SERVICES & FACILITIES ANNUAL REPORT - FY April 2004 to March 2005
SERVICE
Life Sciences Mass Spectrometry Facility
(LSMSF)
FUNDING
Block
AGREEMENT ESTABLISHED as S&F
Bristol-R8/12/15 Lancaster 1984
East Kilbride 1994
Bristol 1992
TERM
5 years
TYPE OF SERVICE PROVIDED:
Under the auspices of the LSMSF, NERC maintains three facility nodes for provision of organic and light stable isotope mass spectrometry to the UK life sciences community, namely the East Kilbride node located at SUERC, the Lancaster node located within CEH-Lancaster and the Bristol node housed within the School of Chemistry at the University of Bristol. By adopting this more integrated approach accessibility to said Facility nodes has been increased as has been the organisation between
Facilities. Whilst this move has not resulted in the physical amalgamation of the three Facilities, the ‘one-stop-shop’ exists as a single point of contact for users, with increased efficiency and synergistic operation thus making better use of the resources currently used in the maintenance of the mass spectrometry services offered. All three Facilities are contracted by NERC to provide
‘free-at-point-of-delivery’ support according to their respective service level agreements (SLAs) or contracts and the NERC
LSMSF steering committee oversees their operation. Each of the three nodes offers a different portfolio of analytical techniques for which UK based researchers may apply to use through NERC Services and Facilities (S&F) by standard peer review procedure.
East Kilbride has strong associations with migration, agro-ecology and conservation studies whilst the primary remit of Lancaster is terrestrial and fresh-water studies (including those deriving from NERC programmes). Bristol fields a wide range of projects to which a compound specific approach is essential, e.g. palaeoclimatic reconstruction (alkenone U K
37
), organic matter sourcing
(biomarkers), faunal population and dietary studies (isotopic PLFA and FFA profiling) and palaeodietary reconstruction.

Isotopically enriched water (D
2
18 O to energy expenditure studies

Natural abundance e.g. 13 C/ 12 C, 15 N/ 14 N, 18 O/ 16 O and 34 S/ 32 S analyses of bulk animal organic matter & molecular oxygen to study food webs & element cycling

Enriched & natural abundance analyses of bulk terrestrial organic matter e.g. 13 C/ 12 C, 15 N/ 14 N

Enriched & natural abundance analyses of gases: e.g. 13 C/ 12 C, 15 N/ 14 N 18 O/ 16 O of CO

Natural & near natural abundance compound specific 13 C/ 12 C, 15 N/ 14
2
, CH
4
, N
2
0 & N
2
N, D/H analyses of biochemical extracts

Organic mass spectrometric analyses of complex mixtures of compounds e.g. volatiles, functionalised molecules etc
NB. Nodes are designated as follows: L = Lancaster, EK= East Kilbride, B = Bristol
ANNUAL TARGETS AND PROGRESS TOWARDS THEM : Lancaster node to: a) consolidate its resources and resume a full service for customers, b) commission stand-alone trace gas and GC-C-IRMS instruments, c) Three methods to be accredited to
UKAS ISO17025 (external audit recommended UKAS status by mid 2005).Status: All three targets achieved. EK Node to: develop sulphur isotope ratio mass spectrometry and dual hydrogen-oxygen analysis in organic materials: Status: Both techniques have advanced in terms of progress made by node staff. Bristol node to: a) make compound specific 15 N and 18 O a viable routine service technique. Good progress has been made but the techniques require further development before becoming routine methods. b)
Promotion of ESI/APCI-LC/MS techniques & initiate further staff training: Both these targets have been achieved.
SCORES AT LAST REVIEW (each out of 5)
Need
5
Uniqueness
5
Date of Last Review:
Quality of Service Quality of Science & Training

