- Email: [email protected]
In the news February
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
In the news
In the news Lab-in-a-drop promises analytical results Chemists at the University of California, San Diego, USA have developed a ‘‘lab-in-a-drop’’ technology that promises nanoscale analytical chemistry and spectroscopy. Michael Sailor and colleagues have been able to manipulate liquid droplets using ‘‘chaperones’’ comprising amphiphilic magnetic onedimensional photonic crystals that allow them to handle smaller and smaller samples without loss to the walls of a container. They saw the controlled manipulation of small volumes of liquids as a challenging problem in microfluidics and a key requirement for many highthroughput analyses and microassays, because samples of DNA, bacteria, viruses or other minute particles can be lost to surfaces that they contact. These losses can reduce the accuracy of analysis and give rise to misleading results. As sample volumes become smaller, the molecules that stick to the surfaces in analytical equipment, such as pipettes and microchannels, become a significant proportion of
the total number of molecules in the sample. However, with Sailor’s ‘‘labin-a-drop’’, the spherical droplet has the lowest ratio of surface area to volume, so the chance of any particle of sample coming into contact with the surface is reduced. This minimizes the amount of material lost. Lead author Jason Dorvee constructed one-dimensional photonic crystals made from porous silicon with amphiphilic properties. When prepared in the form of lm-sized particles and placed in a two-phase liquid, such as dichloromethane/ water, these materials accumulate and spontaneously align at the interface. Dorvee and colleagues have shown [1] that superparamagnetic nanoparticles of Fe3O4 can be incorporated into the porous nanostructure, allowing the materials to chaperone the ll-scale liquid droplets when an external magnetic field is applied. The optical reflectivity spectrum of the photonic crystal displays a peak that serves to identify the droplet.
Figure 1. Magnetic particles surround a water drop immersed in oil (octadecene) demonstrate the potential of ‘‘lab-in-a-drop’’.
0165-9936/$ - see front matter doi:10.1016/j.trac.2004.12.001
They have demonstrated two simple microfluidics applications: filling and draining a chaperoned droplet; and, combining two different droplets to perform a chemical reaction. Their method therefore provides a general means for manipulating and monitoring small volumes of liquids without the use of pumps, valves or a microfluidic container. Reference [1] J.R. Dorvee, A.M. Derfus, S.N. Bhatia, M.J. Sailor, Nat. Mater. 3 (2004) 896.
Contact: Michael J. Sailor, Department of Chemistry and Biochemistry, University of California, San Diego, USA. E-mail: [email protected]
Molecular analysis: right first time With DNA-based analysis increasingly being used in clinical, forensic and industrial applications, LGC is offering a two-day workshop on core skills for molecular biology on 3 and 4 March 2005 at its headquarters in Teddington, Middlesex, UK. The wider uptake of molecular biological methods requires the assurance of the accuracy, reliability and comparability of the techniques, to permit the technology to be used with confidence, according to LGC, the leading independent provider of analytical and diagnostic services in Europe. iii
In the news
This course provides ways to ensure that analytical results can be used with confidence and that experiments are designed appropriately, to produce results first time.
Who should attend? The workshop is aimed at those starting their first job or PhD in a molecular biology laboratory. It will build a strong foundation for a career in bio-analysis, by instilling an awareness of measurement quality and traceability. The main focus of the seminars and practical sessions will be core skills commonly used in molecular analysis, including pipetting, experimental design, set-up and interpretation.
Aims and techniques The workshop aims to: • supply information on the importance of quality in the molecular biology laboratory and examples of how this may be achieved in practice • provide practical guidance on PCR design and performance to ensure reliable results Techniques covered during the workshop will include: • principles and applications of PCR analysis; • practical PCR and gel analysis; • use of controls and reference materials; • statistical analysis; • experimental design; • recording and reporting of results; and, • quality systems and accreditation. Contact: Jacquie Keer, LGC, Teddington, Surrey, UK. Tel.: +020 8943 7449; Fax: +020 8943 2767; E-mail: [email protected] iv
http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
Mycotoxins absent from most UK baby foods Mycotoxins (the toxic/carcinogenic/ mutagenic metabolites of certain moulds within the food and feed industries) were not detectable in 90% of the products analyzed, when the UK Food Standards Agency (FSA) surveyed aflatoxins, ochratoxin A and patulin in foods for infants and young children between November 2003 and April 2004. The FSA sampled 199 foods, including breakfast/rusk products, baby rice, savory products and desert/cereal bar/biscuits. Highperformance liquid chromatography (HPLC) was used to analyze 169 samples for aflatoxins and ochratoxin A. A further 14 products were also analyzed for patulin, and 16 products, including applebased drinks and apple-fruit products, were tested for patulin only, as patulin is much more likely to occur in these products compared with the other mycotoxins studied. Where mycotoxins were detectable, levels were very low and regulatory limits were not exceeded in any of them. The FSA designed the survey to assess the exposure of infants to mycotoxins, and therefore concluded that the results do not raise any concerns for infant health. The limits for mycotoxins on a dry matter basis are set out in EC Regulation 683/2004: • 0.1 lg/kg for aflatoxin B1 and 0.5 lg/kg for ochratoxin A for baby foods and processed cereal-based foods for infants and young children, and dietary foods for special medical purposes intended specifically for infants; and, • 0.025 lg/kg for aflatoxin M1 for infant formulae and followon formulae, including infant milk and follow-on milk and dietary foods for special medical
purposes intended specifically for infants. Aflatoxins were unquantifiable in the majority of samples (99.5%) with only one sample containing detectable levels of aflatoxin B1, but below the limits stipulated in EC Regulation 683/2004. Ochratoxin was not detected in 89% of the samples and was below the legal limits in the remainder of the samples, when measurement uncertainty was taken into account. Patulin was not detected in any of the 30 samples tested. Since none of the samples exceeded legal limits for mycotoxins, when measurement uncertainty was taken into account, the FSA did not take any action. ‘‘The low levels observed during the survey suggest that the babyfood industry is aware of the existing and new legal limits for mycotoxins in baby foods’’, concluded the FSA, ‘‘and have been diligent in their sourcing of highquality ingredients to ensure that levels remain low’’.
