Publications
publications in Volatile organic compounds
1. Belluomo I, Whitlock SE, Myridakis A, Parker AG, Converso V, Perkins MJ, Langford VS, Španěl P, Hanna GB. Combining Thermal Desorption with Selected Ion Flow Tube Mass Spectrometry for Analyses of Breath Volatile Organic Compounds. Anal Chem. 2024 Jan 30;96(4):1397-1401. doi: 10.1021/acs.analchem.3c04286. PMID: 38243802.
Research aim:
To develop and demonstrate a novel instrument integrating thermal desorption (TD) with selected ion flow tube mass spectrometry (SIFT-MS) for analysing volatile organic compounds (VOCs) in human breath, particularly for large-scale multicentre clinical studies.
Main findings:
Integration of SIFT-MS and TD for practical use: The combination of SIFT-MS and TD enables rapid analysis of VOCs while also facilitating the collection of breath samples across multiple locations, making the instrument suitable for use in clinical practice.
Development of a targeted analytical method: A validated method was created to analyse 21 VOCs with diverse chemical classes, chosen for their clinical and biological relevance.
Successful validation across laboratories: The method was validated in two independent laboratories using both calibration standards and real breath samples from healthy volunteers.
2. Myridakis A, Wen Q, Boshier PR, Parker AG, Belluomo I, Handakas E, Hanna GB. Global Urinary Volatolomics with (GC×)GC-TOF-MS. Anal Chem. 2023 Nov 28;95(47):17170-17176. doi: 10.1021/acs.analchem.3c02523. PMID: 37967208.
Research aim:
To develop an improved methodology for global volatolomic profiling using urinary samples for non-invasive disease detection and monitoring.
Main findings:
Reduction in required urine volume and clinical applicability: The required urine volume was reduced by five times (from 2 mL to 0.4 mL), and the methodology was successfully applied to a pancreatic ductal adenocarcinoma cohort, confirming previous findings and discovering additional VOCs with diagnostic potential.
Improved VOC extraction method: A multiphase sorbent was used for VOC extraction, achieving wide coverage and significantly increasing the number of monitored VOCs compared to the previously reported single-sorbent method.
Enhanced biomarker discovery: Multidimensional gas chromatography (GC×GC) further improved biomarker discovery, and data analysis was simplified using a tile-based approach.
3. Wen Q, Myridakis A, Boshier PR, Zuffa S, Belluomo I, Parker AG, Chin ST, Hakim S, Markar SR, Hanna GB. A Complete Pipeline for Untargeted Urinary Volatolomic Profiling with Sorptive Extraction and Dual Polar and Nonpolar Column Methodologies Coupled with Gas Chromatography Time-of-Flight Mass Spectrometry. Anal Chem. 2023 Jan 17;95(2):758-765. doi: 10.1021/acs.analchem.2c02873. PMID: 36602225.
Research aim:
To develop and optimise a complete pipeline for untargeted urinary volatolomic profiling, addressing barriers to wider clinical adoption, and demonstrating its potential for non-invasive disease detection and biomarker discovery.
Main findings:
Clinical applicability and biomarker discovery: The pipeline was successfully applied to a clinical cohort of pancreatic ductal adenocarcinoma patients, identifying four urinary biomarkers (2-pentanone, hexanal, 3-hexanone, and p-cymene) that can distinguish pancreatic ductal adenocarcinoma from control subjects, demonstrating the pipeline’s clinical utility and potential for future biomarker studies.
Optimised extraction and analysis techniques: A novel HiSorb sorptive extraction method was developed, offering high analytical performance and throughput, combined with complementary chromatographic methods and time-of-flight mass spectrometry for broader VOC coverage.
Data pre-processing and quality control: A data pre-processing strategy was established, including internal standard normalisation, batch correction, and strict quality control measures to remove nonreproducible or contaminated features, ensuring high-quality data acquisition.
4. Woodfield G, Belluomo I, Laponogov I, Veselkov K; COBRA1 WORKING GROUP; Cross AJ, Hanna GB. Diagnostic Performance of a Noninvasive Breath Test for Colorectal Cancer: COBRA1 Study. Gastroenterology. 2022 Nov;163(5):1447-1449.e8. doi: 10.1053/j.gastro.2022.06.084. PMID: 35803311.
Research aim:
To develop and evaluate a breath test based on detecting volatile organic compounds (VOCs) for detecting colorectal cancer, aimed at serving as a non-invasive triage tool for identifying patients at risk.
Main findings:
Multicentre feasibility and quality control: The study demonstrated the feasibility of multicentre breath collection and centralised analysis, with robust quality control measures and the selection of endogenous VOCs for model development, further supporting the potential for the breath test as a triage tool in colorectal cancer screening.
Diagnostic model for CRC detection: A breath test model based on 14 endogenous VOCs and body mass index successfully predicted colorectal cancer with an area under the ROC curve (AUC) of 0.87, achieving a sensitivity of 79%, specificity of 86%, and a negative predictive value (NPV) of 97%.
High performance in symptomatic patients: In a subset of symptomatic patients, the model showed even better performance, with an AUC of 0.91, sensitivity of 83%, specificity of 88%, and NPV of 96%, suggesting the breath test’s potential for accurately identifying colorectal cancer in those presenting with symptoms.
5. Belluomo I, Boshier PR, Myridakis A, Vadhwana B, Markar SR, Spanel P, Hanna GB. Selected ion flow tube mass spectrometry for targeted analysis of volatile organic compounds in human breath. Nat Protoc. 2021 Jul;16(7):3419-3438. doi: 10.1038/s41596-021-00542-0. PMID: 34089020.
Research aim:
To provide a standardised protocol for online and offline breath analysis using selected ion flow tube mass spectrometry (SIFT-MS) for non-invasive disease detection and monitoring.
Main findings:
Custom breath collection bags: A procedure for making custom breath collection bags was outlined, facilitating the standardised collection of breath samples, which is essential for reproducibility and broader application of breath analysis in clinical research.
Comprehensive workflow for breath analysis: A detailed protocol was developed for the analysis of 50 human breath samples in less than 3 hours, leveraging the advantages of SIFT-MS for real-time results and direct compound quantification without calibration curves.
Targeted analysis of disease-specific VOCs: The protocol includes methods for analysing key disease-specific volatile organic compounds, such as short-chain fatty acids, aldehydes, phenols, alcohols, and alkanes, providing insight into their potential for disease detection.
6. Antonowicz S, Bodai Z, Wiggins T, Markar SR, Boshier PR, Goh YM, Adam ME, Lu H, Kudo H, Rosini F, Goldin R, Moralli D, Green CM, Peters CJ, Habib N, Gabra H, Fitzgerald RC, Takats Z, Hanna GB. Endogenous aldehyde accumulation generates genotoxicity and exhaled biomarkers in esophageal adenocarcinoma. Nat Commun. 2021 Mar 5;12(1):1454. doi: 10.1038/s41467-021-21800-5. PMID: 33674602.
Research aim:
To investigate the mechanisms behind the accumulation of volatile aldehydes in oesophageal adenocarcinoma (EAC) and explore their potential for improving early diagnosis and understanding their genotoxicity.
Main findings:
Therapeutic implications of aldehyde accumulation: Increased levels of decanal in EAC tissues were linked to reduced ALDH3A2 expression and TP53 deletion, while metformin, an aldehyde scavenger, was shown to reduce aldehyde-induced genotoxicity, suggesting a potential therapeutic approach.
Weak aldehyde detoxification in EAC: The study revealed that weak aldehyde detoxification in EAC leads to the accumulation of endogenous aldehydes, which contributes to the formation of genotoxic DNA-adducts.