5 5
28/01/03
Average
5
CAPACITY of HOST ENTITY
FUNDED by S&F
Lancaster 66%
East Kilbride 100%
Bristol 13%
Staff & Status
Dr AW Stott (SSO), Mr D. Sleep (HSO), Miss H. Grant (SO)
Dr. J Newton (RA2) & Dr. R. McGill (RA1A)
Next
Review
(January)
2008
Contract
Ends
(31 March)
2009
Dr ID Bull (OR2; 100% University of Bristol funded) & Dr R Berstan
(RA1A)
FINANCIAL DETAILS: CURRENT FY
Recurrent
Allocation £k
L 88.04
EK 117.33
B 65.17
½ day unit
L 0.364
EK 0.377 n/a
Unit Cost £k
Analysis n/a n/a
B 0.2
Training/Labwork n/a n/a
B 0.1
FINANCIAL COMMITMENT (by year until end of current agreement) £k
2004-05 L 88.04
EK 122.4
B 75.1
2005-06 L 107.8
EK 128.17
B 75.2
2006-07 L c. 111
EK c. 134
B c. 77.4
2007-08
Capital
Expend £k
0
5
30
L c. 116
EK c. 138.9
B c. 77.7
STEERING COMMITTEE Independent Members Meetings per annum
LSMSF 5 2
Income
£k
0
0
0
2008-09
Full cash cost £k
155.24
160.72
89.01
L c. 121
Other S&F Overseen
0
EK c. 144.5
B c. 80

APPLICATIONS: DISTRIBUTION OF GRADES (Current FY — 2004/05)

5

4

3

2
NERC Grant projects
Other academic
Students
8
4
4
1
1
7

1

R*/Pilot
1
1
5
Pilot
TOTAL 0 16 9 0 0 0 7
APPLICATIONS: DISTRIBUTION OF GRADES (per annum average previous 3 years —2001/2002, 2002/2003 & 2003/2004)

5

4

3

2

1

R*/Pilot
NERC Grant projects
Other Academic
Students
2.3
0.3
10
2
4.3
2.7
1
4.7
0.7
0.3
1
1
0.3
8.7 0.7 0.3 0
2
4
Pilot
TOTAL 2.6 16.3
PROJECTS COMPLETED (Current FY)
NERC Grant projects

5
2

4
5

3
4

2
1

1

Other Academic
Students
Pilot
1
1
Reject
1
1
Reject
0.3
0.3
R*/Pilot
1
USER PROFILE (current FY)
Infrastructure
Grand
Total Supplement to NERC Grant *
Student
Total NERC
52 6 32 23
USER PROFILE (per annum average previous 3 years)
Grand
Total
Infrastructure
Student
Supplement to NERC Grant *
Total NERC
56.7
14 29 15.7
NERC C/S
4
NERC C/S
4
Other
10
Other
9.7
NERC Grant*
NERC Grant*
USER PROFILE (current FY)
Academic
19
USER PROFILE (per annum average previous 3 years)
Academic
28.3
Centre/Survey
4
Centre/Survey
2.7
NERC Fellows
1
NERC Fellows
1.3
OUTPUT & PERFORMANCE MEASURES (current FY)
Publications (by science area & type)
PhD
25
PhD
28
*Combined non-Directed and Directed
PAYG
Student
Total NERC
NERC C/S Other
*Combined non-Directed and Directed
PAYG
Student
Total NERC
NERC C/S Other
Commercial
Commercial
1.7
Non-Ref/ Conf Proc
11
PhD Theses
5
SBA
5
ES
3
SBA ES
9.3 3.7
MS
7
MS
5
AS
0
TFS
43
AS TFS
0 22.3
EO
0
EO
0
Polar
0
Polar
0
Grand Total
58
Grand Total
40.3
Refereed
42
SBA
2
ES
0
Distribution of Projects (by science areas)
MS
14
AS
0
OUTPUT & PERFORMANCE MEASURES (per annum average previous 3 years)
TFS
Publications (by science area & type)
36
Refereed
30
EO
0
5.3
Non-Ref/ Conf Proc
Polar
0
PhD Theses
5
Earth’s life support systems
33
SBA
8.7
ES
5.3
Distribution of Projects (by science areas)
MS AS TFS
8.3 0 34.3
Distribution of Projects (by NERC strategic priority)
Climate Change Sustainable Economies Underpinning Science
EO
0
4.66 3 0
Specific Research
0.66
Polar
0
OVERVIEW & ACTIVITIES IN FINANCIAL YEAR ( 2004/05) :
LSMSF General:
The launching of the LSMSF website featuring the newly designed NERC LSMSF logo, has meant that potential applicants now have a simple and readily accessible information portal facilitating directional guidance to the specific node in question. This mechanism has greatly improved communications between the user community and the node staff, in addition to providing a wealth of useful