Reference [1] Food Standards Agency, Survey of baby foods for mycotoxins, Food Survey Information Sheet 68/04, 2004 (http:// www. food.gov.uk/multimedia/pdfs/fsis 6804.pdf).
UPLC reaches levels that HPLC never did Ultra Performance Liquid Chromatography (UPLC), which Waters Corporation launched at Pittcon 2004 in Chicago last March, can achieve much better performance than high-performance liquid chromatography (HPLC) when combined with mass spectrometry (MS), according to AstraZeneca Principal Scientist Professor Ian Wilson.
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
He was reporting the results of a direct comparison between HPLCMS and UPLC-MS in the analysis of complex biofluid samples for metabonomics, which he described as the quantitative measurement of the dynamic multi-parametric response of living systems to pathophysiological stimuli or genetic modification [1]. AstraZeneca, the major pharmaceutical company, wants metabonomics to deliver biomarkers for disease, genetic difference, pharmacology, efficacy and toxicity. Biomarker ions Prof. Wilson showed data illustrating how UPLC can detect as many as 10,000 potential endogenous biomarker ions in rat urine as compared to only 2000 by conventional HPLC. He predicted that separation-based metabonomics is going to play an important role in the future of drug discovery because it is fast, efficient and very costeffective, and this type of analysis is potentially widely available. HPLC-MS is still at an early stage of development, with less than a dozen papers published, according to
Prof. Wilson. In an experiment, which used the Zucker rat model in comparing urinary HPLC-MS profiles of ‘‘control’’ versus ‘‘Zucker’’ rats from 4 to 20 weeks, he and colleagues showed that HPLC-MS can detect difference (e.g. in toxicity, age, strain, gender, disease and diurnal effects). However, he described the chromatography as only ‘‘OK’’ with the potential for ion suppression being huge and leaving the question of just how many peaks were being missed. When UPLC-MS was compared with HPLC-MS, the improvements were striking (Figs. 2 and 3). He therefore concluded that UPLC-MSbased metabonomics will have an important part to play in the clinic to support drug development, and saw this as a striking example of Sir Humphrey Davy’s maxim in 1840: ‘‘Nothing advances human knowledge so much as the application of a new instrument’’. Reference [1] I. Wilson, Envisioning the potential of UPLC, Waters Media Day, 2 December 2004, Waters Corporation, 2004 (www.waters.com).
In the news
Reference standards can be customized LGC Promochem, Europe’s leading supplier of certified reference materials, has introduced bespoke standard solutions. By talking to scientists in all fields of analysis, LGC Promochem found that high on their wish lists were cost-effective reference standard solutions tailored to their specific requirements. Analysts said they would prefer to use standard mixes containing the correct combination of components at the right levels and in the right solvents, enabling them to save time, effort and money in preparing solutions, and simplifying quality control by having just one solution with a single expiry date. LGC Promochem is meeting this demand by offering custom solutions. By drawing on the expertise and facilities of its leading suppliers of chemical standards, including ULTRA Scientific, LGC Promochem’s custom solutions range offers a large selection of analytes, all mixed according to the customer’s specifications. Components could include:
Figure 2. Three-dimensional maps for (a) HPLC-MS and (b) UPLC-MS of white male mouse urine from (Morning collection) mouse illustrate the much higher information content of the UPLC-generated data with the figure showing retention time, m/z and intensity.
