MassMatters

BMSS42: A Celebration of MS

Editor’s letter

Welcome to the latest edition of Mass Matters! In this issue we feature a recap of our recent 42nd annual meeting, recently held in Manchester, as well as contributions from Tony Bristow, the Females in Mass Spectrometry (FeMS) group as well as two ‘Reflections’ interviews by our Associate Editor, Mervyn Lewis, Hannah Britt and Anyin Li.

We’d also like to take the opportunity to say how saddened we were by the passing of HM Queen Elizabeth II in September 2022 – Her Majesty was a lifelong supported of the sciences via her patronages, and she will be missed by so many.

On a happier note, we welcome new committee members Mike Morris, Aneika Leney and Rhodri Owen following our recent elections, great to have you on board!

Best, Jon.

Jonathan Jones, BMSS Publicity Secretary

Disclaimer: Articles published in Mass Matters are not peer reviewed. Authors are solely responsible for the accuracy and content of their articles. Authors may request that final proofs of their articles are approved by the author prior to publication. If no such request is made, the editorial team will proofread the articles for spelling, grammar and general layout only. Authors must obtain any relevant permission prior to submission, including the permission of the image copyright holder where applicable. The authors retain copyright to articles published in Mass Matters. All images are copyright to their respective owners. Opinions expressed in articles published in Mass Matters are solely those of the original authors and not necessarily represent those of the BMSS or its executive committee.

Chair’s Report

Passport at the ready, toothbrush in hand, mask on face, cabin doors to automatic, and we are ready for departure! This summer the world began to open-up to us all. A brave few ventured across the pond to Minneapolis for ASMS, only to return with COVID from what was termed a super spreader event! That said, UK science was still shared at this international arena, and I’m told by those who attended it was great to be back!

For the more cautious of us, staying closer to home, the BMSS ran two in-person meetings in the spring. The Frank Pullen symposium was a sell-out success. With the help of the Burlington House AV team, the RSC Separations Science Group and BMSS were able to run a synchronous on-line event, enabling many from around the world to participate. After a 2-year hiatus on social events, many of us were clearly out of practice, so there were a few "tired souls” at the train station the next morning, and a strong need to set phone alarms to ensure no one over-shot their destination station. I am absolutely convinced Frank would approve! Shortly after the Frank Pullen Symposium, the EFA SIG and Ambient Ionisation SIG got together for a return to in-person SIG meetings. The synergy between these two SIG’s is wonderful to see, and I certainly hope to see more cross-fertilisation in the future.

The Brits' confidence in international travel had clearly returned by the time IMSC came around in August. After the Dutch and the Germans, UK delegates were the third most abundant in Maastricht which was very pleasing to see. At this meeting, Dr Hannah Britt and I teamed up with Dr Isabelle Kohler from NVMS (the Dutch Mass Spec Society) to deliver a careers workshop, the most popular workshop of the conference! We thank Prof. Alison Ashcroft, who had delivered her Thomson Medal Lecture the day before, and Prof Alexander Makarov for sharing their careers and career choices with us. We were also joined by SistersInScience (https:// sistersinscience.nl/) and I would

encourage you to check out their website to find out more about ways to connect in science. VRS stood by us once again (thank you Bryony Harvey) with a great interactive presentation.

BMSS42 was only a few weeks ago. The author Douglas Adams once told us that 42 is the “Answer to the Ultimate Question of Life, the Universe, and Everything"(1). So, I challenged the delegates to ask their Ultimate Scientific Questions, and to seek their own meanings to Life, the Universe and Everything (Mass Spectrometry). And this they did! It was an invigorating event with over 355 people buzzing around the Royal Northern College of Music. The format this year retained the flash oral presentations, which acted as great adverts to draw delegates to the poster halls to follow up with discussions. In-person posters and the vendor exhibition returned in full force with instruments on display and in one case, in operation. The feedback we have received is heart-warming, with comments that the standard of the presentations, and the level of science delivered, was exceptionally high. I must say that I agree. Some said was better than ASMS. I would like to acknowledge our amazing prize winners. You can read more about this on page 7.

A fabulous meeting, capped by two great plenary lectures: Prof John Langley opened the meeting and kept us awake with his interactive lecture; Prof Michal Sharon travelled all the way from Israel to delight us with her developments in methods for the direct study of proteins from cells. If you missed these, or any of the other talks or poster, then note that the delegates can still access the digital recording from BMSS42, so I encourage you to check out what was going on in the two other parallel sessions.

The BMSS committee is a rolling membership. Having served as Chair for two years, I now hand over to Prof. Neil Oldham, who will be ably supported by Dr Andy Ray as Vice Chair, Dr Mark Barrow as General Secretary and Dr Mark McDowall

as Treasurer. Dr Hannah Britt steps into the Education Officer’s position and Dr Chris Titman will take the reins of the SIG groups as SIG Co-ordinator and Dr Lindsay Harding will be managing future BMSS meetings as the Meetings Secretary. A big thanks must go to Prof Peter O’Connor who completes his tenure as Treasurer, and to Prof Zoltan Takats who completes his tenure as SIG Co-ordinator as both step off the committee. A big thanks also to Dr Ashley Sage who steps off the Advisory Board.

The committee now welcomes three new members aboard. Dr Rhodri Owens, has been a co-opted to the committee for the past couple of years and been very active with our EDI team, so it is great that Rhodri is joining as trustee and General Member. We also welcome Dr Aneika Leney (University of Birmingham) and Dr Mike Morris (Waters Corp.) as trustees and General members. A lot of changes, but I step down knowing that the Society is in excellent hands, and I look forward to relaxing and helping out from the side-lines (well, Advisory Board).

As a personal note, I would like to thank BMSS members, and everyone on committees past and present for supporting me through my various roles as a trustee, and especially as Chair over these past two, somewhat challenging, years.