information regarding LSMSF operations and application protocols. Flyers and full page spreads advertising NERC LSMSF were included in the registration packs given to all attendees at SIMSUG 2005 and Archaeological Sciences conferences. Other advertising activities have included the creation of an entirely new website (http://www.bris.ac.uk/nerclsmsf) for the Bristol node. Publications have increased, as has the overall trend in high quality applications which reflects both an increase in the quality of applications which are accepted by the LSMSFSC and the increase in sample throughput over the last year.
Node overview
Lancaster: 2004/2005 has seen a heavy schedule for the node. CEH resources have focussed upon the node gaining UKAS
ISO17025 accreditation for three of its routine methods by April 2005. Preliminary external audits in January were encouraging and conclusions were reached that the node will become a UKAS accredited laboratory in April for both NERC and CEH analytical work.
All instruments have been fully operational and methods have been re-validated to quality assured ISO17025 standards. Throughput of LSMSF supported work has been exceptionally high, approaching 9,500 samples with all targets met or exceeded. Facility personnel have actively participated in promoting Science awareness to children in local schools and colleges. Dr Stott gave oral presentations to North West school teachers on isotopes in ecology, ran a forensic isotope course for Lancaster and Morecambe college teachers/students and recently, with the support of Mr Sleep and Miss Grant ran a series of ‘Where are you in the food chain’ workshops for over 50 local students as part of National Science week.
East Kilbride: The facility website (http://www.gla.ac.uk/suerc/research/nerclifesci.html) is now in the hands of the node manager rather than SUERC, which means that the pages, including recent projects and developments are updated on a weekly basis. Drs
Newton and McGill gave oral presentations to the 4th International Conference on Applications of Stable Isotope Techniques to
Ecological Studies in Wellington in April, and are organising the 5th Conference to be held in Belfast in 2006. The technical problems of the previous year are now solved with the purchase of a new mass spectrometer and sample throughput has almost doubled as a result. Both staff members undertook a 5 day training course with ThermoElectron in Bremen associated with the purchase of this instrument. We are perfecting a routine technique for measuring sulphur isotopes in (mainly) organic materials, from which two applications are benefiting, and soon hope to develop dual hydrogen-oxygen isotope analysis of organic materials. Last year's explosion of interest in bird migration has only partly subsided, and marine applications are on the increase.
Bristol: The 2004/2005 year has seen a more normal period of activity with service activities no longer being hampered by refurbishment or, compared with previous years, personnel issues. Since his employment on 1 st May Dr Robert Berstan has proven to be an invaluable asset to the node; his high organisational and technical competences helping to ensure a much improved service to users of the Facility. Dr Berstan has already undertaken a 5 day Delta XP GC-EA IRMS training course with ThermoElectron in
Bremen and both Drs Berstan and Bull have received an additional 10 days of onsite training for the ThermoElectron MAT95 XP-
Trap mass spectrometer. Dr Bull gave oral presentations to the 16 th British Organic Geochemical Society meeting in July, the 2005 meeting of the Stable Isotope Mass Spectrometry Users Group in April and 2005 UK Archaeological Sciences conference also in
April. As well as containing all the necessary information and content required by potential users of the Facility the site also aims to promote knowledge transfer by providing explanatory information and educational materials to help non-specialists, who may wish to use Facility services, to better understand the activities of the Bristol node. Such information includes explanations of the hyphenated instrumental techniques provided by the node, examples of projects that have successfully utilised a compound specific approach and, in the near future, ‘wet chemical’ methodologies for sample preparation. The node continued to service applications covering a wide range of NERC science areas whilst concurrently Facility staff have also undertaken their own research activities that have resulted in further outputs, most notably Evershed, R. P., Berstan, R ., Grew, F., Copley, M. S., Charmant, A. J. H., Barham, E., Mottram, H. R. and Brown, G. (2004) Formulation of a Roman Cosmetic. Nature 432, 35-36.
Generic LSMSF training protocol:
One of the primary goals of the LSMSF strategy is to actively underpin Ph.D. student and postdoctoral training. During 2004/05 approximately three quarters of the projects accommodated were student projects which requested visits to the node laboratories to receive ‘hands on’ training from Facility staff. Students have had little if any previous experience of ‘wet chemical’ preparative methodologies, high precision quantitative stable isotope analyses or mass spectrometry and this was their first introduction to vacuum-line cryogenic chemistry and/or isotope-ratio mass spectrometry or related mass spectrometric techniques. The often intensive training they receive within the LSMSF therefore develops skills different and complementary to those required in the field or later in their career development. The combination of laboratory analysis and data interpretation skills is considered an attractive attribute of our 'alumni' and fulfills NERC’s responsibilities to students and fellows as part of their skill development portfolio.
LSMSF capital:
Lancaster : The GV Instruments Isoprime (30k from 03/04 capital) was successfully commissioned and coupled to the GC combustion interface allowing stand alone instrumentation for both compound specific and trace gas techniques. Already applicants are benefiting from this additional resource which has vastly improved instrument capacity, node turnover times and sample throughput.
East Kilbride: A hydrogen gas generator was procured via last year's capital bid. This is used for the production of standard gas in hydrogen isotope analyses and for routine system testing procedures.
Bristol: A PTV injector to improve chromatography on a GC/C/IRMS system and an oil-free compressor to provide pneumatic air pressure for the whole mass spectrometry laboratory was purchased using 9k secured from the 04/05 capital bid round.

