International Journal of Drug Delivery Technology
Volume 15, Issue 2

Salivary VOCs as a Diagnostic Marker for OSCC by Gas Chromatography-Mass Spectrometry Analysis: An Observational Study

Gracelin Ranjitha E1*, Sujatha S2, Sreekanth P Kunjumon3, Vaishnavi Palaniswamy4, Ruchika Choudhary5 

1Department of Oral Medicine and Radiology, Rajas Dental College and Hospital, Kavalkinaru junction, Tirunelveli, 627105, Tamilnadu, India

2Faculty of Dental Sciences, Department of Oral Medicine and Radiology, M.S Ramiah Univeristy of Applied sciences, Bengaluru, Karnataka, India

3Dental department, Aster Women and Child Hospital, Bangaluru, Karnataka, India

4Maharaja Dental Clinic, Coimbatore, Tamil Nadu, India

5Department of Oral Medicine and Radiology, Government Dental College, Jodhpur, Rajasthan, India 

Received: 12th May, 2025; Revised: 25th May, 2025; Accepted: 6thJun, 2025; Available Online: 25th Jun, 2025

ABSTRACT

Background: Head and neck carcinoma is the sixth most common malignancy globally, with oral squamous cell carcinoma (OSCC) being the most prevalent subtype. While histopathological examination remains the diagnostic gold standard, it is invasive and time-consuming. Metabolomic profiling, particularly analysis of volatile organic compounds (VOCs), offers a promising non-invasive diagnostic approach.

Objectives: To identify and compare salivary VOCs between OSCC patients and healthy controls, aiming to explore their potential as diagnostic biomarkers.

Study Design and Setting: A case-control study involving OSCC patients and age- and sex-matched healthy controls.

Participants: A total of 75 subjects were enrolled, including 35 histopathologically confirmed OSCC patients and 40 healthy controls.

Methods: Unstimulated saliva samples were collected and VOCs were extracted using a ZSM-5/PDMS-coated film. Extracts were condensed with 100 μL methanol, and 1.0 μL aliquots were analyzed using gas chromatography–mass spectrometry (GC-MS). Statistical comparisons were performed using the Chi-square test, with significance set at p < 0.05.

Results: Ten VOCs demonstrated statistically significant differences between OSCC and control groups. These included two acids, three alcohols, three ketones, and two alkanes. Notably, butanoic acid, pentanoic acid, and 4-methyl were significantly elevated in OSCC patients (p ≤ 0.001). VOC expression patterns also correlated significantly with age, gender, pTNM staging, and histopathological grade.

Conclusion: Salivary VOC profiling using GC-MS reveals distinct metabolomic signatures in OSCC patients.

Clinical significance: These findings suggest that specific VOCs may serve as non-invasive biomarkers for early diagnosis, prognosis, and therapeutic monitoring of OSCC.

Keywords: Oral Squamous cell carcinoma, Volatile organic compounds, saliva, Gas chromatography mass spectroscopy.

How to cite this article: Gracelin Ranjitha E, Sujatha S, Sreekanth P Kunjumon, Vaishnavi Palaniswamy, Ruchika Choudhary. Salivary VOCs as a diagnostic marker for OSCC by Gas Chromatography-Mass Spectrometry Analysis: An Observational Study. International Journal of Drug Delivery Technology. 2025;15(2):778-84. doi: 10.25258/ijddt.15.2.52

