the relationship of those elements to one another as part of the
systems response to various stimuli is called Systems Biology. In this paper I want to discuss the
importance of metabolomics as a major contributor in any systems biology study and explore an
example from our own research how we extend the potential of metabolomics by combining it with
genetics to understand adaptation and tolerance to drought in wheat, a major food crop.


Keywords: adaptation; bioinformatics tools; metabolomics; tolerance


























12

4
th
Australasian Metabolomics
Symposium 2012



Innovative NMR Metabolomics for Food Safety and Quality Control and
Newborn Urine Screening

F Fang*, B Schuetz, C Cannet, D Krings, H Schaefer, M Moertter, M Spraul

Bruker BioSpin GmbH, Germany.

*Corresponding author: Fang.Fang@bruker-biospin.de

Abstract

NMR is established as an efficient analytical tool for metabolomics analysis based on its highest
reproducibility and transferability. Smallest changes of concentrations of many compounds in
biofluid can then be observed in untargeted and targeted screening with one experiment. Based
on its unmatched reproducibility, NMR based metabolomics is possible to visualise smallest
changes in multiple metabolite concentrations simultaneously, defining the strength of NMR as a
multimarker analytical tool to detect all types of deviations, even those, previously unknown. To
fulfil the stringent requirements of statistical data evaluation, Standard Operation Procedures
(SOPs) from sample handling/preparation to post processing have been developed. The need of
complete standardisation will be shown through a NMR based Metabolomics case study of urine
based newborn screening for inborn errors of metabolism. Given the fact, with minimised sample
preparation NMR can deliver high throughput under full automation. The cost of per sample
analysis is low, however the information content obtained is unique. Such a quality control system
for fruit juices is already available under full automation from measurement to final report. The
same technical procedure has been applied to wine-, milk product-, and edible oil screening.
Examples are explained for every food category mentioned. All applications mentioned are strictly
following standard operation procedures, such allowing to transfer statistical models and
quantification routines between different instruments of the same field strength and different labs.


Keywords: inborn errors; NMR based metabolomics; urine























13

4
th
Australasian Metabolomics
Symposium 2012



Metabolomics and the Nexus with Residue Analysis: the Study of
Health Impacts of Residue Exposure

Bruce Peebles
1,2
, Catherine Rawlinson
1,2
, Joel Gummer
1,2
, Garth Maker
1,2,3
, Ian
Mullaney
2,4
, Robert Trengove
1,2
*


1
Murdoch University Metabolomics Australia Node, Murdoch University, 90 South Street, Murdoch
WA 6150, Australia;
2
Separation Science and Metabolomics Laboratory, Murdoch University, 90
South Street, Murdoch WA 6150, Australia;
3
School of Biological Sciences and Biotechnology,
Murdoch University, 90 South Street, Murdoch WA 6150, Australia;
4
School of Veterinary and
Biomedical Sciences, Murdoch University, 90 South Street, Murdoch WA 6150, Australia.

*Corresponding author: r.trengove@murdoch.edu.au

Abstract

The "omics", particularly proteomics and more recently metabolomics have been drivers in recent
instrumental development, leading to faster chromatography, accurate mass and high resolution
mass analysers, and equally as important robust sample interfaces for minimal sample
preparation. Alongside the hardware developments there have been considerable developments
in software development for mass spectral deconvolution, data visualisation, and mapping of
analytes onto biochemical pathways to identify which pathways are modified in a challenged
sample vs. a control sample. For instance, this allows the mechanisms of disease progression to
be dissected and potential management strategies and potential therapeutic targets to be
identified. These same instrumental developments have been utilised by the residue and
environmental communities with the development of multiresidue methods that screen for in
excess of 1000 analytes in a single analysis. Metabolomics seeks to identify changes in response
to some stimulus, whilst residue and environmental analysis seeks to identify all possible residue
and environmental "stimulus". We are at the emergence of the capability to simultaneously
measure residue and environmental analytes and profiles of primary and secondary metabolites.
We can start to investigate from both directions or investigate "cause" and "effect" simultaneously.
This presentation will address the approach and the challenges that need to be addressed to
facilitate the use of untargeted methods. In particular the different approaches required for
GC/MS based vs. LC/MS based metabolomics. Instrumentation optimisation will be discussed in
the context of combination of targeted compound analysis and metabolite profiling. Collaborative
pathways to advance this approach will be discussed.


Keywords: GC/MS; LC/MS; metabolomics; untargeted methods















14

4
th
Australasian Metabolomics
Symposium 2012



Lipidomics, New Tools and Applications

Markus R. Wenk

Department of Biochemistry, Yong Loo Lin School of Medicine, and Department of Biological
Sciences, National University of Singapore.

Corresponding author: markus_wenk@nuhs.edu.sg

Abstract

Once viewed simply as a reservoir for carbon storage, lipids are no longer cast as bystanders in
the drama of biological systems. The emerging field of lipidomics is driven by technology, most
notably mass spectrometry, but also by complementary approaches for the detection and
characterisation of lipids and their biosynthetic enzymes in living cells. The development of these
integrated tools promises to greatly advance our understanding of the diverse biological roles of
lipids (Wenk 2010 Cell 143(6):888-95). Understanding better the fundamentals of natural variation
in lipidomes as well as specific recognition of individual lipid species are main scientific aims of
SLING, the Singapore Lipidomics Incubator (http://www.sling.nus.edu.sg/). Shaped by a five year
competitive research program supported by the National Research Foundation and the National
University of Singapore, this centre is a major global magnet for collaborating parties in lipidomics
from academia and industry delivering new technologies and intellectual capital.


Keywords: lipidomics; lipids





























15

4
th
Australasian Metabolomics
Symposium 2012



The Application of Mass Profiler Professional in Biomarker Discovery:
A Statistical Analysis & Visualisation Software Tool in PTSD
Metabolomics

G Hamuni
1
, A Karabatsiakis
2
, S Kadereit
3
, R Durairaj
4
, TP Hennessy
5
*, G Currie
6
, IT
Kolassa
2
1
Department of Psychology, Clinical & Neuropsychology, Universitt Konstanz;
2
Institut fr
Psychologie und Pdagogik, Klinische und Biologische Psychologie, Universitt Ulm;
3
Zukunftskolleg,Fachbereich Biologie,Universitt Konstanz;
4
Information Technology & Services,
Strand Life Sciences, Bangalore;
5
Life Sciences Group, Agilent Technologies, Singapore;
6
Life
Sciences Group, Agilent Technologies, Melbourne.

*Corresponding author: thomas.hennesy@agilent.com

Abstract

Biomarker discovery can be performed at all levels of information flow in biological systems.
Integrated Biology then is the discipline compiling and extracting meaningful data to
comprehensibly describe any such system under scrutiny. However, in the quest for biomarkers it
is desirable to obtain and retain only information which shows variance in expression/abundance
levels at the expense of all other information. Metabolite Biomarker discovery is the pursuit to
display differential abundance metabolite levels. MS based differential metabolite profiling is one
way of monitoring up- and down-regulation between samples. Provided any perturbation causes a
response in the system and here in particular at the metabolite level this variation may be
monitored over time. Mass Profiler Professional (MPP) software is used to cluster biological and
technical replicates and differentiate them in terms of the time domain of any variance occurring
between samples where time course experiments are concerned or likewise
expression/abundance levels where differential expression is the result of a treatment or a disease
state. Only the molecular entities that show defined significant differences between samples are
selected and retained by the MPP statistical analysis and visualisation platform and ultimately
identified. The resulting identified metabolites are candidate biomarkers for the response of the
system to the perturbation. The profiling strategy is, hence, ideally suitable to investigate
experimental designs to study disease states in particular and any differential
expression/abundance scenario in general. The analysis strategy on the statistical analysis level
(MPP) will be exemplified via software data (profiling, molecular feature extraction, filtering,
principle component analysis (PCA), condition- and mass-tree generation aka hierarchical
clustering analysis (HCA), fold-change analysis, volcano plot representation) detailing the
workflow to answer questions dedicated to Post Traumatic Stress Disease (PTSD) metabolomics.


Keywords: Mass Profiler Professional (MPP); metabolite biomarker discovery; Post Traumatic
Stress Disease (PTSD)












16

4
th
Australasian Metabolomics
Symposium 2012



Moving Forward with Metabolomics: Scenario in Malaysia

LK Teh*, MZ Salleh*

Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, Universiti Teknologi MARA
Malaysia.

Corresponding authors: tehlaykek@puncakalam.uitm.edu.my/zakisalleh@puncakalam.uitm.edu.my

Abstract

Metabolomics is an important functional genomics tool which received great attention and
emphasis in recent years. Many researchers in Malaysia were working on metabolomics of plant
until 2009 when it was applied into clinical researches. Now, there is a wide range of its
application in plant, microorganisms and human with large metabolic complexity. The benefit of
integrating metabolomics with other omics is believed to provide comprehensive insights for a
variety of cellular metabolic processes in a cell, organ or organism; filling the missing gaps of
genomics, transcriptomics and proteomic analyses. In plant science, metabolomics is useful to
provide clues to understand the metabolic regulatory complex and its physiological state in
response to the environmental stress. Understanding the environmental stressors which control
the production of beneficial metabolites or factors affecting the growth and yield of the plant would
be of great importance in the field of agricultural science. Similarly, the pharmaceutical industry
would cast interest in the mechanism of actions of these valuable metabolites with potential use as
therapeutic agents. While in clinical researches, translating metabolomics to discover biomarkers
for bedside testing is greatly desired as currently available diagnostic tools for many diseases lack
accuracy or are invasive. In PROMISE, we are investigating the metabolic profiles of disease
causing pathogens to understand their pathogenecity and virulence. We are also integrating
genomics with metabolomics in search for biomarkers and understanding the mechanisms
involved in protection and susceptibility to diseases in a study which involves the local aborigines.
Several diseases or health issues would be of focus, which include cancers, cardiovascular
diseases, obesity and intellectual capabilities. In collaboration with other researchers, we are
looking into the use of metabolomics in improving yield and nutrient value of crops/marine
products and cataloguing mechanisms of the biological activities of local herbs.


Keywords: disease markers; metabolomics; pathogenecity; virulence
















4
th
Australasian Metabolomics
Symposium 2012



















ABSTRACTS
Oral Communications























18

4
th
Australasian Metabolomics
Symposium 2012



Oral Communications

Day 1


No. Presenter Title Reference No.
1. Azizan et al.
Footprinting Profiling of L. lactis in Response to
Derivatisation Techniques
MYMS-001po/m
2. Hamzah et al.
Potential Biomarkers for the Monitoring of Kidney
Function in Kidney Transplant Patients
MYMS-002po/m
3. Ibrahim et al.
Partial Least Square-Discriminant Analysis and Youden
Index for Metabolomic Analysis of NMR Spectra of
Exhaled Breath Condensate of Asthmatics and Healthy
Controls
MYMS-003po/m
4. Ismail et al.
Effect of Tocotrienol Rich Fraction (TRF) on Oxidant-
Antioxidant Equilibrium in Reducing Carbamazepine
(CBZ) Induced Lipid Peroxidation
MYMS-004po/m
5. Khalid et al.
Manipulation of In Vitro Cultures of Bosenbergia
rotunda for Enhanced Production of Targeted Bioactive
Compounds in the Phenylpropanoid Biosynthesis
Pathway
MYMS-005po/m
6. Laith et al.
Understanding the Metabolic Response of Catfish
(Clarias garipenus) to Elizabethkingia meningoseptica
Infections via Liquid ChromatographyMass
Spectrometry (LC/MS-QTOF)
MYMS-006po/m
7. Rosli et al.
Metabolites Profile of Tissue and Plasma in Breast
Cancer Patients Using Metabolomic Approach: Role of
Eicosanoid Metabolism
MYMS-007po/m
















19

4
th
Australasian Metabolomics
Symposium 2012



Footprinting Profiling of L. lactis in Response to Derivatisation
Techniques

Kamalrul Azlan Azizan*, Syarul Nataqain Baharum, Normah Mohd Noor

Institute of Systems Biology (INBIOSIS), Universiti Kebangsaan Malaysia, 43600, Bangi,
Selangor, Malaysia.

*Corresponding author: kamalrulazlan@gmail.com

Abstract

In general, most of the known low molecular weight metabolites are non-volatile thus require
chemical derivatisation to increase the volatility of the metabolites especially for GC based
analysis. Here we described the usage of an optimised protocol of conventional technique of TMS
derivatisation with shaking and recently introduced MCF derivatisation to determine the major
groups of metabolite that were produced in the fermented broth cultivated by the bacterium. The
footprinting metabolites of mesophilic L.lactis grown at 30C with shaking and 37C was profiled
using chemical derivatisation prior to gas chromatography mass spectrometry analysis. It was
observed that profiled groups of metabolites were according to the specific chemical derivatisation
used. TMS agent detected ranges of metabolites including alcohols, aldehydes, ketones and
sugars compared to MCF agent which were limited to amino and non-amino acids. In additional,
the major metabolites that contributed towards separation and discrimination in Principal
Component Analysis (PCA) and Partial Least Squares Discriminant Analysis (PLS-DA) is further
demonstrated and validated.


Keywords: L.lactis; MCF derivatisation; TMS derivatisation

























20

4
th
Australasian Metabolomics
Symposium 2012



Potential Biomarkers for the Monitoring of Kidney Function in Kidney
Transplant Patients

S Hamzah
1
*, JKS Shia
1
, G Ahmad
2
, HS Wong
3
, TP Hennessy
1,4
, LK Teh
1
,
MZ Salleh
1
*

1
Pharmacogenomics Research Centre (PROMISE), Faculty of Pharmacy, Universiti Teknologi
MARA, 42300 Puncak Alam, Selangor;
2
Institute of Nephrology and Urology, Hospital Kuala
Lumpur, 50586 Kuala Lumpur;
3
Department of Nephrology, Hospital Selayang, 68100 Batu
Caves, Selangor;
4
Life Sciences Group, Agilent Technologies, Singapore.

*Corresponding authors: sharinahamzah114@yahoo.com/zakisalleh@puncakalam.uitm.edu.my

Abstract

Kidney transplant patients are exposed to various complications such as immunological rejection,
ischemia or reperfusion injury and immunosuppressant induced nephrotoxicity. Currently, the
diagnosis of allograft rejection and toxicity of chronic calcineurin inhibitor relies on invasive
procedure of biopsy. Therefore in this study, a metabolomics approach was explored to search for
compounds that have potential to be further developed into biomarkers to monitor transplanted
patients. Serum was obtained from kidney-transplant patients treated with tacrolimus and healthy
volunteers. Fifty-five biological replicates of patients and 20 biological replicates of controls as well
as technical triplicates were analysed using LC/MS-QTOF (Agilent Tech). Samples were injected
after protein precipitation with acetonitrile and centrifuged for 10 min at 8,500rpm at 4C. The
chromatography was performed on an Agilent 1200 Series using an XDB C18 4.6 M, 1501.8
mm (Agilent Tech.). A total of 9 groups of compound were found to have potential to be developed
into biomarkers. One of the most interesting compounds that were found to be differentially
expressed between the patients and healthy volunteers is Diacylglycerol (DAG). DAG carries the
potential to be used as a marker to monitor the suppression level of the immune system.
Comparison of metabolite profiles between patients with episode of rejection and stable patients
revealed high abundance of Sphingosine-1-phosphate among patients who experienced rejection.
This compound is potentially a biomarker to predict allograft rejection since it has been shown to
carry various immune modulatory functions. Glycerophosphocholines (GPC) is also another
prospective group of compound that exhibited great potential due to its association with tacrolimus
dose-adjusted level and CYP3A5 genotypes demonstrated in this study. The present study
demonstrated the potential use of metabolomics approach for the search of candidate biomarkers
to monitor kidney transplant patients. However, all of these potential compounds need to be
further validated before translating into clinical practices.


Keywords: DAG; glycerophosphocholines (GPC); kidney-transplant patients; potential
biomarkers; rejection; sphingosine-1-phosphate










21

4
th
Australasian Metabolomics
Symposium 2012



Partial Least Square-Discriminant Analysis and Youden Index for
Metabolomic Analysis of NMR Spectra of Exhaled Breath Condensate
of Asthmatics and Healthy Controls

B Ibrahim
1, 2
*, M Nilsson
1, 3
,

SJ Fowler
1, 4

1
University of Manchester, Manchester Academic Health Science Centre, and NIHR Respiratory
and Allergy Clinical Research Facility, University Hospital of South Manchester, Manchester, UK;
2
School of Pharmaceutical Sciences, Universiti Sains Malaysia, Penang, Malaysia;
3
School of
Chemistry, University of Manchester, UK;
4
Lancashire Teaching Hospitals NHS Foundation Trust,
Preston, UK.

