Research News

Raman developments

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Scientists at the Science and Technology Facilities Council (STFC) Rutherford Appleton Laboratory in Oxfordshire, UK, have developed an effective laser-based method for the non-destructive characterisation of the bulk chemical content of pharmaceutical capsules. In collaboration with Pfizer Ltd, the researchers in STFC's Lasers for Science Facility succeeded in quantifying the presence of the active pharmaceutical ingredient in production line relevant capsules to a relative error of 1%. Other established non-invasive methods were unable to reach the same level of accuracy with the same sample.

The technique holds great potential for a range of process control applications in the pharmaceutical industry. The results of the collaborative study are reported in the J. Pharm. Biomed. Anal. (doi: 10.1016/j.jpba.2008.01.013). The development stems from research into Spatially Offset Raman Spectroscopy, which is under development at STFC for a wide range of applications including the detection of explosives in non-metallic containers, the detection of counterfeit drugs through opaque packaging and the non-invasive diagnosis of bone disease and cancer [see Spectrosc. Europe 19(5), 7 (2007)]. The concepts, which are relatively simple to implement, were developed through experiments involving STFC's large scale facilities which provided crucial insight into photon transport processes.

The development is being carried out with STFC's knowledge technology transfer arm and the new techniques are planned for commercialisation through STFC's spin-out company LiteThru Ltd.


NMR identifies marker of bipolar disorder

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Bipolar disorder, or manic depression, is a debilitating psychiatric condition characterised by alternating mania and depression, and affects about one in every hundred people worldwide. Although it is known that the condition can be treated relatively effectively using the moodstabilising drugs lithium and valproic acid, the reasons why these treatments work are poorly understood.

The authors of a new study, from Imperial College London, the University of Cambridge in the UK and the National Institutes of Mental Health in the USA, hope that their research will enable a better understanding of the condition and of how it can be treated.

The researchers compared postmortem brain tissue samples of people with manic depression with those of age and gender matched controls. The samples were taken from the dorsolateral prefrontal cortex, which controls the processes involved in higher cognitive functioning. The researchers analysed these samples using NMR spectroscopy and found that people with manic depression had different concentrations of chemicals in this area of the brain than those without.

The researchers also used rat models to see the effects of lithium and valproic acid on the metabolite makeup of non-bipolar brain tissue. They found that these drugs caused the opposite chemical changes to those seen in the bipolar brain tissue samples. Chemicals that were increased in the bipolar brain tissue were decreased in rats given the mood stabilising drugs, and vice versa.

The researchers' findings lead them to believe that an upset in the balance of different neurotransmitters known as excitatory and inhibitory neurotransmitters, which are involved in sending signals in the brain, may be central to the disorder. The study also suggests that lithium and valproic acid work by restoring the balance of these neurotransmitters in the brain. The work was published in Molecular Psychiatry (doi: 10.1038/


Bakerian lecture on Raman microscopy

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The Bakerian Lecture is the Royal Society’s premier annual lecture in the physical sciences (in practice, physics, chemistry, mathematics, meteorology, metallurgy, astronomy, geology and related subjects) and dates back to 1775. Previous awardees have included Humphrey Davy, Michael Faraday, James Clerk Maxwell, Lord Rayleigh, Ernest Rutherford, Gerhard Herzberg, George Porter etc. Since 1901 when Nobel Prizes were inaugurated, 26 Nobel Laureates have given the Bakerian Lecture.

The 2008 Bakerian Lecture was given by Professor Robin Clark CNZM FRS, University College London, UK, on the subject “Raman Microscopy, Pigments and the Arts/Science Interface” at the Royal Society of London on Thursday 7 February. It was broadcast simultaneously to New Zealand and elsewhere and a recording of the lecture is available from the Royal Society’s webcast archive. Further information may be obtained at the website:




2008 Wolf Foundation Prize in Chemistry

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This has been awarded jointly to William E. Moerner (Stanford University) and Allen J. Bard (University of Texas) for their ingenious creation of a new field of science, single molecule spectroscopy and electrochemistry.

William E. Moerner was the first to perform optical detection and spectroscopy of a single, individual molecule in condensed matter. Allen J. Bard pioneered the development of the scanning electrochemical microscope, allowing high-resolution, chemical imaging of surfaces and the study of chemical reactions at the nanoscopic regime, applied to biological and catalysis systems. By pushing optical detection to the ultimate limit of one molecule, these scientists changed our understanding of the chemistry and physics of individual molecules.

