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PHARMACEUTICAL

CLINICAL

CHEMICAL

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w w w. b i o s c i e n c e w o r l d . c a

JANUARY 2012 Volume 16, Number 1

Active micro and nano-structured

Determining rosuvastatin

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optical spectrometry

ENVIRONMENT

in human plasma by SPE-LC-MS/MS

R&D News.......................... 1 Appointments..................... 6 Pharma Notes..................... 7 New Products................... 15 Calendar........................... 17 Career Spotlight............... 18

NEW MANITOBA AAFC LAB BOLSTERS DEFENCE AGAINST PLANT DISEASES A major upgrade to a laboratory at the Agriculture and Agri-Food Canada Research Station in Morden, MB, has resulted in its certification as a Plant Pest Containment Level 3 (PPC3) facility. Originally established in 1989 as a containment lab, primarily for research on flax rust, it is the first PPC3 facility to be certified by the Canadian Food Inspection Agency under the new ‘Containment Standards for Facilities Handling Plant Pests’ in Canada. PPC3 is the highest containment level for plant pests. Containment is achieved through the use of specialized facilities, stringent operational procedures and state-of-the-art equipment. The lab provides the highest level of safety available for both staff and the public. An example of the kind of work scientists are performing in PPC3 containment at the Morden Research Station (MRS) is their investigation of Ug99 stem rust,

a plant disease that has the potential to devastate wheat crops around the world. AAFC scientists are accelerating their efforts to characterize new sources of resistance and develop new rustresistant varieties. This research will give Canadian farmers the capacity to ward off a national disease outbreak resulting in a drop in wheat production. In 2009, the federal government of Canada allocated $250,000 over two years to begin updating the laboratory at MRS. The funding went towards modernizing the controlled environment facility that is used by scientists to conduct research on exotic plant pathogens. Crucial additional support of the project came from AAFC’s Animal and Plant Health Research Initiative under the Growing Forward policy agreement. The renovation was completed in March 2011 and certified in fall.

FLORIDA SET TO HOST PITTCON 2012 Inside the PPC3 lab at Morden, AAFC’s Dr. Tom Fetch rates wheat plants for their reaction to the pathogen causing Ug99 stem rust.

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The annual Pittcon Conference & Exposition will be held in Orlando at the Orange County Convention Center March 11-15, 2012. Long known as the gathering place for top minds in science, this worldclass conference attracts leaders in scientific and technical innovation. “Pittcon is a most dynamic venue for scientists and researchers to come together in one place to see the newest equipment and learn the latest analytical and measure-

ment techniques in areas such as food and product safety; environmental hazards; homeland security; medical; and drug research and development - all of which impact our everyday lives,” says Jon Peace, president of Pittcon 2012. Organizer’s say the event is the world’s largest annual laboratory science conference and exposition. This year’s event will feature an estimated 1,000 exhibitors, Continued on page 3


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January 2012 Laboratory Focus www.bioscienceworld.ca

NEWS MOUNT SINAI RESEARCHERS UNCOVER NEW MEANS FOR DEVELOPING CANCER THERAPIES The Samuel Lunenfeld Research Institute’s Drs. Frank Sicheri, Tony Pawson and Sebastian Guettler, in collaboration with Dr. Robert Rottapel at the Ontario Can-

cer Institute, have uncovered the detailed architecture of a crucial component of Tankyrase, a protein linked to the bone development disorder cherubism and involved in

a myriad of cellular processes. The discovery is the first structural insight into precisely how the enzyme correctly identifies its targets, or substrates. The work provides

researchers with a greater understanding of Tankyrase’s cellular control processes, and may also lead to the development of new designer drugs to treat cancer. “Until now, we did not understand, from a structural perspective, how Tankyrase Continued from page 3

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January 2012

NEWS Continued from page 2 identifies its substrates,” said Dr. Sicheri, Lunenfeld senior investigator and one of the lead authors of the study. “At atomic resolution, we now have a clearer picture of what these substrates may be, and have new insight into possible novel functions of Tankyrase.” The findings are available online and were published in December in the biomedical journal Cell. Tankyrase is a poly(ADPribose)polymerase (PARP) – one protein of a family of enzymes that modify other proteins with chains of ADPribose and affect many cellular processes. The modification reactions carried out by Tankyrase can directly alter some proteins’ functions,

bring proteins together in protein complexes, or can mark others for degradation. Initially intrigued by Tankyrase because of its involvement in cherubism (a rare genetic disorder caused by mutations in the signaling protein 3BP2), the researchers built upon the findings of Dr. Rottapel’s laboratory. This laboratory found that Tankyrase normally recognizes 3BP2 and targets it for destruction. The amino acids mutated in cherubism coincide with precisely the region in 3BP2 that is recognized by Tankyrase, or the “Tankyrase binding motif.” Cherubism mutations in 3BP2 prevent binding of Tankyrase and therefore result in the accumulation of 3BP2 protein in the cell. Dr. Rottapel’s findings

also appear in the same issue of Cell. The goal of Dr. Sicheri and his team’s work was to uncover the exact mechanism by which Tankyrase recruits its substrates, to explain why cherubism mutations in 3BP2 disrupt Tankyrase binding and thereby learn more about how the enzyme works. Using x-ray crystallography, the team determined the structures of the portion of Tankyrase responsible for substrate binding, bound to a range of different substrates including 3BP2. Using a technique known as fluorescence polarization the researchers then determined the essential signature of the Tankyrase binding motif by which Tankyrase identifies its substrates.

Continued from page 1

FLORIDA SET TO HOST PITTCON 2012

Alan G. Marshall, Florida State University

2,000 presentations and attendees from more than 90 countries. Topics ranging from pharmaceutical, environmental, food safety, energy and other sciences of interest will be covered via networking sessions, technical presentations and short courses where attendees and journalists will be able to meet with the top experts on subjects of their choice and hear what’s coming for the consumer. Additionally, the Pittcon 2012 Program Committee has announced the recipients of its 11 prestigious

awards honoring scientists who have made outstanding contributions to analytical chemistry and applied spectroscopy. Alan G. Marshall from Florida State University will receive the 2012 Pittsburgh Analytical Chemistry Award. This award acknowledges Marshall’s contributions to the field of analytical chemistry through his continuing development of Fourier transform ICR mass spectrometry. His current research spans FTICR instrumentation development, fossil fuels and environmental analysis, and

mapping the primary and higher-order structures of biological macromolecules and their complexes. Other awards and recipients are as follows: Pittsburgh Spectroscopy Award, W.E. Moerner, Stanford University Pittcon Heritage Award, honoring Genzo Shimadzu, Sr. (1839-1894) and Genzo Shimadzu, Jr. (1868-1951), Accepting the award will be Shijehiko Hattori, Shimadzu chairman of the board. The Pittsburgh Conference Achievement Award goes to Christy L. Haynes, University of Minnesota. The ACS Division of Analytical Chemistry Award for Young Investigators in Separation Science Bomem-Michelson Award goes to Jared L. Anderson, University of Toledo. The Dal Nogare Award goes to Purnendu K. Dasgupta, University of Texas. The Charles N. Reilley Award goes to Debra Rolison, Naval Research Laboratory. Winning the Young Investigator Award was Lane Baker, Indiana University. The Ralph N. Adams Award went to Jonathan V. Sweedler, University of Illinois and winning the Williams Wright Award was Richard Crocombe of Thermo Fisher Scientific.

With work by Dr. Tony Pawson’s laboratory, the researchers scanned the entire inventory of human proteins, searching for the signature sequence that is recognized by Tankyrase, correctly predicting many possible new substrates for the enzyme. The result: a deeper understanding of the biology behind Tankyrase’s cellular activities. “Our work provides answers to two big questions. Firstly, we obtained a visual snapshot of how Tankyrase recognizes its substrates and how mutations characteristic of cherubism lead to illness,” said Dr. Guettler, a post-doctoral Fellow in Dr. Sicheri’s and Dr. Pawson’s labs and first author of the study. “Secondly, we learned more about

the possible cellular tasks performed by Tankyrase. The apparent abundance of potential Tankyrase targets and the variety of cellular functions they perform suggests that the complexity of Tankyrase’s biological functions has been underappreciated to date.” Inhibitors of PARPs, and among them Tankyrase, have gained considerable attention recently as potential new anticancer agents. Inhibition of Tankyrase function may hold promise for treating certain breast cancers as well as other cancers, and therefore the present study may help refine treatment strategies for blocking Tankyrase. The study was supported by the Canadian Institutes of Health Research.

