Chemical biomarkers in human breath and oral fluid: Method development and applications in non-invasive clinical analyses

UNIVERSITÀ DEGLI STUDI DI PISA

Doctoral School in

Chemistry and Materials Science

3

rd

year PhD Report

PhD candidate

Francesca G. Bellagambi

Title of PhD Thesis

Chemical biomarkers

in human breath and oral fluid:

Methods development and their applications

in non-invasive clinical analyses

Supervisors

External referees

Prof. Roger Fuoco

University of Pisa, Italy

Prof. Abdelhamid Errachid el Sahli

Claude Bernard University Lyon 1, France

Dr. Fabio Di Francesco

University of Pisa, Italy

Prof. Kim King-Tong Lau

Table of Contents

Attended internal courses

1

Main attended seminars

1

Didactic seminar for students

3

Auxiliary teaching

3

Publications and manuscripts

4

Publications

4

Manuscripts submitted to scientific journals

4

Manuscripts in preparation

5

Participation to national and international conferences (poster, oral presentation)

6

Schools and international stages

8

1

Attended internal courses

 Metals in Medicine (238CC), Prof.ssa Chiara Gabbiani

 Preparazione di progetti di ricerca in inglese. Dr. Adrian Wallwork; 29 Novembre, 4, 6

Dicembre.

 Stesura di articoli scientifici in inglese. Dr. Adrian Wallwork. 5, 8, 15 November 2013.

Main attended seminars



3 December 2013, National Research Council (CNR), Pisa (Italy)



Sicurezza - Conoscere e Condividere. Seminario di Informazione e Formazione - QUARTA

EDIZIONE.



11 December 2013, Department of Chemistry and Industrial Chemistry, University of Pisa

(Italy)



Seminari di passaggio d’anno - Dottorato in scienze chimiche.



20 December 2013, Department of Chemistry and Industrial Chemistry, University of Pisa

(Italy)



Seminari di passaggio d’anno - Dottorato in scienze chimiche.



18 December 2013, Consiglio Nazionale delle Ricerche (CNR), Pisa (Italy)



ICT e eHealth

, Integrazione dell’esistente IFC/IIT/ILC/ISTI.



3 February 2014, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Elementi chimici nei capelli: aspetti analitici e diagnostici, Prof. Sergio Caroli, Istituto Superiore

di Sanità (Italy)



10 February 2014, Consiglio Nazionale delle Ricerche (CNR), Pisa. Rete dono Day,

Crowdfunding - Rete del dono.



20 February 2014, Consiglio Nazionale delle Ricerche (CNR), Pisa (Italy)



How to write a successful proposal in HORIZON 2020. Dr. Katia Insogna – NCP MSCA.



24 February 2014, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



L'Energia dai Fondamenti della Termodinamica allo Sviluppo Sostenibile, Il Work Programme

Energia nella Ricerca EU in Horizon 2020. Prof. Riccardo Basosi, Dept. Department of

Biotechnology, Chemistry and Pharmacy, University of Siena (Italy)



18 March 2014, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Seminari di presentazione I anno. Dottorato in scienze chimiche



9 May 2014, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Electroactive Materials for Photovoltaics and Molecular Electronics. Prof. Ryan C. Chiechi,

University of Groningen (the Netherlands)



23 May 2014, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Light Harvesting and photoprotection in photosynthetic chlorophyll-binding proteins. Prof.

Roberto Bassi, University of Verona (Italy)



29 September 2014, Department of Chemistry and Industrial Chemistry, University of Pisa

(Italy)



Molecular/Nano Wires in Chemical Sensing, Prof. Timothy M. Swager, Massachusetts Institute

of Technology of Boston (USA)



6 October 2014, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Coherent energy transfer in light-harvesting systems, Prof. Jianshu Cao, Massachusetts

Institute of Technology di Boston (USA).