*Corresponding author: baharudin.ibrahim@usm.my

Abstract

Multivariate data analysis and chemometrics are keystone statistical techniques used to identify
markers of diseases in metabolomic studies. Previously, using principal component analysis
(PCA), logistic regression and receiver operating characteristic (ROC), we have shown the ability
of nuclear magnetic resonance (NMR) spectroscopy of exhaled breath condensate (EBC) in
discriminating asthmatics from healthy controls. The aim of this analysis is to employ partial least
square-discriminant analysis (PLS-DA) and Youden Index to re-evaluate this data. The NMR
spectra of EBC were binned to 0.02 ppm and PCA was applied initially to identify outliers.
Multivariate modelling was performed on a training set (70% of the total sample) in order to
produce a discriminatory model classifying asthmatics from healthy controls, and the model tested
in the remaining subjects. PLS-DA identifies a linear regression model between predictors and
binary outcome while Youden Index (sensitivity+specificity-1) is a summary measure of ROC
which represents the maximum potential effectiveness of a marker. In the training set, a model
(R
2
Y=0.43 and Q
2
Y=0.27) discriminating asthmatics (n=57) and controls (n=22) was developed
with an area under the receiver operating curve (AUROC) of 0.93. The highest Youden index
(0.744) was observed at a predicted value of 0.72, which corresponded to a sensitivity of 77% and
a specificity of 96%. In the test set (asthmatics, n=22; controls, n=12) the AUROC was 0.85, thus
providing external validation for the model. The sensitivity and the specificity were 77% and 92%
respectively. Response permutation test (100 permutations) proved internal validity of the model
(R
2
Y-intercept <0.3 and Q
2
T-intercept <0.05).The results were superior to previous analysis of the
same data set. PLS-DA and Youden Index are robust methods for metabolomic analysis of NMR
spectral data to discriminate asthmatics from healthy controls. These techniques also provide for
internal and external validation.


Keywords: asthma; exhaled breath condensate; metabolomics; partial least square-discriminant
analysis; Youden Index











22

4
th
Australasian Metabolomics
Symposium 2012



Effect of Tocotrienol Rich Fraction (TRF) on Oxidant-Antioxidant
Equilibrium in Reducing Carbamazepine (CBZ) Induced Lipid
Peroxidation

MI Ismail
1
*, ES Rosli
1
, A Derahman
1
, M Nornazliya
1
, Z Bannur
1
, MS Rofiee
1
, ZA Zakaria
2
,
TP Hennesy
1,3
, MZ Salleh
1
, LK Teh
1
*

1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Department of Biomedical
Sciences, Faculty of Medical and Health Sciences, Universiti Putra Malaysia, Malaysia;
3
Life
Sciences Group, Agilent Technologies, Singapore.

*Corresponding authors: ikhwanismail86@gmail.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Carbamazepine (CBZ) is commonly used to treat epilepsy but it ranked as one of the top 10 drugs
that caused fatal cutaneous drug reactions which include Steven Johnson Syndrome (SJS) and
Toxic Epidermal Necrolysis (TEN). CBZ induced toxicities has been hypothesised to be related to
the oxidant-antioxidant imbalances. Therefore, we aim to evaluate the potential role of Tocotrienol
Rich Fraction (TRF) in balancing the oxidant-antioxidant impairment induced by CBZ using 48
male Sprague Dawley (SD) rats. Seven groups of rats comprising of 6 rats per cages were
studied. A control group and 6 different groups of rats treated with 3 different dosages of CBZ (50,
100 and 200 mg/kg) and CBZ + TRF were conducted. Organs which include liver, kidney, brain
were collected from each rat on day 7 post treatment. All organs were weighed then freeze
snapped using liquid nitrogen. The organs were immersed into 0.9% NaCl and stored at -80C
before subjected to 6 series of extraction solvent which include ethyl acetate, hexane, ethanol,
methanol, dichloromethane and distilled water. Supernatant were pooled, dried, reconstituted with
mobile phase and injected to the LC/MS-QTOF for analysis. A total of 389 metabolites were
identified, 61 metabolites were related to CBZ treatment. The biochemical pathways including
tryptophan metabolism, purine metabolism and fatty acid (FA) metabolism were shown to be
affected in rats treated with CBZ. TRF was found to down-regulate FA metabolism which is
anticipated to have protective role in reducing lipid peroxidation especially in the brain. In addition,
the significant reduced expression of glutamylphenylalanine also suggest protective role of TRF in
nephrotoxicity. However, further study to evaluate the potential use of TRF as health supplement
in attempts to prevent or reduce toxicities is required.


Keywords: antioxidant; carbamazepine; metabolomic; SJS-TEN; TRF












23

4
th
Australasian Metabolomics
Symposium 2012



Manipulation of In Vitro Cultures of Bosenbergia rotunda for Enhanced
Production of Targeted Bioactive Compounds in the Phenylpropanoid
Biosynthesis Pathway

N Khalid*, N Yusof, Md Mustafa N, SM Wong and EC Tan

Centre for Biotechnology in Agriculture Research, Institute of Biological Sciences, Faculty of
Science, University Malaya, 40603 Kuala Lumpur, Malaysia.

*Corresponding author: lani@um.edu.my

Abstract

Boesenbergia rotunda (BR), a medicinal ginger, with bioactive compounds such as alpinetin,
cardamonin, pinostrobin, pinocembrin, panduratin A and 4-hydroxypanduratin A have shown to
have antibacterial, antifungal, anti-inflammatory, anticancer, and antioxidant properties. In our
work, panduratin A and 4-hydroxypanduratin A exhibited anti-dengue NS2B/ NS3 protease
activity. However, naturally these compounds are not feasible for future drug applications. The aim
of this work is to enhance the production of panduratin A and 4-hydroxypanduratin A through
chemical elicitation and genetic engineering. Initially cell suspension cultures of Bosenbergia
rotunda was established for chemical elicitation and genetic manipulation studies. Cultures were
initiated in shake flasks and growth was optimised in 5 litre bioreactors. Phenylalanine was used
as an elicitor to enhance the production of panduratin A and 4-hydroxypanduratin A. proteomic
approaches to identify proteins that may be involved in the biosynthesis of these compounds were
carried out. Following image analysis, protein spots whose expressions were found to be
regulated were identified using Matrix Assisted Laser Desorption-Ionisation tandem mass
spectrometry. Transcriptomic expression on cell cultures treated with phenylalanine was also
studied. Embryogenic callus from cell suspension was regenerated for genetic transformation
studies. In this work, the effects of initial inoculation volume, temperature and speed of agitation
on cell growth, total and selected flavonoid in suspension cultures of B. rotunda were determined.
The high performance liquid chromatography analysis showed that 2% inoculation volume induced
significantly high accumulation of biomass and flavonoid in the cells. For proteomics studies, thirty
four proteins were identified. Eleven of the proteins involved in the phenylpropanoid biosynthetic
pathway are related to the biosynthesis of cyclohexenyl chalcone derivatives. Transcriptomics
data analysis has provided more information on the phenylpropanoid pathway. As for plant
regeneration, embryogenic callus derived from cell suspension has been successfully optimised at
a high frequency. This study has provided information on the possible enzymes involved in the
phenylpropanoid pathway leading to the production of panduratin A and 4-hydroxypanduratin A.
Growth of suspension cultures were optimised both in small and large scale production.
Regenerable cell suspension cultures have also been established which will allow the expression
of targeted genes for enhanced accumulation of the bioactive compounds using genetic
transformation.


Keywords: 4-hydroxypanduratin; biosynthesis pathway; Bosenbergia rotunda; panduratin A;
phenylpropanoid








24

4
th
Australasian Metabolomics
Symposium 2012



Understanding the Metabolic Response of Catfish (Clarias garipenus)
to Elizabethkingia meningoseptica Infections via Liquid
ChromatographyMass Spectrometry (LCMS QTOF)

Laith AA
1
*, Najiah M
1
, Sifzizul TTM
2
, LK Teh
3
, MZ Salleh
3


1
Department of Fisheries and Aquaculture, Faculty of Fisheries and Aqua Industry,
2
Institute of
Marine Biotechnology, Universiti Malaysia Terengganu, Kuala Terengganu 21030 Terengganu
Malaysia
3
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi
MARA (UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia.

*Corresponding author: saiflaith3@yahoo.ca

Abstract

Safety and quality fish product is an important subject in aquaculture as the global demand for fish
products has doubled and is still rising. Catfish (clarias carepinus) is one of the worlds most
important staple meats, providing food to more than a billion peoplen but the pathogenicity of
Elizabethkingia meningoseptica, a significant bacterial group for human clinical infection, in fish
has not been established. In order to better understand the mechanism of infection and immunity
of fish to bacteria, a study was undertaken to identify metabolites that are related to infection and
resistance. A two-step approach using LC/MS Q-TOF system was employed. Differential
expression analysis of metabolite profiles for samples was conducted using Mass Profiler
Professional (Agilent Tech.) software. The differentially expressed metabolites were further
analysed using quadrupole time-of-flight (Q-TOF) MS/MS. Two metabolic molecules, Alpah-
Amyrin and Guanosine, were examined. The fish metabolic profiles were studied by top-down
chemometric analysis. The LD
50
was estimated at 4x10
5
cfu mL
-1
. Experimental infection of
E.meningoseptica on juvenile catfish was carried out via intraperitoneal (i.p.) route and immersion
(i.m.). E.meningoseptica injected via i.p. route was found to be more potent than i.m. in causing
mortality to juvenile catfish. There was a significant difference (p < 0.05) in the rate of mortality
between i.p. and i.m. group. A significant difference (p < 0.05) was also observed within
immersion groups. Fish in i.p. group showed subcutaneous haemorrhages and lesion in kidney,
while the i.m. group showed hypertrophy, necrosis and removal of dermal layer. In the present
study, clear differences in the metabolite profiles of the different isolates (skin and kidney) were
detected. Guanosine was down-regulated in the serum of catfish infected by E.meningoseptica
isolated from kidney, but it was up-regulated in catfish isolated from skin. On the other hand,
alpha-amyrin was up-regulated in serum of catfish infected by E.meningoseptica isolated from the
skin, but it was down-regulated in catfish isolated from kidney. Alpha-amyrin has strong anti-
inflammatory activity, is a PKA inhibitor as well as a selective protease inhibitor. It has been
reported to prevent molecule oxidation due to UV-B irradiation and prevent skin damage through
prevention of lipid peroxidation and inhibition of PGE
2
release. It also has broad-spectrum
analgesic properties. Guanosine, a purine nucleoside, has important functions in the cellular
metabolism and is used to synthesise enzyme inhibitors, antiviral agents, and anticancer agents.
We conclude that damage in the skin of catfish infected with E.meningoseptica increased its
alpha-amyrin molecules as a response to repair skin damage. Increase in guanosine levels
indicates that there was severe tissue damage in the kidney.


Keywords: catfish; LC-MS QTOF; LD
50
; metabolomics; serum





25

4
th
Australasian Metabolomics
Symposium 2012



Metabolites Profile of Tissue and Plasma in Breast Cancer Patients
Using Metabolomic Approach: Role of Eicosanoid Metabolism

ES Rosli
1
*, NI Mohamed
1
, MZ Salleh
1
, JKS Shia
1
,
M Rohaizak
2
, NS Shahrun
2
,
JJ Saladina
2
, H Roslan
3
, TP Hennessy
1,4
, LK Teh
1
*


1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Department of Surgery,
Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Kuala Lumpur,
Malaysia;
3
UKM Medical Biology Institute (UMBI) and Department of Medicine, Universiti
Kebangsaan Malaysia Medical Centre, (UKMMC), Kuala Lumpur, Malaysia;
4
Life Sciences Group,
Agilent Technologies, Singapore.

*Corresponding authors: syerena@gmail.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Breast cancer is one of the leading causes of death among women worldwide. Many studies had
utilised genomics, transcriptomics and proteomics approaches to understand the regulation and
pathogenesis of the disease. The information is hoped to provide insights to better therapy
outcomes and to help strategise preventive measures. Biomarkers useful for clinical monitoring of
the efficacy of drug treatment among breast cancer patients are heavily searched by researchers.
In this study, we used metabolomics platform to help identify the metabolite profiles which could
be relevant biomarkers in understanding the variation of disease. Ethics approvals from Research
Ethics Committee of UiTM and HUKM were obtained. For this study, tumour and normal tissues of
breast cancer patients were collected from the same patient. Blood was collected from patients
prior to treatment and healthy volunteers based on the inclusion and exclusion criteria. Differential
expressions of metabolites between the two tissues and serum for the patients and healthy
volunteers were investigated. Samples were injected into LC/MS QTOF and the data was
analysed using Mass Profiler Professional (MPP) software for differential analysis. These
metabolic datasets were mined using a range of pattern recognition techniques, including
hierarchical cluster analysis, principal component analysis, partial least squares and neural
networks. Increment of several metabolites in tumour samples has highlighted lipid and eicosanoid
metabolic pathways as potential perturbation in the metabolism of tumour cells in comparison to
normal. Further validation on the metabolites is required before they can be used as breast cancer
biomarkers.


Keywords: breast cancer; biomarker; LC/MS QTOF; lipid and eicosanoid metabolic pathway;
metabolomics














26

4
th
Australasian Metabolomics
Symposium 2012



Oral Communications

Day 2


No. Presenter Title Reference No.
8.
Mohamad Yusof
et al.
Activity-Guided Isolation of the Chemicals Constituents
of Muntingia calabura using Formalin Test
MYMS-008po
9. Aqilah et al.
Isolation and Identification of Aeromonas hydrophila
Isolated from Tilapia, Oreochromis niloticus
MYMS-009po
10. Izwan et al.
Soapdenovo Provides Lower Number of Scaffolds and
Contigs: An Example of Optimum Assembly for Low
Yield Sequencing Data of Acinetobacter baumannii
MYMS-010po
11. Lee et al.
Utilising a Genomics Approach to Investigate the
Mechanism of Antibiotic Resistance in Staphylococcus
aureus
MYMS-011po
12. Hanif et al.
Comparison between r2cat and OSLay Software for
Contigs Arrangement
MYMS-012po
13. Rose Ismet et al.
Whole Genome Sequencing Report of a Malaysian
Aborigine Individual and Annotation of Medico-Genomic
Association Using In-House Genomic Bioinformatics
Workflow
MYMS-013po
14. Hatta et al.
The Patterns of Four Natural Variations in Human
CYP2C9 Genes across Three Populations
MYMS-014po






















27

4
th
Australasian Metabolomics
Symposium 2012



Activity-Guided Isolation of the Chemicals Constituents of Muntingia
calabura Using Formalin Test

MI Mohamad Yusof
1
*, N Ahmad
2
, ZA Zakaria
3
*, MZ Salleh
1
, LK Teh
1


1
Pharmacogenomics Centre, Faculty of Pharmacy, Universiti Teknologi MARA, Puncak Alam,
42300, Selangor, Malaysia;
2
Faculty of Applied Sciences, Universiti Teknologi MARA, Shah Alam,
40450, Selangor, Malaysia;
3
Department of Biomedical Sciences, Faculty of Medicine and Health
Science, Universiti Putra Malaysia, Serdang, 43400, Selangor, Malaysia.

*Corresponding authors: mohd_izwan86@yahoo.com/zaz@medic.upm.edu.my

Abstract

Muntingia calabura is known locally as Kerukup Siam. It has been claimed by the Peruvian folklore
to possess medicinal values which include soothing gastric ulcers, relieving headache and cold,
reducing swelling of the prostate gland and antiseptic. This study focuses on understanding the
antinociceptive effects of the extract using Sprague Dawley (S.D.) rats. In the present study,
activity-guided of methanol extract of Muntingia calabura collected in Shah Alam, Malaysia were
evaluated for their antinociceptive property using the formalin test. Seven fractions of petroleum
ether extract labelled with A, B, C, D, E, F and G were separately treated (orally) with distilled
water or 10 % DMSO as negative controls and morphine and aspirin as the positive control.
Fraction D showed most significant antinociceptive activity when compared to another fraction. At
the dose of 300 mg/kg, fraction D and morphine showed no significant difference in first phase,
while in second phase, fraction D and aspirin showed no significant different. One new compound
together with three known compounds namely 8-hydroxy-6-methoxyflavone, 5-hydroxy-3, 7, 8-
trimethoxyflavone (methylgnaphaliin), 3, 7-dimethoxy-5-hydroflavone and 2, 4-dihydroxy-3-
methoxychalcone were isolated from fraction D.


Keywords: antinociceptive; formalin test; methanol extract; Muntingia calabura


















28

4
th
Australasian Metabolomics
Symposium 2012



Isolation and Identification of Aeromonas hydrophila Isolated from
Tilapia, Oreochromis niloticus

NI Aqilah
1
*, YS Yong
1
, AI Zamani
1
, HH Ruhil
3
, M Nadirah
2
, M Najiah
1


1
Department of Aquaculture Science;
2
Department of Fisheries Science, Faculty of Fisheries and
Aqua-Industry, Universiti Malaysia Terengganu, 21030, Mengabang Telipot, Terengganu,
Malaysia;
3
Faculty of Veterinary medicine, Universiti Malaysia Kelantan, Locked Bag 36,
Pengkalan Chepa 16100 Kota Bharu,Kelantan, Malaysia.

*Corresponding author: n_aqilah610@yahoo.com.my

Abstract

This study was conducted to isolate and identify Aeromonas hydrophila from 30 tilapia fry in
Terengganu, Malaysia. Isolated bacteria were preliminarily identified using GSP agar and thirteen
isolates that showed yellow colony were selected and sub-cultured for further identification using
morphological, biochemical and physiological tests. The phenotypic relationships among isolates
were studied using numerical taxonomy and performed by Unweighted Pair Group Method with
Arithmetic Mean (UPGMA). Data coefficient (S
D
) was used to determine the similarity between the
strain and the data was then compared with previous studies according to Holt el al. (1994) and
Buller (2004). Results show the yellow colony isolates on GSP agar were identified as Aeromonas
hydrophila. Based on numerical taxonomy, isolates show 93.75% and 92.86% similarities from
identification done by Holt et al. and Buller, respectively. From the result, UPGMA and Dice
coefficient were used to estimate the similarities between the samples based on phenotypic
characteristics.