REFERENCES

  1. Gupta B, Bray F, Kumar N, Johnson NW. Associations between oral hygiene habits, diet, tobacco and alcohol and risk of oral cancer: A case–control study from India. Cancer Epidemiol. 2017;51:7–14.
  2. Sharma S, Satyanarayana L, Asthana S, Shivalingesh KK, Goutham BS, Ramachandra S. Oral cancer statistics in India on the basis of first report of 29 population-based cancer registries. J Oral Maxillofac Pathol. 2018;22(1):18–26.
  3. Veluthattil AC, Sudha SP, Kandasamy S, Chakkalakkoombil SV. Effect of hypofractionated, palliative radiotherapy on quality of life in late-stage oral cavity cancer: A prospective clinical trial. Indian J Palliat Care. 2019;25(3):383–90.
  4. Nagi R, Reddy-Kantharaj YB, Rakesh N, Janardhan-Reddy S, Sahu S. Efficacy of light-based detection systems for early detection of oral cancer and oral potentially malignant disorders: systematic review. Med Oral Patol Oral Cir Bucal. 2016;21(4):e447–55.
  5. Feinberg T et al. Cancer metabolism: the volatile signature of glycolysis—in vitro model in lung cancer cells. J Breath Res. 2017;11(1):016008.
  6. Haick H, Broza YY, Mochalski P, Ruzsanyi V, Amann A. Assessment, origin, and implementation of breath volatile cancer markers. Chem Soc Rev. 2014;43(5):1423–49.
  7. Brusselmans L, Arnouts L, Millevert C, Vandersnickt J, van Meerbeeck JP, Lamote K. Breath analysis as a diagnostic and screening tool for malignant pleural mesothelioma: a systematic review. Transl Lung Cancer Res. 2018;7(5):520–36.
  8. Yu JS et al. Saliva protein biomarkers to detect oral squamous cell carcinoma in a high-risk population in Taiwan. Proc Natl Acad Sci U S A. 2016;113(41):11549–54.
  9. Schweitzer A, Knauer SK, Stauber RH. Nuclear receptors in head and neck cancer: current knowledge and perspectives. Int J Cancer. 2010;126(4):801–9.
  10. Goerner M, Seiwert TY, Sudhoff H. Molecular targeted therapies in head and neck cancer—an update of recent developments-. Head Neck Oncol. 2010;2(1):8.
  11. Bouza M, Gonzalez-Soto J, Pereiro R, de Vicente JC, Sanz-Medel A. Exhaled breath and oral cavity VOCs as potential biomarkers in oral cancer subjects. J Breath Res. 2017;11(1):016015.
  12. Wang X, Kaczor-Urbanowicz KE, Wong DTW. Salivary biomarkers in cancer detection. Med Oncol. 2017;34(1):7.
  13. Calenic B, Miricescu D, Greabu M, Kuznetsov AV, Troppmair J, Ruzsanyi V, Amann A. Oxidative stress and volatile organic compounds: interplay in pulmonary, cardio-vascular, digestive tract systems and cancer. Open Chem. 2015;13(1):1–11.
  14. Conte M, et al. The smell of longevity: a combination of volatile organic compounds (VOCs) can discriminate centenarians and their offspring from age-matched subjects and young controls. GeroScience. 2020;42(1):201–16.
  15. Monedeiro F, Monedeiro-Milanowski M, Zmyslowski H, Spinosa B. Evaluation of salivary VOC profile composition directed towards oral cancer and oral lesion assessment. Clin Oral Investig. 2021:1–16.
  16. Ochiai K, Kurita-Ochiai T. Effects of butyric acid on the periodontal tissue. Jpn Dent Sci Rev. 2009;45(2):75–82.
  17. Pulukuri SMK, Gorantla B, Rao JS. Inhibition of histone deacetylase activity promotes invasion of human cancer cells through activation of urokinase plasminogen activator. J Biol Chem. 2007;282(49):35594–603.
  18. Shigeyama H, Wang T, Ichinose M, Ansai T, Lee SW. Identification of volatile metabolites in human saliva from subjects with oral squamous cell carcinoma via zeolite-based thin-film microextraction coupled with GC–MS. J Chromatogr B. 2019;1104:49–58.
  19. Bonuccelli G, et al. Ketones and lactate “fuel” tumor growth and metastasis: evidence that epithelial cancer cells use oxidative mitochondrial metabolism. Cell Cycle. 2010;9(17):3506–14.
  20. Smith D, Wang T, Spanĕl P. On-line. On-line, simultaneous quantification of ethanol, some metabolites and water vapour in breath following the ingestion of alcohol. Physiol Meas. 2002;23(3):477–89.
  21. Phillips M, Gleeson K, Hughes JM, Greenberg J, Cataneo RN, Baker L, McVay WP. Volatile organic compounds in breath as markers of lung cancer: a cross-sectional study. Lancet. 1999;353(9168):1930–3.
  22. Raman M, Ahmed I, Gillevet PM, Probert CS, Ratcliffe NM, Smith S . Fecal microbiome and volatile organic compound metabolome in obese humans with nonalcoholic fatty liver disease. Clin Gastroenterol Hepatol. 2013;11(7):868–75.