2



First Principle Modeling of optical, spectroscopic and charge generation properties in

molecule/semiconductor heterointerfaces, Dr. Mariachiara Pastore, Università di Perugia

(Italy)



20 February 2015, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Rhodium and iridium complexes with aminophosphano ligands: synthesis, structural

characterisation and reactivity, Prof. Vincenzo Passarelli, Centro Universitario de la Defensa di

Saragozza (Spain)



1 April 2015, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy).



The dawn of chemistry: the making of early universe molecules, Prof. Franco A.

Gianturco, University of Innsbruck (Austria).



11 May 2015, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Towards artificial photosynthesis: light harvesting antenna systems and photochemical water

splitting, Prof. Sebastiano Campagna, University of Messina (Italy).



27 May 2015, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Stimuli-Responsive Materials and Biomimetic Fluidics: Key Building Blocks of Futuristic

Autonomous Chem/Bio-Sensing Platforms, Prof. Dermot Diamond della Dublin City University

(Ireland)



18 June 2015, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Electrochemistry and spectroelectrochemistry of bioactive compounds, Dr. Romana Sokolova,

Academy of Sciences of the Czech Republic - Institut J. Heyrovsky (Czech Republic)



24 June 2015 - Scuola Normale Superiore, Pisa



Soft chemistry for functional nanostructures: new opportunities for energy, environmental and

biomedical applications, Dr. Maria Lucia Curri, National Research Council of Bari (Italy)



18 January 2016, National Research Council (CNR), Pisa.



Magnetic Nanoparticles: tuning physical properties for biomedical applications, Prof.

Alessandro Lascialfari, university of Milan (Italy)



26 January 2016, Department of Chemistry and Industrial Chemistry, University of Pisa (Italy)



Impatto delle tecniche microfluidiche in (radio)chimica, Dr. Giancarlo Pascali, Australian

Nuclear Science & Technology Organization, Camperdown Campus, Sidney (Australia)



7‒9 September 2016, Department of Chemistry and Industrial Chemistry, University of Pisa

(Italy)

3

Didactic seminar for students

 Bio-

Chemiluminescenza nell’analisi degli alimenti.

Doctoral School in Chemistry and Materials Science.

Department of Chemistry and Industrial Chemistry, University of Pisa (Italy). 29 October 2014.

 UHPLC-ESI-QQQ

‒ Introduzione alla strumentazione e all’esperienza di laboratorio.

Course of Analytical Chemistry V, Prof. Francesca Modugno.

Department of Chemistry and Industrial Chemistry, University of Pisa (Italy). 10 February 2016

 Introduzione al laboratorio di chimica analitica strumentale.

Course of Laboratory of Instrumental Analytical Chemistry, Dr. Fabio Di Francesco.

Department of Chemistry and Industrial Chemistry, University of Pisa (Italy). 19 April 2016

Auxiliary teaching

o Laboratorio di Chimica Analitica Strumentale, Dr. Fabio Di Francesco. Academic year

2013/2014

o Laboratorio di Chimica Analitica Strumentale, Dr. Fabio Di Francesco. Academic year

2014/2015

Publications and Manuscripts

4

Publications and manuscripts

Publications

 Martelli A, Testa L, Citi V, Marino A, Bellagambi FG, Ghimenti S, Breschi MC, Calderone V.

Pharmacological characterization of the vascular effects of aryl isothiocyanates: Is hydrogen sulfide the real player? Vascular Pharmacology 2014;60:32‒41.

 Ghimenti S, Lomonaco T, Bellagambi F, Tabucchi S, Onor M, Trivella MG, Ceccarini A, Fuoco R, Di Francesco F. Comparison of sampling bags for the analysis of volatile organic compounds in breath. J Breath Res 2015;9:047110.

 Ghimenti S, Tabucchi S, Bellagambi FG, Lomonaco T, Onor M, Trivella MG, Fuoco R, Di Francesco F. Determination of sevoflurane and isopropyl alcohol in exhaled breath by thermal desorption gas

chromatography-mass spectrometry for exposure assessment of hospital staff. J Pharm Biomed

Anal 2015;106:218‒23.