Keywords: Aeromonas hydrophila; data coefficient; Oreochromis niloticus; UPGMA

























29

4
th
Australasian Metabolomics
Symposium 2012



Soapdenovo Provides Lower Number of Scaffolds and Contigs: An
Example of Optimum Assembly for Low Yield Sequencing Data of
Acinetobacter baumannii

Izwan I*, LK Teh, LS Lee, Hanif AK, Irma SZ, MZ Salleh*

Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia.

*Corresponding authors: mohamadizwanbinismail@yahoo.com/zakisalleh@puncakalam.uitm.edu.my

Abstract

Low sequencing yield has been a bane for bioinformaticians, as it limits the output of the data
analysis that follows. This paper aims to provide a comparison between two well-received de novo
assemblers in order to aid in maximising the use of such low yield data. Sequencing data of an
Acinetobacter baumannii strain was selected with data size approximately half of the expected
output, and an overall coverage of at least 50X. The 36 bp short reads were sequenced using the
Illumina GA IIx. The quality scores of the reads were analysed via FastQC, and the sequences
were assembled using both SOAPdenovo (V1.05) and VelvetOptimiser (V2.2.2). The number of
contigs/scaffolds produced using the two software were compared. The optimal syntenic layout
and number of ORFs were compared using OSLay and PRODIGAL, respectively. The assembly
using SOAPdenovo produced 320 scaffolds, whereas VelvetOptimiser yielded 3091 contigs.
Analysis using OSLay revealed better syntenic layout between SOAPdenovo scaffolds and
references A.baumannii ACICU, AYE, ATCC 17978, and SDF. Using the gene prediction script
PRODIGAL, 4497 ORFs for SOAPdenovo-assembled sequences and 4208 ORFs for
VelvetOptimiser-assembled sequences were generated. It is concluded that SOAPdenovo is more
suited for handling low yield data for tertiary analysis.


Keywords: Acinetobacter baumannii; Soapdenovo; VelvetOptimiser























30

4
th
Australasian Metabolomics
Symposium 2012



Utilising a Genomics Approach to Investigate the Mechanism of
Antibiotic Resistance in Staphylococcus aureus

LS Lee
1,2
*, LK Teh
1
, ZF Zainuddin
2
, MZ Salleh
1
*


1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
School of Health
Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia.

*Corresponding authors: lianshien@gmail.com/zakisalleh@puncakalam.uitm.edu.my

Abstract

With the rapidly decreasing cost of whole genome sequencing, genomics has become a viable
approach in examining various facets of the mechanism of antibiotic resistance of pathogenic
bacteria. We seek to examine the utility of this approach by sequencing a methicillin-resistant
Staphylococcus aureus (MRSA) strain isolated from a patient with septicaemia in Kuala Lumpur.
The strain was typed using multi-locus sequence typing (MLST) and disk susceptibility tests were
performed to identify its resistance pattern. The genome sequence was obtained using 104 Mb of
paired-end (300 bp spacing) data from the Illumina GA IIx platform with 36-bp reads. Sequence
data were assembled using the CLCBio Genomics Workbench. The resultant 195 contigs were
overlaid with the Mu50 reference sequence using OSLay to generate fourteen scaffolds. Gaps
were closed using Sanger sequencing. The genome was annotated using BASys and RAST. The
clinical isolate, termed MRSA PR1, has been identified as sequence type 239 (ST239), a health
care-associated MRSA lineage prevalent in Asia, South America, and Eastern Europe that is
typically defined by its extended antibiotic and antiseptic resistance pattern. This is consistent with
the findings of the disc susceptibility tests that show that MRSA PR1 is resistant to oxacillin,
cefuroxime, gentamicin, erythromycin, ciprofloxacin and co-trimoxazole. The MRSA PR1 genome
consists of a 2,725,110-bp circular chromosome with a GC content of 32.6%. It contains 2638
CDS and 19 rRNA features. MRSA PR1 harbours SCC mec type III, which consists of a
composite element comprising SCCmec and SCCmercury. This region harbours ccrC, pI258 and
Tn554 as well as several genes involved in cadmium resistance. Several resistance features were
identified such as the qacA gene which confers resistance to certain antiseptics via an export-
mediated mechanism, and norA which confers resistance to hydrophilic quinolones. This study
provided the whole genome sequence of a locally isolated MRSA strain and identified several key
features that are involved in conferring antibiotic resistance.


Keywords: antibiotic resistance; genomics; methicillin resistance; MRSA; resequencing;
Staphylococcus aureus; whole genome sequencing











31

4
th
Australasian Metabolomics
Symposium 2012



Comparison between r2cat and OSLAY Software for Contigs
Arrangement

Hanif AK*, LS Lee, Izwan I, MH Zulkifli, Azwin Ismail, Irma SZ, LK Teh, MZ Salleh*

Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia.

*Corresponding authors: ikhmalhanif87@yahoo.com/zakisalleh@puncakalam.uitm.edu.my

Abstract

With the high throughput sequencing technology, it has become easier and faster in obtaining
accurate sequences of a genome. This results in tremendous short reads of the sequenced
genome. The data will be assembled using de novo assembler in acquiring novel genes or
variants of the organisms. The contigs that are assembled would be mapped to related species of
high homology before annotation is performed. In this study, the characteristics of the contigs
obtained using r2cat and OSLay will be compared. Proteus mirabilis, Klebsiella pneumonia, and
Streptococcus agalactiae were isolated from samples obtained from septicaemic patients at
Hospital Putrajaya and Hospital Universiti Sains Malaysia. DNA was extracted and libraries were
prepared for whole genome sequencing using Genome Analyser IIx. Output of sequencing data
was assembled using CLCBio version 4.9 and Velvet Optimiser. The contigs of P. mirabilis, K.
pneumonia, and S. agalactiae was assorted using r2cat and OSLay then validated using CLCBio
in map to reference method. The characteristics compared using r2cat and OSLay include contigs
arrangement, number of scaffold produced, total contigs used. For P. mirabilis strain PM001,
OSLay produced nine scaffolds using 96 contigs; the remaining 30 contigs were not able to be
mapped to reference. Meanwhile, r2cat produced one scaffold using 125 contig and leaving one
contig unmapped. For K. pneumonia, OSLay produced twelve scaffolds using 78 contigs out of
122 contigs while r2cat produced one scaffold without any contig unused. Lastly, for S. agalactiae,
OSLay produced six scaffolds and 10 out of 76 contigs were left unused. However, using r2cat
software, scaffolds produced was invalid as more than 80% of the reads were not mapped due to
wrong contigs arrangement. Therefore, OSLay is the preferred choice and provide users the ability
to change parameters for the assembly to achieve optimum result.


Keywords: K. pneumonia; OSLAY; P. mirabilis; r2cat; S. agalactiae















32

4
th
Australasian Metabolomics
Symposium 2012



Whole Genome Sequencing Report of a Malaysian Aborigine
Individual and Annotation of Medico-Genomic Association Using In
House Genomic Bioinformatics Workflow

Rose Ismet
1
*, Husaini I
1
, M Nornazliya
1
, Shafiq A
1
, Z Bannur
1
, LS Lee
1
, LK Teh
1
, Zafarina
Z
2
, Norazmi MN
2
, BP Hoh
3
, Aminuddin A
3
, Fadzilah MN
3
, T Rahman
3
, R Razali
3
, Adzrool
Idzwan Ismail
4
, Kamarudzaman Md. Isa
4
, Mustaffa Halabi Azahari
4
, MZ Salleh*

1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
School of Health
Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia;
3
Faculty of Medicine, UiTM Sungai
Buloh, Selangor, Malaysia;
4
Faculty of Art & Design, UiTM Shah Alam, Selangor, Malaysia.

*Corresponding authors: rose_ismet@yahoo.com/zakisalleh@ puncakalam.uitm.edu.my

Abstract

We believed that the Malaysian Aborigine is a unique group of population, especially the studied
sub-tribes who are decreasing in population number. It is known that the Aborigines genetic
prehistory is affected by genetic drift and natural selection and their gene pool is richly textured by
biocultural and evolutionary processes as well as gene flow or diffusion. However, there is no
whole genome sequencing data of the Malaysian Aborigine available to date. Having the genetic
blueprint of the aborigine could facilitate the understanding of disease and xenobiotic association
as every disease carries its own genetic element be it as an inherited trait or by a response to
environmental stressors. This in turn could lead to the understanding of why these aboriginal
populations decrease in numbers. What is observed at present is that the Malaysian Aborigine is
presented with conditions including G6PD deficiency, hepatosplenomegaly as well as skin
infections. Resistance to malaria has also been observed among the Temuan presented with
G6PD deficiency and elliptocytosis. By performing whole genome sequencing, these findings
could be further confirmed. Thus, we aim to understand the disease risk associated with the
Malaysian Aborigine which could contribute to their fragility and endangerment. We have
sequenced a Che Wong subject using Next-Generation Sequencing (NGS) platform where the
data was then subjected to primary analysis utilising the CASAVA v1.8.2 software. Data
generated for the sample is at a satisfactory quality with 39.3x coverage. A total of 3,786,707
SNPs was successfully called. Short INDELs (< 300 bp) called include 337,269 insertions and
349,602 deletions. Larger structural variants (> 300 bp) detected include 3 insertions, 2,306
deletions and 1,282 CNVs. Further downstream analysis is warren in order to identify the
structural variants associated with diseases among the Che Wong sub-tribe. The ability to identify
the genetic variants that contribute to disease and variable pharmacological responses would aid
the understanding of disease risk and susceptibility.


Keywords: disease risk and susceptibility; Next-Generation Sequencing; pharmacological
responses; structural variants








33

4
th
Australasian Metabolomics
Symposium 2012



The Patterns of Four Natural Variations in Human CYP2C9 Genes
across Three Populations

Fazleen HM Hatta
1, 2
*, LK Teh
1
, MZ Salleh
1
, M Goktas
4
, U Yasar
4
, HK Roh
3
, E Aklillu
2
, L
Bertilsson
2


1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Division of Clinical
Pharmacology, Department of Laboratory Medicine, Karolinska InstitutetKarolinska University
Hospital, Huddinge 141 86 Stockholm, Sweden;
3
Division of Clinical Pharmacology, Department of
Internal Medicine, Gachon University Hospital, Incheon, Korea;
4
Department of Pharmacology,
Hacettepe University Faculty of Medicine, Sihhiye, 06100 Ankara, Turkey.

*Corresponding author: fazleenhaslinda@gmail.com

Abstract

With more than 30 variant alleles, there are at least eight known single nucleotide polymorphisms
(SNPs) that can cause poor drug metabolism by CYP2C9 enzymes. But interestingly, previous
studies revealed unique interindvidual and interethnic variations observed within a population and
among populations of selected individuals that are not carrying a defective allele. In this study, we
aim to search for a relationship between inter-population metabolism of CYP2C9 and its
genotypes. DNA from 85 Swedish, 128 Korean and 51 Turkish healthy volunteers were assessed
for four natural variations [SNP 1 (IVS1+83T>C), SNP 2 (IVS2+73T>C) and SNP 3 (IVS6+95A>G)
and SNP 4 (IVS8109A>T)] of the CYP2C9 gene. PCR/RFLP and allele specific PCR methods
were developed to screen the healthy volunteers excluding individuals carrying CYP2C9*2 or *3
alleles. Metabolic Ratio of losartan/metabolite E-3174 was used as a phenotyping measure. We
found a significant relationship between SNP 4 (IVS8109A>T) and CYP2C9 activity (
2
test,
p=0.011) in the Swedes. No significant relationship between any of the SNPs and MR was
established for Korean and Turkish individuals (
2
test, p=0.154 and 0.355 respectively). Although
a majority of 14% of Swedes and 16% of the Turks are wild type for all four variants, a higher
number of 40% of the Korean are wild type for these variations. We also found that SNP 1, 2 and
3 are very rare in the Korean population (1.6%, complete linkage). We found that SNP 4 IVS8 -
109 T allele is associated with a higher MR of CYP2C9 in healthy Swedish subject, but further
investigations need to be carried out to established a molecular explanation for inter-population
differences of CYP2C9 catalysed metabolism. Haplotypes based on SNPs 1-4 did not seem to
contribute to variation in the MR of the Koreans nor play a role in determining the MR of the
Korean or Turkish individuals.


Keywords: CYP2C9; Koreans; metabolic ratio; natural variations; Swedes; Turkish















Designed by: Mohd Izwan Ismail, PROMISE




4
th
Australasian Metabolomics
Symposium 2012



















ABSTRACTS
Poster Communications























36

4
th
Australasian Metabolomics
Symposium 2012



Poster Communications

No. Presenter Title Reference No.
1. Ashrafzadeh et al.
Non-labelled Relative Comparison of Kedah Kelantan
(Bos indicus) and Mafriwal (Bos taurus Bos indicus)
Sperm Proteins for Fertility Molecular Marker Discovery
MYMS-001pt
2. Hashim et al.
Metabolite Profiles of Agarwood Leaves Aqueous
Extracts
MYMS-002pt
3. Mei et al.
Lipid Profiling At Three Main Stages of Oil Palm Fruit
Mesocarp Development
MYMS-003pt
4. Bannur et al.
Identifying Potential Biomarkers for Colorectal Cancer
Using Metabolomics Approach
MYMS-004pt
5. Derahman et al.
Search of Serum Markers for Aspirin Resistance in
Cardiovascular Disease (CVD) Patients Using
Metabolomics Approach
MYMS-005pt
6. Ebrahimi et al.
Reproducibility and Stability of NMR Spectroscopy and
the Method Development for Urine Metabolomic
Analysis
MYMS-006pt
7. Nornazliya et al.
Comparison of the Metabotypes of the Malaysian
Aborigines with the Urban Healthy Individuals and
Cardiovascular Patients: Signature of Longer Life
Span?
MYMS-007pt
8. Rofiee et al.
Alteration of Metabolite Profile in Acetaminophen
Induced Hepatotoxic Rats
MYMS-008pt
9. Sundram et al.
(E)-labda-8(17),12-diene-15,16-dial Production from
Curcuma mangga (Mango ginger) In Vitro Cultures
MYMS-009pt
10.
Nurul Shahfiza et
al.
Discrimination between Infected Dengue Patient and
Healthy Individual Based On Chemometry of
1
H NMR
Spectroscopy
MYMS-010pt
11. Ali et al.
The Use of LC-MS QTOF-Based Metabolomics to
Define the Potential of Probiotic against Colorectal
Cancer Xenograft in the Mouse Model
MYMS-011pt
12. Mohamed et al.
Patients with Combined Variant of CYP2D6 and Wild-
type ABCB1 C3435T Developed Early Recurrence and
Metastasis: Role of Sphinganine
MYMS-012pt
13. Enche Ady

et al.
Metabolic Profiling for Early Detection of Alzheimers
Disease
MYMS-013pt
14.
Noorzaid Muhamad
et al.
Some Aspect of Urea Metabolism in Parasites Helminth
Teladorsagia circumcincta
MYMS-014pt
15. Shafiq et al.
Molecular Characterisation of Previously Untypeable
Rotavirus
MYMS-015pt


37

4
th
Australasian Metabolomics
Symposium 2012


16. Irma et al.
Insight into Genome and Pathogenicity of Opportunistic
Human Pathogen, Streptococcus agalactiae
MYMS-016pt
17. Husaini et al.
ANNOVAR: A Useful Tool for Functional Annotation of
Genetic Variants of a Human Genome
MYMS-017pt
18. Zulkifli et al.
Whole Genome Sequencing and Comparative Analysis
of Local Klebsiella pneumoniae Strain against Klebsiella
pneumoniae Ntuh-K2044
MYMS-018pt
19. Azwin Ismail et al.
Identification of Interesting Genomic Properties of
Clinical Isolates of Mycobacterium tuberculosis
MYMS-019pt
20. Norfatimah et al.
Comparative Study Using Bioinformatics Pipeline for
Malaysian Mahseer (Tor tambroides) Mitogenome
Assembly
MYMS-020pt
21. Hasbullani et al.
HLA-B*1502 is a Significant Marker for Carbamazepine-
induced Stevens Johnson Syndrome (SJS): Pre-
Implementation Trial of HLA-B*1502
Pharmacogenotyping in a Local Hospital in Malaysia
MYMS-021pt
22. Muda et al.
Genetic Polymorphism of CYP3A4*18 and CYP3A5*3 in
Malaysian Kidney Transplant Patients
MYMS-022pt
23. Abdul Jalil et al.
The Implication of the Polymorphism of UGT1A6 among
Cardiovascular Disease (CVD) Patients treated with
Aspirin
MYMS-023pt
24. Che Omar et al.
Genetic polymorphism of CYP2C19 among the
Cardiovascular Patients of Three Ethnic Groups in
Malaysia
MYMS-024pt
25. Mohamed Ali et al.
Development of a Pharmacogenotyping Method for
Detection of HLA Polymorphism in Personalising
Allopurinol Therapy
MYMS-025pt
26. SS Mamat et al.
Hepatoprotective Activity of Methanol Extract of
Melastoma malabathricum Leaves on Paracetamol-
Induced Liver Injury in Rat
MYMS-026pt
27. Samuel Oii et al.
Chemopreventive Potential of Melastoma
malabathricum Leaves Extract in DMBA/Croton Oil-
Induced Mouse Skin Carcinogenesis
MYMS-027pt
28. Bendt et al.
Mycolic Acids as Diagnostic Markers for Tuberculosis
Case Detection and Drug Efficacy
MYMS-028pt
29.
Narayananaswamy
et al.
A Sensitive and Efficient Method for the Quantification
of Phosphorylated Sphingoid Bases in Biological
Samples
MYMS-029pt






38

4
th
Australasian Metabolomics
Symposium 2012



Non-Labelled Relative Comparison of Kedah Kelantan (Bos indicus)
and Mafriwal (Bos taurus Bos indicus) Sperm Proteins for Fertility
Molecular Marker Discovery

Ali Ashrafzadeh
1
*, Sheila Nathan
1
, Iekhsan Othman
2
, Ting Yee Tee
2
, Saiful Anuar
Karsani
3

1
School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti
Kebangsaan Malaysia, Selangor, Malaysia;
2
School of Medicine and Health Sciences, Monash
University, Sunway Campus, Kuala Lumpur, Malaysia;
3
Institute of Biological Sciences, Faculty of
Science, & University of Malaya Centre for Proteomics Research, University of Malaya, Kuala
Lumpur, Malaysia.

