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Article | Open Access

HPTLC-fluorescent densitometry for screening aflatoxin B1 in millet and buckwheat

Xudong Shia,1, Xingjun Xib,1, Yuetao Jiaa, Zhijian Wanga, Jiawei Guob, Shiyao Wanga, Xiaoqian Tangc,d(), Yisheng Chena()
College of Food Science and Engineering, Shanxi Agricultural University, Taigu 030801, China
China National Institute of Standardization, Beijing 100191, China
Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, China

1 These authors contributed equally to this work.

Peer review under responsibility of Beijing Academy of Food Sciences.

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Highlights

• A novel HPTLC-FLD method was developed for the rapid screening of AFB1 in millet and buckwheat, offering high efficiency and low cost.

• The method enables specific identification and quantitative analysis of AFB1 with high sensitivity, achieving an LOD of 3 μg/kg, an LOQ of 10 μg/kg, and R² of 0.999.

• It enables the simultaneous screening of 17 samples within 1 h, outperforming LC-MS/MS in efficiency and cost, ideal for resource-limited labs.

• The HPTLC image acquisition system enables visual determination of AFB1 presence and preliminary content assessment, simplifying the detection process through this visual screening approach.

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Abstract

Given severe health-hazardous effects of aflatoxin B1 (AFB1) widely occurring in cereal grains and animal feeds, it is highly urgent to develop analytical methods for its rapid screening. In this work, we proposed a simple and high-throughput method for the determination of AFB1 in millet and buckwheat samples using high performance thin layer chromatography (HPTLC) linked to fluorescence densitometry. The first step was to optimize the solid-liquid extraction for the crude clean-up of the samples. The QuEChERS (Quick, Easy, Cheap, Effective, Robust and Safe) extraction strategy was used and different solvent systems for their extraction efficiency of AFB1 from the samples were evaluated. Then, trichloromethane:ethyl acetate (7:3, V/V) was used as the mobile phase to realize the separation of the targeted compound from background noises on silica gel plates. Quantification was readily performed with densitometry in fluorescence mode. In order to fix the optimal excitation wavelength, spectra scanning ranging 250−400 nm was carried out, revealing that 364 nm light gave the highest signal. With the optimized optical system, high sensitivity to AFB1 was achieved, with a limit of detection (LOD) at 3 μg/kg. Apart from that, good linearity (0.999) was obtained within the range of 1−80 ng/band of AFB1. To assess the analysis accuracy, 2 levels of AFB1 (50 and 100 μg/kg) were spiked into real grain samples. The obtained results showed that the recovery rates were within the range of 81.6%−114.0%. The proof-of-concept results of this work evidenced that HPTLC is a promising analytical tool for the screening of mycotoxin in difficult samples.

Keywords

Aflatoxin B1High performance thin layer chromatography (HPTLC)ScreeningMilletBuckwheat

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Food Science and Human Wellness
Volume 14 Issue 5,
May 2025
Article number: 9250229
DOI: 10.26599/FSHW.2024.9250229
Cite this article:
Shi X, Xi X, Jia Y, et al. HPTLC-fluorescent densitometry for screening aflatoxin B1 in millet and buckwheat. Food Science and Human Wellness, 2025, 14(5): 9250229. https://doi.org/10.26599/FSHW.2024.9250229
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