 Salvo P, Ferrari C, Persia R, Ghimenti S, Lomonaco T, Bellagambi F, Di Francesco F. A dual mode

breath sampler for the collection of the end-tidal and dead space fractions. Med Eng Phys

2015;37:539-44.

 Lomonaco T, Salvo P, Ghimenti S., Biagini D., Bellagambi F., Fuoco R., Di Francesco F. A breath

sampling system assessing the influence of respiratory rate on exhaled breath composition. Conf

Proc IEEE Eng Med Biol Soc 2015;2015:7618‒21.

 Salvo P, Lomonaco T, Ghimenti S, Bellagambi F, Fuoco R, Di Francesco F. Saliva and breath

analysis for healthcare. Conf Proc IEEE Eng Med Biol Soc 2016; in press.

Manuscripts submitted to scientific journals

 Bellagambi FG, Degano I, Ghimenti S, T. Lomonaco T, V. Dini V, Romanelli M, Mastorci F, Gemignani A, Fuoco R, Di Francesco F. Determination of salivary -amylase and cortisol in psoriatic subjects undergoing the Trier Social Stress Test. Submitted to Microchemical Journal.

 Lomonaco T, Ghimenti S, Biagini D, Bramanti E, Onor M, Bellagambi FG, Fuoco R, Di Francesco F.

The effect of sampling procedures on the urate and lactate concentration in oral fluid. Submitted to

Microchemical Journal.

 Ghimenti S, Lomonaco T, Biagini D, Bellagambi FG, Onor M, Trivella MG, Ruocco L, Pellegrini G, Di Francesco F, Fuoco R. Determination of warfarin and warfarin alcohols in dried blood spots by

ultra-high performance liquid chromatography coupled to electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS). Submitted to Microchemical Journal.

 Bellagambi FG, Baraket A, Longo A, Vatteroni M, Zine N, Bausells J, Fuoco R, Di Francesco F, Karanasiou GS, Fotiadis DI, Menciassi A, Errachid A. Electrochemical biosensor platform for TNF-

cytokines detection in both artificial and real human saliva: Heart Failure. Submitted to Sensors &

Publications and Manuscripts

5

 Tripoliti E, Papadopoulos T, Karanasiou G, Kalatzis F, Goletsis Y, Ghimenti S, Lomonaco T, Bellagambi F, Trivella MG, Fuoco R, Marzilli M, Scali MC, Abdelhamid E, Fotiadis DI. A

computational approach for the estimation of heart failure patients status using saliva biomarkers.

Conference paper submitted for the 39th Annual International Conference of the IEEE Engineering in Medicine and Biology Society of the IEEE Engineering in Medicine and Biology Society in Jeju Island, Korea during July 11‒15, 2017.

Manuscript in preparation

 Bellagambi F, Ghimenti S, Lomonaco T, Biagini D, Barletta V, Scali MC, Marzilli M, Trivella MG, Fuoco R, Di Francesco F. Determination of aldosterone, cortisol, and 8-iso-PGF2 in oral fluid

samples by UHPLC-MS/MS. Manuscript in preparation.

 Bellagambi FG, Baraket A, Zine N, Bausells J, Ghimenti S, Lomonaco T, Fuoco R, Di Francesco F, Karanasiou GS, Fotiadis DI, Errachid E. A new highly sensitive electrochemical biosensor platform

for cortisol detection in human saliva. Comparison with an LC-MS/MS analytical method. Manuscript

in preparation.

 Bellagambi FG, Baraket A, Longo A, Vatteroni M, Zine N, Bausells J, Fuoco R, Di Francesco F, Karanasiou GS, Fotiadis DI, Menciassi A, Errachid E. A highly sensitive electrochemical biosensor

for TNF- detection in human saliva: a non-invasive monitoring tool for heart failure. Manuscript in

preparation.

 Baroumi L, Baraket A, Bellagambi FG, Ben Ali M., Errachid E. A novel amperometric biosensor for

TNF- detection in human saliva. Manuscript in preparation.