*Corresponding author: ali_ash_3000@yahoo.com

Abstract

Malaysia is still reliant on importation of up to 80% of the nations dairy and beef requirements. In
an attempt to improve local beef and dairy production, cross breeding with high producing breeds
is generally favoured. However, fertility of the crossbred progenies is low as a result of the high
humidity and temperature. This study was undertaken to identify differences in sperm proteome
between the high fertile Malaysian indigenous breed (Kedah Kelantan) and the low fertile
crossbred cattle (Mafriwal) to identify proteins that may be potentially fertile and heat tolerance
molecular marker(s). Frozen semen of three high performance bulls from each breed was
processed to obtain live and pure sperm. Protein separation was conducted and gel bands were
processed for in-gel digestion. For each breed, mass spectrometry data was acquired over 11
replicates. Analysed data identified compounds of different expression levels (99% confidence
level) and protein identification was determined by targeted MS/MS. Among the identified proteins,
fifteen motility and fertility related proteins were up-regulated in Kedah Kelantan. Sperm motility
evaluation by CASA (Computer Assisted Semen Analysis) confirmed significantly higher motility in
Kedah Kelantan sperm compared to Mafriwal. Further analysis of these proteins will allow us to
evaluate their potential as selective markers indicating highly fertile and more environmentally
compatible bulls for breeding purposes in tropical areas.


Keywords: Bos indicus; Bos Taurus; mass spectrometry; molecular marker(s); sperm proteome

















39

4
th
Australasian Metabolomics
Symposium 2012



Metabolite Profiles of Agarwood Leaves Aqueous Extracts

YZH-Y Hashim
1,2
*, Ismail I
1
, Salleh F
1, 2


1
Bioprocess and Molecular Engineering Research Unit (BPMERU), Department of Biotechnology
Engineering, Kulliyyah of Engineering, International Islamic University Malaysia,
P.O. Box 50728, Kuala Lumpur, Malaysia;
2
International Institute for Halal Research and Training
(INHART), Ground Floor, Block E0, Kulliyyah of Engineering Building, International Islamic
University Malaysia, P.O. Box 50728, Kuala Lumpur, Malaysia.

*Corresponding author: yumi@iium.edu.my

Abstract

Agarwood has been known for its invaluable aromatic resin while other parts of the plant are not
fully utilised. Although there are various ethnopharmacological evidences describing the health
beneficial effects of other parts of the plant, very few scientific studies were undertaken to confirm
these claims. This study aims to investigate the metabolites present in water extract of agarwood
leaves. Fresh agarwood leaves from uninoculated trees were collected, washed and dried at room
temperature for two weeks prior to grinding. The powdered leaves (1 g) were then extracted in a
volume of distilled water (30 ml, 40 ml and 50 ml) at 120 rpm, 30 C for 16 hours. Filtrate was
centrifuged at 22000 g for 15 min and supernatant collected. For gas chromatography mass
spectrometry analysis, 150 l of supernatant was transferred to 1.5 ml reaction tube, dried using
vacuum concentrator (30C) for 2 hours and derivatised. The derivatised sample was transferred
to the GC vial prior to analysis in Agilent 7890A GC system coupled to 5975C Inert XL MSD using
HP-5MS column. Data was then subjected to Partial Least Square Discriminant Analysis (PLSDA)
using SIMCA-P+ v12.0 (Umea, Sweden). Although all extracts shared compounds such as
propanoic acid, aspartic acid, proline and fructose oxime, PLSDA clearly separated the
compounds extracted based on ratio of solid to solvent with R2 of 0.698 and Q2 of 0.743.
Samples with 1:40 ratio gave the highest metabolite number ranging from 39 to 64 and were
dominated by gluconic acid, hexadecanoic acid, mannonic acid and dodecyl acrylate. The
metabolites revealed in the leaves water extract may further be isolated for use as raw
ingredients. This would add value to the agarwood leaves which otherwise would be considered
as waste.


Keywords: agarwood; aqueous extract; gas chromatography mass spectrometry; metabolites










40

4
th
Australasian Metabolomics
Symposium 2012



Lipid Profiling At Three Main Stages of Oil Palm Fruit Mesocarp
Development

Theresa Ng Lee Mei*, Tiong Soon Huat, Neoh Bee Keat, Teh Huey Fang, Thang Yin
Mee, Mohd Amiron Ersad,

David R. Appleton

1
Sime Darby Technology Centre Sdn. Bhd., 1
st
Floor, Block B, UPM-MTDC Technology Centre III,
Lebuh Silikon, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia

*Corresponding author: theresa.ng.lee.mei@simedarby.com

ABSTRACT
Oil palm is well known for its effectiveness as oil storage whereby more than 90% of its oil is
stored in fruit mesocarp. The oil accumulates over time at different developmental stages starting
from the lag stage [11 to 14 week after anthesis (WAA)] to lipid biosynthesis stage [15 to 19 WAA]
and finally to the ripening stage [20 to 24 WAA]. In this study, an extensive lipid research is carried
out using targeted metabolomics approach on all the three main stages of oil palm fruit mesocarp
development. Lipids analysis comprised of the total oil content, fatty acid composition,
triacylglycerols (TAG) and phospholipids. The metabolites were extracted and profiled by multiple
platforms such as the Gas Chromatography Mass Spectrometry (GCMS) and Liquid
Chromatography Mass Spectrometry (LCMS). Results showed that the total oil content
increases from first to the third phase with palmitic acid recorded the highest composition followed
by oleic and stearic acids respectively. At the same time, the amount of phospholipids decreases
while TAG accumulation increases, as oil is deposited in the form of TAG across the three main
stages. In all, the different lipid class of oil palm fruit mesocarp development has been
successfully classified and would definitely be useful for palm genotypic studies.

Keywords: metabolites; oil palm; triacylglycerols
























41

4
th
Australasian Metabolomics
Symposium 2012



Identifying Potential Biomarkers for Colorectal Cancer Using
Metabolomics Approach

Z Bannur
1
*, H Hashim
1
, MZ Salleh
1
, TP Hennessy
1, 2
, JKS Shia
1
, Azmi MN
3
, Zailani MH
3
,
Ramasamy P
3
, ZA Zakaria
1,4
, LK Teh
1
*


1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Life Sciences Group,
Agilent Technologies, Singapore;
3
International Islamic University Malaysia, Kuantan 25710
Malaysia;
4
Department of Biomedical Science, Faculty of Medicine and Health Sciences,
University Putra Malaysia (UPM), 43400 Serdang, Selangor, Malaysia.

*Corresponding authors: knight_ly@yahoo.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Colorectal cancer (CRC) is one of the most common malignant diseases among men and women
in Malaysia. Management of CRC among different individuals is complicated as it depends on
many factors such as stage of cancer, environmental factors and type of regimen used to treat
colorectal cancer. Systematic approaches using specific biomarkers to evaluate the risk for
individual patients are therefore desired. The emerging field of LC/MS based global metabolomics
approach provides a platform to help unravel the mechanisms involved in the diseases and is
useful to understand various biological processes in the living system as they are the
intermediates and the end products of metabolism.

This study aims to identify potential biomarkers
for CRC by comparing the metabolites profiles between colorectal cancer patients treated with 5-
fluorouracil (CRC-5-FU) and healthy control using metabolomics approach. Several classes of
metabolites that were down regulated in the CRC-5-FU group include dicarboxylic acid (DA),
polyunsatureated fatty acids (PUFA) and amino acids and conjugates. Conversely, metabolites
that were up-regulated in CRC-5-FU include uremic toxins, bilirubin, bile acid and its conjugates
and fatty acids. Our results confirmed the alteration of metabolic profile in patients with colon
carcinoma which involve the alteration of pathways in the metabolism of bile acid and fatty acid as
well as glycolysis pathways. The distinctive metabolite profiles of CRC-5-FU patients and healthy
control provides clues of several biomarkers or pattern for detection of cancer.


Keywords: 5-FU; biomarkers; CRC; glycolysis pathway; metabolomics


















42

4
th
Australasian Metabolomics
Symposium 2012



Search of Serum Markers for Aspirin Resistance in Cardiovascular
Disease (CVD) Patients Using Metabolomics Approach

A Derahman
1
*, MZ Salleh
1
, O Maskon
2
, NJ Abdul Jalil
1
, RN Che Omar
1
, MI Ismail
1
, ES
Rosli
1
, M Nornazliya
1
, MS Rofiee
1
, Z Bannur
1
, TP Hennessy
3
, JKS Shia
1
, LK Teh
1
*

1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Department of Cardiology,
Hospital Universiti Kebangsaan Malaysia, 56000 Cheras, Kuala Lumpur, Malaysia;
3
Life Sciences
Group, Agilent Technologies, Singapore.

*Corresponding authors: gurlz_aida@yahoo.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Cardiovascular disease (CVD) is the number one cause of death which covers a wide array of
disorders, including diseases of the cardiac muscle and vascular system. Aspirin has been used
as an analgesic-antipyretic for a long time and with lower doses, it is useful to prevent blood clots.
However, in some patients, aspirin fails to protect them from thrombotic complications; this
phenomenon is known as aspirin resistance. Current study aims to identify biomarkers involved
in aspirin resistance in which they can be used to identify who would response better to aspirin
and those who may not response, so alternative drugs can be given at initiation of drug treatment
for prevention of thrombotic complications. Resistance was defined in this study as occurrence of
secondary cardiovascular events despite initiation of aspirin therapy in patients. Ethics approval
were obtained from Research Ethics Committee of Universiti Teknologi MARA (UiTM) and
Hospital Universiti Kebangsaan Malaysia (HUKM) for the conduct of this project. Blood serum was
clean up using cold acetonitrile. Samples were then reconstituted with mobile phase and were
injected into LC/MS-QTOF. Data was acquired using Agilent MassHunter Data Acquisition
software. Data was analysed using software which includes Agilent MassHunter Qualitative
Analysis, Agilent Mass Profiler Professional and Agilent METLIN Professional Metabolite
Database. A total of 106 metabolites were detected. Dendrogram of Hierarchical Clustering
Analysis (HCA) was used to visualise the up- and down-regulation of the expression of
metabolites. The browser of the Human Metabolome Database (HMDB) pathway identified ten
pathways and eight metabolites of interest. Of these metabolites, bilirubin and glycocholic acid
were down-regulated while deoxycholic acid glycine conjugate and prostaglandin G2 were up-
regulated. Prostaglandin G2 which is involved in arachidonic acid pathway may possibly have
relationship with aspirin resistance. However, this requires further validation before it can be used
as biomarker in monitoring aspirin resistance.


Keywords: aspirin resistance; biomarkers; cardiovascular disease (CVD); metabolomics













43

4
th
Australasian Metabolomics
Symposium 2012



Reproducibility and Stability of NMR Spectroscopy and the Method
Development for Urine Metabolomic Analysis

F Ebrahimi
1
*,

KL Chan
1
, F Fang
2
, P Sprenger
2
, V Murugaiyah
1
, CH Teh
2
, B Ibrahim
1
*


1
School of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800 Penang, Malaysia;
2
Bruker
Malaysia.

*Corresponding authors: ebrahimi.foroogh@gmail.com/baharudin.ibrahim@usm.my

Abstract

Accuracy and stability of NMR spectroscopy are vital in urine metabolomics study. Meticulous
method development further ensures the achievement of reliable results. The clustering of sample
groups on the NMR spectra in Orthogonal Partial Least Squares (OPLS) graph could provide
some insight on NMR stability and accuracy. The present study determined the stability and
reproducibility of the applied methodology including the use of a 500 MHz NMR instrument for
urine metabolomics analysis. Twenty urine samples of healthy volunteers were collected and
stored in a -80C freezer. Prior to NMR analysis, the samples were thawed and eight samples
were pooled together and vortex-mixed. Eight samples of 900 l were taken from the pooled
sample and mixed with 100 l of urine buffer (containing phosphate buffer, sodium azide and
trimethylsilyl propanoic acid) in eppendorf tubes. After 10 min of centrifugation, a 600 l of the
supernatant was transferred to a 5 mm NMR tube. Another 12 individual samples (unpooled) were
also prepared in the same manner. The temperature of the NMR spectrometer was calibrated at
300K, and phase correction, baseline correction, tuning and matching, shim optimisation, water
suppression and acquisition parameter optimisation were done. The NMR spectra were binned to
0.04 ppm and the data were analysed using OPLS-Discriminant Analysis (OPLS-DA) to determine
the clustering and separation between samples. The NMR spectra exhibited peaks of high
reproducibility. OPLS-DA analysis yielded a two-component model with R
2
Y=0.99 and Q
2
Y=0.75.
A tight clustering was observed in the pooled samples whilst the unpooled samples were loosely
clustered. A clear separation between pooled and unpooled groups was also observed. The
results showed that the applied urine metabolomics technique and the NMR spectrometer were
stable and reproducible to differentiate between groups. Future works on medicinal plants and
their effects on rat urine metabolomics are on-going.


Keywords: method development; Orthogonal Partial Least Squares; reproducibility; urine
metabolomics
















44

4
th
Australasian Metabolomics
Symposium 2012



Comparison of the Metabotypes of the Malaysian Aborigines with the
Urban Healthy Individuals and Cardiovascular Patients: Signature of
Longer Life Span?

M Nornazliya
1
*, MZ Salleh
1
, MI Ismail
1
, MS Rofiee
1
, ES Rosli
1
, A Derahman
1
, Z Bannur
1
,
Rose Ismet
1
, Husaini I
1
, Shafiq A
1
, Aminuddin A
2
, Fadzilah MN
2
, T Rahman
2
, R Razali
2
,
Adzrool Idzwan Ismail
3
, Kamarudzaman Md. Isa
3
, Mustaffa Halabi Azahari
3
, TP
Hennessy
1,4
, LK Teh
1
*

1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Faculty of Medicine, UiTM
Sungai Buloh, Selangor, Malaysia;
3
Faculty of Art & Design, UiTM Shah Alam, Selangor,
Malaysia;
4
Life Sciences Group, Agilent Technologies, Singapore.

*Corresponding authors: nornazliyamohamad@yahoo.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Orang Asli are believed to have different metabotypes compared to the urban people based on
their lifestyles, habitat, culture, and nutrition behaviour. The urban people are faced with many
diseases such as cardiovascular diseases and cancer, while the Orang Asli was commonly
reported to have skin infection, worm infestations and tuberculosis. These diseases could be due
to the differences in lifestyles. Currently, data about cardiovascular diseases in Orang Asli are
lacking and this lead to the hypothesis that the Orang Asli may be protected from these diseases
due to their lifestyles and diet behaviour. This study is preliminary and aims to investigate the
differences in the metabotypes of the Orange Asli and a group of cardiovascular patients using
global metabolomics approaches. Protein precipitation of serum sample was done using
acetonitrile and the samples were reconstituted with mobile phase prior analysis using LC/MS-
QTOF. The metabolic profiles of the Orang Asli were compared with the profiles of the urban
healthy individuals and cardiovascular patients. Principal component analysis (PCA) showed an
obvious clustering of metabolites in the PCA score plot. ANOVA results showed a significant
difference in the expression of 92 out of 338 metabolites detected among the Orang Asli, healthy
urban and urban patients with cardiovascular diseases (p<0.001). One particular metabolite of
interest is indoleacrylic acid which was down-regulated in both healthy urban and Orang Asli and
up-regulated in CVS patients. Indoleacrylic acid has been reported to inhibit tryptophan synthetase
and thus resulting in lower level of tryptophan and its metabolites such as serotonin and
melatonin. Hence this may cause depression and sleep disorders in those with lower serotonin
and melatonin respectively. Interestingly, lower tryptophan signifies longer life span, and potential
protection from insulin resistance. This result is preliminary and requires further validation of its
significance in protecting Orange Asli from life-styles related diseases, but it provides insight for
the researchers to navigate deeper for more answers.