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In the news
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
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Figure 3. HPLC Vs. UPLC for m/z 401. An extracted ion chromatogram for the ion m/z = 401 for a urine sample obtained from a white female mouse (Morning collection) using either (a) a 2.1 mm x 10 cm Waters Symmetry 3.5 `ım C18 column or (b) 2.1 mm x 5 cm Waters Acquity 1.7 lm C18 column. The columns were eluted with 0–95% aqueous formic acid : acetonitrile (0.1% formic acid) gradient over 10 min at a flow rate of 500 lL/min. The column eluent was monitored by ESI oa-TOF-MS from 50 to 850 m/z in positive ion mode.
• inorganic species, such as metals or anions in aqueous solution; • metals in mineral oil; and, • organic compounds, such as pesticides, polyaromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), volatile organic compounds (VOCs), semi-VOCs and benzene, toluene, ethylbenzene, and xylenes (BTEX), in a variety of organic solvents. The solvent is the customer’s choice – provided, of course, there are no problems with solubility or the mix of analytes. The materials are prepared gravimetrically, with the option of quantitative confirmation, and each is accompanied by a certificate detailing the concentration of each component. There is also the option of an additional DATApak with full analytical and gravimetric data for many of the solutions. Custom solution reference materials can be produced in a variety of pack sizes and types of packaging, including the Certan vial, which can store standards in volatile solvents. vi
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Indian analysis industry supports Analytica-Anacon
tries that use analysis equipment and services in India’’. At the opening, German Consul General Hans-Heinrich Freiherr von Stackelberg and Tata Institute of Fundamental Research Director Subir Bhattacharya underscored Analytica-Anacon’s significance for the Indian analysis and biotechnology market, which they expect to see rapid growth during the next few years.
With 87 exhibitors from 11 countries, the second Analytica-Anacon attracted more than 2244 visitors to the Nehru Center in Mumbai between 25 and 27 November 2004. ‘‘Analytica-Anacon 2004 was probably the largest exhibition for analysis equipment ever to be held in India’’, said Nitrin Kabbin from Indian Analytical Instruments Association (IAIA). ‘‘Given the quality and the range of exhibitors, there is little doubt that Analytica-Anacon has established itself as the event of choice for decision-makers in indus-
Exhibitors pleased The exhibitors were very pleased with the results of AnalyticaAnacon, and the response among the visitors – especially from the pharmaceuticals industry – was excellent, according to Munich Trade Fairs International Group Director Kurt Schraudy, who organized Analytica-Anacon. ‘‘Participation at the exhibition was outstanding’’, stated Ganapathy Ramakrishnan from the Thermo Electron Corporation, India. ‘‘The exhibits that German companies
Contact: LGC Promochem Tel.: +44 (0)20 8943 7682; Fax: +44 (0)20 8943 7554; E-mail: [email protected]
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
had at the national pavilion was an excellent idea and one that should be used for exhibitors from other countries, such as France, Great Britain and the United States. The Analytica-Anacon Conference was also a complete success’’. International exhibitors returning from Mumbai were also satisfied. ‘‘The market for our products and the purchasing power are there – the Indian market is future-oriented and its growth prospects are good’’, concluded Peter Kind from German trade association Spectaris on behalf of the exhibitors at the joint German exhibit. ‘‘Visitor interest was good, and we had a number of highly qualified visitors at our stand’’. More than 400 delegates from throughout India followed the lectures of more than 20 prominent scientists, technical experts and researchers at the accompanying Analytica-Anacon Conference, which consisted of five half-day sessions. It featured the latest analysis methods and techniques in the sectors for pharmaceuticals, environment, materials testing, plastics and polymers, and petrochemicals. There were renowned speakers from the Indian analysis sector, such as R.T. Sane (Director of Ruia College), M. Sundaraman (Head of the Analytical Chemistry Division at the Bhabha Atomic Research Center), and V. Soundararajan (Reliance, the largest petroleum and energy group in India). There were also presentations by international manufacturers, such as Agilent and the Thermo Electron Corporation. The conference focused on the latest developments in analysis equipment and techniques for process-oriented applications. ‘‘We used the conference to present the solutions and innovations that our industry needs to hold its own in global competition and tap local market potential,’’ explained IAIA Director Kabbin. The next Analytica-Anacon will take place at Nehru Center. Mumbai, in November 2006.
Contact: Messe Mnchen GmbH, Munich, Germany. Tel.: +49 (89) 949 11488; Website: http://www.analytica-world.com
LGC launches pharmaceutical and chemical services LGC has launched a Pharmaceutical and Chemical Services (PCS) division, demonstrating the breadth of its specialist services to the biopharmaceutical community, as Europe’s leading independent analytical laboratory providing chemical, biochemical and forensic analysis. PCS evolved by combining LGC’s existing Analytical and Cell-based services in Teddington, UK, with the former Chemical and Process Industries division, based at Runcorn, UK. It also includes Mikromol, the pharmaceutical impurities company that LGC acquired in 2003, and encompasses LGC’s portfolio in drug discovery. ‘‘This new division means that LGC can now offer a comprehensive drug-discovery package to assist our clients through every stage of the drug-development lifecycle, making LGC a uniquely positioned service provider in the UK,’’ explained Derek Craston, Head of Pharmaceutical and Chemical Services at LGC. ‘‘By combining our strengths and skills with the companies we are collaborating with, we are able to deliver a new breadth and depth of service, enabling our customers to achieve their own strategic aims. The creation of our new division at LGC reflects our commitment to providing a world-class service to the pharmaceutical and biotechnology community.’’ Contact: Derek Craston, LGC.