 Lomonaco T, Ghimenti S, Biagini D, Onor M, Bellagambi FG, Fuoco R, Di Francesco F.

Determination of volatile organic compounds in exhaled breath using needle trap micro-extraction technique coupled with tandem mass spectrometry. Manuscript in preparation.

Participation to national and international conferences (poster, oral presentation)

6

Participation to national and international

conferences (poster, oral presentation)

XXV Congresso Nazionale della Società Chimica Italiana (SCI), 7‒12 September 2014, Arcavada di

Rende, Cosenza (Italy)



Lomonaco T, Ghimenti S, Detti F, Bellagambi F, Scali MC, Simioniuc A, Di Francesco F, Fuoco. R.

Determination of volatile organic compounds in human breath samples for heart failure monitoring.

Oral presentation.



Ghimenti S, Salvo P, Lomonaco T, Detti F, Bellagambi F, Ferrari C, Di Francesco F, Fuoco. R. A

breath sampler prototype to collect end-tidal and dead space fractions. Poster.

30th International Symposium on Chromatography - ISC 2014, 14‒18 September 2014, Salzburg

(Austria)



Lomonaco T, Ghimenti S, Detti F, Onor M, Bellagambi F, Fuoco R, Di Francesco F. A purge and trap

method for the analysis of volatile organic compound in human oral fluid. Poster.

37th Annual International Conference of the IEEE Engineering in Medicine and Biology Society, 25‒

29 August 2015, Milan (Italy)



Ghimenti S, Salvo P, Lomonaco T, Detti F, Bellagambi F, Ferrari C, Di Francesco F, Fuoco. R. A

breath sampler prototype to collect end-tidal and dead space fractions. Poster.



Lomonaco T, Salvo P, Ghimenti S, BiaginiD, Bellagambi F, Fuoco R, Di Francesco F. A breath

sampling system assessing the influence of respiratory rate on exhaled breath composition. Oral

presentation.

Giornata di Bioanalitica 2015 – Società Chimica Italiana, Gruppo Divisionale di Bioanalitica, 26 June

2015, Firenze (Italy)



Ghimenti S, Tommaso Lomonaco T, Bellagambi F, Onor M, Trivella MG, Fuoco R, Di Francesco F.

The influence of the sampling procedure on the concentration of uric acid in oral fluid. Poster.

EuroAnalysis 2015 - 18th edition of The European Conference on Analytical Chemistry, 6–10

September 2015, Bordeaux (France)



Bellagambi FG, Degano I, Ghimenti S, Lomonaco T, Dini V, Romanelli M, Mastorci F, Fuoco R, Di Francesco F. Variations of salivary free cortisol levels determined by liquid chromatography with

tandem mass spectrometry (LC-MS/MS) in psoriatic patients undergoing a stress test. Poster.



Ghimenti S, Lomonaco T, Bellagambi FG, Onor M, Di Francesco F, Fuoco R. Influence of the

sampling procedure on the concentration of uric acid in oral fluid. Poster.



Lomonaco T, Ghimenti S, Biagini D, Bellagambi FG, Onor M, Di Francesco F, Fuoco R. Alveolar

breath analysis by needle trap micro extraction technique coupled with gas chromatography/tandem mass spectrometry. Poster.

XXV Congresso Nazionale della Divisione di Chimica Analitica della Società Chimica Italiana (SCI),

Participation to national and international conferences (poster, oral presentation)

7



Bellagambi FG, Degano I, Ghimenti S, Lomonaco T, Dini V, Romanelli M, Mastorci F, Fuoco R, Di Francesco F. Differences in salivary alpha-amylase and cortisol responsiveness in psoriatic patients

undergoing to the trier social stress tests. Poster.



Ghimenti S, Lomonaco T, Bellagambi FG, Onor M, Di Francesco F, Fuoco R. Influence of the

sampling procedure on the concentration of uric acid in oral fluid. Poster.