Keywords: cardiovascular; metabolomics; lifespan; Orang Asli; tryptophan









45

4
th
Australasian Metabolomics
Symposium 2012



Alteration of Metabolite Profile in Acetaminophen Induced Hepatotoxic
Rats

MS Rofiee
1
*, MI Ismail
1
, ES Rosli
1
, JKS Shia
1
, MZ Salleh
1
, ZA Zakaria
2
, TP Hennesy
1,3
,
LK Teh
1
*

1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Department of Biomedical
Sciences, Faculty of Medical and Health Sciences, Universiti Putra Malaysia, Malaysia;
3
Life
Sciences Group, Agilent Technologies, Singapore.

*Corresponding authors: bio_salleh82@yahoo.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Acetaminophen (N-acetyl-p-aminophenol) or paracetamol is usually used as an analgesic and
antipyretic. Acetaminophen is safe at standard recommended dose, but it may produce acute
centrilobular hepatic necrosis, when given at high doses in humans and experimental animals.
This study aims to further understand the mechanism and metabolic changes occur due to hepatic
necrosis induced by acetaminophen. LC/MS-QTOF was used to profile the characteristic changes
in the endogenous metabolites in response to acetaminophen overdose in rats. Hepatotoxicity
was induced in a group of rats (6 rats) by oral administration of acetaminophen (300 mg/kg). Body
weight, liver weight of the rats and gross picture of the liver were documented at 48 hours after
administration of acetaminophen to confirm hepatotoxicity. Serum was collected pre and post
toxicity event. Protein precipitation of serum sample was performed using cold acetonitrile.
Sample were dried and reconstituted with mobile phase prior analysis using LC/MS-QTOF. A total
of 133 compounds satisfying a p-value with cut off 0.05 and fold change cut off 2 were identified.
Principal component analysis (PCA) showed an obvious clustering of metabolites for pre and post
toxicity event. Compounds up-regulated in post toxicity event include p-acetamidophenyl
glucoronide, p-acetamidophenol, 5-hydroxyhydrodolasetron, 2-hydroxy-4 methlvaleric acid and
Stearoylcarnitine; while two compound (cytosine and 5,8-heptadecadiynoic acid) was down
regulated in the treated group. However, further study is required to validate the function of each
of these compounds.


Keywords: APAP; liver toxicity; metabolomics


















46

4
th
Australasian Metabolomics
Symposium 2012



(E)-labda-8(17),12-diene-15,16-dial Production from Curcuma mangga
(Mango ginger) In Vitro Cultures

Tamil CM Sundram
1
*, Lee Guan Serm
2
, Sri Nurestri Abdul Malek
2
, M Suffian M Annuar
1
,
Norzulaani Khalid
1
*

1
CEBAR, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala
Lumpur, Malaysia ;
2
Biochemistry Department, Faculty of Science, University of Malaya, 50603,
Kuala Lumpur, Malaysia.

*Corresponding author: lani@um.edu.my/tgm_tam84@yahoo.co.uk

Abstract

Curcuma mangga Val., (Mango ginger) has been used traditionally as a seasoning for food and
treatment for stomach aches, fever, and cancer related diseases. (E)-labda-8(17),12-diene-15,16
dial is one of the major compounds isolated from rhizomes of Curcuma mangga. Malek et al.
(2011) has demonstrated that this bioactive compound showed cytotoxic effects against six
human cancer cell lines. In this research, callus and cell suspension cultures of Curcuma mangga
were established to be tested as an alternative source of (E)-labda-8(17), 12-diene-15,16 dial. In
vitro cultures will be advantageous compared to field grown rhizomes since in vitro sources is
possible to ensure continuous supply of consistently high quality compounds. Friable calli were
initiated from small and active shoot buds of the rhizomes, when cultured on MS (Murashige and
Skooq) basal medium supplemented with 1 mg/L 2.4-D and 3% sucrose under dark condition. The
growth of callus culture was assessed and harvested fortnightly at different growth stages. After 4
to 8 weeks, rapidly growing friable callus were also transferred and maintained in MS liquid
medium with 1 mg/l 2.4-D and 30 g/L sucrose to obtain liquid suspension culture. Rapidly
proliferating and dense cells were harvested every three days. (E)-Labda-8(17), 12-diene-15,16
dial was extracted from cells, callus, rhizomes and shoots using solvent extraction method. Gas
Chromatography (GC) and Gas Chromatography Flame Ionisation Detector (GCFID) was
performed to examine and compare the quantity of (E)-labda-8(17), 12-diene-15,16 dial production
in cells and callus induced at different growth period with field grown rhizomes and shoot buds.
The results revealed the presence of (E)-labda-8(17), 12-diene-15,16 dial in all materials tested
and were in comparable amounts. With this findings, further work on the enhancement of the
compound could be carried out through elicitation and metabolic engineering.


Keywords: (E)-labda-8(17); 12-diene-15,16 dial; callus; cancer; cell suspension; gas
chromatography; in vitro; mango ginger















47

4
th
Australasian Metabolomics
Symposium 2012



Discrimination between Infected Dengue Patient and Healthy Individual
Based On Chemometry of
1
H NMR Spectroscopy

Nurul Shahfiza
1
, Maulidiani
2
, Hasnah Osman
3
, Khozirah Shaari
2
, TS Chow
4
, TH Tang
1
,
Abdel-Hamid AZ
1
*


1
Infectious Disease Cluster, Advanced Medical and Dental Institute, Universiti Sains Malaysia,
Penang, Malaysia;
2
Laboratory of Natural Product, Institute of Bioscience, Universiti Putra
Malaysia, 43400 UPM, Serdang, Selangor, Malaysia;
3
School of Chemical Sciences, Universiti
Sains Malaysia, Penang, Malaysia;
4
Infectious Disease Unit, Hospital Besar Pulau Pinang,
Penang, Malaysia.

*Corresponding author: abdelhamid@amdi.usm.edu.my

Abstract

Dengue is a major health and pressing threat to Malaysia. The increasing incidence makes the
early diagnosis important as supportive treatment can substantially lower the risk of developing
severe disease. Therefore, the metabolomics approach was applied to investigate the metabolite
candidates that are potentially useful as indicators for dengue disease. Forty-seven patients
diagnosed with dengue fever at Hospital Pulau Pinang and forty-one healthy individuals were
included in this study. Medical record was used to characterise the disease and provisional
diagnosis. Urine samples were collected from confirmed dengue patients. A combination of
1
H
NMR spectrometry and multivariate statistical analysis (SIMCA) was used to differentiate the
metabolic profiles of dengue infected and non-dengue infected subjects. The metabolic
compounds responsible for the differentiation were identified based on
1
H NMR chemical shifts
and compared to Human Metabolome database (HMDB). Orthogonal partial least square
discriminant analysis (OPLS-DA) of the
1
H NMR spectra showed a clear discrimination between
the dengue infected and non-dengue infected subjects with the total R
2
Y (cum) value of 0.935,
and the total Q
2
Y (cum) value is 0.832. The S-plot visualises that carnitine, O-phosphocholine, O-
acetylcholine were the metabolites contribute most to the separation. This study demonstrated
that the metabolites derived from the urinary metabolite profiling approach holds a potential of
using a combination of
1
H NMR spectroscopy and multivariate data analyses in differentiating
dengue infected and non-dengue infected subjects. However, further validation of the metabolites
is required for confirmation.


Keywords:
1
H NMR spectroscopy; dengue; metabolomics; multivariate statistical analysis;
orthogonal partial least square discriminant analysis















48

4
th
Australasian Metabolomics
Symposium 2012



The Use of LC-MS QTOF-Based Metabolomics to Define the Potential of
Probiotic against Colorectal Cancer Xenograft in the Mouse Model

Azidah Ali
1
, Kalavathy Ramasamy
1
*, LK Teh
2
, Vasudevan Mani
3

MZ Salleh
2
, Abu Bakar Abdul Majeed
3

1
Collabrative Drug Discovery Research (CDDR) Laboratory, Faculty of Pharmacy, Uitm Puncak
Alam, 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Pharmacogenomics Centre
(PROMISE), Faculty of Pharmacy, University Teknologi MARA (UiTM), 42300 Bandar Puncak
Alam, Selangor Darul Ehsan, Malaysia;
3
Brain Research Laboratory, Faculty of Pharmacy, Uitm
Puncak Alam, 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia.

*Corresponding author: kalav922@puncakalam.uitm.edu.my

Abstract

Numerous studies have shown that there exists a potential for foods that contain probiotics to
affect the host by changing the colonic microbiota in a way that might prevent colorectal cancer
(CRC). Probiotic is defined as live microbial food ingredients which upon ingestion may be
beneficial to the health of the host. Most of the earlier studies of probiotic effects on CRC were
with regards to the reduction of cancer cell viability or tumour size and monitored only specific end
products such as amino acids or bile acids. The interrelationship between the cellular effects of
probiotics and the metabolites produced have not been well documented and is clearly a
challenge. Metabolomics utilises fundamental analytical techniques to probe the chemical
fingerprint of the sample and has been shown to be effective tool for disease diagnosis, biomarker
screening and characterisation of biological pathway. In this study, metabolic profiling of serum
male mice was carried out to assess the effects of probiotics on human colorectal cancer
xenograft model using liquid chromatography-mass spectrometry quadrupole time of flight
(LC/MS-QTOF). Mice were divided into 6 groups and administered with normal saline (control
group); fermented and unfermented soymilk with probiotics, lactic acid bacteria (LAB 12 and LAB
PC) for 4 weeks before tumour induction and the treatment was continuously given until the
experiment was terminated at the end of 7 weeks. Data were analysed using principle component
analysis (PCA) and clustering analysis. PCA showed the separation between the control group
and treatment group. In addition, four metabolites (docosatriynoic acid, nanodecanoic acid,
ladderane and octadecadienal) were found to be higher in treatment group with p<0.005. This
study highlights the potential of metabolomics for assessing the probiotic effect in an animal model
of xenograft models and exploring other potential metabolic biomarkers.


Keywords: lactic acid bacteria; metabolomics; probiotics; serum; xenograft model











49

4
th
Australasian Metabolomics
Symposium 2012



Patients with Combined Variant of CYP2D6 and Wild-type ABCB1
C3435T Developed Early Recurrence and Metastasis: Role of
Sphinganine

NI Mohamed
1
*, MZ Salleh
1
, Rohaizak M
2
, Shahrun NS
2
, Saladina JJ
2
, Roslan H
3
, Sood
S
4
, Rajoo TS
5
, Muniandy SP
5
, Henry G
5
, Ngow HA
6
, KT Hla U
7
Din J
7
, TP Hennessy
1,8
,
LK Teh
1
*


1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Department of Surgery,
Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Kuala Lumpur,
Malaysia;
3
UKM Medical Biology Institute (UMBI) and Department of Medicine, Universiti
Kebangsaan Malaysia Medical Centre, (UKMMC), Kuala Lumpur, Malaysia;
4
Faculty of Medicine,
Universiti Teknologi MARA (UiTM), Selangor Darul Ehsan, Malaysia;
5
Department of Surgery,
Selayang Hospital, Kuala Lumpur, Malaysia;
6
Department of Internal Medicine, Kulliyah of
Medicine, International Islamic University Malaysia (IIUM)/Tengku Ampuan Afzan Hospital
(HTAA), Pahang Darul Makmur, Malaysia;
7
Department of Surgery, International Islamic
University Malaysia (IIUM), Pahang Darul Makmur, Malaysia;
8
Life Sciences Group, Agilent
Technologies, Singapore

*Corresponding authors: szwanie@gmail.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Tamoxifen has been widely used as the standard adjuvant therapy for breast cancer patients who
are estrogen receptor positive. However, resistance towards tamoxifen therapy was found to
underlies the cause of treatment failure. Earlier findings by our group reveal that patients with
combination of variant alleles of CYP2D6 and wild-type of ABCB1 C3435T developed early
recurrence and metastasis. Therefore, analysis on the differential expression of the metabolites
was performed to understand the role of CYP2D6 and P-glycoprotein in regulation of metabolism
among the patients which affect the risks of recurrence and metastasis. Plasma samples from 31
breast cancer patients treated with tamoxifen were analysed using LC/MS-QTOF (Agilent Tech).
Samples were injected after protein precipitation with acetonitrile and centrifugation for 10 min at
10,000rpm at 4C. The chromatography was performed on Agilent 1200 Series using an XDB
C18 4.6 M, 1501.8 mm. The MS data was analysed using Mass Profiler Professional (MPP)
and Metlin Personal Metabolites Database. Combined partial least square discriminant analysis
(PLS-DA) and hierarchical clustering with Bonferroni correction was done. Patients were grouped
according to the variation of genotypes. Analysis on metabolite clustering revealed that patients
with variant alleles of CYP2D6 and wild-type of ABCB1 C3435T have low level of vitamin D, bile
acid glycine conjugate as well as several sphingoid bases. Sphinganine has been reported to
inhibit growth and induce apoptosis of tumourigenic human breast epithelial cells (HBEC)
compared to normal HBEC cells. Sphinganine is also a substrate of P-glycoprotein which was
pumps out from the cells. The correlation of the lower level of sphinganine and higher risk of
recurrence and metastasis in breast cancer patients however require further analysis using larger
sample size. Proven the theory, supplementation with sphinganine for patients who carry variant
alleles of CYP2D6 and wild-type of ABCB1 C3435T may improve the therapy outcome.


Keywords: ABCB1 C3435T; breast cancer; CYP2D6; LC/MS-QTOF; tamoxifen




50

4
th
Australasian Metabolomics
Symposium 2012



Metabolic Profiling for Early Detection of Alzheimers Disease

Che Nor Adlia Enche Ady
1
,

Kalavathy Ramasamy
1
, Chin Ai-Vyrn
2
, Mohd Nazif Darawi
1
,
Vasudevan Mani
1
, Shahrul Bahyah Kamaruzzaman
2
, Tan Maw Pin
2
,
LK Teh
3
, MZ Salleh
3
, Abu Bakar Abdul Majeed
1

1
Brain Research Laboratory, Faculty of Pharmacy, UiTM Puncak Alam, 42300 Bandar Puncak
Alam, Selangor Darul Ehsan, Malaysia;
2
Department of Medicine, Faculty of Medicine, University
Malaya, 50603 Kuala Lumpur;
3
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy,
University Teknologi MARA (UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan,
Malaysia.


*Corresponding author: kalav922@puncakalam.uitm.edu.my

Abstract

Currently there is no cure for AD. Early diagnosis and intervention are the best options but the
current neurophysiological evaluation to identify patients with AD is usually for those at their
advanced stage and is still defined by signs and symptoms. Convenient non-invasive blood based
test would clearly be valuable in the early detection of patients with memory deficits. A need for
better biomarkers is also critical to provide insight into disease progression and for the
development and testing of drugs. In the present preliminary study, metabolomics is employed to
determine the metabolite profile of healthy and probable Alzheimers disease (AD) patients.
Serum metabolite profiles from patients with probable Alzheimers disease (n=6) and age-matched
healthy control (n=6) were obtained using liquid chromatography-mass spectrometry quadrupole
time of flight (LC/MS-QTOF) platform. Data were analysed by using principle component analysis
(PCA) and unpaired t-test. PCA showed the separation between AD patients and healthy control.
Several metabolites (1-linoleoylphosphatidylcholine and docosatriynoic acid) were found to be
significantly (P<0.05) lower in AD patients. However, the study is just at the initial stage and we
hope to identify potential new blood-based biomarkers indicative of AD using metabolomic
approach.


Keywords: Alzheimers disease; biomarkers; diagnosis; metabolomics; serum




















51

4
th
Australasian Metabolomics
Symposium 2012



Some Aspect of Urea Metabolism in Parasites Helminth Teladorsagia
circumcincta

Noorzaid Muhamad
1*
, S Brown
2
, HV Simpson
3


1
Royal College of Medicine Perak , Universiti Kuala Lumpur, Malaysia;
2
School of Human Life
Sciences, University of Tasmania, Launceston, Australia;
3
Institute of Food, Nutrition & Human
Health, Massey University, New Zealand.

*Corresponding author: noorzaid@rcmp.unikl.edu.my

Abstract

Infective L
3
T. circumcincta secreted or excreted urea at one quarter of the rate of NH
3
/NH
4
+
. The
rate of urea accumulation in the medium was about 84 pmol h
-1
(10
3
larvae)
-1
over 4 hours,
corresponding to about 11 nmol h
-1
mg
-1
protein. We were unable to detect urease activity, but
urea production by both creatine amidinohydrolase and arginine amidinohydrolase could be
detected. The apparent K
m
and V
max
of creatine amidinohydrolase were 1.1 mM and 48 nmol h
-1

mg
-1
proteins, respectively, and the activity was greatest at pH 8. The apparent K
m
and V
max
of
arginine amidinohydrolase were 0.7 mM and 62 nmol h
-1
mg
-1
proteins, respectively, and the
activity was greatest at pH 7.9. The activity of creatine amidinohydrolase and arginine
amidinohydrolase was sufficient to account for the rate of urea secretion or excretion.