Exciting plans are already afoot for 2023 so watch this space! I will be!

Dr Jackie Mosely BMSS Chair

(1) Hitchhikers Guide to the Galaxy by Douglas Adams. New York: Harmony Books, 1980.

Jackie Mosely, BMSS Chair

2023 BMSS membership subscription

Renew your BMSS membership subscription for 2023!

You can renew quickly and easily on-line by visiting the BMSS Website: www.bmss.org.uk.

Why not also encourage colleagues to join the BMSS? As a member of the BMSS they will:

• Be part of a community of people with similar interests

• Have the opportunity to keep up to date with recent technological developments, learn more about MS and share knowledge and experience e.g. by joining Special Interest Groups (SIG’s)

• Be eligible to apply for grants towards small items of equipment, summer studentships and conference travel

• Be entitled to discounts for BMSS meetings

• Benefit from reduced subscription fees to the European Journal of Mass Spectrometry

• Receive copies of Mass Matters, the official benefit from 20% off relevant book titles when purchased via Wiley Publishing (discount code required) publication of the BMSS, published three times a year

Students: Can benefit from our education programme by applying for travel grants, presenting research at meetings and by taking part in the Barber oral prize and Bordoli poster prize competitions. All student members also receive a general mass spectrometry text book upon joining.

New & Established spectrometrists: Can benefit from courses, careers events, links with other societies (such as ChromSoc. & RSC) and networking via the BMSS Annual Conference and Special Interest Group Meetings.

Any membership queries should be directed to: Lisa Sage, BMSS Administrator T: (01606) 810562 E: admin@bmss.org.uk

Recent publications by BMSS members...

Native Ambient Mass Spectrometry of an Intact Membrane Protein Assembly and Soluble Protein Assemblies Directly from Lens Tissue

O. J. Hale, H. J. Cooper, Angew. Chem. Int. Ed. 2022, 61, e202201458; Angew. Chem. 2022, 134, e202201458.DOI: 10.1002/ anie.202201458

Intact protein assemblies with molecular weight 90-113 kDa were analysed directly from eye lens tissue by nanospray-desorption electrospray ionization (nanoDESI). Detergent micelles in the solvent solubilised and preserved the assemblies of the tetrameric membrane protein Aquaporin-0 (113 kDa). Mass spectrometry imaging under non-denaturing conditions revealed the spatial distribution of these protein assemblies, and in situ top-down analysis identified their constituents.

Versatile, Cheap, Readily Modifiable Sample Delivery Method for Analysis of Air-/ Moisture-Sensitive Samples Using Atmospheric Pressure Solids Analysis Probe Mass Spectrometry

Kerry A. Strong, Peter Stokes, David Parker, Amy K. Buckley, Jackie A. Mosely, Claire N. Brodie, and Philip W. Dyer. Anal. Chem. 2022, 94, 32, 11315–11320. DOI: 10.1021/acs.analchem.2c02039

Some chemical compounds are air- and/or solvent-sensitive. This presents a huge problem in mass spectrometry: how to get a sample into the vacuum system of a mass spectrometer without exposing into environmental conditions that would compromise sample integrity. A number of solutions have been

presented over the years, but this solution is simple and has the potential to be adapted for a wide range of mass spectrometers capable of vertically loading Atmospheric pressure Solids Analysis Probe analyses (ASAP) or equivalent.

A reusable glass manifold is used to protect the sample while it is loaded into the mass spectrometer. Recent modifications have enabled the higher temperatures, required by many air- and solvent-sensitive metal-ligand complexes, to be successfully employed. In essence – it does what it says on the tin!

Sizing up DNA nanostructure assembly with native mass spectrometry and ion mobility

Jeroen F. van Dyck, Jonathan R. Burns, Kyle I. P. Le Huray, Albert Konijnenberg, Stefan Howorka & Frank Sobott. Nat Commun 13, 3610 (2022).DOI: https://doi. org/10.1038/s41467-022-31029-5

What can native MS & ion mobility do for DNA nanostructures? The story is here @NatureComms In collaboration with @howorka_lab, we used 6 different DNA strands (50mers) which are designed to pair with each other via 21 bp long duplex regions. Complete assembly results in hexameric barrel-like structures (6 helix bundle, 6HB) but requires sufficient ionic strength. We find assembly intermediates up to the tetramer with ion mobility data suggesting linear growth (2D "sheets"). The pentamer is absent due to the high cooperativity of forming the 6HB. The 6HB falls below the linear trend line due to its barrel-like 3D structure.

Next to 6HB we found unexpected

larger assemblies, 12HB & 18HB. We considered stacked multiple hexamers but found the best match, by ion mobility, to be larger diameter barrels. During gas-phase measurement they compact to the "squished" 12HB-SQ and 18HB-SQ structures.

In summary, MS identified previously undetected intermediates in the assembly process to the hexamer model structure; the oligomerization process was strongly dependent on the ionic strength of the buffer solutions. The shape of the assembly products was tracked with ion mobility and showed transition from planar to cylindrical structures between the tetramer and 6HB. MS also detected previously unknown minor byproducts of the assembly processes, larger diameter barrels with double and triple the mass of the 6HB nanostructure.

We believe that native mass spectrometry combined with ion mobility is poised to make significant contributions to our understanding of the formation and structural diversity of natural and synthetic oligonucleotide assemblies which are relevant in science and technology.

Metabolomics Markers of COVID-19 Are Dependent on Collection Wave

Lewis H, Liu Y, Frampas CF, Longman K, Spick M, Stewart A, Sinclair E, Kasar N, Greener D, Whetton AD, Barran PE, Chen T, Dunn-Walters D, Skene DJ, Bailey MJ. Metabolites. 2022 Jul 30;12(8):713. DOI: 10.3390/ metabo12080713.