International Association of Breath Research (IABR) Summit 2015, 6–10 September 2015, Wien

(Austria)



Lomonaco T, Ghimenti S, Biagini D, Bellagambi FG, Onor M, Di Francesco F, Fuoco R. Alveolar

breath analysis by needle trap micro extraction technique coupled with gas chromatography/tandem mass spectrometry. Poster.



Di Francesco F, Lomonaco T, Ghimenti S, Biagini D, Salvo P, Bellagambi FG, Politino C, Onor M, Fuoco R. Impact of the breathing pattern on the measured concentrations of volatiles in breath. Oral presentation.

XV Italian-Hungarian Symposium on Spectrochemistry, IHSS, 12‒16 June, Pisa (Italy)



Ghimenti S, Lomonaco T, Bellagambi FG, Biagini D, Di Francesco F, Fuoco R. Determination of

warfarin and its active metabolites in dried blood spots. Poster.

38th Annual International Conference of the IEEE Engineering in Medicine and Biology Society, 16‒

20 August 2016, Orlando (Florida, USA)



Salvo P, Lomonaco T, Ghimenti S, Bellagambi F, Fuoco R, Di Francesco F. Saliva and Breath

Analysis for Healthcare. Poster.

XXVI Congresso Nazionale della Divisione di Chimica Analitica della Società Chimica Italiana (SCI),

18‒22 September 2016, Giardini Naxos, Messina (Italy)



Bellagambi FG, Lomonaco T, Ghimenti S, Biagini D, Barletta V, Scali MC, Marzilli M, Trivella MG, Di Francesco F, Fuoco R. Determination of biomarkers in breath and saliva for monitoring heart failure

patients. Oral presentation.



Bellagambi FG, Lomonaco T, Ghimenti S, Biagini D, Barletta V, Scali MC, Marzilli M, Trivella MG, Di Francesco F, Fuoco R. Determination of aldosterone, cortisol, and 8-iso-PGF2 in oral fluid samples

by UHPLC-MS/MS. Poster.



Lomonaco T, Ghimenti S, Bramanti E, Biagini D, Bellagambi FG, Onor M, Fuoco R, Di Francesco F.

The effect of exercise on salivary lactate, alpha-amylase and uric acid in healthy volunteers. Oral

presentation.



Ghimenti S, Lomonaco T, Bellagambi FG, Biagini D, Di Francesco F, Fuoco R. Determination of

warfarin and its active metabolites in dried blood spots. Poster.



Biagini D, Lomonaco T, Ghimenti S, Bellagambi FG, Di Francesco F, Fuoco R. Needle trap

micro-extraction: an innovative approach for the one-step sampling and pre-concentration of volatile organic compounds in exhaled breath. Poster.

Chemistry for the Future, CFF 2016, 7‒9 September 2016, Pisa (Italy)



Bellagambi FG, Lomonaco T, Ghimenti S, Biagini D, Degano I, Trivella MG, Baraket A, Errachid A, Franzini M, Paolicchi A, Dini V, Romanelli M, Marzilli M, Di Francesco F, Fuoco R. Chemical

biomarkers in human breath and saliva: Method development and clinical applications. Oral

presentation.



Lomonaco T, Ghimenti S, Biagini D, Salvo P, Bellagambi FG, Onor M, Trivella MG, Di Francesco F, Fuoco R. The sampling issue for the chemical analysis of breath and oral fluid in clinical applications. Oral presentation.

8

Schools and international stages

 LC-MS 2014. Corso avanzato sull'accoppiamento della spettrometria di massa con tecniche di separazione in fase liquida. 23‒26 June 2014, Lucca (Italy)

 Scuola Nazionale di Chimica Analitica per Dottorandi 2014. Società Chimica Italiana – Divisione di Chimica Analitica. 22‒26 September 2014, Roma (Italy)

 International stage at the Institut Des Sciences Analytiques (ISA), Villeurbanne, Lyon (France). June, July, and October 2016. Supervisor: Prof. Abdelhamid Errachid el salhi.