Professor William E. Moerner´s ingenious contributions to science have centred around two recurrent themes, which on one hand, address the development of a novel and revolutionary spectroscopic tool, single molecule spectroscopy; and on the other, its applications to problems in physics and analytical chemistry, biochemistry and biophysics. Since their pioneering steps in 1987, Moerner and his team have demonstrated a variety of sparking new subfields, including spectral diffusion of individual emitters, lifetime-limited line widths, temperatureinduced dephasing, non-linear saturation of a single molecule, photo-induced Poisson kinetics, blinking and switching of a single emitter, photon anti-bunching and optically-detected magnetic resonance of a single molecular spin. Thus, Moerner’s work trail-blazed a path for the measurement of individual molecules, having broad implication in the investigation of proteins, enzymes, DNA and RNA, and defects in solids or complex materials. Furthermore, this path enables the achievement of super-resolution imaging at the molecular level and endows scientists with the possibility to control the nanoscopic regime and to build molecular- scale devices.

Professor Allen J. Bard’s contributions to fundamental science are remarkably broad and include invention of the scanning electrochemical microscope, now used worldwide in investigations of electro-catalytic mechanisms, in identification of cancerous cells, in mapping transport paths in the skin for trans-dermal drug delivery, and for lithographic patterning of surfaces. His efforts are largely responsible for the development of electrochemically generated electroluminescence reactions, and immunoassay commercial technology, used worldwide in medical diagnosis and research.


Metrology website

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The new UK National Measurement System (NMS) Chemical and Biological Metrology website, www.nmschembio. is now online. It contains a wide range of information and material which will help laboratories ensure the quality, reliability and comparability of measurements.


  • Comprehensive publications database with papers, books, reports, guides and more; Information on key topics such as mass spectrometry, method validation and measurement uncertainty;
  • Training and education resources for schools, universities and laboratories;
  • Events listing of relevant training courses and conferences worldwide;
  • Access to help and expert advice on measurement topics;
  • Projects section detailing work being carried out under the NMS Chemical and Biological Metrology programme.

Registration is required to gain access to free downloadable material such as the Laboratory Skills Training Handbook and the Best Practice Guide for Generating Mass Spectra.


EU/US RM agreement

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A collaborative agreement between the European Commission (EC) Joint Research Centre's (JRC) Institute for Reference Materials and Measurements (IRMM) and the USA's National Institute of Standards and Technology (NIST) was signed on 17 December 2007 with the aim of enhancing trade between the US and the EU, while helping ensure the safety and quality of goods sold in both markets. The agreement advances the development and availability of international measurement standards in chemistry, life sciences and emerging technologies.

Under the agreement, the JRC and NIST will work to better coordinate their research and development programmes in metrology. This will include collaborative research on new measurement methods and their quality assurance, including but not limited to cooperation in the preparation and value-assignment of certified reference materials. The JRC and NIST also plan to share resources and harmonise their respective regional and national responsibilities for chemical metrology, biometrology and international measurement standards.

A number of workshops and conferences are planned as part of this initiative, and the following topics have already been identified:

  • Measurement methods and standards for hazardous substances in electrical and electronic equipment (i.e. RoHs)
  • Reference materials for the analysis of potentially dangerous polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs)
  • New measurement methods, technologies and standards for biofuels, multiplex biological measurements and the health and environmental effects of engineered nanomaterials.

MIMS helps show that animals can fix nitrogen

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It has been thought that only plants can fix nitrogen: convert nitrogen from the air into compounds that can be made into protein. However, researchers at Ocean Genome Legacy in Ipswich, MA, USA, and Harvard Medical School, have used multi-isotope imaging mass spectrometry (MIMS) to show that shipworms (marine clams) can also perform this feat. They burrow into and eat wood, causing massive damage to ships and piers every year. Wood contains very little protein, but the shipworms rely on bacteria living symbiotically in their gills which can fix nitrogen.

MIMS was used to image directly and measure the nitrogen fixation by individual bacteria in host cells, using nitrogen enriched with 15N, and to demonstrate that the fixed nitrogen is used for metabolism by the shipworm.


Grant for NIR work to help athletes

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Scientists from the Department of Biological Sciences at the University of Essex have received a grant of £28,571 from the UK’s Engineering and Physical Sciences Research Council to help in their studies of the use of near infrared (NIR) spectroscopy to determine the levels of oxygen in muscle. This helps athletes to optimise their training schedules and racing strategies. The funding will help design and develop a lightweight, portable device that can be worn comfortably in training and feed information wirelessly in real time to the coach. This will allow optimal targeting of training sessions in the field.