In the news
Tel.: +44 (0)20 8943 7000; E-mail: [email protected]
Scientists report real benefits from applying Lab-on-a-Chip Scientists involved in genomics, basic life sciences and drug discovery have revealed the first results of research that illustrate the benefits of the Agilent 5100 Automated Lab-on-a-Chip Platform (ALP) in providing unattended analysis of thousands of DNA and protein samples a day. In December, Christian Maercker (RZPD Resource Center for Genome Research, Heidelberg, Germany) and Paul Hawtin (AstraZeneca, Macclesfield, Cheshire, UK) described their work at a press briefing at Agilent’s European headquarters in Waldbronn, Germany [1,2]. The 5100 ALP replaces gel electrophoresis in large-scale genomic and proteomic applications by automating sample handling and analysis, and lowering sample-analysis time by a factor of 10. It minimizes sample and reagent use, therefore reducing hazardous waste and cost per sample. The 5100 ALP can simultaneously process up to 4608 samples from industry-standard 96-well or 384-well plates. An automated, robotic handling mechanism loads the sample, with reagents and internal reference standards, onto a microfluidic chip for separation and detection. System software automatically analyzes the results, generating high-quality digital data as well as a gel-like image and electropherogram. All resources required for analysis, including samples, reagents, chips, and pipette tips, are stored within the bench-top instrument. Each chip can be reused several thousand times before replacement.
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In the news
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
Figure 4. Agilent 5100 Automated Lab-on-a-Chip Platform.
The instrument infrastructure (e.g., power supplies, and pneumatic and reagent systems) has been de-
signed to support the launch of further assays that will take advantage of the capabilities of the 5100 ALP.
Checking quality RZPD needs to provide clone-based resources (e.g., cDNAs, proteins and siRNAs), which are indispensable for genome research, explained Maercker. However, until now, each product has had to be checked by gel analysis. This has been a rate-limiting step because RZPD could process a maximum of 400 samples a day as the gels had to be analyzed laboriously by eye. However, with the 5100 ALP, RZPD has achieved high-throughput analysis of 800 samples a day of DNA fragments. The whole process of sample loading and analysis is automated. ‘‘The machine and the software are easy to handle,’’ said Maercker. ‘‘Results are accurate and reliable, the representation of data is clear, and data stored in a database.’’ He particularly highlighted the accuracy of the 5100 ALP with the following results that showed deviations for: • Sizes of PCR fragments ALP vs. ALP (32 samples) 1.3% • Concentration of PCR fragments ALP vs. ALP (34 samples) 2.8% • Concentration of PCR fragments ALP vs. Fluorescence (picogreen dye) 23.9%
Figure 5. Screen shots of the 5100 ALP software show analysis of single PCR products (courtesy of C. Maercker, RZPD).
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Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
In the news
Figure 6. Purification screening using the 5100 ALP software showing (a) gel-like images, and (b) images indicating concentration thresholds of purified protein in 96-well plates (courtesy of P. Hawtin, AstraZeneca).
As a result of RZPD’s experience, Maercker strongly recommended the 5100 ALP for the highthroughput analysis of PCR fragments, and looked forward to the possibility of extending its use to DNA fragments up to 12 kb, proteins and RNA.
High throughput With AstraZeneca, Paul Hawtin’s role as senior research scientist is to assess the potential usefulness of the 5100 ALP for high-throughput protein analysis. He reported a number of benefits of the 5100 ALP: • an increasing the number of samples could be analyzed; • automated, hands-free analysis;
• qualitative and quantitative data; • ease of use; • electronic data format for storage/retrieval; and, • safety and reduced environmental waste. Applications for which he found the 5100 ALP suitable included expression screening, refolding screen analysis, and purification screening (to give sufficient protein for crystallography and NMR studies). Expressing satisfaction with the 5100 ALP’s performance and its applications so far, Hawtin recommended that, for the future, Agilent should increase its flexibility, further integrate it to fully automate the expression/purification pipeline,
offer a relational database, and make reagents cheaper.
References [1] C. Maercker, ‘‘DNA applications of the 5100 platform’’, Press Briefing, Agilent Technologies, Waldbronn, Germany, 1 December 2004. [2] P. Hawtin, ‘‘High-throughput protein analysis using the 5100 ALP’’, Press Briefing, Agilent Technologies, Waldbronn, Germany, 1 December 2004.