SIEMENS HEALTHCARE DIAGNOSTICS AWARDED CONTRACT FOR COAGULATION LABORATORY SOLUTIONS Siemens Healthcare Diagnostics will supply its Sysmex® CS analyzer coagulation testing solutions to the Eastern Ontario Regional Laboratory Association (EORLA), a group of 16 hospitals at 19 sites in the Ottawa region. The deal, which includes Siemens and Sysmex® CS2100i analyzers, will see instrumentation being purchased over the next 24 months based on available funding at each of the sites. The Ottawa Hospital and Pembroke Regional Hospital are currently in the process of implementing the new analyzers. “ The EORLA had very unique needs and criteria for choosing a vendor, from performance of the analyzer to innovation of the assays and expert support,” says Dr. Anthony Giulivi, Division head of Hematopathology and Transfusion Medicine at the Ottawa Hospital. “The Siemens portfolio met all of our needs with comparable results across the product lines, and I was very impressed with the calibre of experts and the manufacturing facilities they offered.” The Sysmex® CS-2100i analyzer is the newest genera-

tion of coagulation systems from Siemens Healthcare Diagnostics Ltd. It is also the first coagulation analyzer to perform Preanalytical Sample Integrity (PSI) checks on every sample. Samples are scanned for haemolysis, icterus, and lipemia prior to analysis. Clotting assays are measured using five simultaneous wavelengths that can adjust for HIL in samples. This feature helps to minimize errors and the need for repeat testing as a result of those interferences. A primary tube sample volume check identifies potential inaccuracies caused by improper sample collection, accepting or flagging sample results based upon fill volume which is a user-programmable setting.

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January 2012 Laboratory Focus www.bioscienceworld.ca

NEWS CQDM INVESTS $1.5 MILLION IN TWO COLLABORATIVE PROJECTS WITHIN THE QUEBEC AND ONTARIO CORRIDOR

Rimouski

Québec City

Qu ébec Ontario

Saint-Hyacinthe Laval

Sherbrooke

Montréal

Ottawa Kingston

Lake Huron

Markham

Toronto

Guelph Kitchener/ Waterloo London

Windsor

HIV/AIDS VACCINE DEVELOPED AT THE UNIVERSITY OF WESTERN ONTARIO PROCEEDING TO HUMAN CLINICAL TRIALS one of only a few in the world. tragic health crisis affecting The first and only preventative HIV vaccine based on a genetically modified killed whole virus has received approval by the United States Food and Drug Administration (FDA) to start human clinical trials. Developed by Dr. Chil-Yong Kang and his team at The University of Western Ontario, with the support of Sumagen Canada, the vaccine (SAV001) holds tremendous promise, having already proven to stimulate strong immune responses in preliminary toxicology tests with no adverse effects or safety risks. It is the only HIV vaccine currently under development in Canada, and

“FDA approval for human clinical trials is an extremely significant milestone for our vaccine, which has the potential to save the lives of millions of people around the world by preventing HIV infection,” said Kang. Western president Amit Chakma said, “This joint venture between Sumagen and Western is a prime example of what collaboration between private industry and university researchers can achieve. Dr. Kang and his team are to be commended for their exceptional talent and remarkable persistence in developing a vaccine that addresses a

millions of people around the globe.” Before it can be commercialized, the SAV001 vaccine must go through human clinical trials with Phase 1, set to begin this month. Through WORLDiscoveries, Western’s technology transfer office, Sumagen Canada has secured patents for the SAV001 vaccine in more than 70 countries, including the U.S., the European Union, China, India and South Korea. The vaccine has been manufactured at a bio-safety level 3 (BSL3) good manufacturing practice (GMP) facility in the U.S.

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The Québec Consortium for Drug Discovery (CQDM) has announced it will fund two joint Quebec/Ontario research projects in biomedical research for a total of $1.5 million. The news was released at the conference “Connecting Life Sciences Across the Ontario-Québec Corridor” that was held in Toronto. “This investment provides an opportunity to strengthen biomedical research in Québec and Ontario by the alignment of scientific and business resources,” said Max Fehlmann, president and CEO of CQDM. Collaborating with MaRS Discovery District and MaRS Innovation, CQDM has established a new pilot competition with se-

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lected projects orientated toward the development of new tools that will accelerate the discovery of more efficient and safer drugs. The projects were evaluated by an international panel of experts for scientific excellence, innovation and impact on biopharmaceutical research. Each project will be funded with a $750K grant from CQDM. The awarded investigators are: Ram Mishra (McMaster University), Joseph Gabriele (CRP40 Inc./McMaster University), Thérèse Di Paolo (Université Laval) and Pierre Blanchet (Université de Montréal) who will validate the use of Catecholamine Regulated Protein (CRP40) as a biomarker for the diagnosis of Parkinson’s Disease. Andrei Yudin (Encycle/ University of Toronto) and Eric Marsault (Université de Sherbrooke) who will develop a new platform for rapid production of medium sized macrocycles as protein-protein interaction probes. “Ontario and Québec are well recognized for the excellence of their research and their capacity to innovate. The collaboration between the two provinces will allow to capitalize on our strengths, to push biopharmaceutical research and to increase our international competitiveness” mentioned Diane Gosselin, vice-president Research and Business Development at CQDM.


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Laboratory Focus January 2012

NEWS 2011 STEACIE PRIZE AWARDED FOR NANOTECHNOLOGY-ENABLED DISEASE DIAGNOSIS

Professor Shana Kelley

Professor Shana Kelley of the Leslie Dan Faculty of Pharmacy is the winner of the 2011 Steacie Prize, marking the third consecutive year that a University of Toronto professor has received this prestigious award. The Steacie Prize recognizes outstanding Canadian research in science and engineering. Winners are selected by a panel appointed by the E.W.R. Steacie Memorial Fund, a private fund dedicated to advancing science and engineering in Canada. “It’s wonderful and at the same time humbling to get this type of recognition – which is really recognition of the talents of all of the past and present members of my research group,” Kelley said.

Kelley’s research centres on the development of nanomaterial-based detection systems that can track miniscule quantities of biomolecular analytes [components]. The highly sensitive DNA and RNA detection systems developed by Kelley are powerful new tools for cancer and infectious disease identification, and represent a major advance over current methods. Using small, non-invasive samples, Kelley’s diagnostic tool is able to identify minute levels of the biomarkers of disease. This technology is able to provide disease diagnosis at a fraction of the costs and in a fraction of the time of current methods, and is able to do so 10 times earlier than current practices allow.

GAMMA-DYNACARE ACQUIRES WARNEX MEDICAL LABORATORIES Laval’s Warnex Inc., provider of lab services to the pharmaceutical sector, has sold the assets of its Medical Laboratories Division for $7.5 million to Ontario’s Gamma-Dynacare Medical Laboratories and will use part of the proceeds to repay most of its debenture debt. Gamma-Dynacare, one of Canada’s largest providers of lab services, will assume certain liabilities of Warnex, taking over 25 employees. Warnex could get up to an additional $2 million in an earnout based on the performance of its Medical Laboratories Division in the year following the takeover. The sale and agreement with the debenture holders “are key steps in our efforts to strengthen the balance sheet and position the company for the future,” said Michael Singer, chairman of Warner’s executive committee. Gamma-Dynacare has more than 2,000 employees, four labs in Ontario, one in Montréal and three in Winnipeg. It also operates 200 patient services centres in Ontario, Québec, Manitoba, Saskatchewan and Alberta. Warnex Medical Laboratories’ biochemistry services include the Prenatest® prenatal screening test – which combines a blood test with ultrasound imaging to assess the risk of Down syndrome

and other fetal chromosomal abnormalities – and the PCA3 test for the detection of prostate cancer. The laboratory also performs a variety of microbiology, virology and serology tests using advanced techniques to provide data to diagnose and evaluate infectious diseases, such as the Influenza A H1N1 virus. Its molecular diagnostics services include DNA-oriented genetic predisposition and tracking tests in various fields of medicine, including oncology, endocrinology and hematology. Through its PRO-DNA Services subsidiary, Warnex

Medical Laboratories is also a leading provider of DNA identification tests for forensic and parentage testing purposes. Gamma-Dynacare plans to operate Warnex Medical Laboratories on a “status quo” basis, with no immediate changes to services or operations. Dr. Yvan Côté, currently the vice-president and general manager of Warnex Medical Laboratories and Warnex PRO-DNA Services, will be assuming a new role with Gamma-Dynacare and will continue to oversee dayto-day operations in Laval and Thunder Bay.

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January 2012 Laboratory Focus www.bioscienceworld.ca

APPOINTMENTS

Dr. Claude Lazure has been appointed interim scientific director and Dr. Michel Bureau has been appointed as an adviser to the Fonds de recherche du Québec - Santé (FRQS), effective January 2012. The appointments were recently announced by Rémi Quirion, chief scientist of Québec and chairman of the board of directors of the FRQS. A researcher at the Institut de recherches cliniques de Montréal, Dr Lazure has been a scientific adviser to the Fonds’ research centre program for the past 10 years. He has also served on several of the Fonds’ evaluation committees. President and CEO of the Fonds from 1996 to 2003, Dr Bureau will bring his clinical research expertise to the organization, especially in matters pertaining to the Québec Biopharmaceutical Strategy and the development of clinical research in Québec in partnership with the Canadian Institutes of Health Research. The FRQS reports to Quebec’s Minister of Economic Development, Innovation and Export Trade and is devoted to promoting and

 Continuing

supporting health research in the province. Zymeworks announces the appointment of Dr. Gordon Ng to the position of vice-president, Preclinical Research and Development. Dr. Ng joins Zymeworks with over fifteen years of biotechnology and pharmaceutical drug discovery experience from the Myelin Repair Foundation, Amgen, and the Merck Frosst Center for Therapeutic Research. During his nine years at Amgen he served as a Scientific director and was instrumental in advancing multiple preclinical candidates towards the clinic, as well as managing multiple internal and partnered research initiatives. Isotechnika Pharma Inc. appoints Donald W. Wyatt to to its board of directors. Wyatt will serve as 3SBio’s representative on the board replacing Dr. David Chen, chief operating officer of 3SBio. Wyatt brings over 20 years of experience in technology transactions and legal practice, including both private and corpo-

