Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Forensic Toxicology

Open Access 22-03-2024 | Intoxication | Original Article

Development of a simple screening method for analyzing cereulide toxin in fried rice using liquid chromatography-tandem mass spectrometry

Authors: Hiroshi Koike, Maki Kanda, Chie Monma, Souichi Yoshikawa, Hiroshi Hayashi, Yoko Matsushima, Yumi Ohba, Momoka Hayashi, Natsumi Furuta, Wakaba Okada, Chieko Nagano, Keiko Yokoyama, Tomoko Yokoyama, Takeo Sasamoto

Published in: Forensic Toxicology

Login to get access

Abstract

Purpose

The presence of cereulide, an emetic toxin produced by Bacillus cereus, in fried rice samples is critical evidence of food poisoning even in situations where B. cereus could not be detected. This study aims to develop a screening method for analyzing cereulide in fried rice using the QuEChERS procedure and liquid chromatography-tandem mass spectrometry (LC–MS/MS).

Methods

Cereulide was identified and quantified in fried rice samples using the QuEChERS extraction method and LC–MS/MS. The accuracies of the methods were determined by analyzing fortified blank samples at two concentrations (10 and 50 µg/kg) conducted on three samples daily for five days.

Results

The QuEChERS procedure removed matrix compounds from fried rice. Characteristic MS/MS spectra enabled the identification of cereulide. As the matrix effects in seven fried rice samples were within ± 6%, an external solvent calibration curve could be used for quantification. This method exhibited good accuracy ranging from 88 to 89%. The relative standard deviations for both repeatability and intra-laboratory reproducibility were < 4%. These standard deviations satisfied the criteria of the Japanese validation guidelines for residues (MHLW 2010, Director Notice, Syoku-An No. 1224–1). The limit of quantification was 2 μg/kg. The applicability of this method was confirmed using the analysis of cereulide in fried rice samples incubated with emetic Bacillus cereus.