This work demonstrates, for the first time, that metabolic dysregulation has partially changed over the course

of the pandemic, reflecting changes in variants, clinical presentation and treatment regimes. It also shows that some metabolic changes are robust across waves, and these can differentiate COVID-19-positive individuals from controls in a hospital setting. This research also supports the hypothesis that some metabolic pathways are disrupted several months after COVID-19 infection.

Sizing up DNA nanostructure assembly with native mass spectrometry and ion mobility

Jeroen F. van Dyck, Jonathan R. Burns, Kyle I. P. Le Huray, Albert Konijnenberg, Stefan Howorka & Frank Sobott. Nat Commun 13, 3610 (2022).DOI: https://doi. org/10.1038/s41467-022-31029-5

Colocation of Lipids, Drugs, and Metal Biomarkers Using Spatially Resolved Lipidomics with Elemental Mapping

Lewis H, Costa C, Dartois V, Kaya F, Chambers M, de Jesus J, Palitsin V, Webb R, Bailey MJ. Anal Chem. 2022 Aug 30;94(34):1179811806. DOI: 10.1021/acs. analchem.2c01940.

We demonstrate how elemental imaging and a new method for spatially resolved lipidomics can be used in combination to probe the relationship between metals, drugs, and lipids in discrete areas of tissues. This new method for spatial lipidomics has been applied to rabbit lung tissues containing a lesion caused by tuberculosis infection. We demonstrate how these methods can be used to probe the association between ion accumulation and lipid profiles and verify local drug distribution.

IN PICTURES: BMSS 42 @ Manchester

Barber Prize

Birmingham University

B.N. Green Prize

Swansea University

Special Mention.

Oxford University

Bordoli Prize Delegates Choice Prize

Manchester University

A special mention was also made to Morgan Johnson, one of our Summer Studentship recipients, who as an undergraduate gave what the judges felt was a quality presentation worthy of special merit. – Jackie Mosely

Kish Adoni

Manuela Pacciarini

Maria Elena Castellani

Emily Hicks

Tony has recently completed an anthology of poems documenting support of a young person through difficult times. In the months ahead, the anthology will go on sale with all proceeds going to charity. He kindly provided this summary for us.

An introduction

I started to write the poems as a way to articulate and understand my emotions as a parent supporting a young person with anorexia.

The poems tell mine and my family’s story through the dark to better times. Our journey is not complete, but we see a brighter future.

I wish that this short anthology can give hope and inspiration to the other families at different stages of the journey we have faced.

An exert from the poem titled Vapour trail: It happened first secretly and slowly, Small changes through days. Then suddenly extreme, Stared through a fog, preventing recognition.

I hope this shows, that even at the saddest times, love and hope remain.

All profits from the sale of the anthology will be shared amongst a number of relevant charities.

A copy of the anthology can be purchased for £8.99 via the link in my Instagram account (awtbristow_ author) or contact me directly at tonywtbristow@icloud.com

Finally, I would like to thank the BMSS Committee for their amazing support in promoting the anthology.

Tony Bristow Poems Managing Your Mental Health During Your PhD: A Survival Guide

Written by Dr Zoë Ayres

Book recommendation by

Krisztina Radi

As we just passed the World Mental Health Day (10th October ) it is more than timely that we also bring some very relevant reading to the attention of our members, especially PhD students, as this book is addressed to them.

The book we are wholeheartedly recommending for our Mass Matters autumn copy readers is written by Dr Zoë Ayres, the newly elected President of the Royal Society of Chemistry’s Analytical Science Community, and Head of Research and Development at Figura Analytics.

“This book is an exploration of PhD experience as never before and provides a “survival guide” for current and

prospective PhD students.

It investigates why mental health issues are so common among the postgraduate population, going beyond the statistics, looking at lived experience of both Zoe herself and as well as a number of current PhD students, who have found balancing mental wellness with the PhD endeavour challenging.

Zoe discusses tips and tricks she wished she had known at the start of her PhD process for managing mental health, such as managing imposter feelings, prioritising workload, and self-care strategies to help others throughout their own journey.

The book goes beyond typical mental health discussions (where the focus for improving mental health is placed on PhD students to become “more resilient”) and explores some of the often-unspoken environmental factors that can impact mental health. These include the PhD studentsupervisor relationship, the pressure

to publish, and systemic problems in academia, such as racism, bullying and harassment.

The book is very much a call to action, providing tangible improvements from Zoe’s perspective that university institutions can make to ensure that academia is a place for all to thrive"

The topics discussed in the book although written with the challenges of the PhD journey in mind will be very useful for all to dive in, as surely can use those tips and tricks in our post PhD life. It is never late to learn to be more aware, find ways to become more resilient, to understand how to manage relationships with someone we welcome guidance from, or deal with the pressure to deliver while navigating our feelings in competitive work environments.

Footnote:

Released in September 2022, the

book is available in both print and e-book formats, with the e-book link largely accessible for free for university students and staff here: https://link.springer.com/ book/10.1007/978-3-031-14194-2

✪ LEEDS ✪ 14-15 NOVEMBER 2022

The Biomacromolecular Structure SIG coordinators invite you to participate in a 2-Day event 14-15 November 2022. The first day will offer hands-on training in HDX-MS and Native MS. The second day will be a classic SIG meeting. Prospective delegates may register for one or both of these options. Please visit the Society’s website for details…

FEATURE: AI & FASIG REVIEW

In review: AI/EFA SIG Combined Meeting

By Peter Ball

A report on the AI/EFA SIGs Combined meeting which took place at the University of Surrey, Guildford, April 27, 2022.

Introduction.