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rate law. He is the founder of The Wyatt Group, a firm that provides consulting on intellectual property acquisition, management and divestiture, including strategic alliances, establishment of subsidiaries and licensing, focusing particularly on US/China technology transactions. Wyatt is currently an advisor to 3SBio. His background in public company legal affairs spans a wide range of responsibilities, including primary management of general legal affairs, transactions, SEC filings, litigation, and an emphasis on worldwide patent management, strategic counseling, regulatory compliance, contracting, litigation management and personnel management. He has extensive biotech and pharmaceutical industry legal experience, including time at Cell Therapeutics and Schering-Plough. Mr. Wyatt received his Juris Doctor and Master of Intellectual Property from the New Hampshire School of Law. Kane Biotech Inc. announces the appointment of Mark Matthewson as the company’s chief financial officer. Matthewson’s services are provided through a Management Services Agreement with MKM Management Services Ltd., a company founded by Matthewson in 1999. He also acts as chief financial officer for Biomedical Commercialization Canada Inc. He brings a wealth of knowledge and expertise to the business and was previously CFO for Monteris Medical Inc., and IMRIS Inc., where he led all aspects of financial management and reporting. Matthewson is a chartered accountant and holds a Bachelor of Science (Mathematics) degree from the University of Manitoba. Miraculins Inc. announces it has appointed Dr. Milan Gupta, MD, FRCPC, FACC to its PreVu medical advisory board. Dr. Gupta is an associate clinical professor of Medicine at McMaster University in Hamilton, ON. He is also an assistant professor of Medicine at the University of Toronto, adjunct scientist in the Division of CV Surgery and an affiliate scientist at the Li Ka Shing Knowledge Institute at St. Michael’s Hospital in Toronto. He is a cofounder and past-chair of the Greater Toronto Area Community Cardiovascular Specialists (GTACC), and past Board member of the Cardiac Care Network of Ontario. He is also the Medical codirector of the Canadian Cardiovascular Research Network. Dr. Gupta received his MD from the

University of Toronto in 1988. Following completion of internal medicine and cardiology residencies in Toronto, he received certification by the Royal College of Physicians and Surgeons of Canada, as well as by the American Board of Internal Medicine. He holds additional certification status by the American Society of Nuclear Cardiology and by the North American Society for Pacing and Electrophysiology. BioTalent Canada has broadened its leadership team with the appointment of François Schubert as chair of the BioTalent Canada Board of Directors effective immediately. He succeeds Bob Ingratta of Fast-Trak Strategies who will remain on the Board as past chair. Schubert is the general manager and chief administrative officer at The Research Institute-McGill University Health Centre. Having already served as a Board Member for BioTalent Canada since 2008 and on BioTalent Canada working groups, Schubert brings more than 30 years of national and international experience in academia, research, biotechnology and Big pharma to BioTalent Canada. MethylGene Inc. announces the appointment of Dr. Rachel Humphrey to the post of executive vice-president and chief medical officer. Dr. Humphrey is board certified in Medical Oncology and has led the development of multiple oncology agents, two of which are currently commercialized (Nexavar® and YERVOY®. Dr. Humphrey joins MethylGene from BristolMyers Squibb where she was most recently vice president, Global Development Lead, Immuno-Oncology. From 2003 to 2011, Dr. Humphrey supervised a number of in-licensed and internal development programs including the Phase 2 to 4 development of ipilimumab (YERVOY®), which achieved global approval for the treatment of metastatic melanoma in 2011. From 1997 until 2003 Dr. Humphrey held increasingly senior clinical development positions at Bayer. Dr. Humphrey received her B.A. degree in biochemistry from Harvard University, her M.D. degree from Case Western Reserve University, and completed her Medical Residency at the Johns Hopkins Hospital. From 1992 until 1997, Dr. Humphrey worked at the U.S. National Cancer Institute (NCI), first as a Clinical Oncology Fellow and later as a staff physician/ scientist in the NCI’s HIV and AIDS Malignancy Branch.


www.bioscienceworld.ca Laboratory Focus

January 2012

PHARMA NOTES Bioniche Life Sciences Inc. (Belleville, ON) has entered into two distribution agreements to expand its product offerings in North America. The first agreement is with MedTrade Products Limited, a UK-based company. The agreement provides a number of equine and companion animal health products based on MedTrade’s CeloxTM technology for exclusive distribution by Bioniche in Canada. The CeloxTM technology uses chitosan, a natural polysaccharide, as a medical device in the animal health industry. The CeloxTM product line includes gauzes and granules that are utilized in wound healing. When mixed with blood, CeloxTM forms a gel-like clot in less than one minute, independent of the body’s normal clotting processes. The second agreement is with Mueller Medical International LLC, a U.S.-based company. The agreement provides an equine product – Equine Gastrafate® - for exclusive distribution in Canada and the U.S. by Bioniche. Equine Gastrafate® is a patented saccharide composition that is used to expedite management of gastrointestinal syndromes characterized by nausea, vomiting, diarrhea, colic and mucosal erosions. This product is often used by equine veterinarians who suspect ulcer colic in horses.

lieved to develop as a result of dysfunction in the protective epithelial layer lining the bladder.

Trillium Therapeutics Inc. (Toronto, ON), a privately-held biopharmaceutical company developing proprietary and innovative biologic therapies begin a Phase 1 clinical trial of its experimental drug, TTI-1612, in patients with interstitial cystitis/bladder pain syndrome (IC/BPS). The company has recently received a No Objection Letter to its Clinical Trial Application from Health Canada’s Biologics and Genetic Therapies Directorate. The single ascending dose trial will be conducted at multiple sites across southern Ontario. IC/BPS, also known as Painful Bladder Syndrome, is a chronic, debilitating and poorly treated bladder disease affecting millions of people. The disease is be-

Xenon (Vancouver, BC) announces a strategic al-

Oncolytics Biotech Inc. (Calgary, AB) has released interim data from its Phase 2 clinical trial using intravenous administration of REOLYSIN® in combination with gemcitabine (Gemzar®) in patients with advanced pancreatic cancer (REO 017) indicating that the clinical study has successfully reached its primary endpoint, and that the drug combination is active. To date, eight patients of 13 evaluable patients in the study had stable disease (SD) for 12 weeks or longer, for a clinical benefit rate (complete response (CR) + partial response (PR) + SD) of 62 per cent. An additional patient had an unconfirmed PR of less than six weeks. The study is using a one sample, two-stage design. In the first stage, 17 patients were to be enrolled, and best response noted. If less than three responses (defined as CR or PR or SD for 12 weeks or more) were observed, the study would have concluded that the combination was inactive and been terminated. If three or more responses were observed among the 17 patents, the study would enroll an additional 16 patients for a total of 33 evaluable patients.

liance with Genentech, a member of the Roche Group, to discover and develop compounds and companion diagnostics for the potential treatment of pain. Under the terms of the agreement, Genentech has an exclusive license to compounds and a non-exclusive license to diagnostics from Xenon for development and commercialization of products. Xenon will receive an undisclosed upfront payment, research funding and is eligible to receive research, development and commercialization milestone payments, totaling up to $646 million for multiple products and indications. In addition, Xenon will receive royalties on sales of products resulting from the collaboration. Aeterna Zentaris Inc. (Québec, QC) announces that the U.S. Food and Drug Administration (FDA) has granted Jose M. Garcia, MD, PhD an Investigational New Drug (IND) approval for the initiation of a Phase 2A trial to assess the safety and efficacy of repeated doses of the company’s ghrelin agonist, AEZS-130 (macimorelin), in patients with cancer cachexia. The study is a double-blind, randomized, placebo-controlled Phase 2A trial to test the effects of different doses of the ghrelin agonist, AEZS-130, in 18 to 26 patients with cancercachexia. The study will be conducted under a cooperative research and develop-

ment agreement (CRADA) with the Michael I. DeBakey Veterans Administration Medical Center which will be funding the study. AEZS-130 will be provided by Aeterna Zentaris. Allon Therapeutics Inc. (Vancouver, BC) has been granted a U.S. patent covering the use of Allon’s drug candidates, including its lead product davunetide, and other pipeline products for the treatment of laserinduced retinal damage. This new patent strengthens Allon’s intellectual property estate, which includes 15 patent families, 60 issued patents and over 30 pending applications worldwide. This patent relates to findings recently published in Acta Ophthalmologica which reported that davunetide has neuroprotective effects in an animal model of retinal laser injury. The results suggest the potential of davunetide as a treatment for retinal damage following retinal laser photocoagulation or other types of ophthalmic laser surgeries. Helix BioPharma Corp. (Toronto, ON) announces that the “clinical hold” on its investigational new drug application for its Topical Interferon Alpha-2b, Phase 2/3, low-grade cervical lesion efficacy trial has been removed by the U.S. Food and Drug Administration (FDA). The proposed Phase 2/3 trial is planned to be a

7

randomized, double-blind, vehicle-controlled study in patients with cervical intraepithelial neoplasia grade 1 or 2 lesions (CIN 1 or CIN 2 respectively). The intended sample size is 492 female subjects to be randomized in a 2:1 ratio of active to control. Eligible women will be premenopausal subjects aged 18 - 55 years at screening, with histologicallyconfirmed CIN 1 or CIN 2 on colposcopic directed biopsy at screening and high risk human papillomavirus (HPV) infection upon the Hybrid Capture® 2 HPV-DNA test. The proposed primary study endpoint will be the resolution of CIN 1 or CIN 2 at month 12, determined by colposcopic directed cervical biopsy together with Pap smear cytology free of ASC-H (atypical squamous cells that cannot exclude high-grade squamous status), AGUS (atypical glandular cells of undetermined significance), LSIL (low-grade squamous intraepithelial lesions), HSIL (high-grade intraepithelial lesions), and adenocarcinoma in situ (AIS) or adenocarcinoma. The study is designed with a 12-month overall duration per patient, including treatment and follow-up. Helix plans that the results of its U.S. Phase 2/3 trial and the results of its European Phase 3 trial, if successful, will be submitted together in order to seek U.S. and European marketing authorizations for the product for this indication.