Conclusions

The QuEChERS extraction procedure described herein showed substantial promise as a reliable screening tool for cereulide in fried rice sample.
Literature
1.
Agata N, Ohta M, Mori M, Isobe M (1995) A novel dodecadepsipeptide, cereulide, is an emetic toxin of Bacillus cereus. FEMS Microbiol Lett 129:17–20PubMed
2.
Ehling-Schulz M, Fricker M, Grallert H, Rieck P, Wagner M, Scherer S (2006) Cereulide synthetase gene cluster from emetic Bacillus cereus: structure and location on a mega virulence plasmid related to Bacillus anthracis toxin plasmid pXO1. BMC Microbiol 6:20CrossRefPubMedPubMedCentral
3.
Marxen S, Stark TD, Frenzel E, Rütschle A, Lücking G, Pürstinger G, Pohl EE, Scherer S, Ehling-Schulz M, Hofmann T (2015) Chemodiversity of cereulide, the emetic toxin of Bacillus cereus. Anal Bioanal Chem 407:2439–2453CrossRefPubMed
4.
Rajkovic A, Grootaert C, Butorac A, Cucu T, De Meulenaer B, van Camp J, Bracke M, Uyttendaele M, Bačun-Družina V, Cindrić M (2014) Sub-emetic toxicity of Bacillus cereus toxin cereulide on cultured human enterocyte-like Caco-2 cells. Toxins 6:2270–2290CrossRefPubMedPubMedCentral
5.
Yamaguchi M, Kawai T, Kitagawa M, Kumeda Y (2013) A new method for rapid and quantitative detection of the Bacillus cereus emetic toxin cereulide in food products by liquid chromatography-tandem mass spectrometry analysis. Food Microbiol 34:29–37CrossRefPubMed
6.
Rønning HT, Asp TN, Granum PE (2015) Determination and quantification of the emetic toxin cereulide from Bacillus cereus in pasta, rice and cream with liquid chromatography-tandem mass spectrometry. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 32:911–921CrossRefPubMed
7.
8.
Schreiber N, Hackl G, Reisinger AC, Zollner-Schwetz I, Eller K, Schlagenhaufen C, Pietzka A, Czerwenka C, Stark TD, Kranzler M, Fickert P, Eller P, Ehling-Schulz M (2022) Acute Liver failure after ingestion of fried rice balls: a case series of Bacillus cereus food poisonings. Toxins (Basel) 14:12. https://​doi.​org/​10.​3390/​toxins14010012CrossRef
9.
Dierick K, Van Coillie E, Swiecicka I, Meyfroidt G, Devlieger H, Meulemans A, Hoedemaekers G, Fourie L, Heyndrickx M, Mahillon J (2005) Fatal family outbreak of Bacillus cereus-associated food poisoning. J Clin Microbiol 43:4277–4279CrossRefPubMedPubMedCentral
10.
Shiota M, Saitou K, Mizumoto H, Matsusaka M, Agata N, Nakayama M, Kage M, Tatsumi S, Okamoto A, Yamaguchi S, Ohta M, Hata D (2010) Rapid detoxification of cereulide in Bacillus cereus food poisoning. Pediatrics 125:e951-955CrossRefPubMed
11.
Naranjo M, Denayer S, Botteldoorn N, Delbrassinne L, Veys J, Waegenaere J, Sirtaine N, Driesen RB, Sipido KR, Mahillon J, Dierick K (2011) Sudden death of a young adult associated with Bacillus cereus food poisoning. J Clin Microbiol 49:4379–4381CrossRefPubMedPubMedCentral
12.
Andersson MA, Hakulinen P, Honkalampi-Hämäläinen U, Hoornstra D, Lhuguenot JC, Mäki-Paakkanen J, Savolainen M, Severin I, Stammati AL, Turco L et al (2007) Toxicological profile of cereulide, the Bacillus cereus emetic toxin, in functional assays with human, animal and bacterial cells. Toxicon 49:351–367CrossRefPubMed
13.
14.
Andersson MA, Mikkola R, Helin J, Andersson MC, Salkinoja-Salonen M (1998) A novel sensitive bioassay for detection of Bacillus cereus emetic toxin and related depsipeptide ionophores. Appl Environ Microbiol 64:1338–1343CrossRefPubMedPubMedCentral
15.
Rowan NJ, Deans K, Anderson JG, Gemmell CG, Hunter IS, Chaithong T (2001) Putative virulence factor expression by clinical and food isolates of Bacillus spp. after growth in reconstituted infant milk formulae. Appl Environ Microbiol 67:3873–3881CrossRefPubMedPubMedCentral
16.
Beattie SH, Williams AG (1999) Detection of toxigenic strains of Bacillus cereus and other Bacillus spp. with an improved cytotoxicity assay. Lett Appl Microbiol 28:221–225CrossRefPubMed
17.
Stark T, Marxen S, Rütschle A, Lücking G, Scherer S, Ehling-Schulz M, Hofmann T (2013) Mass spectrometric profiling of Bacillus cereus strains and quantitation of the emetic toxin cereulide by means of stable isotope dilution analysis and HEp-2 bioassay. Anal Bioanal Chem 405:191–201CrossRefPubMed
18.
Kawamura-Sato K, Hirama Y, Agata N, Ito H, Torii K, Takeno A, Hasegawa T, Shimomura Y, Ohta M (2005) Quantitative analysis of cereulide, an emetic toxin of Bacillus cereus, by using rat liver mitochondria. Microbiol Immunol 49:25–30CrossRefPubMed
19.
Ulrich S, Gottschalk C, Dietrich R, Märtlbauer E, Gareis M (2019) Identification of cereulide producing Bacillus cereus by MALDI-TOF MS. Food Microbiol 82:75–81CrossRefPubMed
20.
Doellinger J, Schneider A, Stark TD, Ehling-Schulz M, Lasch P (2020) Evaluation of MALDI-ToF mass spectrometry for rapid detection of cereulide from Bacillus cereus cultures. Front Microbiol 11:2483CrossRef
21.
Biesta-Peters EG, Reij MW, Blaauw RH, In’t Veld PH, Rajkovic A, Ehling-Schulz M, Abee T (2010) Quantification of the emetic toxin cereulide in food products by liquid chromatography-mass spectrometry using synthetic cereulide as a standard. Appl Environ Microbiol 76:7466–7472
22.