Professor Chris Cooper, Professor Ralph Beneke and Dr Caroline Angus of Essex’s Medical Optics Group will be working with physicists, engineers and computer scientists at University College London who will design the instrumentation. The Essex team will be carrying out testing and optimisation.

Professor Cooper explains that “exercise uses up oxygen and therefore how much oxygen is in the muscle is a measure of whether the oxygen being delivered is keeping up with its consumption. The key to the project is to take this data from the scientists to the coaches so they can use it to help optimise the way athletes warm up, or to design pacing strategies telling athletes when it’s the right time for them to speed up or conserve energy during a race.”

The team aims to have a working prototype in trials by spring 2008. For more information see and an article by Caroline Angus in NIR news last year (


Reactions observed in a single cell

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Bioengineers at the University of California, Berkeley, have discovered a technique that for the first time enables the detection of biomolecules’ dynamic reactions in a single living cell. By taking advantage of the absorptions of organic and inorganic molecules, the team of researchers, led by Luke Lee can determine in real time whether specific enzymes are activated or particular genes are expressed, all with unprecedented resolution within a single living cell.

The technique, described in the 18 November 2007 issue of Nature Methods (doi: 10.1038/nmeth1133), could lead to a new era in molecular imaging with implications for cell-based drug discovery and biomedical diagnostics. The researchers point out that other techniques, such as nuclear magnetic resonance, can at best provide information about a cluster of cells. But to determine the earliest signs of disease progression or of stem cell proliferation, it’s necessary to drill down deeper to the molecular dynamics within a single cell.

“Until now, there has been no non-invasive method that exists that can capture the chemical fingerprints of molecules with nanoscale spatial resolution within a single living cell,” said Lee. “There is great hope that stem cells can one day be used to treat diseases, but one of the biggest challenges in this field is understanding exactly how individual cells differentiate. What is happening inside a stem cell as it develops into a heart muscle instead of a tooth or a strand of hair? To find out, we need to look at the telltale chemical signals involved as proteins and genes function together within a cell.”

The researchers tackled this challenge by improving upon conventional optical absorption spectroscopy. “For conventional optical absorption spectroscopy to work, a relatively high concentration of biomolecules and a large volume of solution is needed in order to detect these subtle changes in frequencies and absorption peaks,” said Lee. “That’s because optical absorption signals from a single biomolecule are very weak, so you need to kill hundreds to millions of cells to fish out enough of the target molecule for detection.”

The researchers came up with a novel solution to this problem by coupling biomolecules, the protein cytochrome c in this study, with tiny particles of gold measuring 20–30 nm long. The electrons on the surface of metal particles such as gold and silver are known to oscillate at specific frequencies in response to light, a phenomenon known as plasmon resonance. The resonant frequencies of the gold nanoparticles are much easier to detect than the weak optical signals of cytochrome c, giving the researchers an easier target.

Gold nanoparticles were chosen because they have a plasmon resonance wavelength ranging from 530 nm to 580 nm, corresponding to the absorption peak of cytochrome c.

“When the absorption peak of the biomolecule overlaps with the plasmon resonance frequency of the gold particle, you can see whether they are exchanging energy,” said study co-lead author Gang Logan Liu, who conducted the research as a UC Berkeley PhD student in bioengineering. “This energy transfer shows up as small dips, something we call ‘quenching,’ in the characteristic absorption peak of the gold particle.”

A relatively small concentration of the molecule is needed to create these quenching dips, so instead of a concentration of millions of molecules, researchers can get by with hundreds or even dozens of molecules. The sensitivity and selectivity of the quenching dips will improve the molecular diagnosis of diseases and be instrumental in the development of personalised medicine, the researchers said.

The researchers repeated the experiment matching the protein haemoglobin with silver nanoparticles and achieved similar results. “Our technique kills two birds with one stone,” Lee said. “We’re reducing the spatial resolution required to detect the molecule at the same time we’re able to obtain chemical information about molecules while they are in a living cell. In a way, these gold particles are like ‘nano-stars’ because they illuminate the inner life of a cellular galaxy.”

Other researchers on the UC Berkeley team are Yi-Tao Long, colead author and postdoctoral scholar in bioengineering; Yeonho Choi, a PhD student in mechanical engineering; and Taewook Kang, a postdoctoral scholar in bioengineering.