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January 2012 Laboratory Focus www.bioscienceworld.ca

FEATURE

BY DR. GUERMAN PASMANIK

Active micro- and nano-structured

optical spectrometry (AMOS) General description of AMOS, technologies to be used and advantages of ps-UV pulses

Introduction Sensors for the discovery and identification of both simplest inorganic molecules and organic compounds (including macro- and bio-molecules in living organisms) are mainly used in pharmacology, bio-medicine and chemistry. In general, the progress in sensor technology is determined by our ability to decompose different molecules contained in a “big” volume over elementary “cubicles”, containing identical molecules with equal mass, mobility, optical spectrum, etc. Thus, in particular we are able to determine the presence of low concentration of analytes given the information about processes, say, in human cells. The most advanced current method of sensing is associated with mass spectrometry. This method has become established as the primary means for protein identification from complex mixtures of biological origin, uncovering the set of proteins that make up human cells. Moreover, due to new approaches based on MALDI imaging, the mapping of protein distribution inside of cells becomes routine procedure. Prospectively measuring the dynamics of map variations make predicting a living cells lifetime achievable. However, this technique is rather expensive and can’t be applied for very big molecules (with sizes of tens and hundreds nanometers). As an alternative, Ion mobility spectrometers allow classification of very big mol-

ecules, but to use this method one usually has to know in advance what kind of analytes need to be found. Laser induced fluorescence gives detailed information about analyte spectra, but strong background emission usually masks the low concentrated molecules of interest. Raman spectroscopy can also provide very useful information about molecular structure and outlines features of molecules with close chemical performances, but it too is masked by laser induced fluorescence which usually exceeds Raman emissions on several orders. As such, there is a need for new methods in detecting molecular structure, and for sensing inorganic molecules and organic compounds. This article proposes a new electrically-controlled (active) filter for the trapping of molecules with high polarizabilities. This new method of molecular sensing (active micro/nano-structured spectroscopy - AMOS) is based on our proprietary technology. The important features of AMOS are its ability to trap bio-molecules based on their polarizability. According to our estimation it is feasible achieving of sensitivity for detection of admixed molecules up better than part per trillion. Likewise, this filter can be a core element of ultra-sensitive analytical spectrometers, filtration of organic compounds in pharmacology, water extraction from air and oil, etc.

Among the different methods of sensing, there are separately staying adsorption methods. These methods are characterized by the physical adherence or bonding of ions and molecules onto the surface of another phase. The typical example is a solgel plate, which absorbs water from air. Absorbed material is analyzed, for example, following ionization (in IMS technology) or optical spectra analysis (Laser Induced Fluorescence, Raman or molecular spectroscopy). However the absorber used for molecules of interest accumulation usually is passive. Recently we have developed a new method of sensing based on a lasermade active filter consisting of dense packaging of micro-holes in dielectric plate matches to vias in structured metal or Indium Tin Oxide electrodes (see Figure 1). By controlling voltage on these electrodes and temperature of the filter it is possible to accumulate molecules and organic compounds near (or inside) of the holes. The physical reason for such accumulation is that the electrostriction interaction of neutral molecules and organic compounds occurs with a high voltage electrical field having a strong non-uniform distribution near and inside of the holes. Especially if such holes are blind (not drilled through a dielectric plate) the bottom of the holes has quasi-periodical spikes (like “stalagmites”) with a typical distance between them achieving up to λ/3, where λ is the laser wavelength in the UV band. For 213 nm, that distance can achieve 71 nm and the transverse size of correspond-

ing spikes is two to three times less (about 25 nm). Due to high gradient of electrical field (sharp spatial change with nanometer scale) the electrostriction forces attract neutral molecules and organic compounds (in particular proteins) inside of the holes to allow bonding them around spikes. As more polarizability α has molecule as stronger interaction occurs. If the molecule velocity induced by electrostriction force (~ τα E2/m) is comparable to its thermal velocity (~ kT/m), then a molecules bonding becomes strong enough to fix that molecules to the filter. Here we use denotations: k is the Boltzmann constant, T is the filter temperature, τ is the time between molecules collisions (free running time), m is the molecule mass. However, if polarizability α is not very high the molecules can’t be fixed on the filter surface or inside of the hole. From the above, one will notice that the selective attraction of analyte to the filter’s holes facilitates optical spectral analysis with relatively low background emission. Thus, one of the important features of AMOS is its ability to trap the bio-molecules based on their polarizabilities, and furthermore the larger the polarizability of the molecule, the better the trapping performances of the AMOS as an analytical device. One important thing to note is that the number of big bio-molecules trapped into each micro-hole (spot) is discrete; correspondingly the molecular performances in individual spots, say, their dipole moments or scattering cross-sections, are also discrete (proportional to the number of molecules). If in the results obtained


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9

January 2012

FEATURE Figure 1

Electrically controllable (active) filter

This is a dielectric plate (100 um silica or lavsan) with 2 orthogonal metal (or Indium Tin Oxide) stripes (marked by yellow color) with 50-100 lines/mm deposited on the opposite sides of the plate and having tiny (5-10 um) holes drilled between the stripes.

there are non-discrete numbers of molecules, then this means that there are some unusual features of trapped molecules. Thus, analyzing all spots (pixels), say, via measuring of response on electrical field scanned over filter cross-section or spectrum of scattered light, we will be able to obtain additional information about the properties of the analyte molecules. Definitely in the case when there is a relatively low concentration of those molecules, vast majority of micro-holes (pixels) don’t have analyte molecules. Nevertheless there may be present spots with analyte molecules. In other words, if analyte molecules are available in any solution, they will also be trapped in certain micro-holes, which make their recognition easier. The most suitable instrument to execute the investigation of analyte molecules in these considered conditions is Raman scattering of the laser light. Raman lines unlike fluorescence will provide more information about the geometrical shape of the molecule. In particular, the organic compounds containing the same elementary groups of molecules, but orientated in different directions with respect to the rest of molecules, have different Raman vibration and rotational modes. Recognition of these modes allows for the discovery of organic compounds containing certain defects, for example, a minor group of molecules originated from a parent bio-molecule, but bonded to a molecular body in an unusual manner (from the side or from anoth-

Figure 2

In the second technology process, a high energy UV laser beam focused by cylindrical optics ablates the surface of the substrate with a thin metal film deposited on one of its faces. The period of patterning is about 5 - 1 20 μm. From another face of the same plate the patterning is to be made with the stripes having orthogonal orientation. Then the UV laser beam drills holes between the stripes in each spot. The voltage given to the structured metal film provides an electrical field up to several kV/cm. That voltage can be variable from spot to spot allowing variable electrical field over the surface. Thus, one can control conditions and create traps for molecules that have different polarizabilities.

Features of AMOS and ways to the improve AMOS design

Set of electrical circuits to excite magnetic field attracting atoms with high magnetic polarizability

er end). The processing of obtained results may uncover details of reproductive living organisms including cancer and other diseases at earlier stages. The use of molecular scattering, in particular so-called Rayleigh wing scattering where the broadening of frequency spectrum and polarization features of scattered light characterize the molecules, is extremely important. Also, we have mentioned the possibility of using surface enhanced Raman scattering (for some molecule complex having a conduction band). And finally regarding Coherent Anti-Stokes Raman Scattering, this method may provide a very high sensitivity especially if we know in advance what kind of analyte is expected. According to our estimation, this means achieving the sensitivity of about one part per trillion.

Technology of AMOS fabrication The most suitable technology for AMOS fabrication is based

on UV picoseconds lasers where the lasers are used in two technology processes: 1) Controllable hole drilling in metal and dielectrics with an accuracy of ~0.1 μm. 2) Controllable removal of thin metal layers deposited on dielectric substrates. Using these two technology processes we are able to deposit a thin metal film on a fused silica surface and drill 5 to 10 μm holes though that film at a depth of about 10100 μm. In the first technology process, due to its relatively high aspect ratio (the depth of the hole to its diameter) the short wavelength UV laser beam (λ = 266 nm or 213 nm) touches the walls of the hole and is reflected towards the hole bottom initiating light-plasma interaction causing the spatial instability of material removal process. The result of that instability is the development of a pattern with a period of about λ/2n (here n is reflective index of dielectric substrate).