SCIENCE SUPPORTED IN FY ( 2004/05) : QUALITY OF SCIENCE
The quality of applications to the Facility has remained consistently high. The LSMSF received a total of 33 applications this reporting year, of which 16 were

4 grade, 9 were

3 grade, 7 resubmissions (R*)/pilot studies and 1 rejected outright. The peer review mechanism ensures that only high quality science is supported. 42 manuscripts have been published in peer-reviewed journals this year, 5 PhD dissertations have been awarded. Facility staff have made presentations at Archaeological Sciences, British
Ecological Society, British Organic Geochemistry Society, Stable Isotope Ecology, Stable Isotope Mass Spectrometry Users Group,
National Science Week & North West Teachers conference.
Lancaster: There is evidence of increased levels of DOC entering rivers that run through peat lands and this excess release of carbon from may lead to these important carbon stores becoming unstable. This increase was originally thought to reflect climate warming and increased river flow. However, by following the fate of a 13 C enriched CO
2
tracer, NERC funded researchers from the University of Bangor in collaboration with node staff showed that elevated CO
2
stimulated plant growth, and larger amounts of DOC was released from plants into soils and waters (published Nature, 2004). NERC funded research by Lancaster University and node staff demonstrated for the first time a direct link between the responses of a wide range of tree species, increased heterotrophic respiration and a resulting decrease in sequestration of root-derived C at elevated CO
2
concentrations. Results suggest that the incorporation of root-derived C into stable, medium- or long-term forest soil C pools may be substantially reduced as the atmospheric CO
2 concentration exceeds much more than 100 μmol mol -1 above current ambient. This would have the potential to trigger a huge positive feedback on the rate of increase in global atmospheric CO
2
concentration and associated climate change (submitted to
Science).
East Kilbride: The use of isotopes in bird migration proved a hot topic in 2003, and the same principles are being applied to other animals, notably sperm whales, in an application from Graham Pierce and Sonia Mendes of the University of Aberdeen. C and N isotope profiles of teeth from stranded animals are giving us information on life history changes in the trophic ecology and latitudinal migrations of male sperm whales. On similar lines, Clive Trueman at the University of Portsmouth is measuring the isotopic spacing between diet and tissue in Atlantic salmon reared on an isotopically constant diet under varying conditions of temperature and photoperiod, designed to modify growth rates. At the conclusion of the project, the effect of growth rate on diet-tissue isotope spacing in C and N will be determined, allowing the use of archived salmon scales to reconstruct dietary and ecological patterns. The longterm goal is to produce temporal records of salmon diet and ocean conditions in order to identify causal mechanisms responsible for the declining wild populations of Atlantic salmon.
Bristol: A successful (