Enrichment of aldehyde groups in EAC tissue: Two groups of aldehydes; short-chain alkanals and medium-chain alkanals, including decanal, were found to be significantly enriched in EAC biopsies and adjacent tissue, with decanal levels correlating with adverse clinical features.
7. Aksenov AA, Laponogov I, Zhang Z, Doran SLF, Belluomo I, Veselkov D, Bittremieux W, Nothias LF, Nothias-Esposito M, Maloney KN, Misra BB, Melnik AV, Smirnov A, Du X, Jones KL 2nd, Dorrestein K, Panitchpakdi M, Ernst M, van der Hooft JJJ, Gonzalez M, Carazzone C, Amézquita A, Callewaert C, Morton JT, Quinn RA, Bouslimani A, Orio AA, Petras D, Smania AM, Couvillion SP, Burnet MC, Nicora CD, Zink E, Metz TO, Artaev V, Humston-Fulmer E, Gregor R, Meijler MM, Mizrahi I, Eyal S, Anderson B, Dutton R, Lugan R, Boulch PL, Guitton Y, Prevost S, Poirier A, Dervilly G, Le Bizec B, Fait A, Persi NS, Song C, Gashu K, Coras R, Guma M, Manasson J, Scher JU, Barupal DK, Alseekh S, Fernie AR, Mirnezami R, Vasiliou V, Schmid R, Borisov RS, Kulikova LN, Knight R, Wang M, Hanna GB, Dorrestein PC, Veselkov K. Auto-deconvolution and molecular networking of gas chromatography-mass spectrometry data. Nat Biotechnol. 2021 Feb;39(2):169-173. doi: 10.1038/s41587-020-0700-3. PMID: 33169034.
Research aim:
To develop a machine learning-based algorithm (MSHub) for auto-deconvolution of gas chromatography-mass spectrometry (GC-MS) data, facilitating reproducible, high-quality spectral analysis and molecular networking of GC-MS data within the GNPS platform.
Main findings:
Enhanced molecular networking and annotation: MSHub’s integration with GNPS enables global molecular networking and data co-analysis, allowing for better annotation at the molecular family level and providing a broader understanding of structural relationships in mass spectrometry data.
Automated spectral deconvolution with MSHub: MSHub utilises unsupervised non-negative matrix factorisation and Fourier transformation to automate GC-MS spectral deconvolution, significantly improving reproducibility and spectral quality without manual parameter setting.
Scalability and efficiency: MSHub scales efficiently with large datasets, processing files in an out-of-core manner to avoid memory overload, allowing for repository-scale operations and faster deconvolution as the number of files increases.
8. Kamal F, Kumar S, Edwards MR, Veselkov K, Belluomo I, Kebadze T, Romano A, Trujillo-Torralbo MB, Shahridan Faiez T, Walton R, Ritchie AI, Wiseman DJ, Laponogov I, Donaldson G, Wedzicha JA, Johnston SL, Singanayagam A, Hanna GB. Virus-induced Volatile Organic Compounds Are Detectable in Exhaled Breath during Pulmonary Infection. Am J Respir Crit Care Med. 2021 Nov 1;204(9):1075-1085. doi: 10.1164/rccm.202103-0660OC. PMID: 34319857.
Research aim:
To determine whether volatile organic compound (VOC) measurements could distinguish viral from bacterial infections in patients with chronic obstructive pulmonary disease (COPD) during acute exacerbations.
Main findings:
No induction by bacterial infections: The VOC signature was not induced by bacterial infections, highlighting its potential as a specific biomarker for viral infection and distinguishing viral from bacterial causes of exacerbations in COPD.
Viral infection VOC signature identified: A specific VOC signature, including decane and other long-chain alkanes, was induced during rhinovirus infection in airway epithelial cells. This signature was also observed in the exhaled breath of healthy subjects experimentally challenged with rhinovirus and COPD patients experiencing viral exacerbations.
Correlation with immune response and exacerbation severity: The VOC signature correlated with the magnitude of antiviral immune responses, viral burden and the severity of exacerbations, suggesting a link between these compounds and the viral infection process.
9. Lin GP, Vadhwana B, Belluomo I, Boshier PR, Španěl P, Hanna GB. Cross Platform Analysis of Volatile Organic Compounds Using Selected Ion Flow Tube and Proton-Transfer-Reaction Mass Spectrometry. J Am Soc Mass Spectrom. 2021 May 5;32(5):1215-1223. doi: 10.1021/jasms.1c00027. PMID: 33831301.
Research aim:
To assess the relationship between two online mass spectrometry techniques, selected ion flow tube mass spectrometry (SIFT-MS) and proton-transfer-reaction time-of-flight mass spectrometry (PTR-ToF-MS), in the quantification of target breath metabolites.
Main findings:
Agreement in VOC detection and cross-platform compatibility: The majority of tested metabolites showed a high degree of agreement between the two MS techniques, with most compounds falling within 95% of the limits of agreement in Bland-Altman analysis, demonstrating strong linear regression and compatibility for data transfer between platforms.
Strong correlation between SIFT-MS and PTR-ToF-MS: The concentration of abundant breath metabolites, acetone and isoprene, showed a strong positive linear correlation between both mass spectrometry techniques (r = 0.97, r = 0.89, respectively; p < 0.001).
High correlation with thermal desorption tube method: Strong positive correlations were also observed between the direct injection and thermal desorption (TD) tube sampling methods for both acetone and isoprene (r = 0.97, r = 0.92, respectively; p < 0.001).
10. Woodfield G, Belluomo I, Boshier PR, Waller A, Fayyad M, von Wagner C, Cross AJ, Hanna GB. Feasibility and acceptability of breath research in primary care: a prospective, cross-sectional, observational study. BMJ Open. 2021 Apr 13;11(4):e044691. doi: 10.1136/bmjopen-2020-044691. PMID: 33849851.
Research aim:
To examine the feasibility and acceptability of breath research in primary care.
Main findings:
Effective collection and analysis of breath samples: 95% of breath samples collected across multiple sites met the quality criteria for mass spectrometry analysis, highlighting the effectiveness of the methodology for sample collection, handling, and analysis.
Feasibility of patient enrolment and breath testing: Both ‘single practice’ and ‘hub and spoke’ recruitment models were effective for patient enrolment, with an average of 5.3 and 4.3 patients enrolled per day, respectively. SMS messaging combined with the ‘hub and spoke’ model was the most efficient method of patient accrual.
High acceptability of the test: Both patients and general practitioners (GPs) demonstrated high acceptability of the breath test and the testing process, indicating that the approach is well-received in primary care settings.
11. Hanna GB, Cross AJ. Editorial: volatile organic compound analysis to improve faecal immunochemical testing in the detection of colorectal cancer. Aliment Pharmacol Ther. 2021 Aug;54(4):504-505. doi: 10.1111/apt.16471. PMID: 34331792.
Research aim:
To evaluate the potential of combining faecal immunochemical tests (FIT) with volatile organic compounds (VOC) analysis as a triage tool for colorectal cancer detection in patients with lower gastrointestinal symptoms.
Main findings:
Need for further prospective studies and validation: The results of the meta-analyses should be viewed optimistically but cautiously, as they were based on unpaired FIT and VOC tests. There is a need for large-scale prospective clinical studies to validate the effectiveness of combined FIT and VOC tests, especially for detecting early-stage colorectal cancer in FIT-negative patients.
Combination of FIT and VOCs may reduce missed cancers: The meta-analyses concluded that combining FIT and VOC tests could potentially be a more effective triage tool for colorectal cancer detection compared to FIT alone. Testing FIT-negative patients for VOCs could reduce the number of missed cancer cases.
Challenges with sample collection and variability: Caution was advised due to significant variability in the VOC data, resulting from the inclusion of multiple sample matrices (breath, urine, faeces) analysed using different instruments. The lack of standardisation in sample collection, transport, and VOC analysis also contributed to this variability.