Keywords: arginine amidinohydrolase; creatine amidinohydrolase; Teladorsagia circumcincta;
urea




























52

4
th
Australasian Metabolomics
Symposium 2012



Molecular Characterisation of Previously Untypeable Rotavirus

Shafiq A
1,*
, Zuridah Hassan
1
, Carl Kirkwood
2

1
Faculty of Health Sciences, UiTM Puncak Alam, 42300
Puncak Alam, Selangor Malaysia;
2
Enteric Virus Research Group, Murdoch Childrens
Research Institute, Royal Childrens Hospital, Parkville,
Victoria, Melbourne.

*Corresponding author: shafiq_aazmi@yahoo.com

Abstract

Group A rotavirus (RV) is a causative agent for acute gastroenteritis among children under the
age of five. Due to genetic drift and shift on VP7 and VP4 gene of rotavirus, some strains were
untypeable using currently available primers. Thus molecular strategy for genotyping needs to
be revised. Newly designed VP7 and VP4 primer pairs to identify uncommon strains will
minimise percentage of untypeable cases. Surveillance study conducted in 1996 using rotavirus
positive stool samples from Kuala Lumpur General Hospital identified eight samples which were
G untypeable and five samples were P untypeable. In this study, these untypeable samples
were re-examined using RNA-PAGE and re-typed using the same VP7 and VP4 primer pairs
with different PCR optimisation. RNA-PAGE profiles using mini gel electrophoresis revealed all
the eight (100%) G untypeable samples were Long e-type. For P untypeable, three samples
(60%) were Long e-type and another two samples (40%) were degraded. Even though all eight
G untypeable RNAs were intact in RNA-PAGE, only six (75%) were G1 and two (25%) samples
were still untypeable. For P typing, three (60%) samples were P [8] and another two (40%)
samples were negative when tested with RT-PCR. Using new VP7 primer pair, two G
untypeable samples could be retyped as G1. An alternative new VP4 primer pair that was
designed to retype all the three (60%) reconfirmed all as P [8] for the second time. All these
untypeable samples were G1P [8] strains. Different brands of reagents, new primer sets,
optimised two steps RT-PCR and direct genogroup assignation were able to circumvent
untypeable problem. The new VP7 and VP4 primer pairs designed in this study could be used
as alternative primers for characterisation of VP7 and VP4 gene as part of continuous
surveillance activity to differentiate co-circulating strains.


Keywords: G1; Rotavirus; RNA-PAGE; RT-PCR; P [8]

















53

4
th
Australasian Metabolomics
Symposium 2012



Insight into Genome and Pathogenicity of Opportunistic Human
Pathogen, Streptococcus agalactiae

Irma SZ
*
, LS Lee, Azwin Ismail, Hanif AK, Izwan I, MH Zulkifli, MZ Salleh and LK Teh

Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia.

*Corresponding author: irmasyakina89@yahoo.com

Abstract

Streptococcus agalactiae is well adapted, asymptomatic coloniser of the human intestinal tract
population. However, S. agalactiae has subsequently emerged as a significant cause of human
diseases. Now it is one of the most common causes of life-threatening invasive bacterial infections
such as septicaemia, pneumonia and meningitis in neonates. It is also an important cause of
morbidity and mortality in pregnant women and adults with underlying chronic diseases. However,
the mechanism and genetic elements that are responsible for the virulency of these bacteria may
vary in different geographical areas. Therefore, this study aims to uncover the genetic elements
that contribute to acquisition of virulency and panthogenicity of Streptococcus agalactiae by using
whole genome sequencing approach. Whole genome sequences of a clinical isolate of
Streptococcus agalactiae was obtained using Illumina sequencing platform. The open reading
frames were defined using Prodigal while their positions were visualised via Artemis. Genome
annotation was done by CLCbio Genomics Workbench 5 and BASys. The assembly result
obtained a total of 52,832,486 reads which cover a total of 1.9 Mbp. The genome has a GC
content of 37% and functional genome analysis annotated more than 2699 of total genes which
may be important in the pathogenecity of Streptococcus agalactiae. The genome of Streptococcus
agalactiae was successfully sequenced, assembled and annotated using next generation
sequencing strategy established in Pharmacogenomics Centre (PROMISE). We believed that
whole genome sequencing is a way forward in development of countermeasures against these
pathogenic bacteria.


Keywords: pathogenicity; Streptococcus agalactiae; virulence; whole genome sequencing




















54

4
th
Australasian Metabolomics
Symposium 2012



ANNOVAR: A Useful Tool for Functional Annotation of Genetic Variants
of a Human Genome

Husaini I*, Rose Ismet, Shafiq A, LK Teh, MZ Salleh*

Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia.

*Corresponding authors: husainix@gmail.com/zakisalleh@puncakalam.uitm.edu.my

Abstract

High-throughput sequencing platforms produce huge amounts of data from diverse genomes
including human, animals and plants. The data include genetic diversity of the genomes such as
single nucleotide variations (SNVs) and other structural variations (SVs). Above all that,
characterising the genetic variations and pinpointing functionally important variants are necessary
to further assist in understanding the evolution and disease susceptibility. Here we described the
usage of ANNOVAR to functionally annotate the genetic variants of the Malay genome data.
Variant calls file (.vcf) of the Malay genome was generated using Burrows-Wheeler Alignment and
SAMtools programs prior to annotation using ANNOVAR. A total of 3,847,934 genetic variants
including mainly single-nucleotide variations (SNVs) and insertions-deletions (indels) were
identified. The variants were functionally annotated using three main options in ANNOVAR which
are; gene-based, region-based and filter-based annotation. Our analysis identified 20,070 variants
in exonic regions using gene-based annotation. Region-based annotation identifies variants in
specific genomic regions through mapping against the Databases of Genomic Variants and
GWAS catalogue. A total of 1,357,949 and 2867 variants were found to match against each
database respectively. Filter-based annotation was used to annotate all variants against dbSNP
(SNP database) and SIFT (Sorting Tolerant From Intolerant). A total of 3,471,680 SNPs matched
against the dbSNP and 375,239 were found to be novel. SIFT annotation on the other hand was
performed to assign functionally important scores to non-synonymous SNPs (ns-SNPs). SIFT
score more than the threshold value of 0.05 is regarded as benign. The annotation resulted in
SIFT scores for 10,609 ns-SNPs. In conclusion, ANNOVAR is a useful tool to characterise genetic
variations and provide insights into further analyses of human genome. The novel variants
discovered will be further analysed to identify amino acid changes, to elucidate their effects on
protein functions and association with diseases.


Keywords: annotation; ANNOVAR; functional; non-synonymous SNPs; SNP
















55

4
th
Australasian Metabolomics
Symposium 2012



Whole Genome Sequencing and Comparative Analysis of Local
Klebsiella pneumoniae Strain against Klebsiella pneumoniae Ntuh-
K2044

MH Zulkifli
*
, LS Lee, Hanif AK, Izwan I, Azwin Ismail, Irma SZ, Shafiq A , LK Teh,
MZ Salleh*

Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia.

*Corresponding authors: sherlock_hafiq@yahoo.com/zakisalleh@puncakalam.uitm.edu.my

Abstract

Klebsiella pneumoniae is a Gram-negative bacterium which can cause several of diseases such
as pneumoniae, urinary tract infection, bacteraemia, liver abscess and meningitis. Some studies
reported that the clinical pattern of community-acquired K. pneumoniae infection varies with
geographical areas. The virulence and pathogenic properties and mechanisms of different
bacterial strains might vary in different strains. The aim of this study was to perform whole genome
sequencing of a locally isolated K. pneumoniae and its comparative analysis against K.
pneumoniae NTUH-K2044. K. pneumoniae which had caused septicaemia in a patient was
identified and cultured. DNA was extracted and sequenced using Genome Analyser IIx (Illumina).
The quality of the sequencing output was determined using FastQC. Ambiguous sequences were
trimmed before assembly and mapped to K. pneumoniae NTUH-K2044. Then, the assemblies of
trimmed-sequences were performed using CLCBio: CLC Genomic Workbench 5.1. The annotation
was performed using Rapid Annotation Subsystem Technology (RAST) and then the genome
sequence was visualised using Arthemis. The sequencing produced an output of 6,271,798 reads.
FastQC revealed 56% of GC content for this sample while the reference genome has 57%. After
trimming, 6,228,844 reads were produced with 99.32% of trimming efficiency. The assembly result
showed that 122 contigs have been produced with size of 5,305,345 bp. Annotation results
showed 4,876 protein-encoding genes, 4819 coding sequences and 56 RNAs for local sequence
meanwhile 4,983 protein-encoding genes, 4871 of coding sequences and 111 RNAs in reference
sequence. We conclude that there are significant differences between this particular strain and the
reference sequence in terms of genetic contents. Hopefully, this could provide better knowledge
regarding the virulence and pathogenicty of Klebsiella pneumoniae.


Keywords: CLCBio; Klebsiella pneumonia; next-generation sequencing; Velvet; whole genome
sequence













56

4
th
Australasian Metabolomics
Symposium 2012


Identification of Interesting Genomic Properties of Clinical Isolates of
Mycobacterium tuberculosis

Azwin Ismail
1
*, LS Lee
1,2
, Irma SZ
1
, MH Zulkifli
1
, Izwan I
1
, Hanif AK
1
, Shafiq A
1
, LK Teh
1
,
Norazmi MN
2
, Zainuddin F. Zainul
2
, Tang TH
3
, Najimuddin N
4
, Ngeow YF
5
, MZ Salleh
1
*

1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
School of Health
Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia;
3
Advanced Medical and Dental
Institute, Universiti Sains Malaysia, Penang, Malaysia;
4
School of Biological Sciences, Universiti
Sains Malaysia, Penang, Malaysia;
5
Faculty of Medicine Building, University of Malaya, Malaysia.

*Corresponding authors: nuwinismail@gmail.com/zakisalleh@puncakalam.uitm.edu.my

Abstract

Tuberculosis meningitis (TBM) is the most severe extrapulmonary complication of Tuberculosis
caused by the obligate human pathogen, Mycobacterium tuberculosis (MTB). It has distinctive rod
shaped with rich lipid cell wall. It is widely studied due to its virulence and high prevalence which
had contributed to expanding global health crisis and re-emergence of TB cases. A whole genome
sequencing study using clinical isolates of Mycobacterium tuberculosis was conducted with the
aim to investigate and provide new insight to the pathogenicity and virulence of this pathogen.
Mycobacterium tuberculosis LRGS_TB008 was isolated from human cerebrospinal fluid in a
patient diagnosed with tuberculosis. The DNA was sequenced using a Genome Analyser IIx
(Illumina) sequencer. The reads were assembled using CLCBio (CLC Genomics Workbench
version 5.1) and annotated using BASys and RAST. The whole genome data generated
approximately 41X depth of coverage assuming a genome size of 4 megabases. The genome
sequence assembled comprised of a circular chromosome of 4,411,532 bp, with high G+C content
of 67%, 4,231 potential protein-coding sequences and 48 RNAs genes. Annotation of
LRGS_TB008 genome analysis was performed using the reference genome sequence of M.
tuberculosis H37Rv. The TB008 genome was most similar to strain H37Rv with 99% nucleotide
sequence identity over the alignable sequences. A total of 672 new SNPs were identified when
LRGS_TB008 were compared with M. tuberculosis H37Rv genomes. However, further validation
on the functionality of the SNPs is required.


Keywords: Mycobacterium tuberculosis, pathogenicity; virulence; whole genome sequencing


















57

4
th
Australasian Metabolomics
Symposium 2012



Comparative Study Using Bioinformatics Pipeline for Malaysian
Mahseer (Tor tambroides) Mitogenome Assembly

Norfatimah MY
1,2
*, LK

Teh
3
, MZ Salleh
3
, Mat Isa MN
4
and Siti Azizah MN
2

1
Faculty of Applied Sciences, Universiti Teknologi MARA, Shah Alam, 40450, Malaysia;
2
School of
Biological Sciences Universiti Sains Malaysia Penang 11800, Malaysia;
3
Pharmacogenomics
Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA (UiTM), 42300 Bandar
Puncak Alam, Selangor Darul Ehsan, Malaysia;
4
Malaysia Genome Institute, Kajang, 43000,
Malaysia.

*Corresponding author: norfatimah778@salam.uitm.edu.my

Abstract

Animal mitochondrial DNA has been studied extensively to address problems in population
genetics and systematic of closely related species and individuals within a species as well as to
detect heteroplasmy levels (mitochondrial diseases) in human. The goal of this study is to develop
computational methods to analysed NGS data generated for Tor tambroides (Malaysian Mahseer)
mitochondrial genomes (mitogenome). Two assembly approaches which is Velvet 1.2.07 (Linux
based platform) and CLC Genomics Workbench 4.9 (automated commercial software) for the
mitogenomic references mapping assemblies were developed, applied and compared. Total
Genomic DNA of a single individual sample was isolated from the caudal fin using a tissue DNA
extraction kit (Biotools, Madrid, Spain) and produced at least 5g in 50l for the concentration and
must be in the ratio of 1.8-2.0 at A260/280. The DNA sample was then prepared using Illumina
Truseq kit and sequenced by Illumina Genome Analyser IIX with 2x100 bp read length (Paired-
End). The sample was sequenced in one lane to obtain a high amount of data. Before the
sequence reads were assembled, the reads were subjected to quality control procedures to avoid
the presence of low quality bases. The total number of reads after trimmed by using both
approach is 75 million from the total reads of 80 million with the average length of 94.85 and the
total bases are 7.1 Gb. The most related cyprinidae family, Cyprinus carpio with ID NC_001606
was choose and mapped to Tor tambroides data to perform assembly. Results shows only 2.2 Mb
were mapped to the reference mitogenome by CLC and 1.6 Mb by Velvet. Total reference
coverage for both approach are slightly difference with 134 X compared 95 X for CLC and Velvet
respectively. A detailed study on the bioinformatics pipeline for mitogenomic study would allow
comparison to be made with various bioinformatics tools which could generate high-quality
mitogenome.


Keywords: CLC Bio; mitogenome; NGS; Tor tambroides; Velvet














58

4
th
Australasian Metabolomics
Symposium 2012



HLA-B*1502 is a Significant Marker for Carbamazepine-Induced Stevens
Johnson Syndrome (SJS): Pre-Implementation Trial of HLA-B*1502
Pharmacogenotyping in a Local Hospital in Malaysia

Hasbullani Z
1
, SNA Syeith
1
, RN Che Omar
1
, LK Teh
1
, Fazleen HMH
1
, Sharina H
1
, Syafirul
MS
1
, Shanthi Datuk Puvana Raja
2
, Md Hanip Rafia
2
, MI Ismail
1
, MZ Salleh
1

1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Institute of Neurology,
Hospital Kuala Lumpur, Wilayah Persekutuan, Malaysia.

*Corresponding authors: hasbullanizakaria@yahoo.com/zakisalleh@puncakalam.uitm.edu.my

Abstract

Stevens Johnson Syndrome (SJS) and Toxic Epidermal Necrolysis (TEN) are two fatal adverse
drug reactions (ADR) encountered in some patients prescribed carbamazepine (CBZ) or
phenytoin. FDA USA had published a medical alert which recommended HLAB*1502 screening
before these 2 drugs were prescribed to patients of Asian ancestry as the likelihood of SJS/TEN
were highly associated with HLAB*1502. The aim of this study is to investigate the relevance of
carbamazepine-induced severe cutaneous adverse drug reaction with HLA-B*1502 in a cohort of
patients. Seventy five (75) patients attending clinics at Department of Neurology, Hospital Kuala
Lumpur (HKL) were recruited. DNA was extracted from blood samples obtained from each patient.
Patients genotypes were determined by Alleles Specific Polymerase Chain Reaction (AS-PCR)
developed in-house. Among the 75 patients, 41 were newly registered patients and genotype
screening was conducted before patients were prescribed with CBZ. Among the patients in this
group, 21.95% were positive for HLAB*1502. In another cohort in which CBZ had been withdrawn
due to SJS/ TEN; all of them (6) were positive for HLAB*1502. One patient with positive
HLAB*1502 however did not develop SJS/TEN and is therefore tolerant. The calculated ratio of
patients at risk of developing SJS/TEN based on this small samples size is 238.33 (95% CI :
8.6783 6545.42; p =0.0012). We conclude that the implementation of HLA-B*1502 screening is
necessary to avoid patients at risk of SJS/TEN from being prescribed CBZ due to the high odd
ratio of observed in this study.


Keywords: ADR; CBZ; HLA-A*1502; SJS-TEN

















59

4
th
Australasian Metabolomics
Symposium 2012



Genetic Polymorphism of CYP3A4*18 and CYP3A5*3 in Malaysian
Kidney Transplant Patients

MR Muda, S Hamzah, JKS Shia, LK Teh, MZ Salleh

Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia.