Contact: Claudia Hachter, Agilent Technologies. Tel.: +49 (0)7243 602 100; E-mail: claudia_hachtel@agilent. com
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In the news
In the news Lab-in-a-drop promises analytical results Chemists at the University of California, San Diego, USA have developed a ‘‘lab-in-a-drop’’ technology that promises nanoscale analytical chemistry and spectroscopy. Michael Sailor and colleagues have been able to manipulate liquid droplets using ‘‘chaperones’’ comprising amphiphilic magnetic onedimensional photonic crystals that allow them to handle smaller and smaller samples without loss to the walls of a container. They saw the controlled manipulation of small volumes of liquids as a challenging problem in microfluidics and a key requirement for many highthroughput analyses and microassays, because samples of DNA, bacteria, viruses or other minute particles can be lost to surfaces that they contact. These losses can reduce the accuracy of analysis and give rise to misleading results. As sample volumes become smaller, the molecules that stick to the surfaces in analytical equipment, such as pipettes and microchannels, become a significant proportion of
the total number of molecules in the sample. However, with Sailor’s ‘‘labin-a-drop’’, the spherical droplet has the lowest ratio of surface area to volume, so the chance of any particle of sample coming into contact with the surface is reduced. This minimizes the amount of material lost. Lead author Jason Dorvee constructed one-dimensional photonic crystals made from porous silicon with amphiphilic properties. When prepared in the form of lm-sized particles and placed in a two-phase liquid, such as dichloromethane/ water, these materials accumulate and spontaneously align at the interface. Dorvee and colleagues have shown [1] that superparamagnetic nanoparticles of Fe3O4 can be incorporated into the porous nanostructure, allowing the materials to chaperone the ll-scale liquid droplets when an external magnetic field is applied. The optical reflectivity spectrum of the photonic crystal displays a peak that serves to identify the droplet.
Figure 1. Magnetic particles surround a water drop immersed in oil (octadecene) demonstrate the potential of ‘‘lab-in-a-drop’’.
0165-9936/$ - see front matter doi:10.1016/j.trac.2004.12.001
They have demonstrated two simple microfluidics applications: filling and draining a chaperoned droplet; and, combining two different droplets to perform a chemical reaction. Their method therefore provides a general means for manipulating and monitoring small volumes of liquids without the use of pumps, valves or a microfluidic container. Reference [1] J.R. Dorvee, A.M. Derfus, S.N. Bhatia, M.J. Sailor, Nat. Mater. 3 (2004) 896.
Contact: Michael J. Sailor, Department of Chemistry and Biochemistry, University of California, San Diego, USA. E-mail: [email protected]
Molecular analysis: right first time With DNA-based analysis increasingly being used in clinical, forensic and industrial applications, LGC is offering a two-day workshop on core skills for molecular biology on 3 and 4 March 2005 at its headquarters in Teddington, Middlesex, UK. The wider uptake of molecular biological methods requires the assurance of the accuracy, reliability and comparability of the techniques, to permit the technology to be used with confidence, according to LGC, the leading independent provider of analytical and diagnostic services in Europe. iii
In the news
This course provides ways to ensure that analytical results can be used with confidence and that experiments are designed appropriately, to produce results first time.
Who should attend? The workshop is aimed at those starting their first job or PhD in a molecular biology laboratory. It will build a strong foundation for a career in bio-analysis, by instilling an awareness of measurement quality and traceability. The main focus of the seminars and practical sessions will be core skills commonly used in molecular analysis, including pipetting, experimental design, set-up and interpretation.
Aims and techniques The workshop aims to: • supply information on the importance of quality in the molecular biology laboratory and examples of how this may be achieved in practice • provide practical guidance on PCR design and performance to ensure reliable results Techniques covered during the workshop will include: • principles and applications of PCR analysis; • practical PCR and gel analysis; • use of controls and reference materials; • statistical analysis; • experimental design; • recording and reporting of results; and, • quality systems and accreditation. Contact: Jacquie Keer, LGC, Teddington, Surrey, UK. Tel.: +020 8943 7449; Fax: +020 8943 2767; E-mail: [email protected] iv
http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
Mycotoxins absent from most UK baby foods Mycotoxins (the toxic/carcinogenic/ mutagenic metabolites of certain moulds within the food and feed industries) were not detectable in 90% of the products analyzed, when the UK Food Standards Agency (FSA) surveyed aflatoxins, ochratoxin A and patulin in foods for infants and young children between November 2003 and April 2004. The FSA sampled 199 foods, including breakfast/rusk products, baby rice, savory products and desert/cereal bar/biscuits. Highperformance liquid chromatography (HPLC) was used to analyze 169 samples for aflatoxins and ochratoxin A. A further 14 products were also analyzed for patulin, and 16 products, including applebased drinks and apple-fruit products, were tested for patulin only, as patulin is much more likely to occur in these products compared with the other mycotoxins studied. Where mycotoxins were detectable, levels were very low and regulatory limits were not exceeded in any of them. The FSA designed the survey to assess the exposure of infants to mycotoxins, and therefore concluded that the results do not raise any concerns for infant health. The limits for mycotoxins on a dry matter basis are set out in EC Regulation 683/2004: • 0.1 lg/kg for aflatoxin B1 and 0.5 lg/kg for ochratoxin A for baby foods and processed cereal-based foods for infants and young children, and dietary foods for special medical purposes intended specifically for infants; and, • 0.025 lg/kg for aflatoxin M1 for infant formulae and followon formulae, including infant milk and follow-on milk and dietary foods for special medical
purposes intended specifically for infants. Aflatoxins were unquantifiable in the majority of samples (99.5%) with only one sample containing detectable levels of aflatoxin B1, but below the limits stipulated in EC Regulation 683/2004. Ochratoxin was not detected in 89% of the samples and was below the legal limits in the remainder of the samples, when measurement uncertainty was taken into account. Patulin was not detected in any of the 30 samples tested. Since none of the samples exceeded legal limits for mycotoxins, when measurement uncertainty was taken into account, the FSA did not take any action. ‘‘The low levels observed during the survey suggest that the babyfood industry is aware of the existing and new legal limits for mycotoxins in baby foods’’, concluded the FSA, ‘‘and have been diligent in their sourcing of highquality ingredients to ensure that levels remain low’’.