The performance of the AMOS depends on diameter and surface density of the holes, their depth, the ability of making spikes on the bottom, type of dielectric material and the polarization of the laser beam making the holes. We can separate several types of AMOS technology: 1. Simplified version of AMOS based on metal-dielectric-metal “sandwich”. In that case the holes through metal and dielectric are made from opposite sandwich faces. The holes from one face are blind, and holes from opposite side are drilled only through metal. It allows collecting analytes on one side and provides optical spectral analysis on the other; 2. A Structured version of AMOS based on the ability to control voltage inside of individual holes; 3. A variation of the metaldielectric-metal “sandwich” with simpler (no blind) holes. The main goal of such a device is filtering molecules with high polarizability and ensuring their separation in a gaseous or liquid mixture from other molecules.

One new and interesting structures made by the UV laser ablation is the set micro circuits around the vias or blind holes (see Figure 2) The electrical current passes these micro-circuits with a typical diameter of 10-20 μm excites the magnetic field inside of the hole. This creates a field that selectively attracts atoms, molecular complex and micro-particles with high magnetic polarizability. This method can open up new possibilities for analysis of metal micro-particles in oil or cleaning oil from such micro-particles, by trapping of atoms of less-common metals, for isotope separation, etc. One could call it “Ω-technology” because each micro-circuit looks like the Ancient Greek letter Ω.

Expected applications and market prospects for AMOS There are several important applications associated with the use of active filters including the filtering of analytes which would prove useful to the bio-medical analysis, pharmaceutical industry. The filters can also be adapted for ion mobility spectrometry, in separating selected molecules or organic compounds such as those required for the growing of crystals including organic ones. The filters can assist in water extraction from oil to provide improved performance of gasoline or kerosene. Likewise they can be used for high efficient water extraction from air or alternatively air extraction from water.

Author thanks Dr. Sabatino Nacson, Adolf Kleiner and Mrs. Maria Konchalina for fruitful discussion and help. Dr. Guerman Pasmanik is the lead researcher for Amos Photonics Research Technology, a small technology company, developing and selling optical equipment globally.

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January 2012 Laboratory Focus www.bioscienceworld.ca

FEATURE

B Y : W ILLIA M FA ULKNER , KIMB ERLEY PHIPPS , THERMO F IS HER S CIENTIFIC, RU NCORN, CHESHIRE, U K

Determination of Rosuvastatin in Human Plasma by SPE-LC-MS/MS using SOLA and Accucore RP-MS column

A SIMPLE, RAPID AND SENSITIVE PROCEDURE for the determination of rosuvastatin in human plasma by liquid chromatography-tandem mass spectrometry was developed and evaluated. The drug was isolated from within the plasma matrix using a Thermo Scientific SOLA 96 well plate, and the components of the resultant extracts were separated on a Thermo Scientific Accucore RP-MS column under reversed-phase, gradient conditions. Detection was performed on a triple quadrupole mass spectrometer under positive polarity, heated electrospray ionisation (HESI) conditions operating in selected reaction monitoring (SRM) mode. The analytical procedure is both accurate and precise, and is characterized by high levels of recovery and an absence of any significant matrix interfering effects. Introduction Rosuvastatin [(3R, 5S, 6E)-7 - [4- (4 fluorophenyl) -2- (N-methylmethanesulphonamido) -6- (propan-2-yl) pyrimidin-5-yl] -3, 5 - dihydroxyhept -6-enoic acid] is a synthetic, orally administered member of the ‘statin’ class of cholesterol lowering drugs. This particular statin is marketed by Astra Zeneca as ‘Crestor’. Employed as an adjunct to dietary modification, the drug is used to treat primary hypercholesterolaemia, mixed dyslipidaemia and hypertriglyceridaemia in an attempt to reduce the risk of atherosclerosis and poor cardiovascular health. In terms of the mechanism of its action, rosuvastatin is a selective and competitive inhibitor of the enzyme 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase. This particular enzyme catalyses the conversion of HMG-CoA to mevalonate, a precursor of cholesterol. Accucore™ columns are packed with silica particles that have been manufactured using a core enhanced technology. These particles comprise a solid core upon which a porous layer is deposited. This configuration minimizes diffusional (mass trans-

fer) pathways and permits the use of shorter columns and higher flow rates to achieve remarkably fast, highly efficient and highly resolved separations. In contrast to totally porous sub-2 μm particles, columns containing the solid core particles generate a competitive number of theoretical plates, but, with greatly reduced backpressures. Accucore RP-MS columns employ an alkyl chain of an optimized length in order to provide a more effective coverage of the silica surface. The immobilised phase exhibits a slightly lower retentivity than that offered by traditional C18 phases, but, the resultant high efficiencies and excellent peak symmetries make this the phase of choice for those chromatographic procedures which demand detection by mass spectrometry. SOLA™ is a revolutionary new solid phase extraction (SPE) product. This first in class SPE product range introduces next-generation, innovative, technological advancements which offer unparalleled performance in comparison to conventional SPE and products intended for the precipitation of proteins and phospholipids. The ben-

Figure 1

Figure 2

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Assessment of accuracy and precision Procedural accuracy and precision were evaluated by replicate (n = 6) examination of extracted QC samples at three levels of concentration. A summary of the results is shown in Table 2. The accuracy and precision of the analytical procedure were found to fall comfortably within the limits of acceptance generally applied to bioanalytical methods.

90

Relative Abundance

Relative Abundance

Area Ratio

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Linearity of response The relationship between analytical response and concentration was investigated over the range 1-1000 ng/ mL. A calibration line was constructed using data derived from extracted matrix-fortified samples at eight levels of concentration (excluding blanks). Calibration lines were run at the beginning and end of the analytical sequence with the bottom and top standards being extracted in duplicate in both lines. All data were used to assess the degree of linearity. A graphical plot of relative response (Astd/Aistd) as a function of the concentration of rosuvastatin is shown in Figure 1. Calibration data are summarised in Table 1. The analytical response was found to be linear (using a 1/x weighted regression algorithm) with a coefficient of determination (r2) of 0.9984 in the range 1 - 1000 ng/mL.

100

SRM derived from examination of extracted standard S1 (1 ng/mL)

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Figure 3

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Linearity of response over the dynamic range 1 – 1000 ng/mL r2 = 0.9984

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efits of using SOLA products include: (1) higher levels of reproducibility, (2) improvements in the cleanliness of extracts, (3) reduction in elution volumes (4) increases in procedural sensitivity. SOLA products have significant advantages for the analyst when isolating compounds from complex matrices, particularly in high throughput bioanalytical and clinical laboratories where higher speed of analysis and the ability to use smaller quantities of sample and solvent are critical. The increased performance of SOLA products gives greater confidence in the accuracy of analytical results and lowers cost without compromising ease of use or requiring complex method development. A number of researchers have reported the measurement of rosuvastatin in human plasma and pharmaceutical formulations.1,2 Typical chromatographic approaches include separation using C18 phases (via hydrophobic interactions) and retention of the ionized molecule via an ion-exchange mechanism. The purpose of this particular study is to demonstrate the effectiveness of a combination of SOLA products (reversed

phase) solid phase extraction material and an Accucore RP-MS column for the determination of rosuvastatin in human plasma by liquid chromatography tandem mass spectrometry.

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Laboratory Focus January 2012

Evaluation of recovery The recovery of analyte was assessed by comparison of the measured concentrations of rosuvastatin in matrix extracted QC samples with those concentrations found in post-extraction spiked samples which had been fortified at the same level (See Table 3). The level of analyte recovery (99.3 %) and the precision (% RSD = 4.88) between replicates demonstrate that both the efficiency of the extraction procedure and its repeatability are substantially more than satisfactory. Statistical assessment is based upon data derived from the replicate examination of both pre- (n = 6) and postextracted (n = 3) plasma samples. Evaluation of matrix effect The existence of a matrix effect was determined by comparison of the measured concentrations of rosuvastatin in post-extracted fortified plasma samples with those concentrations found in the reference standards that had been fortified at the same level. The results, summarized in Table 4, indicate that there is no significant matrix effect. Statistical assessment is based upon data derived from the replicate examination of both post-extracted plasma samples (n = 3) and reference standards (n = 6). Specificity and sensitivity SRM chromatograms derived from the examination of the unfortified and fortified plasma samples are shown in Figures 2 and 3. It is evident that the unfortified plasma sample contains an impurity (Tr = 1.92 minutes). However, under the adopted chromatographic condi-

tions, the separation is sufficient to prevent any overlap of the response from this endogenous plasma species upon the principal analytical response (Tr = 1.49 minutes). The SRM derived from the zero blank is shown in Figure 4. The mass spectral evidence suggests that there is no proton exchange and concomitant conversion of the deuterated to the non-deuterated form.