The first in person meeting of these SIGs post-pandemic was well attended by more than 60 delegates with strong support from vendors both exhibiting and presenting. The local organisers are to be complimented for their input to enable the meeting to run smoothly. David Megson commented that the venue had to be changed at short notice and he was thankful that the University of Surrey and Dr Patrick Sears, in particular, had stepped in to enable the meeting to go ahead as planned. Oral and poster presentations were featured with the keynote presented by Dr Simon Hird (Waters Corporation) and the ‘Peter Ryan prize’ was awarded to Simone Mathias, a postgraduate at the University of Surrey for the best ECR poster presentation. The introduction to the meeting was given by Professor Paul Townsend.

A brief description of the nine orals and twelve posters is given in the following paragraphs. It is worth noting that one of the features of the meeting related to the posters, which were introduced by the respective presenters prior to the poster session.

Oral Presentations.

The keynote presentation by Dr Simon Hird was on Direct Mass Spectrometry for High Throughput Food Analysis. Dr Hird described a mass spectrometry method, using ambient ionisation techniques, combined with multivariate statistical analysis, as an approach for rapid characterization of

food and animal tissues, either with no requirement for sample preparation or with just a simple dilution/extraction step. Here the focus was on two of techniques; Rapid Evaporative Ionization Mass Spectrometry (REIMS) and Atmospheric Solids Analysis Probe (ASAP) mass spectrometry.

‘Automated Headspace Analysis of Selective Fragrance Binding and Release from Cucurbiturils using Selected Ion Flow Tube Mass Spectrometry’ was jointly presented by Mark Perkins (Anatune) and Ben Cheesman (Aqdot Ltd). Cucurbiturils are barrel-shaped molecules that can selectively bind and hold fragrance (or other) molecules. The bound molecules can then be released in response to triggers such as humidity, heat, pH, or displacement by other molecules such as malodours. Analysis of the headspace above aqueous mixtures of fragrances can yield important information regarding differential binding of compound classes. However, this requires the analytical technique used for investigation to be both fast and largely immune to humidity. Selected Ion Flow Tube Mass Spectrometry (SIFT-MS) is a soft ionisation mass spectrometric technique that offers real-time, targeted measurement of volatile and semi-volatile compounds without the need for chromatographic separation, or the removal of residual water. By integrating SIFTMS with autosampler technology it is possible to analyse dynamic headspace conditions over long periods of time.

‘Native LESA mass spectrometry of Intact Proteins and Protein Complexes Directly from Living Bacterial Colonies’ was presented by a first-year

Chinese postgraduate student, Yuying Du, from Professor Helen Cooper’s group at Birmingham University. Previously, it had been demonstrated that liquid extraction surface analysis mass spectrometry (LESA- MS) could be utilised for the analysis of denatured intact proteins directly from living colonies of bacteria. In this presentation, combined electroporation was employed in conjunction with LESA MS using native-like solvents to perform native MS of proteins directly from bacteria.

‘Advanced applications of the SICRIT ion source: From fast classification and fraud detection to detailed aroma profiling and emission monitoring’ was a topic presented by Jan-

Christoph Wolf & Markus Weber (Plasmion GmbH). Soft Ionisation by Chemical Reaction in Transfer (SICRIT) is an ambient ionisation source that enables various new applications of mass spectrometry. Due to its “flow through” operation principle multiple sampling and enrichment techniques can be easily interfaced. Since its introduction 2018 many different applications have evolved around this technology. In this presentation we want to share some insights on recent and advanced applications of this technology in the area of food and environmental analysis. Possible applications of SICRIT range from direct headspace measurements for fast and easy fraud detection or origin classification to solid phase

Simone Mathias receiving The Peter Ryan Award.

extraction – gas chromatography high resolution mass spectrometry (SPME-GC-HRMS) for highly sensitive and selective identification and quantification of individual trace components.

‘The Development of Rapid Evaporative Ionisation Mass Spectrometry for the Detection and Diagnosis of Plant Parasitic Nematode Infections in Crops’ was presented by Alice Flint, Queen's University Belfast, who described plant parasitic nematodes (PPNs, such as, root-knot and cyst nematodes) as major agricultural pests, which exhibit broad host ranges, including members of the Fabaceae, Poaceae and Solanaceae families. PPNs cause wilting, stunting and chlorosis, with resultant yield losses up to 25% costing an estimated £100 billion annually. However, these aboveground PPN symptoms resemble other environmental stresses resulting from root dysfunction and can display slow onset. Current PPN diagnosis requires root and soil analysis; a low throughput, destructive process necessitating specialist expertise. Therefore, the development of non-destructive assay would enable early detection, improving management and monitoring.

‘Identification of β-Carotene Oxidation Products Produced by Bleaching Clay Using UPLC-ESI-MS/MS’ by Andrew Hambly of the University of Bristol. Vegetable oils often undergo some degree of refining before being made into products for the food and pharmaceutical industries. One of the refining steps is known as ‘bleaching’ or decolourising. Here the oil is mixed with an acidactivated clay (AAC) adsorbent and contaminants which are detrimental to the final product quality are removed. The contaminants include lipid oxidation products and plant pigments chlorophyll and β-carotene. While it is known that β-carotene adsorbs readily to AACs, it is not understood whether this removal process is purely physical or involves chemical degradation of the pigment as well. In the presented work the aim was to provide extra mechanistic insight into the interaction between β-carotene and AAC adsorbents by studying the extracts of the spent AAC using UPLC-ESI-MS/MS.

‘LAP-MALDI MS for high-speed sample screening: from 5 to 40 samples/s’ was a topic presented by Henriette Krenkel of Reading University. Fast analyses are in

general desirable for obvious cost and convenience reasons, but particularly large-scale experiments require high sample throughput. Mass spectrometry offers a labelfree alternative for photometric readouts of enzymatic assays used in clinical screening experiments and diagnostics. Although providing less false-positive readouts, the throughput of MS techniques is still not matching traditional methods. Liquid atmospheric pressure matrixassisted laser desorption/ionisation (LAP-MALDI) aims to close this gap by providing contact-free laserbased desorption for fast sample exchange and good compatibility with reagents commonly used in biological assays.