12. Wen Q, Boshier P, Myridakis A, Belluomo I, Hanna GB. Urinary Volatile Organic Compound Analysis for the Diagnosis of Cancer: A Systematic Literature Review and Quality Assessment. Metabolites. 2020 Dec 29;11(1):17. doi: 10.3390/metabo11010017. PMID: 33383923.
Research aim:
To summarise the published literature on cancer-associated urinary volatile organic compounds (VOCs) and evaluate their potential as biomarkers for early cancer detection.
Main findings:
Study design limitations affecting consistency: The heterogeneity in study design, methodological approaches, and reporting quality led to inconsistencies between studies. These limitations need to be addressed to improve the reliability and reproducibility of urinary VOC analysis for cancer diagnosis in future research.
Distinct VOC profiles for cancer types: The review identified 48 urinary VOCs from 13 studies involving 1266 participants. These VOCs belonged to 11 chemical classes and exhibited high diagnostic performance. The VOC profiles were distinctive for each cancer subtype, with limited cross-over between them, suggesting the potential for specific biomarkers for different cancers.
Promising diagnostic performance for various cancers: The studies covered five cancer subtypes; prostate cancer, gastrointestinal cancer, leukaemia/lymphoma, lung cancer, and bladder cancer, demonstrating that urinary VOCs can offer a non-invasive diagnostic method with promising performance for cancer detection.
13. Goh YM, Antonowicz SS, Boshier P, Hanna GB. Metabolic Biomarkers of Squamous Cell Carcinoma of the Aerodigestive Tract: A Systematic Review and Quality Assessment. Oxid Med Cell Longev. 2020 Feb 21;2020:2930347. doi: 10.1155/2020/2930347. PMID: 32685090.
14. Abbassi-Ghadi N, Antonowicz SS, McKenzie JS, Kumar S, Huang J, Jones EA, Strittmatter N, Petts G, Kudo H, Court S, Hoare JM, Veselkov K, Goldin R, Takáts Z, Hanna GB. De Novo Lipogenesis Alters the Phospholipidome of Esophageal Adenocarcinoma. Cancer Res. 2020 Jul 1;80(13):2764-2774. doi: 10.1158/0008-5472.CAN-19-4035. Epub 2020 Apr 28. PMID: 32345674.
Research aim:
To assess whether desorption electrospray ionisation mass spectrometry imaging (DESI-MSI) can objectively identify primary oesophageal adenocarcinoma and unveil mechanisms of phospholipidomic reprogramming in cancer development.
Main findings:
High discrimination for oesophageal adenocarcinoma using DESI-MSI: Multivariate models derived from phospholipid profiles of 117 patients were highly effective in discriminating oesophageal adenocarcinoma, achieving an area under the curve (AUC) of 0.97 in discovery and 1.0 in validation cohorts.
Enrichment of polyunsaturated phosphatidylglycerols in oesophageal adenocarcinoma: Oesophageal adenocarcinoma samples exhibited a marked enrichment of polyunsaturated phosphatidylglycerols with longer acyl chains, with a stepwise increase in premalignant tissues, indicating a link between lipid metabolism and cancer progression.
Link between de novo lipogenesis and phospholipidome reprogramming: Silencing the carbon switch enzyme ACLY in oesophageal adenocarcinoma cells shortened glycerophospholipid chains, demonstrating a mechanistic connection between fatty acid and glycerophospholipid synthesis pathways, influencing the phospholipidomic landscape in cancer.
15. Vadhwana B, Belluomo I, Boshier PR, Pavlou C, Španěl P, Hanna GB. Impact of oral cleansing strategies on exhaled volatile organic compound levels. Rapid Commun Mass Spectrom. 2020 May 15;34(9):e8706. doi: 10.1002/rcm.8706. PMID: 31880852.
Research aim:
To assess the impact of oral cleansing strategies on exhaled volatile organic compound (VOC) levels and provide standardised practices for breath sampling in disease detection.
Main findings:
Effect of water rinse on VOC levels: Oral cleansing with water significantly reduced the concentrations of volatile fatty acids and alcohols (e.g., butanoic acid, pentanoic acid, ethanol), suggesting that water rinsing is effective in minimising oral contamination of VOCs.
Impact of flavoured oral products on VOCs: Toothbrushing and alcohol-free mouthwash increased levels of aldehydes and phenols (e.g., acetaldehyde, menthone, p-cresol), likely caused by flavoured ingredients in these products, which may interfere with VOC measurements.
No effect on sulphur compounds: No significant reductions in sulphur compound concentrations were observed following any of the oral cleansing interventions, indicating that these compounds may be less affected by oral cleansing procedures.
16. Markar SR, Chin ST, Romano A, Wiggins T, Antonowicz S, Paraskeva P, Ziprin P, Darzi A, Hanna GB. Breath Volatile Organic Compound Profiling of Colorectal Cancer Using Selected Ion Flow-tube Mass Spectrometry. Ann Surg. 2019 May;269(5):903-910. doi: 10.1097/SLA.0000000000002539. PMID: 29194085.
Research aim:
To investigate the potential of propanal as a breath biomarker for the diagnosis and recurrence detection of colorectal cancer by analysing exhaled volatile organic compounds (VOCs) in colorectal cancer patients and evaluating its performance in a prospective cohort.
Main findings:
Propanal as a biomarker for colorectal cancer diagnosis: Multivariate analysis identified elevated levels of propanal in the breath of colorectal cancer patients compared to controls. At a threshold of 28 ppbv, propanal showed a sensitivity of 96% and specificity of 76% for colorectal cancer diagnosis.
Validation in a prospective cohort: In a separate cohort, propanal at a threshold of 28 ppbv showed a sensitivity of 83.3% and specificity of 84.7% for distinguishing colorectal cancer patients from controls.
Propanal levels and colorectal cancer recurrence: After surgery, propanal levels returned to control levels in patients with no recurrence, while levels increased significantly in patients with colorectal cancer recurrence. At the same threshold of 28 ppbv, the sensitivity for detecting recurrence was 71.4%, with a specificity of 90.9%.
17. Hanna GB, Boshier PR, Markar SR, Romano A. Accuracy and Methodologic Challenges of Volatile Organic Compound-Based Exhaled Breath Tests for Cancer Diagnosis: A Systematic Review and Meta-analysis. JAMA Oncol. 2019 Jan 1;5(1):e182815. doi: 10.1001/jamaoncol.2018.2815. Epub 2019 Jan 10. Erratum in: JAMA Oncol. 2019 Jul 1;5(7):1070. PMID: 30128487.
Research aim:
To determine the diagnostic accuracy of volatile organic compound (VOC) breath tests for cancer detection and to evaluate sources of methodological variability in test performance.
Main findings:
High diagnostic accuracy for cancer detection: The pooled analysis of 63 studies with 3554 patients revealed a mean area under the receiver operating characteristic curve of 0.94, indicating strong diagnostic accuracy for VOC breath tests in cancer detection.
Effective sensitivity and specificity: VOC breath tests demonstrated a sensitivity of 79% (95% CI, 77%-81%) and specificity of 89% (95% CI, 88%-90%), highlighting their potential for accurate cancer diagnosis.
Methodological variability influencing results: Variability in VOC test results was found to be influenced by factors such as breath collection methods, patient physiological conditions, test environment, and analysis techniques, which may affect the consistency and reliability of the tests.
18. Adam ME, Fehervari M, Boshier PR, Chin ST, Lin GP, Romano A, Kumar S, Hanna GB. Mass-Spectrometry Analysis of Mixed-Breath, Isolated-Bronchial-Breath, and Gastric-Endoluminal-Air Volatile Fatty Acids in Esophagogastric Cancer. Anal Chem. 2019 Mar 5;91(5):3740-3746. doi: 10.1021/acs.analchem.9b00148. PMID: 30699297.