Conclusion An analytical procedure based upon SPE-LC-MS/MS for the determination of rosuvastatin in human plasma was successfully developed and evaluated. The procedure was found to exhibit good linearity (r2 = 0.9984) for concentrations of rosuvastatin in the range 1 - 1000 ng/mL. The accuracy and precision for all samples examined were found to be < 6.1 % and < 5.8 % respectively. At a concentration of 400 ng/mL, the level of recovery of analyte (99.3 %) and its repeatability (% RSD = 4.9 %) were both found to be excellent. There was no significant matrix effect. The performance characteristics of the method combined with its simplicity and rapidity mean that it can be adopted routinely in clinical environments.

References 1. Pelat, M.; Dessy, C.; Massion, P.; Desager, J.-P.; Feron, O.; Balligand, J.-L., J. Circ. 2003, 107, 2480 2. Trivedi, R.K.; Kallem, R.R.; Mullangi, R.; Srinivas, N.R., J. Of Pharm. Biomed. Anal. 2005, 39(3-4), 661-669

FEATURE SPE Apparatus Part Number SOLA 96-well plate (10 mg/2 mL)

60309-001

Calibration Standards A primary standard of rosuvastatin was prepared in 1:4 (v/v) MeOH-water at a concentration of 105 ng/mL. Secondary standards (SS1 – SS8) were prepared by subsequent serial dilution of the primary standard in 1:4 (v/v) MeOH-water. 4.5x10-LabFocus:2

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Page 1

Safeguard Our Aquatic Animal Resources Protect aquatic animals against disease If you find or suspect a reportable disease in finfish, molluscs or crustaceans, you must tell the Canadian Food Inspection Agency (CFIA). Reportable diseases could seriously harm Canada’s aquatic animals, international trade and the economy. The CFIA works to prevent these diseases from being introduced into, and spreading within, Canada. For more information and a listing of reportable aquatic animal diseases, call 1-800-442-2342 or visit www.inspection.gc.ca/aquatic

Experimental Details Chemicals and Reagents

Part Number

Fisher Scientific Water (LC-MS grade)

W/0112/17

Fisher Scientific Methanol (LC-MS grade)

M/4062/17

Fisher Scientific Formic acid (‘Optima’, 90 %, LC-MS grade)

A117-50

Fisher Scientific Propan-2-ol (HPLC grade, 99.5+ %)

P/7507/17

Fisher Scientific Acetonitrile (LC-MS grade)

A/0638/17

Fisher Scientific Acetone (AR grade, 99.8+ %)

A/0600/15

Human plasma (Supplied by Seralab, lithium heparin as anticoagulant) Sample Handling Equipment Fisher Scientific Finnpipette F2 pipettor kit

Part Number PMP-020-220F

10 μL - 100 μL, 100 μL - 1000 μL, 1 mL – 10 mL Fisher Scientific Finntip pipette tips, 10 μL

PMP-107-110W

Fisher Scientific Finntip pipette tips, 200 μL

PMP-107-600F

Fisher Scientific Finntip pipette tips, 1000 μL

PMP-103-206K

Fisher Scientific Finntip pipette tips, 10 mL

PMP-107-040R

Thermo Scientific borosilicate glass vials

60180-600

(2 mL, 12 mm x 32 mm) with 8 mm black screw cap fitted with a silicone/PTFE seal Solid Phase Extraction Hardware Thermo Scientific positive pressure SPE manifold (capable of processing 96-well microplates). Solvent evaporation system (capable of processing 96-well microplates).

Contribuons à la protection de nos ressources animales aquatiques Protégeons les animaux aquatiques des maladies Si vous décelez ou soupçonnez la présence d’une maladie déclarable chez des poissons, des mollusques ou des crustacés, vous devez en informer l’Agence canadienne d’inspection des aliments (ACIA). Une maladie déclarable peut avoir des effets néfastes sur les animaux aquatiques du Canada, le commerce international et l’économie. L’ACIA s’efforce de prévenir l’introduction et la propagation de ces maladies au pays. Pour de plus amples renseignements et pour obtenir une liste des maladies des animaux aquatiques déclarables, appelez le : 1-800-442-2342 ou consultez le : www.inspection.gc.ca/aquatique


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January 2012 Laboratory Focus www.bioscienceworld.ca

FEATURE

Uniguard direct connection guard cartridge holder

852-00

(2.1 mm I.D) A primary standard of d6-rosuvastatin, to be used as internal standard, was

Mobile phase:

prepared in 1:4 (v/v) MeOH/water at a concentration of 5000 ng/mL.

(A)

Water + formic acid at 0.1 % (v/v)

Calibration standards of rosuvastatin were prepared at eight different levels

(B)

MeOH + formic acid at 0.1 % (v/v)

of concentration (i.e., 1, 2, 10, 50, 250, 500, 900 and 1000 ng/mL) by fortification of the plasma matrix (100 μL) with a measured quantity (10 μL)

Gradient:

of appropriate stock standard. The internal standard (d6-rosuvastatin) was

Time (min)

added at the 500 ng/mL level into each of the calibrants.

0 5

Standards S1 and S8 were prepared in duplicate whilst the remaining

0.1 5

standards (S2 – S7) were prepared in singlet

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1.62 5

QC standards of rosuvastatin were prepared in replicate (n = 6) at three

3.60 5

levels of concentration (i.e., 3, 400 and 750 ng/mL) by fortification of the

Flow rate:

0.75 mL/min

plasma matrix (100 μL) with a measured quantity (10 μL) of appropriate

Column temperature:

60 °C

spiking solution. The internal standard (d6-rosuvastatin) was added at

Detection:

MS

500 ng/mL.

Injection volume:

15 μL

Syringe volume:

100 μL

QC Standards for Post-Extraction Fortification (i.e., ‘overspiking’)

Syringe flush:

Strong solvent - 9:9:2 (v/v) MeCN/iPrOH/acetone

Unfortified plasma samples (n = 9) were extracted in triplicate and fortified

Weak solvent - 1:4 (v/v) MeCN-water

post extraction at three levels of concentration, i.e., 1, 400 and 750 ng/mL.

Autosampler temperature - 10 °C Run time:

3.60 minutes

Reference (unextracted) Standards Reference standards (which were not subject to extraction) were prepared in

Mass spectrometric conditions

replicate (n=6) at three different levels of concentration by the introduction

Ionisation parameters:

of a measured quantity (10 μl) of appropriate spiking solution ([Ros.]QCLLOQ

Heated electrospray operating in positive polarity mode (HESI-2 probe)

= 10, [Ros.]QCMED = 4000, [Ros.]QCHIGH = 7500 ng/mL) and internal standard (d6-

Spray voltage:

3000 V

rosuvastatin, 10 μL, [d6-Ros.] = 104 ng/mL) into 1:4 (v/v) MeOH-water (180

Vaporiser temperature:

475 °C

μL).

Sheath gas pressure: Ion sweep gas pressure:

Blank Samples

Capillary temperature:

Double blank samples were prepared in duplicate by the introduction of

Declustering voltage:

water (20 μL) into plasma (100 μL). These were unfortified with respect to

Collision pressure:

65 0.5 300 °C 0V 1.5 mTorr

both rosuvastatin and its deuterated analogue. MS acquisition parameters: Zero Blank Sample

Quantification was performed by selected reaction monitoring (SRM) using

The ‘zero blank’ comprised plasma (100 μL) spiked with internal standard only.

the precursor-to-product combinations shown below:

Isolation of Rosuvastatin using SOLA (Extraction Procedure) The extraction was carried out using a positive pressure SPE manifold capable of processing 96-well microplates. • Condition SOLA with MeOH (1.0 mL at 1.0 mL/minute) • Condition SOLA with water (1.0 mL at 1.0 mL/minute) • Load sample (100 μL) and allow to permeate SOLA material under gravity

Scan type:

• Wash with 0.1 % (v/v) formic acid (500 μL at 1.0 mL/minute)

Peak width:

Q1 - 0.7 (FWHM)

• Wash with 10 % (v/v) MeOH (500 μL at 1.0 mL/minute)

Q3 - 0.7 (FWHM)

• Elute with 90 % (v/v) MeOH (2 x 200 μL at 1.0 mL/minute)

Scan width:

The methanolic extracts were evaporated to dryness under a stream of

Scan time:

nitrogen at 40 °C and reconstituted in 1:4 (v/v) MeOH-water (200 μL).

Divert valve:

SRM

0.2 m/z 0.2 s

Instrumentation Separation was carried out using a Thermo Scientific Accela 600 pump interfaced to both a Thermo Scientific Accela Open Autosampler, and, a Thermo Scientific TSQ Vantage triple stage quadrupole mass spectrometer. Chromatographic Conditions Column:

Thermo Scientific Accucore RP-MS

2.6 μm, 50 x 2.1 mm

Guard column: Accucore Defender (RP-MS)

guard column 2.6 μm, 10 x 2.1 mm

Part Number 17626-052130

MS acquisition time: 3.60 minutes Data Processing

17626-012105

All data were processed using Thermo Scientific LCQuan (v. 2.6) software. Algorithm for integration - ICIS


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FEATURE

SRM derived from examination of the zero blank (upper trace: d6-rosuvastatin, lower trace: undeuterated analogue

Figure 4 100 90

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Table 1. Linearity of response for the determination of rosuvastatin in human plasma

Table 3. Recovery data for the determination of rosuvastatin in human Plasma

Table 2. Accuracy and precision data for the determination of rosuvastatin in human plasma

Table 4. Effect of matrix on the determination of rosuvastatin in human plasma

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Shin-Etsu Silicones Shin-Etsu Silicones of America’s newest HCR (High Consistency Rubber) product line – Sil-XShin™ Silicone Elastomers are specifically engineered for extruded tubing and profiles. SilX-Shin Silicone Elastomers are available in a wide range of Shore A Hardness from 30 to 80 and come in a convenient 10:1 ratio of base compound to catalyst. The most unique advantage of the Sil-XShin product line is that one universal catalyst can be used with each of the six bases, providing ultimate flexibility. The Sil-X-Shin series has a single, universal catalyst that cures any of the six bases with less than 10 per cent of the composition. Sil-X-Shin Silicone Elastomers are USP Class VI compliant, and their platinum, addition-cure chemistry produces a tight surface finish with few residual by-products.