Reference [1] Food Standards Agency, Survey of baby foods for mycotoxins, Food Survey Information Sheet 68/04, 2004 (http:// www. food.gov.uk/multimedia/pdfs/fsis 6804.pdf).
UPLC reaches levels that HPLC never did Ultra Performance Liquid Chromatography (UPLC), which Waters Corporation launched at Pittcon 2004 in Chicago last March, can achieve much better performance than high-performance liquid chromatography (HPLC) when combined with mass spectrometry (MS), according to AstraZeneca Principal Scientist Professor Ian Wilson.
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
He was reporting the results of a direct comparison between HPLCMS and UPLC-MS in the analysis of complex biofluid samples for metabonomics, which he described as the quantitative measurement of the dynamic multi-parametric response of living systems to pathophysiological stimuli or genetic modification [1]. AstraZeneca, the major pharmaceutical company, wants metabonomics to deliver biomarkers for disease, genetic difference, pharmacology, efficacy and toxicity. Biomarker ions Prof. Wilson showed data illustrating how UPLC can detect as many as 10,000 potential endogenous biomarker ions in rat urine as compared to only 2000 by conventional HPLC. He predicted that separation-based metabonomics is going to play an important role in the future of drug discovery because it is fast, efficient and very costeffective, and this type of analysis is potentially widely available. HPLC-MS is still at an early stage of development, with less than a dozen papers published, according to
Prof. Wilson. In an experiment, which used the Zucker rat model in comparing urinary HPLC-MS profiles of ‘‘control’’ versus ‘‘Zucker’’ rats from 4 to 20 weeks, he and colleagues showed that HPLC-MS can detect difference (e.g. in toxicity, age, strain, gender, disease and diurnal effects). However, he described the chromatography as only ‘‘OK’’ with the potential for ion suppression being huge and leaving the question of just how many peaks were being missed. When UPLC-MS was compared with HPLC-MS, the improvements were striking (Figs. 2 and 3). He therefore concluded that UPLC-MSbased metabonomics will have an important part to play in the clinic to support drug development, and saw this as a striking example of Sir Humphrey Davy’s maxim in 1840: ‘‘Nothing advances human knowledge so much as the application of a new instrument’’. Reference [1] I. Wilson, Envisioning the potential of UPLC, Waters Media Day, 2 December 2004, Waters Corporation, 2004 (www.waters.com).
In the news
Reference standards can be customized LGC Promochem, Europe’s leading supplier of certified reference materials, has introduced bespoke standard solutions. By talking to scientists in all fields of analysis, LGC Promochem found that high on their wish lists were cost-effective reference standard solutions tailored to their specific requirements. Analysts said they would prefer to use standard mixes containing the correct combination of components at the right levels and in the right solvents, enabling them to save time, effort and money in preparing solutions, and simplifying quality control by having just one solution with a single expiry date. LGC Promochem is meeting this demand by offering custom solutions. By drawing on the expertise and facilities of its leading suppliers of chemical standards, including ULTRA Scientific, LGC Promochem’s custom solutions range offers a large selection of analytes, all mixed according to the customer’s specifications. Components could include:
Figure 2. Three-dimensional maps for (a) HPLC-MS and (b) UPLC-MS of white male mouse urine from (Morning collection) mouse illustrate the much higher information content of the UPLC-generated data with the figure showing retention time, m/z and intensity.
http://www.elsevier.com/locate/trac
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In the news
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
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Figure 3. HPLC Vs. UPLC for m/z 401. An extracted ion chromatogram for the ion m/z = 401 for a urine sample obtained from a white female mouse (Morning collection) using either (a) a 2.1 mm x 10 cm Waters Symmetry 3.5 `ım C18 column or (b) 2.1 mm x 5 cm Waters Acquity 1.7 lm C18 column. The columns were eluted with 0–95% aqueous formic acid : acetonitrile (0.1% formic acid) gradient over 10 min at a flow rate of 500 lL/min. The column eluent was monitored by ESI oa-TOF-MS from 50 to 850 m/z in positive ion mode.