4) application from Dr Guillaumé Masse of the University of Plymouth utilised data obtained from compound specific

13 C analyses provided at the Bristol node to help ascertain the nature of isoprenoid biosynthesis in marine algae. Isoprenoid biosynthesis in the widespread diatomaceous algae, Rhizosolenia setigera (Brightwell) and Haslea ostrearia (Simonsen), results not only in the production of diterpenoids, triterpenoids, and sterols but, unusually for diatoms, also in the production of sesterterpenoids.
By using 13 C and 2 H isotopic labelling techniques followed by NMR and mass spectrometry, specific inhibition of mevalonate (MVA) and methylerythritol (MEP) pathways, and comparison with the natural 13 C/ 12 C isotope ratios of the lipids, the different biosynthetic pathways of the sesterterpenes and other isoprenoids have now been determined. The observations ostrearia revealed that terpenoid biosynthesis in diatoms is species-dependent and cannot simply made for R. setigera and H. be grouped according to structural type. Triterpenes appeared to be made by the MVA route as in higher plants, whereas sesterterpenes and sterols can be made by either the MVA or MEP routes. In neither organism were the isoprenoids biosynthesized by leucine metabolism. Sesterterpene and triterpene biosynthesis in diatoms has not been investigated previously. This work led to a publication in the Proceedings of the National
Academy of Sciences. Isoprenoid biosynthesis in the diatoms Rhizosolenia setigera (Brightwell) and Haslea ostrearia (Simonsen).
PNAS 101 , 4413-4418).
FUTURE DEVELOPMENTS/STRATEGIC FORWARD LOOK
In 2005/2006, the LSMSF aims to: continue the synergistic operation of the three nodes e.g. sharing of knowledge, expertise and analytical capabilities and provide a level of insurance against catastrophic instrument failure. Each specific node has its own future development strategies aimed at improving the overall quality of LSMSF service to the end user:
Lancaster: Applications received during 04/05 have been predominately TFS requesting bulk

13 C and

15 N analyses and gaseous N
2 and N
2
O analyses falling in the MS science area. A similar trend is expected in 05/06, however it is predicted that more labelled and analyses and N
2
gas analyses will be forthcoming in 2005/2006. Additionally, we intend to develop our trace gas 13 C methods to accommodate lower level carbon concentrations than are currently not measurable using bulk methodologies.
East Kilbride:

13 C and

15 N analysis of organic material remains the predominant requirement of node users, and applications involving D/H analysis are constant. Despite the new Delta Plus XP mass spectrometer, switching between analytical techniques and the development of new ones (e.g.

34 S and dual D/H and

18 O) have considerable detrimental effect on scheduling; hence it is likely that in the next few years we will be looking towards purchasing a second mass spectrometer. Last year we began our first applications involving sulphur isotope analysis, and we expect interest in

34 S to increase, particularly in applications involving marine/terrestrial discrimination. Dual D/H and

18 O is expected to attract interest and we hope to develop this method during the coming year.
Bristol: Applications continue to be received from a diverse range of scientific fields requesting support for both organic and light stable isotopic compound specific analyses. The operation of instrumentation has recently been streamlined to dedicate instruments to specific techniques, e.g. compound specific

D and light stable isotopic analysis of N containing compounds. Extensive methodological development at Bristol of the latter has recently come to fruition and reliable

13 C and

15 N analysis of N containing compounds is expected to become a routine technique over the next few months. The node has received continued applications requesting provision for such analyses and will continue to seek funding for the ideal instrumentation (a ThermoElectron MAT 253) with which to make them. Facility staff are also actively seeking funds for the provision of a ThermoElectron LC/IRMS which will enable the light stable isotopic analysis of highly polar (e.g. phospholipids) and involatile compounds (e.g. proteins), a technique that has the potential of providing critical insights into complex, natural systems that are unobtainable by any other method. Over the ensuing period the node will also investigate possible mechanisms that may provide technical help with the daily operation of instrumentation.