Research aim:
To investigate the production of volatile fatty acids (VFAs) in oesophagogastric cancer through ex vivo and in vivo analysis of various anatomical compartments and assess their potential as biomarkers for earlier cancer detection.
Main findings:
Elevated VFA levels in ex vivo cancer tissues: In ex vivo experiments, VFAs (acetic, butyric, pentanoic, and hexanoic acids) and acetone were found to be elevated in the headspace above oesophagogastric cancer tissues compared to control tissues, suggesting a potential biomarker for cancer detection.
Diagnostic potential of specific VFAs in vivo: Receiver-operating-characteristic analysis showed that butyric acid and pentanoic acid in gastric-endoluminal air from 25 cancer patients demonstrated significant differences from controls, with an area under the curve of 0.80 (95% CI: 0.65–0.93), indicating potential for cancer-specific breath tests.
VFA levels highest in gastric-endoluminal air: VFA concentrations were highest in oesophagogastric-endoluminal air compared to mixed- and bronchial-breath samples, highlighting the importance of analysing gastric-endoluminal air for accurate detection of oesophagogastric cancer.
19. Romano A, Hanna GB. Identification and quantification of VOCs by proton transfer reaction time of flight mass spectrometry: An experimental workflow for the optimization of specificity, sensitivity, and accuracy. J Mass Spectrom. 2018 Apr;53(4):287-295. doi: 10.1002/jms.4063. PMID: 29336521.
Research aim:
To develop a workflow for method development in Proton Transfer Reaction Time of Flight Mass Spectrometry (PTR-ToF-MS) for targeted volatile organic compound (VOC) analysis, focusing on identifying product ions, optimising quantification accuracy, and addressing fragmentation patterns.
Main findings:
Optimised experimental conditions: Experiments conducted under breath-relevant conditions (100% humid air) and varying electric field values (E/N = 48–144 Td) demonstrated that lower electric fields (<90 Td) reduced fragmentation and improved sensitivity and identification, particularly for aldehydes using NO+ ions, resulting in up to a 4-fold increase in sensitivity.
Development of a novel calibration methodology: A new calibration approach utilising diffusion tubes as gravimetric standards was introduced. This method allowed for accurate quantification, achieving experimental errors of 8% or lower between gravimetric measurements and calculated concentrations for the tested compounds.
Fragmentation pattern analysis of target compounds: The fragmentation patterns of 13 selected compounds (including aldehydes, fatty acids, and phenols) were determined, providing insight into how different conditions and ion types (H3O+, NO+, O2+) affect the detection and quantification of VOCs in PTR-ToF-MS.
20. Romano A, Doran S, Belluomo I, Hanna GB. High-Throughput Breath Volatile Organic Compound Analysis Using Thermal Desorption Proton Transfer Reaction Time-of-Flight Mass Spectrometry. Anal Chem. 2018 Sep 4;90(17):10204-10210. doi: 10.1021/acs.analchem.8b01045. PMID: 30106567.
Research aim:
To develop and evaluate a platform combining thermal desorption (TD) and proton transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS) for large-scale breath analysis in clinical practice, focusing on its application for detecting biomarkers of colorectal and oesophagogastric adenocarcinoma.
Main findings:
High sensitivity and recovery for VOCs: The TD-PTR-ToF-MS platform demonstrated excellent sensitivity, with limits of detection ranging from 0.2 to 0.9 ppbV and limits of quantification between 0.3 and 1.5 ppbV. Analytical recoveries from TD tubes were consistently 80% or higher, ensuring reliable detection of target volatile organic compounds (VOCs).
Strong linear response and precision: The system exhibited a linear response in the low- to mid-ppbV range for the seven oxygenated VOCs tested, with high correlation (R² = 0.98–0.99). The coefficients of variation were below 20%, indicating high reproducibility and precision for VOC quantification.
Platform scalability for clinical use: The platform demonstrated the ability to process nearly 100 TD tubes within 24 hours, highlighting its suitability for large-scale clinical studies and real-time breath analysis. This capability supports the potential for breath-based biomarkers to be implemented in routine clinical practice.
21. Markar SR, Wiggins T, Antonowicz S, Chin ST, Romano A, Nikolic K, Evans B, Cunningham D, Mughal M, Lagergren J, Hanna GB. Assessment of a Noninvasive Exhaled Breath Test for the Diagnosis of Oesophagogastric Cancer. JAMA Oncol. 2018 Jul 1;4(7):970-976. doi: 10.1001/jamaoncol.2018.0991. PMID: 29799976.
Research aim:
To determine the diagnostic accuracy of a breath test for the detection of oesophagogastric cancer in a multicentre validation study.
Main findings:
Diagnostic accuracy of the 5-VOCs model: The 5-VOCs model used for breath analysis demonstrated good diagnostic accuracy, with an area under the curve of 0.85, indicating reliable identification of oesophagogastric cancer.
High sensitivity and specificity: The breath test achieved a sensitivity of 80% and a specificity of 81% for diagnosing oesophagogastric cancer, making it a promising non-invasive diagnostic tool for clinical use.
Patient demographics and cancer stage: The oesophagogastric cancer group was older (median age 68 years) and had a higher proportion of men (82%) compared to the control group. The majority of patients with oesophagogastric cancer had advanced tumour stages (T3/4) and nodal metastasis, highlighting the potential of this breath test for detecting more advanced cancer stages.
22. Markar SR, Brodie B, Chin ST, Romano A, Spalding D, Hanna GB. Profile of exhaled-breath volatile organic compounds to diagnose pancreatic cancer. Br J Surg. 2018 Oct;105(11):1493-1500. doi: 10.1002/bjs.10909. PMID: 30019405.
Research aim:
To quantify differences in exhaled breath volatile organic compounds (VOCs) between patients with pancreatic cancer and those without cancer.
Main findings:
Identification of significant VOCs: A total of 66 VOCs were identified, with 12 showing significant differences between cancer and non-cancer groups in the development cohort, suggesting potential biomarkers for pancreatic cancer detection.
Diagnostic performance in the validation cohort: Analysis of the significant VOCs in the validation cohort showed an area under the curve (AUC) of 0.736 (sensitivity 81%, specificity 58%) for differentiating cancer from no cancer, and 0.744 (sensitivity 70%, specificity 74%) for distinguishing adenocarcinoma from non-cancer patients.
Potential for breath VOCs in pancreatic cancer detection: The study indicates that breath VOCs can differentiate patients with pancreatic cancer from those without cancer, highlighting the potential of breath analysis as a non-invasive diagnostic tool.
23. Boshier PR, Fehervari M, Markar SR, Purkayastha S, Spanel P, Smith D, Hanna GB. Variation in Exhaled Acetone and Other Ketones in Patients Undergoing Bariatric Surgery: a Prospective Cross-sectional Study. Obes Surg. 2018 Aug;28(8):2439-2446. doi: 10.1007/s11695-018-3180-5. PMID: 29516396.
Research aim:
To determine the role of exhaled ketones as non-invasive markers of nutritional status in patients undergoing surgical treatment for obesity.
Main findings:
Increase in exhaled acetone after dietary and surgical interventions: Exhaled acetone concentrations significantly increased following a pre-operative low carbohydrate diet (1396 ppb) and bariatric surgery (1693 ppb), compared to baseline concentrations (410 ppb, P < 0.0001), indicating its potential as a marker of metabolic changes.
Decrease in other ketones post-treatment: Concentrations of heptanone (6.5 vs. 4.1 vs. 1.4 ppb, P = 0.021) and octanone (3.0 vs. 1.4 vs. 0.7 ppb, P = 0.021) decreased significantly after the low carbohydrate diet and surgical interventions, highlighting changes in ketone profiles during treatment.
