MALDI-TOF mass spectrometry analysis of small molecular weight compounds (under 10 KDa) as biomarkers of rat hearts undergoing arecoline challenge

Pharm Biol. 2013 Apr;51(4):488-91. doi: 10.3109/13880209.2012.742117. Epub 2013 Jan 22.

Abstract

Context: Statistical and clinical reports indicate that betel nut chewing is strongly associated with progression of oral cancer because some ingredients in betel nuts are potential cancer promoters, especially arecoline. Early diagnosis for cancer biomarkers is the best strategy for prevention of cancer progression. Several methods are suggested for investigating cancer biomarkers. Among these methods, gel-based proteomics approach is the most powerful and recommended tool for investigating biomarkers due to its high-throughput. However, this proteomics approach is not suitable for screening biomarkers with molecular weight under 10 KDa because of the characteristics of gel electrophoresis.

Objective: This study investigated biomarkers with molecular weight under 10 KDa in rats with arecoline challenge.

Materials and methods: The centrifuging vials with membrane (10 KDa molecular weight cut-off) played a crucial role in this study. After centrifuging, the filtrate (containing compounds with molecular weight under 10 KDa) was collected and spotted on a sample plate for MALDI-TOF mass spectrometry analysis.

Results: Compared to control, three extra peaks (m/z values were 1553.1611, 1668.2097 and 1740.1832, respectively) were found in sera and two extra peaks were found in heart tissue samples (408.9719 and 524.9961, respectively). These small compounds should play important roles and may be potential biomarker candidates in rats with arecoline.

Discussion and conclusions: This study successfully reports a mass-based method for investigating biomarker candidates with small molecular weight in different types of sample (including serum and tissue). In addition, this reported method is more time-efficient (1 working day) than gel-based proteomics approach (5~7 working days).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arecoline / toxicity*
  • Biomarkers, Tumor / metabolism*
  • Carcinogens / toxicity*
  • Centrifugation
  • Electrophoresis, Gel, Two-Dimensional
  • Male
  • Molecular Weight
  • Muscle Proteins / chemistry
  • Muscle Proteins / metabolism*
  • Myocytes, Cardiac / drug effects*
  • Myocytes, Cardiac / metabolism
  • Proteomics / methods*
  • Rats
  • Rats, Sprague-Dawley
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization*

Substances

  • Biomarkers, Tumor
  • Carcinogens
  • Muscle Proteins
  • Arecoline