*Corresponding author: rahimi1612@gmail.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Tacrolimus has been widely used in combination with other immunosuppressant to prevent
allograft rejection after transplantation. It is characterised by narrow therapeutic window that
makes the needs for therapeutic drug monitoring essential to prevent rejection or toxicity event.
Extensive researches had shown the involvement of cytochrome P450 (CYP) enzymes in
contributing to interindividual differences of pharmacokinetic and bioavailability of tacrolimus in
kidney transplant patients. CYP3A4 and CYP3A5 are the major enzymes of CYP3A subfamily
that are abundantly expressed in the liver and have the capacity to metabolise approximately up to
50 % of marketed drugs including tacrolimus. Based on previous studies, it has been suggested
that polymorphism of CYP3A5*3 affected the expression of the enzyme by producing cryptic
splicing site while polymorphism of CYP3A4*18 causes amino acid alteration that produce non-
functional enzyme. The objective of this study is to investigate the influence of genetic
polymorphism of CYP3A5*3 and CYP3A4*18 toward tacrolimus level among kidney transplant
patients in Malaysia population. Eighty-kidney transplant patients treated with tacrolimus were
recruited. DNA was extracted from peripheral venous blood using salting out method. They were
genotyped for CYP3A5*3 and CYP3A4*18. The genotypes were correlated with tacrolimus dose
adjusted trough levels (dose/body weight) per at 3-, 6-, 9-, and 12-month post-transplantation. At
3-month post transplantation, tacrolimus dose adjusted trough levels were higher for patients with
CYP3A5*3/*3 genotype compared to those with CYP3A5*1/*1 genotype (2.31; 95% CI 1.6-3.6 vs.
0.75; 95% CI 0.56-1.7) ng/ml/mg/kg, p< 0.0001). Similar pattern were observed for the association
of tarcolimus level with genotypes at 6-, 9-, and 12-months post-transplantation. CYP3A4
polymorphism has no significant correlation with the tacrolimus dose adjusted trough level. In
conclusion, patients who carry CYP3A5*3/*3 require lower doses of tacrolimus to reach target
trough level as compared to those with CYP3A5*1/*1 genotype.


Keywords: tacrolimus; CYP3A polymorphism; kidney transplantation
















60

4
th
Australasian Metabolomics
Symposium 2012



The Implication of the Polymorphism of UGT1A6 among Cardiovascular
Disease (CVD) Patients Treated with Aspirin

NJ Abdul Jalil
1*
, MZ Salleh
1
, O Maskon
2
, A Derahman
1
, RN Che Omar
1
, H Zakaria
1
, JKS
Shia
1
, LK Teh
1*


1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Department of Cardiology,
Hospital Universiti Kebangsaan Malaysia, 56000 Cheras, Kuala Lumpur, Malaysia.

*Corresponding authors: nolin_lynn@yahoo.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

UDP-glucuronosyltransferase 1A6 (UGT1A6) is a major enzyme involved in the glucuronidation of
salicylic acid. UGT1A6 gene which encodes UGT1A6 is polymorphic. The single nucleotide
polymorphisms (SNPs) of UGT1A6 gene that play important role in the metabolism of salicylic acid
include UGT1A6*2 (A541G, rs2070959) and UGT1A6*3 (A522C, rs1105879). The aim of this
study is to determine the allelic frequency of UGT1A6 gene in Malaysian population. The project
was approved by relevant Research Ethics Committee. A total of 151 patients with cardiovascular
disease who were treated with 75 -150 mg daily dose of aspirin were recruited. DNA was
extracted from the blood samples and genotyped for the single nucleotide polymorphisms (SNPs)
using polymerase chain reaction (PCR). Analysis of UGT1A6 genotypes in these patients revealed
that 71 were homozygote UGT1A6*1/*1. Four percent (4%) were heterozygote UGT1A6*1/*2,
14% were heterozygote UGT1A6*1/*3, and 28% were heterozygote UGT1A6*2/*3. Of the 151
patients, only 10 patients were homozygous for UGT1A6*3/*3. No homozygous UGT1A6*2/*2 was
detected. The allele frequencies of UGT1A6*1, UGT1A6*2 and UGT1A6*3 were 0.6 (95% CI
0.08-0.96), 0.2 (95% CI 0.01 0.87) and 0.2 (95% CI 0.01 0.87) respectively. Allele frequencies
of UGT1A6*1 were 0.56, 0.62 and 0.40 in the Malays, Chinese and Indians respectively. The
frequency of 0.2 for all the 3 ethnic groups. The Indians have the highest frequency of UGT1A6*3
[0.40; 95% CI 0.22 0.40) compared to the Malays (0.30) and Chinese (0.20) UGT1A6 is highly
polymorphic in Malaysia and therefore may have important implication in personalised medicine to
ensure safe and cost effective therapy.


Keywords: aspirin; cardiovascular disease; Malaysian population; UGT1A6


















61

4
th
Australasian Metabolomics
Symposium 2012



Genetic polymorphism of CYP2C19 among the Cardiovascular Patients
of Three Ethnic Groups in Malaysia

RN Che Omar
1
*, NJ Abdul Jalil
1
, A Derahman
1
, H Zakaria
1
, O Maskon
2
, JKS Shia
1
, MZ
Salleh
1
, LK Teh
1
*


1
Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia;
2
Department of Cardiology,
Hospital Universiti Kebangsaan Malaysia, 56000 Cheras, Kuala Lumpur, Malaysia

*Corresponding authors: ruhil_ira@yahoo.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Cytochrome P450 is 2C19 is a highly polymorphic enzyme that cause marked interindividual and
interethnic variation in the metabolism and disposition of many therapeutic agents. The goal of this
study was to determine the frequencies of important allelic variant of CYP2C19 in a cohort of
cardiovascular patients attending follow up clinic at Hospital UKM. This project is approved by
local Research Ethics Committee. Patients were explained the study protocol and written
informed consents were obtained before the study. A total of 150 cardiovascular patients were
recruited. They were unrelated Malaysians aged 1865 years. Blood sample were obtained from
antecubital vein for DNA extraction. All samples were genotyped for single nucleotide
polymorphisms (SNPs) of CYP2C19*2 (rs4986893) and CYP2C19*3 (rs4244285) using
polymerase chain reaction (PCR). The allele frequencies for CYP2C19*1 are 81%, 75% and 71%
in the Malays, Chinese and Indian respectively. CYP2C19*2 was present at 16%, 17% and 26% in
the Malays, Chinese and Indian respectively; and carriers of CYP2C19*3 comprised of 3%, 8%
and 3% in the Malays, Chinese and Indian respectively. There were significant differences in the
poor metaboliser genotypes (*2/*2, *2/*3, *3/*3) between the Malays and Indians (Chi square test,
P 0.05). We conclude that frequencies of CYP2C19 genotypes were different among the three
major ethnic groups in Malaysia. This result is useful for clinical practitioners in understanding the
differences in drug responses observed among their patients and the potential of molecular
diagnostics in personalised drug therapy.


Keywords: cardiovascular disease; CYP2C19; poor metaboliser



















62

4
th
Australasian Metabolomics
Symposium 2012



Development of a Pharmacogenotyping Method for Detection of HLA
Polymorphism in Personalising Allopurinol Therapy

N Mohamed Ali*, LK Teh*, Fazleen HM Hatta, S Hamzah, MZ Salleh

Pharmacogenomics Centre (PROMISE), Faculty of Pharmacy, University Teknologi MARA
(UiTM), 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia.

*Corresponding authors: norleenali@gmail.com/tehlaykek@puncakalam.uitm.edu.my

Abstract

Previous studies performed in Europe and Israel had shown that allopurinol is commonly
associated with drug induced Stevens - Johnson syndrome (SJS) and Toxic Epidermal Necrolysis
(TEN). Human leukocyte antigen (HLA) A33 and B58 was the first genetic variants associated with
allopurinol induced skin eruption. On the other hand, researchers in Taiwan (Han Chinese
population), Thailand and Korea showed that there is a strong association of HLA-B*5801 allele
and severe cutaneous adverse drug reaction (SCAR). The pathogenesis of allopurinol induced
cutaneous reaction is consistent with delayed type immune mediated reaction. Observed familial
predisposition in conjunction with the pathogenesis of allopurinol induced cutaneous reaction,
alludes to the potential of genetic based immunological markers. The aim of this study is to
develop an accurate PCR based method to determine the genetic variants of HLA-B*5801 which
can be used for association study of HLA polymorphism with allopurinol induced cutaneous
adverse drug reaction in Malaysian population. Genomic DNA was extracted from the whole blood
sample using salting out method. Allele specific polymerase chain reaction (PCR) method was
developed based on specific 3 end primer mismatch for tagging SNPs of HLA-B*5801. To avoid
any misinterpretation of the PCR result, positive control containing DNA fragment of tagging SNPs
for HLA-B*5801 were cloned into plasmid. Both the positive and negative controls were used for
quality assurance and were included in each PCR run. Method was validated by direct
sequencing. The method developed is useful to study the association of HLA polymorphism with
the susceptibility of allopurinol induced cutaneous adverse drug reactions. The assay developed
can be used in clinical practice to help identify patients at risk of SCAR and thus reduce cost of
patient care. This will help realise personalised medicine and improve patients quality of life.


Keywords: human leukocytes antigen (HLA); polymerase chain reaction (PCR); severe
cutaneous adverse reaction (SCAR); Steven Johnson Syndrome (SJS); Toxic Epidermal
Necrolysis
















63

4
th
Australasian Metabolomics
Symposium 2012



Hepatoprotective Activity of Methanol Extract of Melastoma
malabathricum Leaves on Paracetamol-Induced Liver Injury in Rat

SS Mamat
1
*, N Mohtarrudin
2
, ZA Zakaria
1
*


1
Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra
Malaysia, 43400 UPM Serdang, Selangor, Malaysia;
2
Department of Pathology, Faculty of
Medicine& Health Sciences,Universiti Putra Malaysia, 43400 UPM , Serdang Selangor , Malaysia.

*Corresponding authors: syariah_islamic@yahoo.com/dr_zaz@yahoo.com

Abstract

The aim of the present study was to determine the hepatoprotective effect of methanol extract of
Melastoma malabathricum L. leaves (MEMM; family Melastomaceae) against paracetamol (PCM)-
induced liver toxicity. The dried, grinded leaves of M. malabathricum (40 g) were soaked with 800
ml methanol (1:20 (w/v)) for 72 hours at room temperature. The supernatant was collected, filtered
using cotton wool followed by Whatman No. 1 filter paper, and finally subjected to the rotary
evaporation process. This process was repeated for another two times. The vehicle (10% DMSO),
200 mg/kg silymarin or MEMM (50, 250 and 500 mg/kg) were administered orally once daily for 7
successive days in rats (n=6). Hepatotoxicity was induced by the administration of PCM (3 g/kg) 3
hours after the last MEMM administration. On the 9
th
day, blood was collected followed by the
immediate sacrifice of animals to collect the liver for histopathological examination. Hepatic
enzymes, AST (aspartate aminotransferase), ALP (alkaline phosphatase) and ALT (alanine
aminotransferase), were assessed as indicators of liver damage. The results obtained showed
that pre-treatment with MEMM significantly (p<0.05) reduced the PCM-induced serum levels of
hepatic enzyme markers compared to negative control group (P<0.05). Liver histopathology also
showed that MEMM reduced the incidence of liver lesions including hepatic steatosis, lymphocyte
infiltration and marked necrosis as compared to negative control group. In conclusion, the
methanol extract of leaves of M. malabathricum exerts potential hepatoprotective property that
warrants further investigation.


Keywords: hepatoprotective activity; liver enzyme markers; Melastoma malabathriucm; methanol
extract; paracetamol

















64

4
th
Australasian Metabolomics
Symposium 2012



Chemopreventive Potential of Melastoma malabathricum Leaves
Extract in DMBA/Croton Oil-Induced Mouse Skin Carcinogenesis

Samuel Oii KK*, R Rodzi, F Othman, ZA Zakaria

Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra
Malaysia, 43400 UPM Serdang, Selangor, Malaysia.

*Corresponding author: roy_1906@yahoo.co.uk

Abstract

The ideas that cancer is preventable had contributed to research on plant-based products for
chemoprevention. Melastoma malabathricum known as Pokok Sendudok in Malays community
is a plant used traditionally to treat diarrhoea, dysentery, cuts, wounds, and other ailments.
Previous studies had shown M.malabathricum exhibited in-vitro anti-proliferative activity. The aim
of this study was to investigate the anti-carcinogenic activity of methanol leaves extract of
M.malabathricum (MMLE) in two stage mouse skin carcinogenesis model. The skin tumours were
initiated by a single dose of 7,12-dimethylbenz[a]anthracene (DMBA) topically on shaved dorsal
skin, followed by twice weekly administration of 1% croton oil (100 g/100 l) as tumour promoter.
A varying doses (30, 100 and 300 mg/kg of body weight) of MMLE were tested on DMBA/croton
oil-induced mouse skin carcinogenesis. Tumours were examined and measured weekly for 15
weeks. At the end of the experiment, animals in carcinogen control developed tumour burden of
5.33 1.36, in contrast mice treated with 30, 100 and 300 mg/kg MMLE developed tumour burden
of 4.40 1.68, 1.16 0.41 and 1.00 0.13 with percentage of tumour incidence of 62.5%, 37.5%
and 12.5% respectively. Mice treated with 100 mg/kg and 300 mg/kg showed higher tumour
volume per individual tumour in contrast to carcinogen control, although they are significantly
different (P<0.05). Histopathological findings of skin showed different degree of suppression of
MMLE on DMBA/croton oil-induced hyperplasia and papillomatosis in contrast to carcinogen
group. Results from the study indicated pre-treatment of MMLE 30 minutes prior administration of
croton oil resulted in protection against skin carcinogenesis in a dose-dependent manner.


Keywords: 7, 12-dimethylbenz[a]anthracene; chemopreventive; croton oil; Melastomaceae;
Melastoma malabathricum, methanol extract



















65

4
th
Australasian Metabolomics
Symposium 2012



Mycolic Acids as Diagnostic Markers for Tuberculosis Case Detection
and Drug Efficacy

Guanghou Shui
1
*, Anne K Bendt
1
*, Ignasius A Jappar
1
, Hui Ming Lim
1
, Marie Laneelle
3
,
Maxime Herv
5
, Laura E Via
4
, Gek Huey Chua
1
, Martin W Bratschi
1,10
, Siti Zarina Zainul
Rahim
6
, Ang Lay Teng Michelle
6
, Soo-Hee Hwang
7
, Jong-Soek Lee
8
, Seok-Yong Eum
8
,
Hyun-Kyung Kwak
8
, Mamadou Daff
3
, Vronique Dartois
2,5
, Gerd Michel
9
, Clifton E. Barry
III
4
, Markus R Wenk
1,2,10


1
Yong Loo Lin School of Medicine, National University of Singapore, Department of Biochemistry
and
2
Department of Biological Sciences, National University of Singapore, Singapore;
3
Dpartement Mcanismes Molculaires des Infections Mycobactriennes, IPBS-UMR, Toulouse
Cedex, France;
4
Tuberculosis Research Section, Laboratory of Clinical Infectious Disease, NIAID,
NIH, USA;
5
Novartis Institute for Tropical Diseases, Singapore;
6
Department of Microbiology,
National University of Singapore, Singapore;
7
National Masan Tuberculosis Hospital, Masan,
Republic of Korea;
8
International Tuberculosis Research Center, Masan, Republic of Korea;
9
Foundation for Innovative New Diagnostics (FIND), Geneva, Switzerland;
10
Swiss Tropical and
Public Health Institute, University of Basel, Socinstr. 57, P.O. Box, 4002 Basel.

*Corresponding author: anne_bendt@nuhs.edu.sg

Abstract

Tuberculosis, which is caused by infection with Mycobacterium tuberculosis, remains a major
global health threat, with over nine million new cases and close to two million deaths annually.
Detection of active tuberculosis cases in endemic countries still relies on sputum smear
microscopy, which only has an average sensitivity of 65% and which is highly dependent on the
individual examiner. The more sensitive culture methodology on the other hand takes weeks, thus
further delaying initiation of treatment and potential spread of pathogens. Better diagnostics are
therefore urgently needed. Since a high percentage of patients are co-infected with HIV,
diagnostics should be suitable for immunocompromised individuals. Furthermore, a specific
molecular marker which reflects clearance of bacteria from lungs would be helpful to monitor
treatment response and drug efficacy in pre-clinical and clinical trials.


Keywords: diagnostic marker; lipidomics; mass spectrometry; Mycobacterium tuberculosis;
mycolic acids

















66

4
th
Australasian Metabolomics
Symposium 2012



A Sensitive and Efficient Method for the Quantification of
Phosphorylated Sphingoid Bases in Biological Samples

*Pradeep Narayanaswamy
1
*, Husna Begum
2
, Bowen Li
3
, Federico Torta
4
, Rudolf Grimm
5
, Markus
R. Wenk
1-3


1
Department of Biological Sciences, National University of Singapore,
2
NUS Graduate School,
National University of Singapore,
3
Department of Biochemistry, National University of Singapore,
4
Mechanobiology Institute, National University of Singapore, Singapore, and
5
Agilent
Technologies, Santa Clara, USA.

*Corresponding author: a0078608@nus.edu.sg

Abstract

Sphingosine-1-Phosphate (S1P) is an important bioactive lipid mediator and a normal constituent
of human plasma. S1P is synthesized from sphingosine by the enzyme Sphingosine Kinase and
its biological activity is modulated through binding to specific receptors. S1P plays important
physiological roles in the regulation of cell growth, differentiation, motility and survival; in addition it
participates in pathological conditions like autoimmunity, cancer, angiogenesis and myocardial
infarction. Thus S1P gained clinical importance as biomarker for diseases. For the detection of
S1P in biological matrices we developed a simple, quick, sensitive and reliable assay by using an
Agilent HPLC-Chip MS system after derivatisation of the sample. New method is a valuable tool
for the detection of S1P levels from small amounts of human and mouse plasma samples.