‘Meat Speciation Via Deployable ASAP-QDa’ was reviewed by Valerio Converso (LGC). The aim of this study was to explore the potential use of ambient mass spectrometric techniques in the food industry. In particular, a deployable ASAP-QDa has been used for meat speciation analyses in a non-laboratory environment. The developed process from sample intake to results is quick and user friendly, not requiring scientifically trained personnel to

perform the assay and interpret the data.

‘According to chemistry, whisky is a solution’ as suggested by Nick Molden, Oxford Indices, in his presentation. Whisky comes with much history, folklore and personal tastes, but it is also a big business which thrives as a premium product made with high quality methods. ? The manufacturing process is deep science with a dash of art, which requires advanced skills to create perfect flavours and odours, but also sets up the incentive and possibility for fraud and adulteration. The objective of the presentation was to demonstrate the greater potential for identification and discovery of volatile organic compounds (VOC’s) enabled by twodimensional gas chromatography time-of-flight mass spectrometry.

FeMS: Females in Mass Spectrometry

Charlotte Hutchings, Anne K. Bendt, Caroline Géhin, Ólöf G. Isberg, Bini Ramachandran, Maggie Tam, Sara Zanivan

Who are FeMS?

Females in Mass Spectrometry (FeMS) is a community-led initiative that supports individuals in mass spectrometry and related fields. We spoke to Anne Bendt, co-founder of FeMS, to discover how the movement began:

“At a science advisory board meeting in 2019, the late Michael Wakelam asked me: ‘Are you still the only female faculty in your program?’. I felt a tinge of embarrassment. Despite having a gender-conscious director and myself as deputy, I was the only senior female in our program. How come it is so challenging to recruit and retain female STEM scientists, especially for leadership roles, or for conferences to invite female keynote speakers? If we had a repository of CVs of women in the mass spec field, then the resource could be used to find candidates for job openings, speakers, mentors, chairs, and committee members. That was the main motivation for Amber Herold, Hema Ketha, Margaret Thorsteinsdottir, Grace van der Gugten and myself in starting FeMS, an international network of women in the mass spec field at all career stages, spanning across academia, governmental agencies, and industry.

From a LinkedIn group in 2019, we grew rapidly with new members and the creation of a website and Twitter account. FeMS membership grew substantially during the COVID-19 pandemic; our monthly online happy hours provided a

much-needed avenue for social and technical interaction. We received generous sponsorship to create impact designing awards. In the last two years, FeMS has also added a job board, a speaker’s directory, and mentorship programs. Importantly, FeMS is not only for females. FeMS welcomes every gender - we learn and grow together, and we open doors for one another.

events were hugely appreciated during the COVID-19 pandemic. Asher Newsome, a scientist at Smithsonian Museum Conservation Institute, told us that these virtual networking events were “a real point of relief whilst their research progress was largely stymied”! The popularity of these sessions is attested by others like Ólöf Gerður Ísberg, a PhD student from University of Iceland, who recalls

(Front row: Karolina Sulek, Sofina Begum, Anne Bendt, Lucy Woods. Back row: Margret Thorsteinsdottir, Julie Courraud, Cristina Legido-Quigley, Vanessa Hall.)

FeMS initiatives

FeMS coordinates in-person and virtual events to connect with the community. One great example is the FeMS Virtual Happy Hour, which allows individuals from around the globe to network in an informal environment. These meet-ups also serve to develop soft skills and share advice on scientific careers, including job interviews, mentoring across cultures, building great teams, and addressing systemic racism in STEM. The format of these virtual meet-ups was accommodative to those who find it easier to interact with peers in a less crowded setting, says Bini Ramachandran. Bini identifies as an introvert and looks forward to the breakout room networking discussions. The virtual

looking forward to attending FeMS virtual meetings in the mornings.

In 2021, FeMS introduced their Transition Talks, a structured conversation for graduate students and postdocs transitioning into mass spectrometry-based jobs. During their 10-minute talk, early career researchers introduce themselves, their research, and outline their desired future transition. Several presenters connected with their future employers through the FeMS Transition Talks!

Now, with the world opening back up again, FeMS has begun hosting in-person gatherings, thanks to generous sponsors. There were FeMS activities, such as panel discussions

and networking socials, at ASMS 2021 and 2022, MSACL 2022, IMSC 2022 and NMetC 2022.

FeMS supports the community

FeMS supports early career researchers through mentorship, financial support, and exposure within the field. For example, Charlotte Hutchings, a PhD student from University of Cambridge, used the FeMS spotlight to get acquainted with the community. The FeMS spotlight is a Twitter post that introduces individuals and their research to the FeMS following. Charlotte said that “[she] had a really great response and it really boosted [her] confidence (and follower count)”.

The FeMS mentorship programme provides support in small group settings to mentees at various career stages. Sara Zanivan, a FeMS mentor, says, “I am both a PI of a cancer research lab and head of the MS proteomic facility at the CRUK Beatson Institute. I wanted to share my hybrid experience in academia, and to expand my network in the mass spec field”. Asher Newsome volunteered as a FeMS mentor to “give back” to the community with his perspective as a government scientist. He found being a mentor very rewarding and educational. From a mentee’s perspective, Bini Ramachandran found the virtual workshops extremely helpful during her transition from Postdoc to a Technical Lead at a Proteomics Core Facility.

Get involved with FeMS

Connect with FeMS via mailing list (femalesinms.com/contact/), Twitter (@FemalesInMS), and LinkedIn (www.linkedin.com/ groups/13745861/)!

The above picture is from the in-person FeMS event at the Nordic Metabolomics Conference in Copenhagen, Denmark in September. The picture was taken at the Rosalind Franklin Clinical Metabolomics Award Ceremony. The award recipient was Dr. Sofina Begum. The award was sponsored by Bruker; the event was sponsored by Sciex.