• inorganic species, such as metals or anions in aqueous solution; • metals in mineral oil; and, • organic compounds, such as pesticides, polyaromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), volatile organic compounds (VOCs), semi-VOCs and benzene, toluene, ethylbenzene, and xylenes (BTEX), in a variety of organic solvents. The solvent is the customer’s choice – provided, of course, there are no problems with solubility or the mix of analytes. The materials are prepared gravimetrically, with the option of quantitative confirmation, and each is accompanied by a certificate detailing the concentration of each component. There is also the option of an additional DATApak with full analytical and gravimetric data for many of the solutions. Custom solution reference materials can be produced in a variety of pack sizes and types of packaging, including the Certan vial, which can store standards in volatile solvents. vi
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Indian analysis industry supports Analytica-Anacon
tries that use analysis equipment and services in India’’. At the opening, German Consul General Hans-Heinrich Freiherr von Stackelberg and Tata Institute of Fundamental Research Director Subir Bhattacharya underscored Analytica-Anacon’s significance for the Indian analysis and biotechnology market, which they expect to see rapid growth during the next few years.
With 87 exhibitors from 11 countries, the second Analytica-Anacon attracted more than 2244 visitors to the Nehru Center in Mumbai between 25 and 27 November 2004. ‘‘Analytica-Anacon 2004 was probably the largest exhibition for analysis equipment ever to be held in India’’, said Nitrin Kabbin from Indian Analytical Instruments Association (IAIA). ‘‘Given the quality and the range of exhibitors, there is little doubt that Analytica-Anacon has established itself as the event of choice for decision-makers in indus-
Exhibitors pleased The exhibitors were very pleased with the results of AnalyticaAnacon, and the response among the visitors – especially from the pharmaceuticals industry – was excellent, according to Munich Trade Fairs International Group Director Kurt Schraudy, who organized Analytica-Anacon. ‘‘Participation at the exhibition was outstanding’’, stated Ganapathy Ramakrishnan from the Thermo Electron Corporation, India. ‘‘The exhibits that German companies
Contact: LGC Promochem Tel.: +44 (0)20 8943 7682; Fax: +44 (0)20 8943 7554; E-mail: [email protected]
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
had at the national pavilion was an excellent idea and one that should be used for exhibitors from other countries, such as France, Great Britain and the United States. The Analytica-Anacon Conference was also a complete success’’. International exhibitors returning from Mumbai were also satisfied. ‘‘The market for our products and the purchasing power are there – the Indian market is future-oriented and its growth prospects are good’’, concluded Peter Kind from German trade association Spectaris on behalf of the exhibitors at the joint German exhibit. ‘‘Visitor interest was good, and we had a number of highly qualified visitors at our stand’’. More than 400 delegates from throughout India followed the lectures of more than 20 prominent scientists, technical experts and researchers at the accompanying Analytica-Anacon Conference, which consisted of five half-day sessions. It featured the latest analysis methods and techniques in the sectors for pharmaceuticals, environment, materials testing, plastics and polymers, and petrochemicals. There were renowned speakers from the Indian analysis sector, such as R.T. Sane (Director of Ruia College), M. Sundaraman (Head of the Analytical Chemistry Division at the Bhabha Atomic Research Center), and V. Soundararajan (Reliance, the largest petroleum and energy group in India). There were also presentations by international manufacturers, such as Agilent and the Thermo Electron Corporation. The conference focused on the latest developments in analysis equipment and techniques for process-oriented applications. ‘‘We used the conference to present the solutions and innovations that our industry needs to hold its own in global competition and tap local market potential,’’ explained IAIA Director Kabbin. The next Analytica-Anacon will take place at Nehru Center. Mumbai, in November 2006.
Contact: Messe Mnchen GmbH, Munich, Germany. Tel.: +49 (89) 949 11488; Website: http://www.analytica-world.com
LGC launches pharmaceutical and chemical services LGC has launched a Pharmaceutical and Chemical Services (PCS) division, demonstrating the breadth of its specialist services to the biopharmaceutical community, as Europe’s leading independent analytical laboratory providing chemical, biochemical and forensic analysis. PCS evolved by combining LGC’s existing Analytical and Cell-based services in Teddington, UK, with the former Chemical and Process Industries division, based at Runcorn, UK. It also includes Mikromol, the pharmaceutical impurities company that LGC acquired in 2003, and encompasses LGC’s portfolio in drug discovery. ‘‘This new division means that LGC can now offer a comprehensive drug-discovery package to assist our clients through every stage of the drug-development lifecycle, making LGC a uniquely positioned service provider in the UK,’’ explained Derek Craston, Head of Pharmaceutical and Chemical Services at LGC. ‘‘By combining our strengths and skills with the companies we are collaborating with, we are able to deliver a new breadth and depth of service, enabling our customers to achieve their own strategic aims. The creation of our new division at LGC reflects our commitment to providing a world-class service to the pharmaceutical and biotechnology community.’’ Contact: Derek Craston, LGC.