Doping detection

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One of the most common substances used for doping is EPO (erythropoietin), which is difficult to detect. In an era when there are increasing numbers of “copies” of biotechnologically produced medications (biosimilars), it is also becoming more and more difficult to detect the difference between the body’s own EPO and that made biosynthetically. Chemists at Vienna University of Technology (TU) working jointly with ARC Seibersdorf, are developing a new analytical method, based on MALDI mass spectrometry, to track down the perpetrators of doping.

“With the aid of MALDI mass spectrometry, a method that is used for non-destructive desorption/ionisation of large molecules, especially biopolymers, we compare the deceptively similar ‘humanised’ form of EPO with the body’s own substance. The two samples differ in the structure of the amino acid chains and/or in that of the associated sugar chains. Depending on the structure of these sugar chains and where they bind to, we can recognise whether this is a natural or biosynthetic EPO”, explains Professor Günter Allmaier of the Institute of Chemical Technologies and Analytics at Vienna University of Technology.

Previous methods, for example iso electric focusing, exhibit several weaknesses. First, it takes between two and three days to obtain the test results. Furthermore, the method is regarded as difficult to automate, and is based on antibodies which can detect EPO in urine but sometimes are too nonspecific and do not distinguish the structure sufficiently precisely. Allmaier and his co-workers are concentrating now on a search for suitable analytical strategies that can detect recombinant EPO directly in urine. Lab-on-chip technology is to be combined with laser-based time-of-flight mass spectrometry. Following the testing phase, Allmaier estimates that the method may reach the patentable stage around 2009 and provide a valuable support in the fight against doping. Allmaier points out that “the most essential point in our strategy is that we are developing a method with which the EPO molecule itself is detected. All the other methods used so far have been indirect.”

EPO preparations increase the production of red blood cells, which in turn transport more oxygen in the blood. As a result, the organism’s performance improves. That is why EPO has been misused for doping as far back as the late 1980s, mainly in endurance sports such as cycling. Recently, Günter Allmaier received the John Beynon Prize Award 2007 for the most innovative publication in Rapid Communications in Mass Spectrometry for the period 2005 to 2006 ( PDFSTART). This work was also the starting point for intensive cooperation with Dr Reichel of ARC Seibersdorf’s doping control laboratory.


SPR imaging

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Using a new imaging technique based on surface plasmon resonance (SPR), a fast and accurate profile of autoantibodies present in the blood serum of rheumatic patients can be made. This profile can give valuable information about the progress of the disease. A unique feature of the SPR technique is that it directly tests on blood serum, without complex preprocessing. A special chip will enable many parallel tests. Scientists from the University of Twente and the Radboud University Nijmegen, both in The Netherlands, describe the new imaging technique in the Journal of the American Chemical Society (doi: 10.1021/ja075103x).

The scientists have run tests on the serum of 50 RA patients as well as a control group of 29 people. Direct testing on blood serum is unique: in other techniques fluorescent labels and preprocessing are necessary to visualise the relevant proteins. The diluted serum is led over a special gold coated microchip containing a large number of spots with a specific peptide coating. Whenever these peptides interact with autoantibodies present in the serum, this process can be monitored by Surface Plasmon Resonance Imaging (SPR). Using laser light, all gold spots are scanned: the reflection of light of the spots changes whenever there is a molecular interaction on the spot. At a certain angle of light, there is no reflection at all: this is the so-called SPR dip undergoing a shift caused by the interaction. The technique goes beyond proving that autoantibodies are present: the interaction between the protein and the antibody can be monitored real-time and without any labels.

Autoantibodies are manufactured by the immune system as a reaction on the citrullinated proteins playing a role in rheumatoid arthritis. On a single chip, several types of peptides can be placed, for rapid parallel screening. The next step is to investigate in what way the patient profiles help to monitor the progress of the disease. This could lead to more personalised treatment in the future. The applications are not limited to monitoring rheuma or other autoimmune diseases: SPR imaging can be used for monitoring a wide range of biomolecular interactions.

The research was led by Dr Richard Schasfoort of the BIOS Lab-on-a-chip group, part of the MESA+ Institute for Nanotechnology of the University of Twente. He has cooperated closely with the Biomolecular Chemistry group of Professor Ger Pruijn at the Radboud University Nijmegen. The research has been financed by the Dutch Technology Foundation (STW) within a project called “Proteomics on a chip for monitoring autoimmune diseases”.


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