Get to know

Happy Hour virtual meetings! Sign up for the Mentorship program. Open to everyone, not just females. Apply for our Empowerment Awards.

FeMS Spotlight Check out our Job Board

Females in Mass Spectrometry (FeMS) is a community-led initiative to create a network of support for women in the field of mass spectrometry.

In Memoriam: Friends we have lost in 2022

Holger Cramer

Remembrance words courtesy of Cambridge University

With sadness it is announced that Holger Kramer, Head of the LMB’s Mass Spectrometry facility at the University of Cambridge, died on Tuesday 23rd August 2022, aged 43. He suffered a brain haemorrhage from a ruptured aneurysm. He was a highly interactive, inquisitive and engaged scientist, who had worked in mass spectrometry and proteomics for over 14 years and published around seventy papers on a range of topics.

For a fuller obituary the BMSS encourage you to visit the University’s memorial page: www2.mrc-lmb.cam.ac.uk/holger-kramer-1979-2022/

Bob Finnigan

Remembrance words courtesy of Mark Harrison

It is with great sadness I share with you the news that Bob Finnigan passed away on August 13 2022, aged 95. He passed peacefully surrounded by his loving family.

Bob was not particularly well known around BMSS circles and was certainly more appreciated in the US. However his contribution to commercialising GC/MS is well documented and should not be forgotten. Let’s face it, nearly everyone uses a commercial MS combined with a data system and a chromatograph, for which Bob was a truly a pioneer.

Reflections: Mass Spectrometry as a Career Choice

Hannah Britt, Ph.D. Postdoctoral Researcher, University of Oxford, UK

Interviewed by Mervyn Lewis, Associate Editor

What was your impression of mass spectrometry when you were first introduced to it?

The first time I remember being aware of mass spectrometry was during the first year of my undergraduate degree, although given how widely-used the technique is in the world around us I am surprised that it wasn’t earlier! My impressions were that it was an odd balance of beautiful spectra, especially the way fragmentation let you put together a jigsaw puzzle of what you were looking at, combined with the most complex and confusing gas phase behaviour. I think trying to understand that behaviour seemed like an exciting challenge though, and once I saw how much fun it was to actually

use the instruments, and how many different applications there are, I was hooked.

What persuaded you that mass spectrometry could be a good career option for you? Were you attracted by an application of mass spectrometry or excited by the prospect of developing it as an instrumental technique?

The main reason I knew that mass spectrometry was a good career option for me was the fact that I truly just found it fascinating, and really enjoyed practically performing the technique. When I first started out I was definitely drawn to the application of mass spectrometry, particularly in biology and forensics, as I loved feeling that the skills I was building could be used in so many ways and impact the world around me. My scientific curiosity was peaked, there seemed to be so many questions that I could look into using mass spectrometry. However, as time

goes on I am appreciating the development of mass spectrometry as an instrumental technique more and more, because I have come to realise that it drives the application side. At the moment I think I manage a balance of the two, and that flexibility it just another reason I know that mass spectrometry is a good career for me.

How did you go about finding your first opportunity in mass spectrometry?

To be honest, I was very lucky in finding my way into mass spectrometry as I just sort of fell into it by chance. When I came to choose a project during the final year of my undergraduate degree I knew that I wanted to work on methods to study biological systems, and that ended up being mass spectrometry. During that project I was very fortunate to work with Jackie Mosely, she taught me a lot technically about the instruments

but also introduced me to the BMSS. When it came to the first time I actively searched for an opportunity (applying for a postdoctoral position), that community was key because it meant I already knew people who worked in areas that I was interested in. I reached out to these people and looked at their websites for opportunities that were a good fit for me, and ultimately ended up moving to the Thalassinos Lab at University College London.

In what application fields do you see new opportunities in mass spectrometry?

Even all these years after the foundations of mass spectrometry were laid, it is still a field that is thriving in so many different areas. Personally I work in the area of biological mass spectrometry, and in terms of new opportunities in that field I see lots of exciting things happening. Firstly, there has been wonderful work recently moving

mass spectrometry into cells and organisms so that molecules can be studied in the native context, and I believe that is a field where there is still plenty of scope for innovative new developments. Beyond that, I am excited to see how biological mass spectrometry integrates with other experimental and computational techniques to move forward our understanding of the biological landscape. By working with other disciplines like this I think the mass spectrometry community can learn a lot, and that even more areas for expansion will become clear to us. Ultimately, I believe MS will keep evolving as new challenges and puzzles present themselves, and that makes it a very exciting field to work in.

of mass spectrometry. I believe that automation will expand the potential applications and user pool, which is a great opportunity to welcome new members into our community. The experiences and insights of these users with their varied backgrounds will probably be a big driving force in developing new opportunities in mass spectrometry. With informatics,

I think the developments are fantastic in helping us make the best use of the power of mass spectrometry, both by teasing even more information out of our current data, and by integrating mass spectrometry with other techniques. That being said, I believe there will always be a place in mass spectrometry for innovation, and a need for those with the technical skills and understanding to complement developments in automation and informatics.

Could you describe your views on career prospects in mass spectrometry for young people?

Mass spectrometry is a fantastic career for young people to consider going into, because it’s a field with so much variety. As long as you have the fundamental knowledge of how mass spectrometry works, your career options are anything from sales and marketing through to engineering, research and development. Furthermore, mass spectrometry is applied in so many settings, you can have a career probing fundamental biological questions, solving crimes using forensics, or developing new drugs (plus a lot more). The BMSS conferences are excellent

places to learn more about exciting and unusual career prospects in mass spectrometry, I’ve been lucky enough to hear about mass spectrometry on Mars, in studying the bones of Richard III, and in arson investigation. Not to mention the fact that with new developments in mass spectrometry, additional career options are coming every day, and young people can get involved and drive that!