In the news
Tel.: +44 (0)20 8943 7000; E-mail: [email protected]
Scientists report real benefits from applying Lab-on-a-Chip Scientists involved in genomics, basic life sciences and drug discovery have revealed the first results of research that illustrate the benefits of the Agilent 5100 Automated Lab-on-a-Chip Platform (ALP) in providing unattended analysis of thousands of DNA and protein samples a day. In December, Christian Maercker (RZPD Resource Center for Genome Research, Heidelberg, Germany) and Paul Hawtin (AstraZeneca, Macclesfield, Cheshire, UK) described their work at a press briefing at Agilent’s European headquarters in Waldbronn, Germany [1,2]. The 5100 ALP replaces gel electrophoresis in large-scale genomic and proteomic applications by automating sample handling and analysis, and lowering sample-analysis time by a factor of 10. It minimizes sample and reagent use, therefore reducing hazardous waste and cost per sample. The 5100 ALP can simultaneously process up to 4608 samples from industry-standard 96-well or 384-well plates. An automated, robotic handling mechanism loads the sample, with reagents and internal reference standards, onto a microfluidic chip for separation and detection. System software automatically analyzes the results, generating high-quality digital data as well as a gel-like image and electropherogram. All resources required for analysis, including samples, reagents, chips, and pipette tips, are stored within the bench-top instrument. Each chip can be reused several thousand times before replacement.
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In the news
Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
Figure 4. Agilent 5100 Automated Lab-on-a-Chip Platform.
The instrument infrastructure (e.g., power supplies, and pneumatic and reagent systems) has been de-
signed to support the launch of further assays that will take advantage of the capabilities of the 5100 ALP.
Checking quality RZPD needs to provide clone-based resources (e.g., cDNAs, proteins and siRNAs), which are indispensable for genome research, explained Maercker. However, until now, each product has had to be checked by gel analysis. This has been a rate-limiting step because RZPD could process a maximum of 400 samples a day as the gels had to be analyzed laboriously by eye. However, with the 5100 ALP, RZPD has achieved high-throughput analysis of 800 samples a day of DNA fragments. The whole process of sample loading and analysis is automated. ‘‘The machine and the software are easy to handle,’’ said Maercker. ‘‘Results are accurate and reliable, the representation of data is clear, and data stored in a database.’’ He particularly highlighted the accuracy of the 5100 ALP with the following results that showed deviations for: • Sizes of PCR fragments ALP vs. ALP (32 samples) 1.3% • Concentration of PCR fragments ALP vs. ALP (34 samples) 2.8% • Concentration of PCR fragments ALP vs. Fluorescence (picogreen dye) 23.9%
Figure 5. Screen shots of the 5100 ALP software show analysis of single PCR products (courtesy of C. Maercker, RZPD).
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Trends in Analytical Chemistry, Vol. 24, No. 2, 2005
In the news
Figure 6. Purification screening using the 5100 ALP software showing (a) gel-like images, and (b) images indicating concentration thresholds of purified protein in 96-well plates (courtesy of P. Hawtin, AstraZeneca).
As a result of RZPD’s experience, Maercker strongly recommended the 5100 ALP for the highthroughput analysis of PCR fragments, and looked forward to the possibility of extending its use to DNA fragments up to 12 kb, proteins and RNA.
High throughput With AstraZeneca, Paul Hawtin’s role as senior research scientist is to assess the potential usefulness of the 5100 ALP for high-throughput protein analysis. He reported a number of benefits of the 5100 ALP: • an increasing the number of samples could be analyzed; • automated, hands-free analysis;
• qualitative and quantitative data; • ease of use; • electronic data format for storage/retrieval; and, • safety and reduced environmental waste. Applications for which he found the 5100 ALP suitable included expression screening, refolding screen analysis, and purification screening (to give sufficient protein for crystallography and NMR studies). Expressing satisfaction with the 5100 ALP’s performance and its applications so far, Hawtin recommended that, for the future, Agilent should increase its flexibility, further integrate it to fully automate the expression/purification pipeline,
offer a relational database, and make reagents cheaper.
References [1] C. Maercker, ‘‘DNA applications of the 5100 platform’’, Press Briefing, Agilent Technologies, Waldbronn, Germany, 1 December 2004. [2] P. Hawtin, ‘‘High-throughput protein analysis using the 5100 ALP’’, Press Briefing, Agilent Technologies, Waldbronn, Germany, 1 December 2004.
Contact: Claudia Hachter, Agilent Technologies. Tel.: +49 (0)7243 602 100; E-mail: claudia_hachtel@agilent. com
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