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Slide Imaging System PerkinElmer Inc. launches the VectraTM 2 automated slide imaging system, a turnkey platform that allows true walk-away batch processing of up to 200 tissue sections at a time. Additionally, the Vectra™ 2 addresses the need of scientists focused on biomarker research, pharmaceutical development, clinical research and tissue application test development for validated tissue labeling and automated scoring methodology. The new platform adds HistoRX’s AQUA® technology to Vectra’s unique multispectral and image analysis capabilities. Its unique multispectral capabilities, coupled with the powerful image analysis tool inFormTM, can be used to explore even the most demanding of today’s tissue analysis problems, from multi-analyte in situ cytometric analysis and cell phenotyping to stereology-like analyses of tissue disease states. The technology provides greater than 97 per cent interand intra-assay coefficient of variation, giving researchers assurance in the quality of their results. The system supports both tissue sections and tissue microarrays (TMAs) with ease and offers a slide hotel for batch scanning of up to 200 slides. The Vectra platform is empowered by the same multispectral imaging technology that drives over 400 NuanceTM cameras sold to date, giving researchers the ability to analyze multiple fluors/chromagens in a sample, quantifying each separately and accurately.

www.perkinelmer.com

NEW PRODUCTS Vessels The Thermo Scientific CryoExtra™ vacuum-insulated, stainless steel vessels are available in four different sizes, from 407 litres to 1,630 litres, accommodating both vapour and liquid-phase storage. These containers minimize top-to-bottom temperature variation, preserving the integrity of critical cryogenically frozen samples. Its innovative lid and neck design, combined with advanced temperature monitoring, reduce temperature fluctuations that occur due to lid openings. Furthermore, the microprocessor-based LN2 level control ensures that liquid nitrogen levels are tightly controlled, and the de-fog function allows easy, pushbutton sample identification.With the ability to store up to 90,000 1.2 – 2.0 mL vials, the largest of the four CryoExtra models (1,630 liter) has the same external footprint as the 1,350 liter unit. All four CryoExtra containers can accommodate both vertical and horizontal racks, and the Thermo Scientific racking solutions are specifically designed for this vessel configuration, further maximizing capacity.

www.thermoscientific.com

MILLIPLEX® MAP kit EMD Millipore launches three MILLIPLEX® MAP kits for cellular metabolism. Based on the Luminex® xMAP® assay technology platform and cellular metabolism antibodies from MitoSciences®, the MILLIPLEX® MAP kits enable fast, sensitive, simultaneous quantification of multiple cellular metabolism biomarkers involved in fatty acid oxidation and the pyruvate dehydrogenase complex. Each kit provides highly validated and characterized cellular metabolism immunoassay antibodies, all necessary reagents, and optimized protocols.

www.emdmillipore.com

Ultracentrifuges New Optima X Series preparative ultracentrifuges from Beckman Coulter, Inc. incorporate an array of contemporary technical features to enhance the user experience and increase productivity. With a large touchscreen display, the Optima X Series product offers a selection of nine languages while real-time graphing plots speed and temperature over time as a run progresses. Enhanced remote control capabilities allow the ultracentrifuges to be monitored from a personal computer or a smart phone. When powered on but not running, the unit requires the same energy as a 60-watt incandescent bulb, further increasing energy efficiency. Optima X Series ultracentrifuges reach a maximum speed of 100,000 rpm and generate forces up to 802,000 x g. Optima X Series instruments are available in two models – the XE, which delivers all of the basic features required to quickly set up and complete a run, and the XPN, which includes all features of the XE plus networking capability, on-board simulation tools and a variety of customizable security and tracking features. The XE is available in 90,000 and 100,000 rpm configurations, while the XPN is offered in 80,000, 90,000 and 100,000 rpm configurations.

www.beckmancoulter.com

15


16

NEW PRODUCTS 96-Well DispoEquilibrium DIALYZER™ The Harvard Apparatus 96Well DispoEquilibrium DIALYZER™ provides simple and reliable techniques to analyze up to 96 equilibrium dialysis chambers simultaneously on disposable dialysis plates. These ready to use equilibrium dialysis plates are convenient and cost effective tools for ligand binding experiments including serum protein binding, protein-drug binding, protein-protein binding and protein-DNA binding assays. Each test well in the plate consists of two chambers separated by a regenerated cellulose membrane with a molecular weight cut off of either 5,000 or 10,000 Daltons. Each chamber holds up to 300µl of sample or buffer. To decrease the dialysis time, the plate needs to be rotated through 360° in a vertical position. Harvard Apparatus can also supply a one or two plate rotator or an eight plate rotator with temperature control.

www.harvardapparatus.com

RTR-505V Wireless Logger T&D Corporation introduces its new RTR505V Wireless Logger. The RTR-505V can measure DC voltages from 0 to 22VDC with accuracy of ±0.5mv and can measure voltage directly or from the output of a wide variety of sensors and instruments. The RTR-505V features a large LCD display, capacity for 16,000 readings, IP64 water resistance and up to four years battery life with the “L” version. Compatible with all TandD RTR-500 Series Wireless Data Collectors, the RTR-505V can be seamlessly integrated into a wireless network with other logger types. They can be monitored locally, through a cellular phone system, or anywhere in the world over the Internet. Advanced features include real-time monitoring and warning notifications. The communication range is up to 500 ft. which can be extended with the use of a repeater.

www.tandd.com

January 2012 Laboratory Focus www.bioscienceworld.ca

DNA Barcodes Aids The NEXTflex-96 DNA Barcodes aids next generation sequencing by increasing throughput using barcoded indices allowing users to pool multiple library preparations in a single flow cell lane. The NEXTflex-96 DNA Barcodes accomplish this by providing 96 indexed adapters, each of which contains a unique 8 nt sequence. The indices are designed to be error resistant and allowing for proper differentiation between samples. Using the NEXTflex-96 DNA Barcodes, the index is added to the sample during the adapter ligation step, allowing PCR amplification to be reduced or eliminated, thus minimizing amplification-based bias and preventing poor reads from single base errors introduced during PCR. These barcodes can be used to multiplex genomic DNA, RNA and ChIP sequencing libraries.

www.biooscientific.com

Fume Hoods Air Science introduces its new Purair ECO™ line of Energy-Saving Ductless Fume Hoods designed for both chemical and particulate protection over a broad range of laboratory and industrial applications. The Purair ECO is available with a choice of controllers including the company’s new ECOair™ touchpad control with color display interface. An optional BACnet network interface connects all cabinet control, monitoring and alarm functions to an open-source facility monitoring system. The system is based on an industry-wide, non-proprietary ASHRAE compliant protocol for green building management. The Purair ECO is available in five standard sizes from 30” wide to 69” wide.

www.airscience.com

COMPANY & ADVERTISER INDEX COMPANY

PAGE WEBSITE

BioTalent Canada................................ 6................................www.biotalent.ca Beckman Coulter............................... 15.................. www.beckmancoulter.com Caledon Laboratory Chemicals.......... 4......................www.caledonlabs.com

Titrator JM Science’s AQUACOUNTER® Volumetric Karl Fischer Titrator (AQV-300) is reliable, easy-to-use and has performance features which allows measure of moisture content from low to high concentrations quickly and accurately. Measurements are simple and routine with unsurpassed precision. The AQV-300 has six built-in calculation modes to accommodate solid, liquid and gas samples. It includes a statistics package with one-touch calculations. Four files with preset conditions can be stored in memory and allows instant recall of data for up to 20 samples. A built-in detector monitors titration status and a direct key access allows entry of titration parameters. This compact unit with a very small footprint has balance and computer interfaces for GLP and ISO documentation and it is CE approved. The titrator comes with DVD training movie, plasticized QuickStart guide for use in the lab, paper and pdf manual. Also included, complete accessories, download software for transferring results to a laptop or PC, RS-232 cable, and AQUACOUNTER® KF Reagent starter kit.