Correlation with body weight and metabolic markers: Exhaled acetone and octanone concentrations were correlated with excess body weight (ρ -0.264, P = 0.005), as well as with neutrophil and triglyceride levels (P < 0.05), suggesting their potential use in monitoring nutritional status and metabolic health in obese patients.
24. Antonowicz S, Hanna GB, Takats Z, Bodai Z. Pragmatic and rapid analysis of carbonyl, oxidation and chlorination nucleoside-adducts in murine tissue by UPLC-ESI-MS/MS. Talanta. 2018 Dec 1;190:436-442. doi: 10.1016/j.talanta.2018.08.029. PMID: 30172530.
Research aim:
To develop a rapid and sensitive ultra-performance liquid chromatography (UPLC) method for the analysis and quantification of nucleoside-adducts in DNA samples, addressing challenges in DNA damage quantification and providing a method for high-throughput analysis.
Main findings:
Development of a rapid, sensitive UPLC method: A six-minute UPLC method was developed, effectively separating seven candidate nucleoside-adducts from four unmodified nucleosides, with sensitivity to 1 adduct per 10^8 normal bases using 20 µg DNA input for most targets.
High accuracy and precision: The method demonstrated accuracy ranging from 81% to 119% across five tissue types and a relative standard deviation of 4–13%, confirming reliable quantification of nucleoside-adducts.
Fast and high-throughput processing: The method allowed for an analytical processing time of less than 8 hours, enabling the preparation and analysis of up to 96 samples per day. This throughput capability facilitates large-scale genotoxicity studies.
25. Doran SLF, Romano A, Hanna GB. Optimisation of sampling parameters for standardised exhaled breath sampling. J Breath Res. 2017 Dec 6;12(1):016007. doi: 10.1088/1752-7163/aa8a46. PMID: 29211685.
Research aim:
To investigate the impact of different breath sampling parameters (fraction of exhaled breath, sample volume, and flow rate) on the detection of volatile organic compounds (VOCs) for the identification of biomarkers for oesophagogastric cancer.
Main findings:
Influence of sample volume on VOC detection: Increasing the breath sample volume improved the detection of VOCs, with larger volumes enhancing the levels of acetone and potential biomarkers for oesophagogastric cancer.
Effect of breath fraction and flow rate on VOCs: The fraction of exhaled breath (whole vs. lower airway) did not significantly impact the concentration of potential biomarkers for oesophagogastric cancer. However, the flow rate affected the recovery of certain VOCs, with higher flow rates reducing the recovery of phenols and acetone.
Optimisation of sampling parameters for clinical studies: The study concluded that for clinical studies targeting oesophagogastric cancer biomarkers, the optimal breath sampling parameters are a 500 ml sample volume of whole breath with a flow rate of 200 ml/min, along with a clean air supply to minimise contamination from ambient air.
26. Boshier PR, Knaggs AL, Hanna GB, Marczin N. Perioperative changes in exhaled nitric oxide during oesophagectomy. J Breath Res. 2017 Nov 7;11(4):047109. doi: 10.1088/1752-7163/aa9387. PMID: 29033395.
Research aim:
To investigate the variation in exhaled nitric oxide (NO) levels during oesophagectomy, with a focus on the response to one-lung ventilation (OLV) and the correlation with physiological variables and clinical outcomes.
Main findings:
Exhaled NO levels during two-lung ventilation: Baseline exhaled NO levels (2.9 ppb) increased after lung re-inflation (3.5 ppb) but decreased postoperatively at 2 hours (2.1 ppb) and 12 hours (2.2 ppb), indicating fluctuations in NO levels throughout the perioperative period.
Impact of OLV on exhaled NO: A significant reduction in exhaled NO levels was observed in the ventilated lung during OLV (1.6 ppb vs 3.1 ppb), whereas re-inflation of the collapsed lung resulted in higher NO levels (3.4 ppb vs 2.7 ppb), suggesting ventilation status influences NO concentrations.
Correlation with clinical variables: Exhaled NO levels showed significant correlation with systolic blood pressure and lactate (P < 0.007). Additionally, higher NO levels were observed in patients who developed postoperative respiratory complications, suggesting a potential link between NO and postoperative lung injury.
27. Pabary R, Huang J, Kumar S, Alton EW, Bush A, Hanna GB, Davies JC. Does mass spectrometric breath analysis detect Pseudomonas aeruginosa in cystic fibrosis? Eur Respir J. 2016 Mar;47(3):994-7. doi: 10.1183/13993003.00944-2015. PMID: 26846826.
Research aim:
The study aimed to investigate the potential of exhaled volatile organic compounds (VOCs) as biomarkers to distinguish between cystic fibrosis (CF) patients with and without chronic Pseudomonas aeruginosa infection, using SIFT-MS for VOC profiling.
Main findings:
VOC differences: Significant differences in VOC profiles between CF patients with and without P. aeruginosa infection were observed. Key VOCs, including 2-aminoacetophenone (2-AA), dimethyl disulfide (DMDS), and butanol, were found to be indicative of infection status.
Diagnostic performance: A binary logistic regression model combining these VOCs achieved an area under the curve of 0.842 (sensitivity 82.8%, specificity 70.5%) for distinguishing between CF patients with and without chronic P. aeruginosa infection.
Improvement with full VOC profile: Using all 12 VOCs in the model improved discrimination between the two groups, demonstrating that the full VOC profile holds more potential than individual VOCs alone for accurate diagnosis.
28. Huddy JR, Weldon SM, Ralhan S, Painter T, Hanna GB, Kneebone R, Bello F. Sequential simulation (SqS) of clinical pathways: a tool for public and patient engagement in point-of-care diagnostics. BMJ Open. 2016 Sep 13;6(9):e011043. doi: 10.1136/bmjopen-2016-011043. PMID: 27625053.
Research aim:
The study aimed to evaluate the use of sequential simulation (SqS) as a tool for public and patient engagement (PPE) in the development of a volatile organic compound-based breath test point-of-care test for diagnosing oesophagogastric cancer.
Main findings:
Positive participant engagement: All participants felt able to contribute to discussions and expressed support for the idea of an oesophagogastric cancer breath test, showing the effectiveness of SqS in engaging the public and patients.
Key themes for future development: Five main themes emerged regarding the proposed breath test, including awareness of oesophagogastric cancer, barriers to testing, test device design, the clinical pathway, and test device placement.
Effectiveness of SqS in PPE: SqS facilitated an immersive, participatory experience that allowed participants and researchers to co-construct knowledge, helping guide future research and development activities for point-of-care test adoption.
29. Antonowicz S, Kumar S, Wiggins T, Markar SR, Hanna GB. Diagnostic Metabolomic Blood Tests for Endoluminal Gastrointestinal Cancer–A Systematic Review and Assessment of Quality. Cancer Epidemiol Biomarkers Prev. 2016 Jan;25(1):6-15. doi: 10.1158/1055-9965.EPI-15-0524. PMID: 26598534.
Research aim:
The study aimed to assess the diagnostic accuracy of blood-based metabolomic biomarkers for endoluminal gastrointestinal cancer using endoscopic diagnosis as the reference standard.
Main findings:
Significant differences in metabolites: 246 metabolites were found to be significantly different between cancer patients and controls after multiplicity correction, with incremental metabolic changes observed as disease progressed.
Promising biomarkers identified: Two colorectal cancer biomarkers were validated in independent populations, with one approved for clinical use, highlighting the potential of serum metabolomics for gastrointenstinal cancer diagnosis.
Challenges in clinical applicability: Network analysis revealed that many modulated metabolites had extensive molecular interconnectivity, which could limit their clinical applicability, suggesting the need for more standardisation and high-resolution discovery analytics in future studies.