Delbrassinne L, Andjelkovic M, Dierick K, Denayer S, Mahillon J, Van Loco J (2012) Prevalence and levels of Bacillus cereus emetic toxin in rice dishes randomly collected from restaurants and comparison with the levels measured in a recent foodborne outbreak. Foodborne Pathog Dis 9:809–814CrossRefPubMed
23.
Decleer M, Rajkovic A, Sas B, Madder A, De Saeger S (2016) Development and validation of ultra-high-performance liquid chromatography-tandem mass spectrometry methods for the simultaneous determination of beauvericin, enniatins (A, A1, B, B1) and cereulide in maize, wheat, pasta and rice. J Chromatogr A 1472:35–43CrossRefPubMed
24.
In’t Veld PH, van der Laak LFJ, van Zon M, Biesta-Peters EG (2018) Elaboration and validation of the method for the quantification of the emetic toxin of Bacillus cereus as described in EN-ISO 18465—Microbiology of the food chain—Quantitative determination of emetic toxin (cereulide) using LC-MS/MS. Int J Food Microbiol 288:91–96
25.
Anastassiades M, Lehotay SJ, Stajnbaher D, Schenck FJ (2003) Fast and easy multiresidue method employing acetonitrile extraction/partitioning and “dispersive solid-phase extraction” for the determination of pesticide residues in produce. J AOAC Int 86:412–431CrossRefPubMed
26.
Martínez Vidal JL, Garrido Frenich A, Aguilera-Luiz MM, Romero-González R (2010) Development of fast screening methods for the analysis of veterinary drug residues in milk by liquid chromatography-triple quadrupole mass spectrometry. Anal Bioanal Chem 397(7):2777–2790CrossRefPubMed
27.
Nakajima T, Nagano C, Sasamoto T, Hayashi H, Kanda M, Kanai S, Takeba K, Matsushima Y, Takano I (2012) Development and validation of rapid analysis method for multi-class veterinary drugs in livestock products by LC-MS/MS. Shokuhin Eiseigaku Zasshi 53:243–253CrossRefPubMed
28.
Yogendrarajah P, Van Poucke C, De Meulenaer B, De Saeger S (2013) Development and validation of a QuEChERS based liquid chromatography tandem mass spectrometry method for the determination of multiple mycotoxins in spices. J Chromatogr A 5(1297):1–11CrossRef
29.
Dybowski MP, Dawidowicz AL (2018) Application of the QuEChERS procedure for analysis of Δ9-tetrahydrocannabinol and its metabolites in authentic whole blood samples by GC-MS/MS. Forensic Toxicol 36(2):415–423CrossRefPubMedPubMedCentral
30.
Koike H, Kanda M, Hayashi H, Matsushima Y, Ohba Y, Nakagawa Y, Nagano C, Sekimura K, Hirai A, Shindo T, Kamiie J, Sasamoto T, Hashimoto T (2018) Identification and quantification of cereulide in cow’s milk using liquid chromatography-tandem mass spectrometry. Food Addit Contam Part A 35:2424–2433CrossRef
31.
Ministry of Health, Labour and Welfare (MHLW), Tokyo, Japan (2010) Director notice guideline for the validation of analytical methods for agricultural chemical residues in food. Syoku-An No. 1224–1
32.
Ellison SLR, Williams A (2012) Eurachem guide CITAC quantifying uncertainty in analytical measurement, 3rd ed. Eurachem, Belgique
33.
Bauer T, Stark T, Hofmann T, Ehling-Schulz M (2010) Development of a stable isotope dilution analysis for the quantification of the Bacillus cereus toxin cereulide in foods. J Agric Food Chem 58:1420–1428CrossRefPubMed
34.
SANTE/11312/2021 (2022) Guidance document on analytical quality control and method validation procedures for pesticide residues and analysis in food and feed. (Document No. SANTE/11312/2021).
35.
CAC/GL.71–2009 (2014) Guidelines for design and implementation of national regulatory food safety assurance programme associated with the use of veterinary drugs in food proceeding animals. (Document No. CAC/GL.71–2009)
36.
Lynch KL, Breaud AR, Vandenberghe H, Wu AH, Clarke W (2010) Performance evaluation of three liquid chromatography mass spectrometry methods for broad spectrum drug screening. Clin Chim Acta 411:1474–1481CrossRefPubMedPubMedCentral
37.
Xing Y, Meng W, Sun W, Li D, Yu Z, Tong L, Zhao Y (2016) Simultaneous qualitative and quantitative analysis of 21 mycotoxins in Radix Paeoniae Alba by ultra-high performance liquid chromatography quadrupole linear ion trap mass spectrometry and QuEChERS for sample preparation. J Chromatogr B 1031:202–213CrossRef
38.
Hayashi M, Kanda M, Yoshikawa S, Nakajima T, Hayashi H, Matsushima Y, Ohba Y, Koike H, Nagano C, Otsuka K, Sasamoto T (2023) Single-laboratory validation study and surveillance using an improved multiresidue analytical method for veterinary drugs in livestock products by LC–MS/MS. Shokuhin Eiseigaku Zasshi 64:53–60CrossRefPubMed
39.
Kawano Y, Sato H, Koga M, Matsunaga N, Hidaka C, Nakamura K (2021) Rapid method of cereulide analysis in processed foods using LC-MS/MS. Jpn J Food Chem Safty 28:98–106
Metadata
Title
Development of a simple screening method for analyzing cereulide toxin in fried rice using liquid chromatography-tandem mass spectrometry
Authors
Hiroshi Koike
Maki Kanda
Chie Monma
Souichi Yoshikawa
Hiroshi Hayashi
Yoko Matsushima
Yumi Ohba
Momoka Hayashi
Natsumi Furuta
Wakaba Okada
Chieko Nagano
Keiko Yokoyama
Tomoko Yokoyama
Takeo Sasamoto
Publication date
22-03-2024
Publisher
Springer Nature Singapore
Keyword
Intoxication
Published in
Forensic Toxicology
Print ISSN: 1860-8965
Electronic ISSN: 1860-8973
DOI
https://doi.org/10.1007/s11419-024-00683-3