www.jmscience.com

Canadian Food Inspection Agency..... 11 .................... www.inspection.gc.ca Chemical Institute of Canada............. 6.............................www.cheminst.ca EMD Millipore................................... 15.......................www.emdmillipore.com Eppendorf........................................ 20....................... www.eppendorf.com GE Healthcare................................... 16......................www.gehealthcare.com IsotechnikaPharma Inc........................ 6..........................www.isotechnika.com Kane Biotech Inc................................. 6.........................www.kanebiotech.com Meile Professional.......................... 5, 7............. www.mieleprofessional.ca MethylGene Inc.................................. 6......................... www.methylgene.com Mettler Toledo.................................. 15.....................................www.mt.com Miraculins Inc..................................... 6........................... www.miraculins.com PerkinElmer....................................... 15........................ www.perkinelmer.com Shin-Etsu Silicones............................ 15........www.shinetsu.co.jp/e/index.shtml Thermo Scientific............................... 15................. www.thermoscientific.com VWR................................................ 19................................. www.vwr.com Wyvern Scientific............................. 2........................ www.wyvernsci.com Zymeworks Inc................................... 6..........................www.zymeworks.com


www.bioscienceworld.ca Laboratory Focus

FEBRUARY

January 2012

CALENDAR

March 28-30

ACETECH Symposium February 4-8 Venue: Whistler, BC SLAS 2012 Tel: (604) 683-5852 Venue: San Diego, CA Fax: (604) 683-3879 Tel: (630)256-7527 Web: www.acetech.org Fax: (630) 741-7527 RC_lab_new:Layout 1 1/19/2012 9:27 AM Page 1 Email: slas@slas.org APRIL Web: www.slas.org

February 13-14 Bio CEO Investor Conference 2012 Venue: New York, NY Email: bd_registration@bio.org Web: www.bio.org/events/ conferences/bio-ceo-andinvestor-conference

April 29-May 2

May 20-24

World Congress on Industrial Biotechnology and Bioprocessing Venue: Orlando, Fl Email: info@bio.org Web: www.bio.org/worldcongress

April 14-17 ACRP 2012 Conference & Expo Venue: Houston, TX Web: www.acrp2012.org

MAY

ASMS Conference on Mass Spectrometry Venue: Vancouver, BC Tel: (505) 989-4517 Fax: (505) 989-1073 Email: office@asms.org Web: www.asms.org

February 16-20 AAAS Annual Meeting Venue: Vancouver, BC Tel: (202) 326-6400 Fax: (202) 842-1065 Email: meetings@aaas.org Web: www.aaas.org

February 25 BIOTECanada CEO Whistler Summit Venue: Whistler BC Email: Kira.pejemsky@biotech.ca Web: www.biotech.ca/en/default.aspx

February 26 –28 BioPartnering North America™ Venue: Vancouver, BC Tel: (831) 464-4230 Fax: (831) 464-4230 Web: http://www.techvision.com/bpn/

February 27-March 1 Cold Chain Management & Temperature Control Summit Venue: Toronto, ON Tel: 1-800-882-8684 Email info@iqpc.com Web: www.coldchainpharm.com

MARCH March 5-7 Growing the Margins Venue: London, ON Tel: (416) 426-7029 Fax: (416) 426-7280 Email: info@gtmconference.ca Web: www.gtmconference.ca

March 11-16 PITTCON Conference & Expo Venue: Orlando, FL Tel: (412) 825-3220 Fax: (412) 825-3224 Email: info@pittcon.org Web: www.pittcon.org

March 19-21 Bio-Europe Spring 2012 Venue: Amsterdam, The Netherlands Tel: (760) 930-0500 Email: chundschell@ebdgroup.com Web: www.ebdgroup.com/bes/ index.php

17

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January 2012 Laboratory Focus

CAREER SPOTLIGHT Bio-economy Career Profile

Compiled by BioTalent Canada Position: Director, Discovery Research Name: Dr. Shawn Ritchie Company: Phenomenome Discoveries Inc. Salary Range: As market demands

What I do:

There’s a lot to do and see at Pittcon 2012 Chances are if you’re reading this publication, you’re familiar with the Pittcon Conference and Expo and what it’s all about, but for the uninitiated here’s the scoop. Organized by the Pittsburgh Conference on Analytical Chemistry and Applied Spectroscopy, the conference dates back to the 1950’s and to this day is one of the world’s largest events devoted to laboratory science attracting nearly 20,000 attendees from industry, academia and government from 90 countries worldwide every year. This year’s event runs March 10 to 15, 2012 at the Orange County Convention Center in Orlando. As a scientist or researcher looking for the latest innovations in laboratory equipment – Pittcon is a must attend event. For starters, industry giants such as Thermo Fisher, Agilent, Waters, Wyatt and hundred others will be on hand to showcase their latest and greatest products, services and ideas. Additionally, for those looking to further their own professional development, the Pittcon Organizing Committee has announced 34 new courses for its 2012 Short Course Program. The short courses range from half-day up to two-day classes; including beginner, intermediate, and advanced level curricula in the fields of data analysis, food, industrial hygiene, life science liquid chromatography, lab management, and pharmaceutical sciences, polymers, quality/ regulatory/compliance, and rheology. These courses are great tool for skill-building and to enhance your career options. Moreover, where else can you get face-to-face with all the vendors (close to 950 exhibiting companies), see the latest instrumentation, and participate in product demonstrations? With so much going on, it should be a fantastic event that sets the stage for an exciting year for the industry.

Phenomenome Discoveries Inc. is involved in metabolic profiling and identifying metabolites in various biological samples using technology based on mass spectrometry; we develop technology and informatics to assist large pharmaceutical companies in the drug development pipeline, and use the technology internally to understand human diseases and develop better diagnostic approaches. As Director of Discovery Research, I primarily oversee and manage the contract research projects and internal recovery efforts. My work involves managing and training teams of scientists working on specific projects, mining data, and writing client reports. I have a Ph.D. in Biochemistry focusing on cancer genomics, which serves me well when working on projects related to finding and identifying metabolite markers that are directly related to cancer. Our company also works on developing screening tools that can be used by the general population to predict risk for various diseases, for example, colon cancer.

What education and skills do candidates need for this position?

As a director, you require a Ph.D. in a life science, such as, biochemistry, organic chemistry, analytical chemistry, or possibly physiology. If you prefer laboratory work to management, a master’s degree is probably satisfactory. Whether you decide to go the academic route or work within a company, it is a personal decision and there are many rewards with either choice. New Ph.D. graduates typically continue in academia and then apply for associate positions at universities to continue research. However, I took a different route, as I wanted to work more independently with a smaller group and have an opportunity to move forward within the company. I was drawn to the technology that our company was developing and was excited by its potential in the health sciences. Since I can have up to 10 or 15 projects at one time, my management and people skills are constantly tested. In this position, you need to have solid laboratory skills, be able to think on the fly, troubleshoot, and discover new ways of handling data. A person in my position also needs to be well read and up on current literature in the field. Above all, in this position you need to have a passion for science and the ability to work effectively within a multidisciplinary team of people. For example, I spend significant time working with computer programmers in the development of cutting edge bioinformatics software; synthetic organic chemists; clinicians; business development and marketing people; and even lawyers. It also helps to be an independent thinker and effective problem solver.

What are the best parts of your job?

I enjoy the high-tech, multidisciplinary nature of my work. But overall, it is the rapid translation of the discoveries into new diagnostics or therapeutics for cancer and neurological diseases that motivates me daily. The fact that the work we do will reduce the incidence of diseases and improve current treatments in a relatively short timeframe is very exciting.


Choice Matters CHOICE MATTERS Because No Two Labs are Completely Alike Science should never be limited by a lack of options. Every lab deserves the freedom to find tools perfectly suited to specific needs. VWR is committed to offering broad product choice to a world of accelerating scientific advancement. We give you direct access to the world’s most respected equipment, supplies, chemicals, furniture, and more. Talk to us about the needs of your lab. We are ready to help you succeed.

Contact your VWR Sales Representative, visit VWR.com, or call 1.800.932.5000 today. VWR, forms of VWR and the VWR logo and/or design are either registered trademarks ® or trademarks™, or service marks SM of VWR International, LLC in the United States and/or other countries. ©2011 VWR International, LLC. All rights reserved.


C025.A1.0108.A © 2012 Eppendorf AG

l

Enzyme kinetics and advanced assays

l

Accommodates both standard cuvettes and microliter volumes

l

SpectraZoom® enables data processing directly on the device

NEW!

Light up your day! Eppendorf BioSpectrometer® The new Eppendorf BioSpectrometer expands the current product offering with a broad spectrum of applications and extremely simple operation. Measure and record UV/Vis spectral ranges or just measure individual wavelengths from 200 nm to 830 nm. Two different models make it possible to select the optimal instrument for your research.

Eppendorf BioSpectrometer basic/kinetic: l Xenon flashlamp with extra long life l Pre-programmed methods for a quick start l Guided software procedure minimizes errors l Direct operation on the device, no PC required l Integrated data processing for specific methods BioSpectrometer kinetic only: Integrated, temperature-controlled cuvette shaft for substrate and enzyme kinetics

l

For more information visit www.eppendorf.com/biospectrometer

www.eppendorf.com • Email: info@eppendorf.com In the U.S.: Eppendorf North America, Inc. 800-645-3050 • In Canada: Eppendorf Canada Ltd. 800-263-8715

Laboratory Focus January 2012  

Laboratory Focus is the leading editorial-based laboratory magazine to serve the Canadian lab market. Published bi-monthly, it is the only...

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