30. Abbassi-Ghadi N, Jones EA, Gomez-Romero M, Golf O, Kumar S, Huang J, Kudo H, Goldin RD, Hanna GB, Takats Z. A Comparison of DESI-MS and LC-MS for the Lipidomic Profiling of Human Cancer Tissue. J Am Soc Mass Spectrom. 2016 Feb;27(2):255-64. doi: 10.1007/s13361-015-1278-8. PMID: 26466600.
Research aim:
The study aimed to compare desorption electrospray ionisation-mass spectrometry (DESI-MS) and ultraperformance liquid chromatography-electrospray ionisation-mass spectrometry (UPLC-ESI-MS) platforms for profiling glycerophospholipid (GPL) species in oesophageal cancer tissue.
Main findings:
Correlation of relative abundances: The relative abundance of GPLs, normalised to a common peak, showed a high correlation (0.70, P < 0.001), indicating that the GPL profiles detected by both DESI-MS and UPLC-ESI-MS are congruent and supporting the use of DESI-MS for lipidomic profiling and potential quantification.
Similarities in detected GPLs: Both DESI-MS and UPLC-ESI-MS platforms detected a similar range of GPL species, with differences primarily linked to adduct formation rather than the presence of distinct GPL species.
Adduct formation differences: Phosphatidylcholines as formate adducts in UPLC-ESI-MS and alkali metal adducts of phosphatidylcholines in DESI-MS were the main contributors to differences in mass spectra, especially in negative and positive ion modes.
31. Abbassi-Ghadi N, Golf O, Kumar S, Antonowicz S, McKenzie JS, Huang J, Strittmatter N, Kudo H, Jones EA, Veselkov K, Goldin R, Takats Z, Hanna GB. Imaging of Esophageal Lymph Node Metastases by Desorption Electrospray Ionization Mass Spectrometry. Cancer Res. 2016 Oct 1;76(19):5647-5656. doi: 10.1158/0008-5472.CAN-16-0699. PMID: 27364550.
Research aim:
The study aimed to use desorption electrospray ionisation-mass spectrometry imaging (DESI-MSI) to objectively detect lymph node metastases (LNM) in oesophageal adenocarcinoma and evaluate its diagnostic accuracy compared to histopathological assessment.
Main findings:
Objective detection of LNM: DESI-MSI provided consistent lipidomic profiles of individual tissue types within lymph nodes, allowing for the objective identification of LNM, which were closely associated with the primary tumour tissue compared to benign lymph node tissues.
Correlation with histological images: Spatial mapping of lipidomic profiles in DESI-MSI matched tissue type distribution patterns in corresponding immunohistochemistry images, validating the technique’s accuracy.
High diagnostic performance: The multivariate statistical algorithm developed for LNM detection achieved a sensitivity of 89.5%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 97.2%, when compared to immunohistochemistry.
32. Wiggins T, Kumar S, Markar SR, Antonowicz S, Hanna GB. Tyrosine, phenylalanine, and tryptophan in gastroesophageal malignancy: a systematic review. Cancer Epidemiol Biomarkers Prev. 2015 Jan;24(1):32-8. doi: 10.1158/1055-9965.EPI-14-0980. PMID: 25344892.
Research aim:
The aim of this review was to evaluate the potential of tyrosine, phenylalanine, and tryptophan as biomarkers for gastroesophageal cancer by analysing their concentrations in biofluids and tissues of affected patients.
Main findings:
Decreased serum concentrations: Six studies investigating serum concentrations found a general decrease in tyrosine, phenylalanine, and tryptophan, except one study which reported an increase in phenylalanine.
Increased concentrations in gastric content and urine: Five studies reported elevated concentrations of these aromatic amino acids in the gastric contents of patients with gastroesophageal cancer, and two studies found increased concentrations in urine.
Tissue concentration variations: Three studies showed increased concentrations of these aromatic amino acids in tissue samples from patients, suggesting a potential role in the pathophysiology of the cancer.
33. Markar SR, Wiggins T, Kumar S, Hanna GB. Exhaled breath analysis for the diagnosis and assessment of endoluminal gastrointestinal diseases. J Clin Gastroenterol. 2015 Jan;49(1):1-8. doi: 10.1097/MCG.0000000000000247. PMID: 25319742.
Research aim:
This review aimed to evaluate the current role of volatile organic compound (VOC) breath analysis in the diagnosis and assessment of endoluminal gastrointestinal diseases, including inflammatory bowel disease (IBD), celiac disease, and colorectal and gastroesophageal cancers.
Main findings:
IBD and breath alkanes: IBD was associated with an increase in alkanes in exhaled breath, with the degree of increase correlating with disease activity in some studies.
Colorectal cancer VOC profiling: While colorectal cancer could be distinguished from controls based on VOC profiling, variability in the metabolites analysed across studies prevented the development of a consistent diagnostic model.
Gastroesophageal cancer VOC profiles: Significant differences in the VOC profiles of patients with gastroesophageal cancer were observed, suggesting a potential role for breath analysis in non-invasive diagnosis.
34. Kumar S, Huang J, Abbassi-Ghadi N, Mackenzie HA, Veselkov KA, Hoare JM, Lovat LB, Španěl P, Smith D, Hanna GB. Mass Spectrometric Analysis of Exhaled Breath for the Identification of Volatile Organic Compound Biomarkers in Esophageal and Gastric Adenocarcinoma. Ann Surg. 2015 Dec;262(6):981-90. doi: 10.1097/SLA.0000000000001101. PMID: 25575255.
Research aim:
The study aimed to assess whether volatile organic compounds (VOCs) in exhaled breath analysis using Selected Ion Flow Tube Mass Spectrometry (SIFT-MS) could distinguish oesophageal and gastric adenocarcinoma from non-cancer controls.
Main findings:
VOC differences in cancer vs. controls: Twelve VOCs, including pentanoic acid, hexanoic acid, and various aldehydes, were significantly higher in the cancer groups compared to non-cancer controls (P < 0.05).
High diagnostic accuracy: The area under the curve for differentiating oesophageal and gastric adenocarcinoma from non-cancer controls was 0.97 and 0.98, respectively, indicating very high accuracy.
Prediction model validation: The diagnostic prediction model, which incorporated the significant VOCs, showed good discriminatory ability with an area under the curve of 0.92 ± 0.01 in the model subset and 0.87 ± 0.03 in the validation subset.
35. Hicks LC, Huang J, Kumar S, Powles ST, Orchard TR, Hanna GB, Williams HR. Analysis of Exhaled Breath Volatile Organic Compounds in Inflammatory Bowel Disease: A Pilot Study. J Crohns Colitis. 2015 Sep;9(9):731-7. doi: 10.1093/ecco-jcc/jjv102. PMID: 26071410.
Research aim:
The aim of the pilot study was to identify, quantify, and analyse volatile organic compounds (VOCs) present in the breath of individuals with inflammatory bowel diseases (IBD), specifically Crohn’s disease (CD) and ulcerative colitis (UC), and to distinguish their VOC profiles from healthy controls, potentially providing insights into disease pathogenesis and complementing current diagnostic algorithms.
Main findings:
Distinct VOC profiles for CD, UC, and controls: OSC-PLS-DA modelling successfully distinguished both CD and UC from healthy controls, as well as differentiating between CD and UC, demonstrating good sensitivity and specificity.
Key VOCs identified: Statistically significant VOCs, including dimethyl sulphide, hydrogen sulphide, hydrogen cyanide, ammonia, butanal, and nonanal, were used in the analysis, contributing to the ability to discriminate between the disease groups and controls.