Acknowledgements

Hannah would like to thank her PhD and postdoctoral mentors, Dr John Sanderson, Dr Jackie Mosely, Professor Kostas Thalassinos, and Professor Dame Carol Robinson, for their support. Hannah is a postdoctoral researcher at the University of Oxford, and receives funding support from the Wellcome Trust.

What is your opinion about the impact that automation and informatics will have on mass spectrometry?

This is a very interesting question, I certainly think that developments in automation and informatics will change the field

Anyin Li, Ph.D.

Assistant Professor, University of New Hampshire, USA

Interviewed by

Mervyn Lewis, Associate Editor

What was your impression of mass spectrometry when you were first introduced to it?

Strong magnet winnowing grain. I was first introduced to a sector type mass spectrometer in a textbook during my sophomore year. After that, my true handson experience was with a 3-Tesla FT-ICR mass spectrometer that had EI/CI source. Strong magnet was my very first impression of mass spec. Till this day, I still have the habit of putting away my metal key chain before running a mass spec. experiment. Another of my early impressions with mass spec is the process of separation. I have always been amazed by how the molecules are sorted out in space, which associate me with childhood memory of rice winnowing in the countryside.

Role models and the problemsolving aspects. When starting as a fresh PhD student at Purdue University, I was lucky enough to have run into a few highly talented and dedicated senior graduate students, namely, Frank (Mingkun) Fu, He Wang, and Abraham Badu-Tawiah. With very different backgrounds, these folks joined mass spectrometry research groups. At the time I met them, they were all in the process of trying to figure out something in their research. At Purdue, there is also a “culture” for

mass spec, pioneered by Prof. Cooks. Seeing how these folks succeed and how they do research convinced me mass spectrometry could be a career option for me.

Initially, I was attracted by the gas phase chemistry applications of mass spectrometry. As research goes on, I also became interested in developing instrumental techniques to realize unique gas phase chemistry that may be utilized in applications.

How did you go about finding your first opportunity in mass spectrometry?

Proton affinity of radical ions. My first opportunity in mass spectrometry was to do gas phase chemistry in the lab of Prof. Hilkka Kenttämaa. Measuring proton affinity of highly reactive radicals was like to taste the sweetness of a popping candy. You can do it using mass spec and this still fascinates me. This challenging project was a great first opportunity that helped me to gain in-depth learning for many aspects of mass spectrometry. It also certainly reinforced that I want to study mass spectrometry for my PhD.

In what application fields do you see new opportunities in mass spectrometry?

There are always new opportunities for mass spec because it is “universal” i.e. everything has a mass and can theoretically show up in a mass spectrum. For example, the new developments in the pharmaceutical industry, particularly the recent burst of “new therapeutic modalities” have been providing a lot of problems for analytical chemists. Mass spectrometry is ideal for many of these problems.

There are even more opportunities in research because mass spec is

not yet quantitatively “universal” i.e. analytes are being detected with varying efficiencies. This is an age-old problem that people have gotten used to. People are bypassing this problem by other means such as using isotopically labelled internal standards. Imagine when mass spec is truly quantitative, we will be able to carry out many new analyses in different ways.

What is your opinion about the impact that automation and informatics will have on mass spectrometry?

One great thing about mass spectrometry is that it has been boosted by many technology innovations that may initially seem irrelevant. Automation and informatics are no different. I think they are going to have long lasting impact on how we conduct analysis and data interpretation. Obviously, there will be more automation and even robotics in mass spec. Mass spectrometry is going to produce data in increasing sizes and higher dimensions. These “big data” may be processed in many new ways to allow new interpretations.

Despite getting more automated, these developments will also bring new opportunities for experimentalists and researchers. Automation and big data desire mass spectrometry to be more robust and universally quantitative.

A lot of development is still needed for mass spec to be seamlessly coupled with automation and data technology.

Could you describe your views on career prospects in mass spectrometry for young people?

It is really a good time for a career in mass spectrometry. Mass spectrometry solves many problems so you could be working with lots of disciplines, such as chemistry, biology, pharmacy. The demand from industry is strong so there is job security. The field is also evolving (boosted by technology breakthrough in all fields) so rapidly that things are getting outdated in 5 or 6 years. This is a particularly exciting sign for young people to consider mass spectrometry as a research field.

Acknowledgements

Anyin would like to thank Prof. Hilkka Kenttämaa and Prof. Graham Cooks for their mentoring and support throughout his PhD, and Prof. Facundo Fernandez for mentoring his postdoctoral research. Anyin Li is now an Assistant Professor at the University of New Hampshire, and acknowledges funding supports from NSF.

Assessing Individual Biological Variation in the Urinary Steroid Profile

Anna Cooper

Principal Investigator: Dr. Angela Taylor, University of Birmingham.

Abstract

Background

Urinary steroid quantification is used to diagnose many endocrine disorders. However, there is variability in the steroid excretion related to age, gender, menstrual cycle status, and BMI. In addition, there is variation in the analytical techniques and between different scientists.

Methods

We investigated variation in individual urinary steroid profiles and the variation in steroid extraction and quantification using gas chromatographymass spectrometry (GC-MS) by independent scientists.

Results

Intra- and inter-assay variability between scientists in the lab was minimal. For repeat 24-hour urines from the same individual there is 4.2% to 56.1% (%RSD) variation in 24-hour excretion with DHEA, 16a-OH-DHEA, P’dienol, 18-OH-THA, and 11b-OH-ETIO showing the most variation.

Conclusions

Both the analytical and biological variation must be accounted for when interpreting diagnostic results produced by GC-MS urinary steroid profiling.

Introduction

Urinary Steroids

Steroid hormones are cyclical compounds composed of carbon atom rings with crucial roles across a vast range of physiological functions

and processes. Prior to excretion, steroids are conjugated to produce sulphate or glucuronide conjugates to increase water solubility and therefore permit excretion via the urine.

