Chemical Markers for Short- and Long-Term Areca Nut Exposure

Abstract

Background: Areca nut (AN) chewing causes oral cancer. AN cessation programs are the most effective approach to reduce AN chewing induced cancers but require biomarkers to determine program compliance and success. Objectives: To explore chemical markers for short- and long-term AN exposure using non-invasively collected saliva, buccal cells (BCs), and scalp hair of chewers. Methods: Saliva was collected from a male chewer before and up to 2 days after AN chewing. Saliva was separated into supernatant and pellet (BCs) then analyzed by spectrophotometry and liquid chromatography (LC) with UV/VIS detection. Scalp hair was collected from four chewers and analyzed for areca alkaloids using direct analysis in real time-tandem mass spectrometry (DART-MSMS). Results: The red pigmented saliva after chewing showed no valuable signals when either the saliva supernatant or pellet (BCs) were analyzed by spectrophotometry. Saliva analysis by LC–UV/VIS showed diagnostically valuable signals at 488 nm up to 5 and 24 h post chewing in the supernatant and pellet, respectively. DART-MSMS analysis detected two of the four AN specific alkaloids (arecoline and arecaidine) in male but none in female hair. Conclusions/Importance: LC–UV/VIS analysis of the red pigments extracted from saliva and BCs after AN chewing showed distinct signals up to 24 h post chewing while DART-MSMS analysis in BCs and scalp hair showed selective signals of AN alkaloids for several weeks or months after AN exposure. Chemical hair treatment might prevent detection of areca alkaloids in hair. AN cessation trials and other programs now have essential tools for bioverification.

Keywords:

• Areca nut
• saliva
• buccal cells
• scalp hair
• biomarkers
• cessation program
• HPLC
• UV/VIS spectrophotometry
• DART-MS

Acknowledgment

The authors would like to thank Dr. Kevin Cassel (UHCC) and Srue Wakuk for assistance with hair sample collections, Tricia Debaun for assistance with HPLC and spectrophotometry analysis, and all volunteers for their specimen donations.

Disclosure statement

The authors declare no potential conflicts of interest.

Additional information

Funding

National Cancer Institute awards U54 CA143727 and P30 CA71789.