Diagnostic accuracy: Analyses showed good diagnostic performance, with areas under the curve of 0.86 for CD vs healthy controls, 0.74 for UC vs healthy controls, and 0.83 for CD vs UC, indicating strong potential for breath VOC profiling as a diagnostic tool for IBD.
36. Chadwick G, Groene O, Riley S, Hardwick R, Crosby T, Hoare J, Hanna GB, Greenaway K, Cromwell DA. Gastric Cancers Missed During Endoscopy in England. Clin Gastroenterol Hepatol. 2015 Jul;13(7):1264-1270.e1. doi: 10.1016/j.cgh.2015.01.025. PMID: 25645877.
Research aim:
The aim of this study was to estimate the proportion of gastric cancer cases missed at endoscopy in England and to identify factors associated with missed lesions by performing a population-based retrospective cohort study.
Main findings:
Missed gastric cancers: 8.3% of patients diagnosed with gastric cancer had undergone endoscopy within 3 to 36 months prior to their diagnosis, indicating that these cancers were missed during the procedure.
Association with cancer stage: An endoscopy conducted 3 to 36 months before diagnosis was associated with an earlier cancer stage. Early-stage cancers (stages 0 or 1) were more likely to be detected than later-stage cancers (stage 2 or greater).
Demographic factors: Younger patients (<55 years) and women were more likely to have had an endoscopy within 3 to 36 months before their gastric cancer diagnosis, suggesting that these groups may have more frequent endoscopic evaluations.
37. Boshier PR, Mistry V, Cushnir JR, Kon OM, Elkin SL, Curtis S, Marczin N, Hanna GB. Breath metabolite response to major upper gastrointestinal surgery. J Surg Res. 2015 Feb;193(2):704-12. doi: 10.1016/j.jss.2014.09.004. PMID: 25282400.
Research aim:
The aim of this prospective clinical study was to investigate whether breath analysis could provide insights into the metabolic response following oesophagectomy and gastrectomy, and identify potential biomarkers for postoperative complications, including lung injury.
Main findings:
Metabolic changes in breath metabolites: Breath acetone levels decreased significantly after oesophagectomy, with lower levels observed compared to gastrectomy patients at 48- and 72-hours post-surgery. Conversely, isoprene concentrations increased significantly after oesophagectomy.
Hydrogen cyanide levels in pneumonia: Patients who developed pneumonia post-surgery had significantly lower hydrogen cyanide concentrations in their breath at 72 hours, suggesting it may be a biomarker for postoperative lung injury.
Breath analysis as a non-invasive tool: Breath analysis using selected ion flow-tube mass spectrometry was shown to be a valuable, non-invasive method for monitoring metabolic changes and detecting complications, such as pneumonia, after major upper gastrointestinal surgery.
38. Huang J, Kumar S, Hanna GB. Investigation of C3-C10 aldehydes in the exhaled breath of healthy subjects using selected ion flow tube-mass spectrometry (SIFT-MS). J Breath Res. 2014 Sep;8(3):037104. doi: 10.1088/1752-7155/8/3/037104. PMID: 25190002.
Research aim:
The aim of this study was to investigate the use of selected ion flow tube-mass spectrometry in detecting and quantifying C3 to C10 saturated aldehydes (propanal, butanal, pentanal, hexanal, heptanal, octanal, nonanal, and decanal) in the exhaled breath of healthy human volunteers.
Main findings:
Propanal is the most abundant aldehyde: Among the aldehydes studied, propanal was found to be the most abundant in the exhaled breath of healthy volunteers.
Trace concentrations of C4-C10 aldehydes: The concentrations of C4 to C10 aldehydes (butanal, pentanal, hexanal, heptanal, octanal, nonanal, and decanal) were found to be very low, with their median concentrations being less than 3 ppbv.
Reliability of Nalophan® bag sampling method: The Nalophan® bag sampling method used for breath collection was found to be reliable, with no significant degradation of water levels in the breath sample up to 4 hours after collection.
39. Chadwick G, Groene O, Hoare J, Hardwick RH, Riley S, Crosby TD, Hanna GB, Cromwell DA. A population-based, retrospective, cohort study of esophageal cancer missed at endoscopy. Endoscopy. 2014 Jul;46(7):553-60. doi: 10.1055/s-0034-1365646. Epub 2014 Jun 27. PMID: 24971624.
Research aim:
The aim of this study was to estimate the proportion of oesophageal cancers missed at endoscopy on a national level and to investigate the relationship between miss rates and patient and tumour characteristics.
Main findings:
Missed cancer rate: A total of 7.8% (95% CI 7.1-8.4) of patients with oesophageal cancer had undergone endoscopy in the 3-36 months before their diagnosis, indicating a significant number of cancers were missed at endoscopy.
Stage-dependent miss rates: Patients with early-stage (stage 0/1) cancers were more likely to have had an endoscopy (34.0%) compared to those with later-stage cancers, with only 10.0% of stage 2 cancers and 4.5% of stage 3/4 cancers having been previously investigated.
Common misdiagnosis: The most common diagnosis during prior endoscopies was an oesophageal ulcer, which was found in 48.2% of cases, leading to potential misdiagnosis of early cancers as benign conditions.
40. Abbassi-Ghadi N, Veselkov K, Kumar S, Huang J, Jones E, Strittmatter N, Kudo H, Goldin R, Takáts Z, Hanna GB. Discrimination of lymph node metastases using desorption electrospray ionisation-mass spectrometry imaging. Chem Commun (Camb). 2014 Apr 11;50(28):3661-4. doi: 10.1039/c3cc48927b. PMID: 24407514.
Research aim:
The aim of this research was to investigate the use of desorption electrospray ionisation mass spectrometry imaging (DESI-MSI) for the identification of lymph node metastases in cancer, to assess whether the metabolic profile of the primary tumour can be applied to identify metastases in lymph nodes.
Main findings:
Effective identification of lymph node metastases:
DESI-MSI successfully identified and discriminated between cancerous lymph nodes and normal tissue based on distinct molecular ion spectral patterns, achieving 100% sensitivity and specificity for identifying various tissue types, including metastatic and normal lymphatic tissues.
Metabolic similarity between primary tumour and lymph node metastases:
The metabolic profiles of lymph node metastases were similar to those of the primary tumour, with specific lipidomic markers, particularly in the 600-900 m/z range, showing significant overlap between the two tissue types. This similarity suggests that the molecular ion spectral pattern of the primary tumour can be used to identify metastases in lymph nodes.
Development of molecular ion spectral pattern for intraoperative use:
A molecular ion spectral pattern derived from the primary tumour was used to identify metastases in lymph nodes with high accuracy. This approach allows for the objective detection of lymph node metastases during surgery, providing a potential tool to guide intraoperative decision-making without the need for histopathologist intervention.
41. Kumar S, Huang J, Abbassi-Ghadi N, Španěl P, Smith D, Hanna GB. Selected ion flow tube mass spectrometry analysis of exhaled breath for volatile organic compound profiling of esophago-gastric cancer. Anal Chem. 2013 Jun 18;85(12):6121-8. doi: 10.1021/ac4010309. PMID: 23659180.
Research aim:
The aim of this study was to investigate the potential of exhaled breath analysis of volatile organic compounds (VOCs) for diagnosing oesophagogastric cancer by comparing the VOC profiles of patients with cancer, non-cancerous upper gastrointestinal diseases, and healthy individuals.
Main findings:
Significant differences in VOCs between cancer and control groups: The concentrations of 4 VOCs, hexanoic acid, phenol, methyl phenol, and ethyl phenol, were significantly different between the oesophagogastric cancer group and the positive control group (patients with non-cancerous upper gastrointestinal diseases).
High diagnostic accuracy using VOC profiling: Analyses revealed that a combination of the 4 VOCs could discriminate oesophagogastric cancer from positive controls, with an integrated area under the curve of 0.91, indicating a high level of diagnostic accuracy.