Gas Chromatography-Mass Spectrometry

(GC-MS)

Approximately 30 steroid metabolites are selected for analysis, to include representative metabolites of the major steroid hormones and their precursors. Measurement of individual excretion of these metabolites can enable diagnosis of disorders or facilitate further clinical investigation. Steroids are extracted from the urine and hydrolysed to release them from their sulphate or glucuronide conjugates. Next, the steroids must be chemically derivatised to enhance their stability and volatility at the high temperatures necessary for GC-MS. Subsequently, the steroids were separated by gas chromatography using a fusedsilica column. Due to variations in column liquid phase interaction, the gaseous steroids arrive at the mass spectrometer at varying times after travelling through the heated column. The steroids become ionised after entering the ionisation source of the mass spectrometer. One ion of defined mass/charge (m/z) ratio is selected for the detection of each compound. For steroid identification, a mass spectrum can be generated from each peak, and the mass spectra compared to a reference standard.

Aims

Using GC-MS, we aimed to assess the biological variation in the urinary steroid profiling. We aimed to evaluate the variability in steroid metabolite concentrations in the

same patient, between different patients, and we also assessed variability of steroid quantification when samples were extracted by the same scientist and different scientists.

Methods

Urine is firstly passed through a solid phase extraction (SPE) cartridge to remove both the free and conjugated steroids. Enzyme hydrolysis is necessary to then release the conjugated steroids from their sulphate and glucuronide conjugates. Then SPE is performed again to extract the now deconjugated steroids from the aqueous hydrolysate. After addition of an internal standard (stigmasterol) to enable steroid quantification, the steroids are derivatised. Steroidal carbonyl groups are converted to methyloximes, and hydroxyl groups are silylated. Finally a liquid/

liquid extraction is performed. The final derivative, a methyloximestrimethylsilyl ester, is injected into the GC-MS.

Results

Inter- and Intra-Scientist Variation. Two scientists each extracted and analysed six samples, which consisted of a pool of urine obtained from six healthy males. Inter- and intra-scientist variability was assessed (Figure 1). To evaluate the intra-scientist variability, the percentage relative standard deviation (%RSD) was calculated, by dividing the standard deviation for each steroid by the average concentration and multiplying the result by 100. Ideally, the %RSD value should be below 20% to demonstrate consistency between the concentrations calculated for each steroid within repeats of the same sample (Table 1).

Individual Steroid Variation.

Steroids were extracted and quantified in 24-hour urine samples collected over six consecutive days from a 22-year-old female on the contraceptive patch (Figure 2). We expected little variation between the steroid metabolite concentrations recorded for each sample. Firstly, because the patient collected all samples within a short consecutive time period, meaning physiological variation would be at its minimum. Furthermore, the patient was on a hormonal contraceptive, so consistent levels of synthetic hormones were released into the bloodstream each day. However, 10 of the steroid metabolites were generating a %RSD larger than 20%.

Discussion

Inter- and Intra-Scientist Variation. The inter-scientist variation was minimal (Figure 1). The error bars (standard deviation) overlap for every steroid metabolite between the scientists, indicating low variability between their results. The intra-scientist variation was also negligible, with only one steroid metabolite (6b-OH-Cortisol) producing a %RSD greater than 20% for each scientist (Table 1). As the %RSD was high for this steroid for both scientists, this may imply that

Figure 1 – The average concentration of each steroid metabolite determined by six extractions of the same sample analysed on the GC-MS by two scientists. The error bars (standard deviation) overlap for every metabolite, indicating low inter-scientist variability.

the large variation is not due to their extraction technique but may be analytical issue. For three quarters of the steroid metabolites, the %RSD value was greater for Scientist 1 than for Scientist 2.

Individual Steroid Variation

The individual steroid variation between 24-hour samples collected over consecutive days was larger than anticipated. Notably, DHEA, 16a-OH-DHEA, P’dienol,

18-OH-THA, and 11b-OH-ETIO generated large %RSD values (32.5%-56.1%). This variation can arise due to a multitude of factors, including stress, diet alterations, hydration level changes, and hormonal changes beyond the contraceptive patch. Any of these circumstances could have caused these differences to arise.

Conclusion

Both the analytical and biological variation must be accounted for when interpreting diagnostic results produced by GC-MS urinary steroid profiling. Investigating ratios of steroids can reduce variation. This is a profiling technology and absolute concentrations of a single steroid should not be relied on for diagnostic interpretation. Instead, the profile as a whole must be analysed to look at changes in pathways of steroid metabolism.

References

1. Shackleton CH. Profiling steroid hormones and urinary steroids. J. Chromatogr. 1986 Jun 20;379:91156.

2. Shackleton CH. Mass spectrometry in the diagnosis of steroid-related disorders and in hypertension research. J Steroid Biochem Mol Biol. 1993;45:127-40

3. Shackleton CHL. Genetic disorders of steroid metabolism diagnosed by mass spectrometry. In: Blau N.(Ed.) Laboratory Guide to the Methods in Biochemical Genetics, Chapter 5.3. Springer-Verlag, Berlin. 2008

Personal Statement

I have thoroughly enjoyed taking part in the IMSR Summer School and have found the experience to be hugely valuable. My confidence and skills within the lab have grown massively, hence I feel far more prepared to undertake my third year project in the upcoming months, which I hope will involve mass spectrometry. This project has confirmed to me that a career in the lab would be incredibly desirable for the future, and I have now developed a great interest in analytical chemistry; an area I had never explored previously.

Figure 2 – The mean concentration of each steroid metabolite across 24-hour urine samples collected over six consecutive days from a 22-year-old female on the contraceptive patch.

Table 1 – The %RSD for each steroid by each scientist. The inter-scientist

can be assessed by comparing the %RSD for each steroid, between the scientists.