Bacteria detection based on the evolution of enzyme-generated volatile organic compounds: Determination of Listeria monocytogenes in milk samples

• Published in: Analytica chimica acta 2014-10, Vol.848, p.80-87
• Main Authors: Tait, Emma, Perry, John D., Stanforth, Stephen P., Dean, John R.
• Format: Article
• Language: English
• Subjects: Amidohydrolases - metabolism; Aniline Compounds - analysis; Aniline Compounds - isolation & purification; Aniline Compounds - metabolism; Animals; Bacteria; beta-Glucosidase - metabolism; Contamination; Enzyme substrates; Fluorobenzenes - analysis; Fluorobenzenes - isolation & purification; Fluorobenzenes - metabolism; Food; Food Microbiology - methods; Gas Chromatography-Mass Spectrometry; Headspace solid-phase microextraction gas chromatography–mass spectrometry; Joining; Listeria monocytogenes; Listeria monocytogenes - enzymology; Listeria monocytogenes - isolation & purification; Mass spectrometry; Milk; Milk - microbiology; Nitrophenols - analysis; Nitrophenols - isolation & purification; Nitrophenols - metabolism; Organic compounds; Solid Phase Extraction; Volatile organic compounds; Volatile Organic Compounds - analysis; Volatile Organic Compounds - isolation & purification; Volatile Organic Compounds - metabolism
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2014.07.029

[Display omitted] •Rapid detection of Listeria monocytogenes contamination in food.•Use of VOC liberating enzyme substrates.•Analysis of VOCs by HS-SPME GC–MS.•Use of selective agents to aid detection. The rapid detection of Listeria monocytogenes contamination in food is essential to prevent food-borne illness in humans. The aim of this study was to differentiate non-contaminated milk from milk contaminated with L. monocytogenes using enzyme substrates coupled with the analysis of volatile organic compounds (VOCs). The method is based on the activity of β-glucosidase and hippuricase enzymes and the detection of a specific VOC i.e. 2-nitrophenol and 3-fluoroaniline, respectively. VOCs were extracted, separated and detected by headspace-solid phase microextraction coupled to gas chromatography–mass spectrometry (HS-SPME GC–MS). This approach required the inclusion of the selective agent's cycloheximide, nalidixic acid and acriflavine HCl in the growth medium to inhibit interfering bacteria. The VOCs were liberated by L. monocytogenes provided that samples contained at least 1–1.5×102CFUml−1 of milk prior to overnight incubation. This approach shows potential for future development as a rapid method for the detection of L. monocytogenes contaminated milk.