Keywords: angiogenesis; myocardial infarction; sphingosine-1-phosphate
























Designed by: Mohd Izwan Ismail, PROMISE



4
th
Australasian Metabolomics
Symposium 2012



















APPENDICES
























xi

4
th
Australasian Metabolomics
Symposium 2012



List of Participants

No. Name Email Organisation
1.
Aishah Adam
(Prof. Dr.)
aishah_adam@puncakalam.uitm.ed
u.my
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
2.
Anne Kathrin
Bendt (Dr.)
anne_bendt@nuhs.edu.sg
Department of Biochemistry, Yong Loo Lin School
of Medicine, and Department of Biological
Sciences, National University of Singapore.
3. Ali Ashrafzadeh ali_ash_3000@yahoo.com
School of Biosciences and Biotechnology, Faculty
of Science and Technology, Universiti
Kebangsaan Malaysia, Selangor, Malaysia
4.
Asbiyatulaida
Derahman
gurlz_aida@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
5. Azidah Ali azidah85@yahoo.com.my
Collaborative Drug Discovery Research, Level 7,
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
6. a Azizah Othman akanami@hotmail.com
Faculty of Medicine and Health Sciences,
Universiti Putra Malaysia, Serdang, Selangor,
Malaysia
7. Baharudin Ibrahim baharudin.ibrahim@usm.my
School of Pharmaceutical Sciences, Universiti
Sains Malaysia, Penang, Malaysia
8.
Che Nor Adlia
Enche Ady
che.adlia@gmail.com
Collaborative Drug Discovery Research, Level 7,
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
9.
Chin Siok Fong
(Dr.)
chinsiokfong@gmail.com
Sime Darby Technology Centre Sdn. Bhd., 1st
Floor, Block B, UPM-MTDC Technology Centre III,
Lebuh Silikon, Universiti Putra Malaysia, Serdang,
Selangor, Malaysia
10.
Christopher
Bowen
christopher_bowen@agilent.com Agilent Tech. Australia
11.
David Ross
Appleton
david.ross.appleton@simedarby.co
m
Sime Darby Technology Centre Sdn. Bhd., 1st
Floor, Block B, UPM-MTDC Technology Centre III,
Lebuh Silikon, Universiti Putra Malaysia, Serdang,
Selangor, Malaysia
12.
David Wishart
(Prof. Dr.)
david.wishart@ualberta.ca
Department of Computing Science and Biological
Sciences at the University of Alberta, Edmonton
13.
Dayana Sazereen
Md Hasni
dayanasazereen@yahoo.com
Collaborative Drug Discovery Research, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
14. Delphia Shem delphia@mosti.gov.my

Agro-Biotechnology Institute, Malaysia (ABI),
Serdang, Selangor, Malaysia

15.
Erda Syerena
Rosli
syerena@gmail.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
16. Fang Fang (Dr.) Fang.Fang@bruker-biospin.de
Application Chemist for Bruker BioSpin, GmbH
Silberstreifen 4 76287, Rheinstetten, Germany
17. Fauziah Abdullah fauziahabdullah@frim.gov.my
Phytochemistry Programme, Natural Products
Division, FRIM, Kepong, Selangor, Malaysia


xii

4
th
Australasian Metabolomics
Symposium 2012


18.
Fazleen Haslinda
Mohd Hatta
fazleenhaslinda@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
19. Forough Ebrahimi ebrahimi.foroogh@gmail.com
Department of Clinical Pharmacy, School of
Pharmaceutical Sciences, Universiti Sains
Malaysia, Penang, Malaysia
20.
Gunaletchumy
Selva Perumal
gl_2512@hotmail.com
Department of Medical Microbiology, Faculty of
Medicine, Universiti Malaya, Kuala Lumpur,
Malaysia
21. Hanum Yaakub hanumy@gmail.com
Collaborative Drug Discovery Research, Level 7,
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
22. Hasbullani Zakaria hasbullanizakaria@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
23.
Hemalatha
Visuvalingam
hema@leco.com.my Universiti Sains Malaysia, Penang, Malaysia
24.
Irma Syakina
Mohd Zaki
irmasyakina89@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
25. Izmil Haikal Zainol Milhaikal89@gmail.com
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
26.
Jean-Frdric
Weber
(Prof. Dr.)
jffweber@puncakalam.uitm.edu.my
Atta-ur-Rahman Institute for Natural Product
Discovery, Level 9, Faculty of Pharmacy,
Universiti Teknologi MARA, Puncak Alam,
Selangor, Malaysia
27.
John Shia Kwong
Siew (Dr.)
johnshia@puncakalam.uitm.edu.my
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
28.
Kalavathy
Ramasamy
(Assoc. Prof. Dr.)
kalav922@salam.uitm.edu.my
Collaborative Drug Discovery Research, Level 7,
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
29.
Kamalrul Azlan
Azizan
kamalrulazlan@gmail.com
Institute of Systems Biology (INBIOSIS), Universiti
Kebangsaan Malaysia, Bangi, Selangor, Malaysia
30. Laith Razzak (Dr.) saiflaith3@yahoo.ca
Department of Fisheries and Aquaculture, Faculty
of Fisheries and Aqua Industry, Universiti
Malaysia Terengganu, Kuala Terengganu,
Terengganu, Malaysia
31. Lee Lian Shien lianshien@gmail.com
Pharmacogenomics Centre, Level 7,Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
32. Lim Vuanghao lvghao@amdi.usm.edu.my
Advanced Medical & Dental Institute, Universiti
Sains Malaysia, No 1-8 (Lot 8), Persiaran Seksyen
4/1, Bandar Putra Bertam, Kepala Batas. Pulau
Pinang, Malaysia
33. Loke Mun Fai (Dr.) lmunfai@gmail.com
Department of Microbiology, Faculty of Medicine,
University Malaya, Kuala Lumpur, Malaysia
34.
Lydiatul Shima
Ashari
lyd_84shima@yahoo.co.uk
School of Health Sciences, Universiti Sains
Malaysia, Kubang Kerian, Malaysia
35. Mailina Jamil mailina@frim.gov.my
Herbal Products Development Programme,
Natural Product Division, FRIM, Kepong,
Selangor, Malaysia


xiii

4
th
Australasian Metabolomics
Symposium 2012


36. Markus R. Wenk markus_wenk@nuhs.edu.sg
Department of Biochemistry, Yong Loo Lin School
of Medicine, and Department of Biological
Sciences, National University of Singapore
37.
Matthias Pelzing
(Dr.)
matthias.pelzing@bdal.de
Application Manager for Bruker
Daltonics Asia Pacific/ Japan, Bruker Biosciences
Pty Ltd, Australia
38.
Mohamad Izwan
Ismail
izwanspp@gmail.co
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
39.
Mohd Hafis
Yuswan
use_one86@yahoo.com
Agro-biotechnology Institute, Serdang, Selangor,
Malaysia
40.
Mohd Ikhwan
Ismail
ikhwanismail86@gmail.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
41.
Mohd Ikmal Hanif
Abdul Khalid
ikmalhanif87@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
42.
Mohd Izwan
Mohamad Yusof
mohd_izwan86@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
43.
Mohd Nazri Hj.
Abu

Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
44.
Mohd Rahimi
Muda
rahimi1621@gmail.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
45.
Mohd Salleh
Rofiee
bio_salleh82@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
46.
Mohd Shafiq
Aazmi
shafiq_aazmi@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
47.
Mohd Zaki Salleh
(Prof. Dr.)
zaki0104@yahoo.co.uk
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
48.
Muhamad Hafiq
Zulkifli
sherlock_hafiq@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
49.
Muhammad
Husaini Ismail
husainix@gmail.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
50. Nazrien Kaman nenazr@mosti.gov.my
Agro-biotechnology Institute, Serdang, Selangor,
Malaysia
51.
Noorzaid
Muhamad (Assoc.
Prof. Dr.)
noorzaid@rcmp.unikl.edu.my
Royal College of Medicine Perak , Universiti Kuala
Lumpur, Malaysia
52.
Nor Amalina
Ahmad Alwi
amal_wie89@yahoo.com.my
Collaborative Drug Discovery Research, Level 7,
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
53.
Norfatimah
Mohamed Yunus
norfatimah778@salam.uitm.edu.my
Faculty of Applied Sciences, Universiti Teknologi
MARA, Shah Alam, Selangor, Malaysia
54.
Nor Izwani
Mohamed
szwanie@gmail.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia


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4
th
Australasian Metabolomics
Symposium 2012


55. Nor Nadia Ban nornadia_ban@hotmail.com
Collaborative Drug Discovery Research, Level 7,
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
56.
Norleen Mohamed
Ali
norleenali@gmail.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
57.
Norlelawati
A.Talib
noleata@yahoo.com
Department of Basic Medical Sciences, Kulliyah of
Medicine, International Islamic University
Malaysia, Kuantan Campus, Kuantan, Pahang
58.
Nornazliya
Mohamad
nornazliyamohamad@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
59.
Norzulaani Khalid
(Prof. Dr.)
lani@um.edu.my
Centre for Biotechnology in Agriculture Research,
Institute of Biological Sciences, Faculty of
Science, University Malaya, Kuala Lumpur,
Malaysia
60. Nur Azwin Ismail nuwinismail@gmail.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
61.
Nur Jalinna Abdul
Jalil
nolin_lynn@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
62.
Nur Syafiqah
Rahim
nursyafiqahrahim@gmail.com
Collaborative Drug Discovery Research, Level 7,
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
63. Nur Syahidah syahidah@mosti.gov.my

Agro-Biotechnology Institute Malaysia (ABI)

64.
Nurul Aqilah
Iberahim
n_aqilah610@yahoo.com.my
Department of Aquaculture Science, Faculty of
Fisheries and Aqua-Industry, University Malaysia
Terengganu, Mengabang Telipot, Terengganu
65.
Nurul Aqmar
Mohamad Nor
Hazalin
nurulaqmar@puncakalam.uitm.edu.
my
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
66.
Nurul Ashikin Md.
Hazmi
nurul_ashikin_ck@yahoo.com
Agro-biotechnology Institute, Serdang, Selangor,
Malaysia
67. Nurul Hamiyah
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
68.
Nurul Shahfiza
Noor
nurulshahfiza@gmail.com
Advanced Medical & Dental Institute,
Universiti Sains Malaysia, No. 1-8, Persiaran
Seksyen 4/1, Bandar Putra Bertam, Kepala Batas,
Penang
69.
Pradeep
Narayanaswamy
a0078608@nus.edu.sg
Department of Biochemistry, Yong Loo Lin School
of Medicine, and Department of Biological
Sciences, National University of Singapore
70.
Qamarul Hafiz
Zainul Abidin


Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
71. Rehvathy Vellayan rev_rehva86@hotmail.com
Department of Medical Microbiology, Faculty of
Medicine, University of Malaya, Kuala Lumpur
72.
Richard
Muhammad
richard@puncakalam.uitm.edu.my
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
73. Robert Trengove r.trengove@murdoch.edu.au
Murdoch University Metabolomics Australia Node,
90 South Street, Murdoch WA 6150, Australia


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th
Australasian Metabolomics
Symposium 2012

74. Robin Philp robin_philp@agilent.com
Life Sciences Group, Agilent Technologies,
Singapore
75. Roihanah Rodzi roy_1906@yahoo.co.uk
Department of Biomedical Science, Faculty of
Medicine and Health Sciences, Universiti Putra
Malaysia, Serdang, Selangor, Malaysia
76.
Rose Iszati Ismet
Nayan
rose_ismet@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
77.
Rosmadi Yusoff
(Dr.)
Rhous_y@yahoo.co.uk
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
78. Rudolf Grimm rudolf_grimm@agilent.com Agilent Technologies, Santa Clara, USA
79.
Ruhil Hayati
Hamdan
ruhilhayati1982@gmail.com
Aquatic Animal Health Unit (AAHU), Faculty of
Veterinary Medicine, Universiti Putra Malaysia,
Selangor, Malaysia
80.
Ruhil Nadirah
Che Omar
ruhil_ira@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
81.
Ruzianisra
Mohamed
ruzianisra@salam.uitm.edu.my
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor DE, Malaysia
82. Salfarina Ramli
salfarina2892@puncakalam.uitm.ed
u. my
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor DE, Malaysia
83. Selene Tan selene_tan@agilent.com Agilent Tech. Malaysia
84. Sharina Hamzah sharinahamzah114@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
85.
Siti Azma Jusoh
(Dr.)
sitiazma@puncakalam.uitm.edu.my
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
86.
Siti Nooraishah
Hussin
nooraishah0352@puncakalam.uitm.
edu.my
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor DE, Malaysia
87.
Siti Syariah
Mamat
syariah_islamic@yahoo.com
Department of Biomedical Science, Faculty of
Medicine and Health Sciences, Universiti Putra
Malaysia, Serdang, Selangor, Malaysia
88. Suhana Samat
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
89.
Tamil Chelvan
Meenakshi
Sundram
lani@um.edu.my
CEBAR, Institute of Biological Sciences, Faculty of
Science, University of Malaya, 50603 Kuala
Lumpur, Malaysia
90. Tan Hooi Poay tanhp@frim.gov.my
Phytochemistry Programme, Natural Products
Division, FRIM, Kepong, Selangor, Malaysia
91.
Tang Thean Hock
(Assoc. Prof. Dr.)
tangth.amdi@gmail.com
Advanced Medical & Dental Institute, Universiti
Sains Malaysia, No. 1-8, Persiaran Seksyen 4/1,
Bandar Putra Bertam, Kepala Batas, Penang,
Malaysia
92.
Teh Lay Kek (Prof.
Dr.)
tehlaykek@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
93.
Theresa Ng Lee
Mei
theresa.ng.lee.mei@simedarby.com
Sime Darby Technology Centre Sdn. Bhd., 1st
Floor, Block B, UPM-MTDC Technology Centre III,
Lebuh Silikon, Universiti Putra Malaysia, Serdang,
Selangor, Malaysia
94.
Theresa Yap Wan
Cheng
theresawcyap@gmail.com
Department of Medical Microbiology, Faculty of
Medicine, University of Malaya, Kuala Lumpur


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th
Australasian Metabolomics
Symposium 2012


95.
Thomas P
Hennessy (Prof.
Dr.)
thomas.hennessy@agilent.com
Life Sciences Group, Agilent Technologies,
Singapore
96. Ute Roessner (Dr.) u.roessner@unimelb.edu.au
Australian Centre for Plant Functional Genomics
and Metabolomics Australia, School of Botany, the
University of Melbourne, 3010 Victoria, Australia
97.
Wan Iryani Wan
Ismail (Dr.)
w_iryani@puncakalam.uitm.edu.my
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor, Malaysia
98. Wong Wai Kuan neowwk@yahoo.com Universiti Malaya, Kuala Lumpur, Malaysia
99.
Yumi Zuhanis
Has-Yun Bt.
Hashim (Dr.)
yumi@iium.edu.my
Bioprocess and Molecular Engineering Research
Unit (BPMERU), Department of Biotechnology
Engineering, Kulliyyah of Engineering,
International Islamic University Malaysia, Kuala
Lumpur, Malaysia
100. Zafirah Liyana
zafira2927@puncakalam.uitm.edu.
my
Faculty of Pharmacy, Universiti Teknologi MARA,
Puncak Alam, Selangor DE, Malaysia
101.
Zakaria M Bannur
(Dr.)
knight_ly@yahoo.com
Pharmacogenomics Centre, Level 7, Faculty of
Pharmacy, Universiti Teknologi MARA, Puncak
Alam, Selangor, Malaysia
102.
Zainul Amiruddin
Zakaria
dr_zaz@yahoo.com
Department of Biomedical Science, Faculty of
Medicine and Health Sciences, Universiti Putra
Malaysia, Serdang, Selangor, Malaysia
103.
Zam Zureena
Mohd Rani
zzmr_242153@yahoo.com
UKM Medical Biology Institute (UMBI) and
Department of Medicine, Universiti Kebangsaan
Malaysia Medical Centre, (UKMMC), Kuala
Lumpur, Malaysia
104. Z
u
Zunoliza Abdullah zunoliza@frim.gov.my
Phytochemistry Programme, Natural Products
Division, FRIM, 52109 Kepong, Selangor,
Malaysia






















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4
th
Australasian Metabolomics
Symposium 2012



Floor Plan























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th
Australasian Metabolomics
Symposium 2012



Contact Number


Secretariat of Symposium

Secretariat Team members Contact number
Hospitality
(transport/ hotel)
: Mrs. Nurul Aqmar Mohd Nor Hazalin
: Ms. Sharina Hamzah
: Mr. Mohd Syafirul Shamsuddin
: Ms. Rose Iszati Ismet Nayan
019-3439154
017-6540441
013-9501647
017-3767660
Exhibition
(sponsors)
: Mrs. Lee Lian Shien
: Mrs. Norleen Mohamed Ali
: Mr. Muhammad Husaini Ismail
016-3053405
012-3457994
019-5279610
ICT
(for oral communication)
: Mr. Mohd Izwan Mohamad Yusof
: Mr. Mohd Ikmal Hanif Abdul Khalid
: Mr. Mohamad Izwan Ismail
017-9383035
019-6784205
019-3663859

































4
th
Australasian Metabolomics
Symposium 2012



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