Potential of VOC profiling as a non-invasive diagnostic tool: The study highlights the potential of VOC profiling in exhaled breath as a non-invasive method for identifying individuals with oesophagogastric cancer, offering a promising alternative to more invasive diagnostic procedures.
42. Huang J, Kumar S, Abbassi-Ghadi N, Spaněl P, Smith D, Hanna GB. Selected ion flow tube mass spectrometry analysis of volatile metabolites in urine headspace for the profiling of gastro-esophageal cancer. Anal Chem. 2013 Mar 19;85(6):3409-16. doi: 10.1021/ac4000656. PMID: 23421902.
Research aim:
The aim of this study was to evaluate the potential of urine volatile organic compound (VOC) profiling, using selected ion flow tube mass spectrometry (SIFT-MS), as a non-invasive method to identify gastroesophageal cancer by comparing VOC profiles from cancer patients, non-cancerous upper gastrointestinal disease patients, and healthy individuals.
Main findings:
Significant differences in VOC concentrations: Seven VOCs; acetaldehyde, acetone, acetic acid, hexanoic acid, hydrogen sulfide, methanol, and phenol, showed significant differences in concentration between the cancer, non-cancer disease, and healthy groups.
Increased VOC concentrations in cancer patients: The concentrations of acetaldehyde, acetone, acetic acid, hexanoic acid, hydrogen sulfide, and methanol were higher in the cancer cohort compared to healthy controls, while phenol levels were lower in cancer patients.
High diagnostic accuracy using VOC profiling: Analyses revealed that a combination of 6 VOCs (acetaldehyde, acetone, acetic acid, hexanoic acid, hydrogen sulfide, and methanol) could discriminate cancer patients from non-cancer controls, with an integrated area under the curve of 0.904, indicating strong diagnostic potential for identifying high-risk gastroesophageal cancer patients.
43. Boshier PR, Hanna GB, Marczin N. Exhaled nitric oxide as biomarker of acute lung injury: an unfulfilled promise? J Breath Res. 2013 Mar;7(1):017118. doi: 10.1088/1752-7155/7/1/017118. PMID: 23445570.
Research aim:
The aim of this study was to analyse the factors that have contributed to the success of exhaled nitric oxide (NO) as a biomarker in asthma and contrast these with challenges faced in using exhaled NO as a biomarker for acute lung injury.
Main findings:
Success in asthma management: Exhaled NO has achieved significant success in asthma, with patients using handheld devices to monitor inflammatory aspects of their condition at home. This success has been supported by regulatory bodies, clinical societies, and insurance companies.
Challenges in acute lung injury diagnosis: Despite changes in exhaled NO being observed in acute lung injury, exhaled NO has not yet proven to be a reliable surrogate biomarker for this life-threatening condition.
Proposed framework for acute lung injury evaluation: The study provides a proposed framework for the definitive evaluation of exhaled NO as a clinically useful biomarker in acute lung injury, based on a comparison of the factors that contributed to its success in asthma.
44. Abbassi-Ghadi N, Kumar S, Huang J, Goldin R, Takats Z, Hanna GB. Metabolomic profiling of oesophago-gastric cancer: a systematic review. Eur J Cancer. 2013 Nov;49(17):3625-37. doi: 10.1016/j.ejca.2013.07.004. PMID: 23896378.
Research aim:
This review aimed to identify metabolomic biomarkers of oesophagogastric cancer in human biological samples and to discuss the dominant metabolic pathways associated with the observed changes.
Main findings:
Metabolites associated with oesophagogastric cancer: Significant differences were found in metabolites related to glycolysis, the tricarboxylic acid cycle, anaerobic respiration, and protein/lipid metabolism. Lactate and fumarate were the most commonly identified biomarkers related to cellular respiration, while valine, glutamine, and glutamate were key amino acid biomarkers.
Lipid metabolism products identified as biomarkers: Products of lipid metabolism, such as saturated and unsaturated free fatty acids, ketones, aldehydes, and triacylglycerides, were also identified as significant biomarkers of oesophagogastric cancer.
Need for improved study design: The majority of studies showed an unclear risk of bias in patient selection due to insufficient details about patient recruitment, suggesting the need for clearer methodologies in future research.
45. Kumar S, Huang J, Cushnir JR, Španěl P, Smith D, Hanna GB. Selected ion flow tube-MS analysis of headspace vapor from gastric content for the diagnosis of gastro-esophageal cancer. Anal Chem. 2012 Nov 6;84(21):9550-7. doi: 10.1021/ac302409a. PMID: 23035898.
Research aim:
This study aimed to quantify volatile organic compounds (VOCs) in samples from patients with gastroesophageal cancer, non-cancer diseases of the upper gastrointestinal tract, and healthy controls, in order to identify potential biomarkers for cancer diagnosis.
Main findings:
Significant VOC differences: Seven VOCs: acetone, formaldehyde, acetaldehyde, hexanoic acid, hydrogen sulphide, hydrogen cyanide, and methyl phenol, were found to be significantly different between cancer and healthy groups, indicating their potential as biomarkers for gastroesophageal cancer.
Potential for multi-VOC profiling: Analyses discriminated cancer patients from healthy controls with an area under the curve of 0.9, suggesting that a VOC profile may be more effective than a single biomarker in diagnosing cancer.
Prospective diagnostic tool: The study supports further investigation into the use of VOC profiling as a molecular-oriented diagnostic test for gastroesophageal cancer, which could lead to a non-invasive method for early detection.
46. Boshier PR, Priest OH, Hanna GB, Marczin N. Influence of respiratory variables on the on-line detection of exhaled trace gases by PTR-MS. Thorax. 2011 Oct;66(10):919-20. doi: 10.1136/thx.2011.161208. PMID: 21474496.
Research aim:
The study aimed to investigate the impact of changes in expiratory flow, breath-holding, and paced hyperventilation on the concentrations of selected volatile organic compounds (VOCs) in exhaled breath, using proton transfer mass spectrometry.
Main findings:
Influence of expiratory flow rate: The concentration of acetone increased with higher expiratory flow rates (50, 100, 250 ml/s), demonstrating the influence of flow rate on VOC levels in exhaled breath.
Effect of hyperventilation on VOCs: Levels of methanol, acetaldehyde, ethanol, and dimethyl sulphide decreased significantly after 30 seconds of paced hyperventilation, suggesting that hyperventilation lowers the concentration of certain VOCs in exhaled breath.
Effect of breath-holding on VOCs: After 30 seconds of breath-holding, the concentrations of methanol, acetone, isoprene, and dimethyl sulphide increased significantly, indicating that breath-holding can raise the levels of certain VOCs.
47. Boshier PR, Cushnir JR, Mistry V, Knaggs A, Španěl P, Smith D, Hanna GB. On-line, real time monitoring of exhaled trace gases by SIFT-MS in the perioperative setting: a feasibility study. Analyst. 2011 Aug 21;136(16):3233-7. doi: 10.1039/c1an15356k. PMID: 21717028.
Research aim:
The study aimed to perform real-time, online analysis of exhaled breath during the perioperative period of laparoscopic surgery in anaesthetised patients, using selected ion flow tube mass spectrometry (SIFT-MS) to monitor water vapour, acetone, isoprene, and propofol.
Main findings:
Stable acetone levels: Breath acetone concentrations remained relatively constant throughout the perioperative period, with a slight increase towards the end of the surgery.
Increase in isoprene after abdomen inflation: A clear increase in breath isoprene concentrations was observed following abdominal inflation with carbon dioxide, indicating a physiological response during surgery.
Consistent propofol detection: The concentration of propofol in exhaled breath remained stable throughout the surgery, reflecting the continuous intravenous administration of the anaesthetic without significant fluctuation.